Anti-TTYH2 antibodies and methods of use thereof

Abstract

This application relates to antibodies or antigen-binding fragments thereof that bind to Tweety family member 2 (TTYH2) protein, as well as polynucleotides and vectors that encode for such antibodies or antigen-binding fragments. This application further relates to methods of producing the antibodies or antigen-binding fragments and using the antibodies or antigen-binding fragments for treatment of diseases.

Description

Attorney Docket No: 243735.000428 ANTI-TTYH2 ANTIBODIES AND METHODS OF USE THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 652,565, filedMay 28, 2024, the contents of which is incorporated by reference herein in their entirety. SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submittedelectronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on May 21, 2025, is named 243735_000428_SL.xml and is 78,305 bytes in size. FIELD OF THE INVENTION

[0003] This application relates to antibodies or antigen-binding fragments thereof that bind totweety family member 2 (TTYH2) protein, as well as polynucleotides and vectors that encode for such antibodies or antigen-binding fragments. This application further relates to methods of producing the antibodies or antigen-binding fragments and using the antibodies or antigen-binding fragments for treatment of diseases e.g., cancer or an autoimmune / autoinflammatory disease. BACKGROUND

[0004] Type I interferon (IFN) stands out as a pivotal cytokine bridging innate and adaptiveimmunity, crucial for immune modulation and defense against pathogens and diseases. Despite its efficacy, systemic administration of recombinant Type I IFN or stimulator of interferon genes (STING) agonist that potently induces Type I IFN often yields modest clinical signal, propensity for adverse side effects, and poor tolerability of regimes for patients, prompting the need for more effective approaches to enhance Type I IFN activity, preferentially at the disease site.

[0005] Type I IFNs play a crucial role in the immune system’s response to a broad spectrum ofhuman diseases. Major functions of Type I IFNs include: 1) priming of both innate and adaptive immune responses; 2) regulation of inflammation; and 3) anti-proliferative function and the induction of apoptosis. Overall, the major function of Type I IFNs is to orchestrate an effective immune response against pathogens and malignancies while regulating inflammation to maintain immune homeostasis. 1 314460903v1Attorney Docket No: 243735.000428

[0006] Given the critical role of Type I IFN in immune homeostasis and defense, active clinicalprograms have been initiated to target Type I IFN for various human diseases. Recombinant Type I IFNs (IFN-α or IFN-β) have been utilized in the treatment of various conditions including cancer (e.g., leukemia, kidney cancer, Kaposi’s sarcoma, and melanoma), and autoimmune disorders (e.g., multiple sclerosis (MS), Bechet’s disease) (Helen M. Lazear, John W. Schoggins, Michael S. Diamond. Shared and Distinct Functions of Type I and Type III Interferons. 2019 Apr 16; 50(4):p907-923). While it is easier to understand the role of Type I IFN in anti-cancer responses given its role in immunomodulation and inducing cancer cell apoptosis, the mechanism of recombinant Type I IFNs in protecting against (but not aggravating) multiple sclerosis (MS) is more complex. Type I IFNs are thought to be related to negatively regulate immune responses in the brain, preserve the integrity of the brain-blood barrier (BBB), promote remyelination etc., making Type I IFN an important therapeutic target in the management of MS. While therapies involving recombinant Type I IFN or STING agonists which can potently induce Type I IFN offer broad-spectrum activity against cancer and inflammation, their use is limited by high immuno- toxicity, modest efficacy, and poor tolerability of regimes for patients through systemic administration, necessitating alternative or complementary approaches. Between 15% - 40% of people undergoing Type I IFN treatment develop major depressive disorders12. Prolonged exposure to Type I IFN treatment may lead to liver inflammation and fibrosis in certain viral infections like chronic hepatitis B and C. Less commonly, IFN treatment has also been associated with anxiety, lethargy, psychosis, and parkinsonism. It is thus critical to identify key pathways that modulate Type I IFN specifically in diseased tissue(s) and to design more disease-focused therapies.

[0007] In the cancer setting, advances in immune checkpoint blockade (ICB) immunotherapy,such as by antibodies targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1(PD-1) / programmed death ligand-1 (PD-L1 (PD-(L)1)), have reshaped the cancer treatment landscape3-5. Anti-PD-L1 therapy has demonstrated durable clinical responses with mild adverse effects across many different cancer types4-5. Anti-PD-L1 targets a key immune evasion mechanism in the tumor microenvironment (TME) to repair tumor-specific T cell immunity, and the success of this therapy is generally thought to arise from “hot” tumors with abundant T cell responses critical for eliminating tumor cells5. However, a large subset of patients is not responsive to anti-PD-L1 therapy (primary resistance) or develops an acquired 2 314460903v1Attorney Docket No: 243735.000428 resistance5. Lymphocyte activation gene 3 (LAG-3, also referred to herein as LAG3) is another T cell checkpoint receptor mainly expressed on PD-1 positive T cells, also known as “exhausted” T cells6. Anti-LAG-3 antibody, relatlimab has been recently approved for the treatment of melanoma, but only in combination with anti-PD-1 antibody nivolumab6. Unfortunately, strategies aimed at various other immune checkpoints (either receptors or ligands), such as T cell immunoreceptor with immunoglobulin (Ig) and ITIM (immunoreceptor tyrosine-based inhibitory motif) domain (TIGIT), cluster of differentiation 47 (CD47), etc., have not yielded significant success in human cancer trials5,7. These clinical observations emphasize the need to understand how anti-PD-L1 works and identify new therapies beyond the current strategies, especially for the cold tumors with weak T cell activities. SUMMARY OF THE INVENTION

[0008] As specified in the Background section above, there are unmet clinical needs to overcomeimmunotherapy resistance by identifying novel immune checkpoints and the associated molecular mechanisms. The present application addresses these and other needs.

[0009] In one aspect, provided herein is an antibody, or antigen-binding fragment thereof, thatspecifically binds to tweety family member 2 (TTYH2) protein, comprising:(i) a heavy chain variable region (HCVR) that comprises the heavy chain complementarity-determining region 1 (HCDR1), HCDR2, and HCDR3 of an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1, 9, or 17, or a variant thereof; and / or(ii) a light chain variable region (LCVR) comprising the light chain complementarity-determiningregion 1 (LCDR1), LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, 13, or 21, or a variant thereof.

[0010] In some embodiments, the antibody or antigen-binding fragment comprises:(a) an HCVR that comprises the HCDR1, HCDR2, and HCDR3 of an HCVR that comprises theamino acid sequence set forth in SEQ ID NO: 1, or a variant thereof, and an LCVR that comprises the LCDR1, LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, or a variant thereof;(b) an HCVR that comprises the HCDR1, HCDR2, and HCDR3 of an HCVR that comprises theamino acid sequence set forth in SEQ ID NO: 9, or a variant thereof, and an LCVR that 3 314460903v1Attorney Docket No: 243735.000428 comprises the LCDR1, LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 13, or a variant thereof; and / or(c) an HCVR that comprises the HCDR1, HCDR2, and HCDR3 of an HCVR that comprises theamino acid sequence set forth in SEQ ID NO: 17, or a variant thereof, and an LCVR that comprises the LCDR1, LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 21, or a variant thereof.

[0011] In some embodiments, the antibody or antigen-binding fragment comprises:(a) an HCVR that comprises an HCDR1 that comprises the amino acid sequence set forth in SEQID NO: 2, an HCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 3, and an HCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 4, and an LCVR that comprises an LCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 6, an LCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 7, and an LCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 8;(b) an HCVR that comprises an HCDR1 that comprises the amino acid sequence set forth in SEQID NO: 10, an HCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 11, and an HCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 12, and a LCVR that comprises an LCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 14, an LCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 15, and an LCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 16; and / or(c) an HCVR that comprises an HCDR1 that comprises the amino acid sequence set forth in SEQID NO: 18, an HCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 19, and an HCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 20, and an LCVR that comprises an LCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 22, an LCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 23, and an LCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 24.

[0012] In some embodiments, the antibody or antigen-binding fragment comprises:(a) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1, 9, or 17, or avariant thereof, and / or(b) an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, 13, or 21, or avariant thereof.

[0013] In some embodiments, the antibody or antigen-binding fragment comprises:4 314460903v1Attorney Docket No: 243735.000428(a) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1, or a variantthereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, or a variant thereof;(b) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 9, or a variantthereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 13, or a variant thereof; and / or(c) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 17, or a variantthereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 21, or a variant thereof.

[0014] In another aspect, provided herein is an antibody, or antigen-binding fragment thereof,that competes for binding to TTYH2 protein with the antibody or antigen-binding fragment described above.

[0015] In another aspect, provided herein is an antibody, or antigen-binding fragment thereof,that binds to the same epitope of TTYH2 protein as the antibody or antigen-binding fragment described above.

[0016] In another aspect, provided herein is an antibody, or antigen-binding fragment thereof,that specifically binds to TTYH2 protein, wherein the antibody or antigen-binding fragment thereof binds to the second ectodomain (ECD2) of TTYH2 protein.

[0017] In some embodiments, the antibody or antigen-binding fragment thereof binds to anepitope on TTYH2 protein comprising, consisting essentially of, or consisting of the sequence FAARGDY (SEQ ID NO: 25).

[0018] In some embodiments, the antibody or antigen-binding fragment binds to human TTYH2protein and / or mouse TTYH2 protein.

[0019] In some embodiments, the antibody or antigen-binding fragment is recombinant.

[0020] In some embodiments, the antibody or antigen-binding fragment is a human antibody, ahumanized antibody, a chimeric antibody, a murine antibody, a monoclonal antibody, a single chain antibody, a bispecific antibody or antigen-binding fragment thereof, a bi-epitopic antibody or antigen-binding fragment thereof, a Fab, a Fab’, a F(ab’)2, a Fv, an scFv, a VH domain, or a nanobody. 5 314460903v1Attorney Docket No: 243735.000428

[0021] In some embodiments, the bispecific antibody or antigen-binding fragment thereof is abispecific T- cell engager (BiTE), bispecific NK-cell engager (BiKE), or a bi-specific macrophage engager (BiME).

[0022] In some embodiments, the BiTE comprises an antigen-binding domain that specificallybinds to cluster of differentiation (CD) 3 (CD3), alpha beta T cell receptor (TCR), or gamma delta TCR.

[0023] In some embodiments, the BiKE comprises an antigen-binding domain that specificallybinds to Fc gamma receptor III (FcγRIII or CD16), natural killer group 2, member D (NKG2D), or natural cytotoxicity triggering receptor 3 (NCR3).

[0024] In some embodiments, the BiME comprises an antigen-binding domain that specificallybinds to Fc gamma receptor III (FcγRIII or CD16), dendritic cell-associated C-type lectin 1 (Dectin-1), cluster of differentiation (CD47), signal regulatory protein alpha (SIRPA), or Tyro3, Axl, and MerTK (TAM) receptor(s).

[0025] In some embodiments, the bispecific antibody or antigen-binding fragment thereofcomprises an antigen-binding domain that specifically binds to an immune checkpoint, a cytokine or a receptor thereof, a tumor-associated antigen (TAA), or an immune stimulatory receptor.

[0026] In some embodiments, the immune checkpoint is programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), lymphocyte activation gene 3 (LAG-3 or CD223), cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), cluster of differentiation 47 (CD47), T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3), B7 homolog 3 protein (B7- H3 or CD276), B7-H4, V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)), T cell immunoreceptor with Ig and ITIM Domains (TIGIT), signal regulatory protein alpha (SIRPA), signaling lymphocytic activation molecule family members (SLAMF), poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R), adenosine A2A receptor (A2aR), adenosine A2b receptor (A2bR), G protein-coupled receptor 171 (GPR171), insulin like growth factor binding protein 7 (IGFBP7), cluster of differentiation 93 (CD93), CD96, CD226, natural killer group protein 2A (NKG2A), natural killer group protein 2D (NKG2D), a killer cell lectin like receptor G1 (KLRG1), a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2), a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3), sialic acid-binding immunoglobulin-like lectin (Siglec)-15, cluster of differentiation 24 (CD24), sialic 6 314460903v1Attorney Docket No: 243735.000428 acid-binding immunoglobulin-like lectin (Siglec)-10, P-selectin glycoprotein ligand-1 (PSGL-1), V-set and Ig domain-containing protein 3 (VSIG3, also known as B7 and T cell immunoglobulin domain-containing superfamily member (BT-IgSF) or immunoglobulin superfamily member 11 (IGSF11)), leucine rich repeats and immunoglobulin like domains 1 (LRIG1), fibrinogen-like protein 1 (FGL1), B and T lymphocyte attenuator (BTLA), leukocyte associated immunoglobulin like receptor 1 (LAIR-1), cluster of differentiation 160 (CD160), leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2), leukocyte immunoglobulin-like receptor (LILRB4), angiopoietin 2 (ang2), or vascular endothelial growth factor (VEGF).

[0027] In some embodiments, the immune checkpoint is PD-1, PD-L1, LAG-3, and / or CTLA-4.

[0028] In some embodiments, the cytokine is interleukin-2 (IL-2), IL-6, IL-10, IL-15, IL-21,IFN-α, IFN-β, IFN-γ, chemokine (C-C motif) ligand 19 (CCL19), chemokine (C-C motif) ligand 21 (CCL21), interleukin-18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF), or a receptor thereof.

[0029] In some embodiments, the tumor-associated antigen (TAA) is oncofetal antigen 5T4(5T4), angiopoietin 2 (Ang2), B-cell maturation antigen (BCMA), cluster of differentiation 123 (CD123), cluster of differentiation 19 (CD19), cluster of differentiation 20 (CD20), cluster of differentiation 22 (CD22), cluster of differentiation 33 (CD33), cluster of differentiation 38 (CD38), cluster of differentiation 47 (CD47), carcinoembryonic antigen (CEA), carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), Claudin 6, Claudin 18.2, c-type lectin domain family 12 member A (CLEC12A), delta-like ligand 3 (DLL3), epidermal growth factor receptor (EGFR), epithelial cell adhesion molecule (EpCAM), Fc receptor homolog 5 (FcRH5), fms-like tyrosine kinase 3 (FLT3), disialoganglioside (GD2), Glypican-3, glycoprotein A33 (gpA33), G protein- coupled receptor class C group 5 member D (GPRC5D), human epidermal growth factor receptor 2 (Her2), human epidermal growth factor receptor 3 (Her3), melanoma antigen family A4 (MAGE- A4), mesenchymal-epithelial transition factor (MET), mucin 16 (MUC16), mucin 17 (MUC17), New York esophageal squamous cell carcinoma 1 (NY-ESO-1), placental cadherin (P-cadherin), preferentially expressed antigen in melanoma (PRAME), prostate stem cell antigen (PSCA), prostate-specific membrane antigen (PSMA), somatostatin receptor 2 (SSTR2), six 7 314460903v1Attorney Docket No: 243735.000428 transmembrane epithelial antigen of the prostate 1 (STEAP1), tumor-associated calcium signal transducer 2 (TROP2, also known as tumor-associated calcium signal transducer 2 (TACSTD2)), angiopoietin 2 (Ang2), TEK tyrosine kinase (Tie2), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR), gamma delta T-cell receptor (γδTCR), Kirsten rat sarcoma viral oncogene homolog (KRAS), or rapidly accelerated fibrosarcoma (RAF).

[0030] In some embodiments, the immune stimulatory receptor is cluster of differentiation 28(CD28), inducible T-cell costimulatory (ICOS), transmembrane and immunoglobulin domain containing 2 (TMIGD2), natural cytotoxicity receptor 3 (NCR3), natural cytotoxicity receptor 1 (NCR1), natural cytotoxicity receptor 2 (NCR2), tumor necrosis factor receptor superfamily member 9 (4-1BB, also known as cluster of differentiation 137 (CD137)), tumor necrosis factor receptor superfamily member 4 (OX40), cluster of differentiation 30 (CD30), cluster of differentiation 40 (CD40), death receptor 3 (DR3), cluster of differentiation 226 (CD226, also known as DNAX accessory molecule 1 (DNAM-1)), class I restricted T cell-associated molecule (CRTAM), cluster of differentiation 27 (CD27), herpes virus entry mediator (HVEM), tumor necrosis factor receptor 1 (TNFR1), tumor necrosis factor receptor 2 (TNFR2), cluster of differentiation 2 (CD2), cluster of differentiation 7 (CD7), toll-like receptor 4 (TLR4), toll-like receptor 7 (TLR7), or toll-like receptor 9 (TLR9).

[0031] In some embodiments, the bispecific antibody or antigen-binding fragment thereofcomprises an antigen-binding domain that specifically binds to tumor necrosis factor-alpha (TNF- α), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 13 (IL-13), interleukin 17A (IL-17A), interleukin 17F (IL-17F), chemokine (C-C motif) ligand 2 (CCL2), C-X-C motif chemokine ligand 10 (CXCL10), IFN-α, IFN-β, very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29), cluster of differentiation 20 (CD20), B activating factor (BAFF), transmembrane activator and CAML interactor (TACI), cluster of differentiation 19 (CD19), interleukin-23 receptor (IL-23R), cluster of differentiation 19 (CD22), cluster of differentiation 38 (CD38), B-cell maturation antigen (BCMA), PD-1, BTLA, CD40, CD40 ligand (CD40L), OX40, OX40L, B7, CD28, CTLA-4, tumor necrosis factor-like cytokine 1A (TL1A), DR3, or thymic stromal lymphopoietin (TSLP), or a variant thereof.

[0032] In some embodiments, the antibody or antigen-binding fragment is an IgG antibody.

[0033] In some embodiments, the antibody or antigen-binding fragment is of IgG1, IgG2, IgG3,or IgG4 subclass. 8 314460903v1Attorney Docket No: 243735.000428

[0034] In some embodiments, the antibody or antigen-binding fragment is of IgG1 or IgG4subclass.

[0035] In some embodiments, the antibody comprises an Fc region comprisingL234A / L235A / P329G (LALA-PG) amino acid substitutions, wherein the residues are numbered according to the EU index of Kabat.

[0036] In some embodiments, the antibody or antigen-binding fragment(i) antagonizes the inhibitory function of TTYH2 on a myeloid cell or cancer cell;(ii) stimulates Type I interferon (IFN) response or myeloid function upon binding to TTYH2protein expressed on a myeloid cell or cancer cell; (iii)activates stimulator of interferon genes (STING) / Type I IFN pathway signaling, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway signaling, or TANK-binding kinase 1 / interferon regulatory factor 3 (TBK1 / IRF3) pathway signaling, or a combination thereof, upon binding to TTYH2 protein expressed on a myeloid cell or cancer cell; and / or (iv)controls TNF-α, IL-6, CCL2, IL-10, and / or CXCL10, or myeloid cell differentiation.

[0037] In some embodiments, the myeloid cell is a macrophage or a dendritic cell.

[0038] In some embodiments, the macrophage is an M2 macrophage and / or tumor-associatedmacrophage.

[0039] In another aspect, provided herein is an antibody-drug conjugate comprising the antibodyor antigen-binding fragment described above, conjugated to a heterologous moiety.

[0040] In some embodiments, the heterologous moiety is an immune checkpoint inhibitor, acytokine, a tumor-associated antigen (TAA)-targeting agent, an immune agonist, a cytotoxic agent, an siRNA, or an antisense oligonucleotide.

[0041] In some embodiments of the antibody-drug conjugates, the immune checkpoint inhibitoris a programmed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3, also known as cluster of differentiation 223 (CD223)) inhibitor, a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3, also known as cluster of differentiation 276 (CD276)) inhibitor, a B7-H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as PD-1H) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling 9 314460903v1Attorney Docket No: 243735.000428 lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor- related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a cluster of differentiation 96 (CD96) inhibitor, a cluster of differentiation 226 (CD226) inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat- associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin- like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid- binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as B7 and T cell immunoglobulin domain-containing superfamily member (BT-IgSF) or immunoglobulin superfamily member 11 (IGSF11)) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, or a vascular endothelial growth factor (VEGF) inhibitor, or a combination thereof.

[0042] In some embodiments of the antibody-drug conjugates, the immune checkpoint inhibitoris a PD-1 inhibitor or a PD-L1 inhibitor.

[0043] In some embodiments of the antibody-drug conjugates, the PD-1 inhibitor or PD-L1inhibitor is nivolumab, pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, 10 314460903v1Attorney Docket No: 243735.000428 spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof.

[0044] In some embodiments of the antibody-drug conjugates, the immune checkpoint inhibitoris a LAG-3 inhibitor.

[0045] In some embodiments of the antibody-drug conjugates, the LAG-3 inhibitor is relatlimab(BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA-1953, or a variant or a combination thereof.

[0046] In some embodiments of the antibody-drug conjugates, the LAG-3 inhibitor is relatlimab(BMS-986016), or a variant thereof.

[0047] In some embodiments of the antibody-drug conjugates, the immune checkpoint inhibitoris a CTLA-4 inhibitor.

[0048] In some embodiments of the antibody-drug conjugates, the CTLA-4 inhibitor isipilimumab, tremelimumab, quavonlimab, BNT316 / ONC-392 (gotistobart), or porustobart (HBM4003), or a variant or a combination thereof.

[0049] In some embodiments of the antibody-drug conjugates, the cytokine is interleukin (IL)-2 (IL-2), IL-6, IL-10, IL-15, IL-21, IFN-α, IFN-β, IFN-γ, chemokine (C-C motif) ligand 19 (CCL19), chemokine (C-C motif) ligand 21 (CCL21), interleukin -18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF) or a receptor thereof.

[0050] In some embodiments of the antibody-drug conjugates, the TAA-targeting agent targets5T4, Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY-ESO-1, P- cadherin, PRAME , PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, or RAF, or a combination thereof.

[0051] In some embodiments of the antibody-drug conjugates, the immune agonist is a 4-1BBagonist, an OX40 agonist, a CD40 agonist, a CD30 agonist, a glucocorticoid-induced tumor 11 314460903v1Attorney Docket No: 243735.000428 necrosis factor receptor-related protein (GITR) agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a transmembrane and immunoglobulin domain-containing protein 2 (TMIGD2, also known as CD28H) agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

[0052] In some embodiments, the heterologous moiety is a tumor necrosis factor-alpha (TNF-α)inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL- 4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA-targeted T- cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor,or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

[0053] In another aspect, provided herein is a polynucleotide encoding the antibody or antigen-binding fragment of described above.

[0054] In another aspect, provided herein is a vector comprising the polynucleotide describedabove.

[0055] In another aspect, provided herein is a host cell expressing the antibody or antigen-binding fragment described above, or comprising the polynucleotide described above or the vector described above.

[0056] In some embodiments of the host cell, the antibody or antigen-binding fragment isrecombinantly produced. 12 314460903v1Attorney Docket No: 243735.000428

[0057] In another aspect, provided herein is a method of producing the antibody or antigen-binding fragment described above, wherein said method comprises culturing the host cell described above, and isolating said antibody or antigen-binding fragment.

[0058] In another aspect, provided herein is a pharmaceutical composition comprising theantibody or antigen-binding fragment described above, the antibody-drug conjugate described above, the polynucleotide described above, or the vector described above, and a pharmaceutically acceptable carrier or diluent.

[0059] In some embodiments, the pharmaceutical composition further comprises one or moreimmune checkpoint inhibitors, cytokines, tumor-associated antigen (TAA)-targeting agents, immune agonists, cytotoxic agents, siRNAs, or antisense oligonucleotides.

[0060] In some embodiments of the pharmaceutical composition described herein, the immunecheckpoint inhibitor is a programmed cell death protein 1 (PD-1) inhibitor, a programmed death- ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a B7-H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)- 15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin- like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine 13 314460903v1Attorney Docket No: 243735.000428 rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, or a vascular endothelial growth factor (VEGF) inhibitor, or a combination thereof.

[0061] In some embodiments of the pharmaceutical composition described herein, the immunecheckpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a CTLA-4 inhibitor, or a combination thereof.

[0062] In some embodiments of the pharmaceutical composition described herein, the PD-1inhibitor or PD-L1 inhibitor is nivolumab, pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof.

[0063] In some embodiments of the pharmaceutical composition described herein, the LAG-3inhibitor is relatlimab (BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA-1953, or a variant or a combination thereof.

[0064] In some embodiments of the pharmaceutical composition described herein, the LAG-3inhibitor is relatlimab (BMS-986016), or a variant thereof.

[0065] In some embodiments of the pharmaceutical composition described herein, the immunecheckpoint inhibitor is a CTLA-4 inhibitor. 14 314460903v1Attorney Docket No: 243735.000428

[0066] In some embodiments of the pharmaceutical composition described herein, the CTLA-4inhibitor is ipilimumab, tremelimumab, quavonlimab, BNT316 / ONC-392 (gotistobart), or porustobart (HBM4003), or a variant or a combination thereof.

[0067] In some embodiments of the pharmaceutical composition described herein, the cytokineis interleukin-2 (IL-2), IL-6, IL-10, IL-15, IL-21, IFN-α, IFN-β, IFN-γ, IL-18, CCL19, CCL21, monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte- macrophage colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G- CSF), or a receptor thereof.

[0068] In some embodiments of the pharmaceutical composition described herein, the TAA-targeting agent targets 5T4, Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY-ESO-1, P-cadherin, PRAME, PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, or RAF, or a combination thereof.

[0069] In some embodiments of the pharmaceutical composition described herein, the immuneagonist is a 4-1BB agonist, an OX40 agonist, a CD40 agonist, a CD30 agonist, a GITR agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a CD28H / TMIGD2 agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

[0070] In some embodiments, the pharmaceutical composition further comprises stimulator ofinterferon genes (STING) agonist or toll-like receptor (TLR) agonist, or a combination thereof.

[0071] In some embodiments of the pharmaceutical composition described herein, the STINGagonist is DMXAA, 2’3’-cGAMP, ADU-S100 / MIW815, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING (BI 1387446), E7766, TAK-676, SNX281, or SYNB1891, or a variant or a combination thereof.

[0072] In some embodiments of the pharmaceutical composition described herein, the TLRagonist is a TLR1 / TLR2 agonist, a TLR2 / TLR6 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

[0073] In some embodiments, the pharmaceutical composition further comprises one or more ofa tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 15 314460903v1Attorney Docket No: 243735.000428 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL- 17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, , a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19- targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38- targeted T cell engager, a BCMA-targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

[0074] In another aspect, provided herein is a kit comprising (i) the antibody or antigen-bindingfragment described above, the antibody-drug conjugate described above, the polynucleotide described above, or the vector described above, and (ii) optionally, packaging for the same and / or instructions for use.

[0075] In another aspect, provided herein is a method for stimulating Type I IFN response of amyeloid cell or cancer cell or enhancing myeloid function in a subject in need thereof, comprising administering to the subject an effective amount of an agent that targets TTYH2 such that the Type I IFN response of the myeloid cells or cancer cell is stimulated.

[0076] In some embodiments of the method described herein, the myeloid cell is a macrophageor a dendritic cell.

[0077] In some embodiments of the method described herein, the macrophage is an M2macrophage and / or tumor-associated macrophage.

[0078] In another aspect, provided herein is a method for depleting a cancer cell that expressesTTYH2 protein in a subject in need thereof, comprising administering to the subject an effective amount of an agent that targets TTYH2 such that the cancer cell is depleted. 16 314460903v1Attorney Docket No: 243735.000428

[0079] In some embodiments of the method for depleting a cancer cell that expresses TTYH2protein described herein, the agent that targets TTYH2 comprises the antibody or antigen-binding fragment described above, the antibody-drug conjugate described above, the polynucleotide described above, the vector described above, or the pharmaceutical composition of described above.

[0080] In another aspect, provided herein is a method of treating or preventing a cancer in asubject in need thereof, comprising administering to the subject an effective amount of an agent that targets TTYH2.

[0081] In some embodiments of the above-described methods, the agent that targets TTYH2 isselected from a small molecule, a protein, a protein-drug conjugate, a nucleotide molecule, a gene editing system, an engineered cell system, and any combination thereof.

[0082] In some embodiments of the above-described methods, the protein is selected from apeptide; an antibody or antigen-binding fragment thereof; a monobody; engineered, low-density- lipoprotein-receptor-derived, A domain (LDLR-A) (e.g., Avimers™); a designed ankyrin repeat protein (DARPin) lipocalin (e.g., anticalins); an affibody; engineered, Protein-A-derived, Z domain (Affibodies™) CTLD3 (e.g., Tetranectin); C-type lectin-like domain scaffolds; Sac7d- derived polypeptides (e.g., Nanoffitins®or affitins); engineered, tenascin-derived, tenascin type III domain (e.g., Centyrin™), thioredoxin (e.g., peptide aptamer); KALBITOR®; the β-sandwich (e.g., iMab); gamma-B crystallin-derived scaffold or engineered, ubiquitin-derived scaffold (e.g., Affilins); engineered, protease inhibitor-derived, Kunitz domain (e.g., EETI-II / AGRP, BPTI / LACI-D1 / ITI-D2); engineered antibody mimics; miniproteins; engineered, Fyn-derived, SH2 domain (e.g., Fynomers®); genetically manipulated counterparts of the foregoing that retains its binding functionality, and any combination thereof.

[0083] In some embodiments of the above-described methods, the antibody or antigen-bindingfragment thereof is selected from an intact antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a single-chain variable fragment (scFv), a VH domain, a nanobody, a Bi-specific T-cell engager (BiTE), a bispecific killer cell engager (BiKE), a bi-specific macrophage engager (BiME), a CrossMab, a tri-specific binding partner, and any combination thereof. 17 314460903v1Attorney Docket No: 243735.000428

[0084] In some embodiments of the above-described methods, the nucleotide molecule isselected from an antisense oligonucleotide, a micro RNA (miRNA), a small interfering RNA (siRNA), a short hairpin RNA (shRNA), a single guide RNA (sgRNA), and any combination thereof.

[0085] In some embodiments of the above-described methods, the gene editing systemcomprises a CRISPR-associated protein (Cas) nuclease, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN), a meganuclease, any endo- or exo- nuclease, variants thereof, fragments thereof, or any combination thereof.

[0086] In some embodiments of the above-described methods, the engineered cell systemcomprises a chimeric antigen receptor (CAR) modified cell comprising a fragment that targets TTYH2.

[0087] In some embodiments of the above-described methods, the CAR modified cell is includea CAR-modified T cell (CAR-T cell), a CAR-modified natural killer (NK) cell (CAR-NK cell), or a CAR-macrophage (CAR-M).

[0088] In some embodiments of the above-described methods, the agent is conjugated to adetectable label, a chemotherapeutic agent, a radioisotope, an affinity tag, or a toxin.

[0089] In some embodiments of the method of treating or preventing a cancer described herein,the agent that targets TTYH2 comprises the antibody or antigen-binding fragment described above, the antibody-drug conjugate described above, the polynucleotide described above, the vector described above, or the pharmaceutical composition described above.

[0090] In some embodiments of the method of treating or preventing a cancer described herein,the cancer is resistant to an immunotherapy comprising one or more immune checkpoint inhibitors.

[0091] In some embodiments of the method of treating or preventing a cancer described herein,the immune checkpoint inhibitor is a programmed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a (B7-H4) inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation 18 314460903v1Attorney Docket No: 243735.000428 molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat- associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin- like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid- binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, or a vascular endothelial growth factor (VEGF) inhibitor, or a combination thereof.

[0092] In some embodiments of the method of treating or preventing a cancer described herein,the immune checkpoint inhibitor comprises a PD-1 inhibitor or PD-L1 inhibitor.

[0093] In some embodiments of the method of treating or preventing a cancer described herein,the PD-1 inhibitor or PD-L1 inhibitor is nivolumab, pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof. 19 314460903v1Attorney Docket No: 243735.000428

[0094] In some embodiments of the method of treating or preventing a cancer described herein,the immune checkpoint inhibitor is a LAG-3 inhibitor.

[0095] In some embodiments of the method of treating or preventing a cancer described herein,the LAG-3 inhibitor is relatlimab (BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK- 420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA-1953, or a variant or a combination thereof.

[0096] In some embodiments of the method of treating or preventing a cancer described herein,the LAG-3 inhibitor is relatlimab (BMS-986016), or a variant thereof.

[0097] In some embodiments of the method of treating or preventing a cancer described herein,the immune checkpoint inhibitor is a CTLA-4 inhibitor.

[0098] In some embodiments of the method of treating or preventing a cancer described herein,the CTLA-4 inhibitor is ipilimumab, tremelimumab, quavonlimab, BNT316 / ONC- 392 (gotistobart), or porustobart (HBM4003), or a variant or combination thereof.

[0099] In some embodiments of the method of treating or preventing a cancer described herein,the method further comprises administering to the subject one or more additional anti-cancer therapies.

[0100] In some embodiments of the method of treating or preventing a cancer described herein,the one or more additional anti-cancer therapies comprise an immunotherapy, a chemotherapy, a targeted therapy, a radiotherapy, or a combination thereof.

[0101] In some embodiments of the method of treating or preventing a cancer described herein,the immunotherapy is interferon beta-1a (IFN beta-1a), interferon beta-1b (IFN beta-1b), glatiramer acetate, mitoxantrone, natalizumab, teriflunomide, fingolimod, dimethyl fumarate, or alemtuzumab, or a variant or combination thereof.

[0102] In one aspect, provided herein is a method of treating or preventing anautoimmune / autoinflammatory disease in a subject in need thereof, comprising administering to the subject an effective amount of an agent that targets TTYH2.

[0103] In some embodiments of the method of treating or preventing anautoimmune / autoinflammatory disease described herein, the agent that targets TTYH2 comprises the antibody or antigen-binding fragment described above, the antibody-drug conjugate described 20 314460903v1Attorney Docket No: 243735.000428 above, the polynucleotide described above, the vector described above, or the pharmaceutical composition described above.

[0104] In some embodiments of the method of treating or preventing anautoimmune / autoinflammatory disease described herein, the method further comprises administering to the subject one or more additional anti-autoimmune / autoinflammatory therapies.

[0105] In some embodiments of the method of treating or preventing anautoimmune / autoinflammatory disease described herein, the one or more additional anti- autoimmune / autoinflammatory therapies comprise an immunotherapy or a targeted therapy, or a combination thereof.

[0106] In some embodiments of the method of treating or preventing anautoimmune / autoinflammatory disease described herein, the immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL- 13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20- targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA- targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

[0107] In some embodiments, the immunotherapy is a tumor necrosis factor-alpha (TNF-α)inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, or a combination thereof. 21 314460903v1Attorney Docket No: 243735.000428

[0108] In another aspect, provided herein is a method of increasing effectiveness of animmunotherapy in a subject in need thereof, comprising co-administering to the subject an immunotherapy with an effective amount of an agent that targets TTYH2.

[0109] In some embodiments of the above-described methods, the agent that targets TTYH2 isselected from a small molecule, a protein, a protein-drug conjugate, a nucleotide molecule, a gene editing system, an engineered cell system, and any combination thereof.

[0110] In some embodiments of the above-described methods, the protein is selected from apeptide; an antibody or antigen-binding fragment thereof; a monobody; engineered, low-density- lipoprotein-receptor-derived, A domain (LDLR-A) (e.g., Avimers™); a designed ankyrin repeat protein (DARPin) lipocalin (e.g., anticalins); an affibody; engineered, Protein-A-derived, Z domain (Affibodies™) CTLD3 (e.g., Tetranectin); C-type lectin-like domain scaffolds; Sac7d- derived polypeptides (e.g., Nanoffitins® or affitins); engineered, tenascin-derived, tenascin type III domain (e.g., Centyrin™), thioredoxin (e.g., peptide aptamer); KALBITOR®; the β-sandwich (e.g., iMab); gamma-B crystallin-derived scaffold or engineered, ubiquitin-derived scaffold (e.g., Affilins); engineered, protease inhibitor-derived, Kunitz domain (e.g., EETI-II / AGRP, BPTI / LACI-D1 / ITI-D2); engineered antibody mimics; miniproteins; engineered, Fyn-derived, SH2 domain (e.g., Fynomers®); genetically manipulated counterparts of the foregoing that retains its binding functionality, and any combination thereof.

[0111] In some embodiments of the above-described methods, the antibody or antigen-bindingfragment thereof is selected from an intact antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a single-chain variable fragment (scFv), a VH domain, a nanobody, a Bi-specific T-cell engager (BiTE), a bispecific killer cell engager (BiKE), a bi-specific macrophage nano-engager (BiME), a CrossMab, a tri-specific binding partner, and any combination thereof.

[0112] In some embodiments of the above-described methods, the nucleotide molecule isselected from an antisense oligonucleotide, a micro RNA (miRNA), a small interfering RNA (siRNA), a short hairpin RNA (shRNA), a single guide RNA (sgRNA), and any combination thereof.

[0113] In some embodiments of the above-described methods, the gene editing systemcomprises a CRISPR-associated protein (Cas) nuclease, a zinc finger nuclease (ZFN), a 22 314460903v1Attorney Docket No: 243735.000428 transcription activator-like effector nuclease (TALEN), a meganuclease, any endo- or exo- nuclease, variants thereof, fragments thereof, or any combination thereof.

[0114] In some embodiments of the above-described methods, the engineered cell systemcomprises a chimeric antigen receptor (CAR) modified cell comprising a fragment that targets TTYH2.

[0115] In some embodiments of the above-described methods, the CAR modified cell is includea CAR-modified T cell (CAR-T cell), a CAR-modified natural killer (NK) cell (CAR-NK cell), or a CAR-macrophage (CAR-M).

[0116] In some embodiments of the above-described methods, the agent is conjugated to adetectable label, a chemotherapeutic agent, a radioisotope, an affinity tag, or a toxin.

[0117] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the agent that targets TTYH2 comprises the antibody or antigen-binding fragment of described above, the antibody-drug conjugate described above, the polynucleotide described above, the vector described above, or the pharmaceutical composition described above.

[0118] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immunotherapy and the agent that targets TTYH2 are administered sequentially.

[0119] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immunotherapy and the agent that targets TTYH2 are administered simultaneously in one composition or in separate compositions.

[0120] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immunotherapy comprises one or more immune checkpoint inhibitors, cytokines, tumor-associated antigen (TAA)-targeting agents, or immune agonists.

[0121] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immune checkpoint inhibitor is a programmed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG- 3 or CD223) inhibitor, a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain- containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a B7- H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) 23 314460903v1Attorney Docket No: 243735.000428 inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat- associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin- like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid- binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, a vascular endothelial growth factor (VEGF) inhibitor, or a combination thereof.

[0122] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immune checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a LAG3 inhibitor, a CTLA-4 inhibitor, or a combination thereof.

[0123] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the PD-1 inhibitor or PD-L1 inhibitor is nivolumab, pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, 24 314460903v1Attorney Docket No: 243735.000428 serplulimab, sintilimab, spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof.

[0124] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the LAG-3 inhibitor is relatlimab (BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA-1953, or a variant or a combination thereof.

[0125] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the LAG-3 inhibitor is relatlimab (BMS-986016), or a variant thereof.

[0126] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immune checkpoint inhibitor is a CTLA-4 inhibitor.

[0127] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the CTLA-4 inhibitor is ipilimumab, tremelimumab, or quavonlimab, BNT316 / ONC-392 (gotistobart), or porustobart (HBM4003), or a variant or a combination thereof.

[0128] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the cytokine is interleukin-2 (IL-2), IL-6, IL-10, IL-15, IL-21, IFN-α, IFN-β, IFN-γ, IL-18, CCL19, CCL21, monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF), or a receptor thereof.

[0129] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the TAA targeting agent targets 5T4, Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE- A4, MET, MUC16, MUC17, NY-ESO-1, P-cadherin, PRAME, PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, or RAF, or a combination thereof.

[0130] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immune agonist is a 4-1BB agonist, an OX40 agonist, a CD40 agonist, a CD30 agonist, a GITR agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a 25 314460903v1Attorney Docket No: 243735.000428 CD28H / TMIGD2 agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

[0131] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the immunotherapy comprises stimulator of interferon genes (STING) agonist or toll-like receptor (TLR) agonist or a combination thereof.

[0132] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the STING agonist is DMXAA, 2’3’-cGAMP, ADU-S100 / MIW815, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING (BI 1387446), E7766, TAK-676, SNX281, or SYNB1891, or a variant or a combination thereof.

[0133] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, the TLR agonist is a TLR1 / TLR2 agonist, TLR2 / TLR6 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

[0134] In some embodiments of the method of increasing effectiveness of an immunotherapydescribed herein, immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA-targeted T- cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion 26 314460903v1Attorney Docket No: 243735.000428 protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

[0135] In some embodiments of an above-described method, the subject has a solid cancer.

[0136] In some embodiments of an above-described method, the solid cancer develops as a hottumor with abundant anti-tumor immune responses particularly T cell activities.

[0137] In some embodiments of an above-described method, the solid cancer develops as a coldtumor with weak anti-tumor immune responses particularly T cell activities.

[0138] In some embodiments of an above-described method, the solid cancer is a lung cancer, aglioma, a medulloblastoma, a thyroid cancer, a colorectal cancer, a head and neck cancer, a gastric cancer, a stomach cancer, a liver cancer, a pancreatic cancer, a renal cancer, a urothelial cancer, a prostate cancer, a testis cancer, a breast cancer, a cervical cancer, an endometrial cancer, an ovarian cancer, a gallbladder cancer, a sarcoma, or a melanoma.

[0139] In some embodiments of an above-described method, glioma is a glioblastoma or anastrocytoma.

[0140] In some embodiments of an above-described method, the solid cancer is a colorectalcancer or a melanoma.

[0141] In some embodiments of an above-described method, the solid cancer is a pancreaticcancer or a glioblastoma.

[0142] In some embodiments of an above-described method, the subject has a hematologiccancer.

[0143] In some embodiments of an above-described method, the hematologic cancer is acutemyeloid leukemia (AML), acute lymphoblastic leukemia (ALL), acute leukemia of ambiguous lineage, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, chronic myeloid neoplasm, non-Hodgkin lymphoma (e.g., follicular non-Hodgkin lymphoma), Hodgkin lymphoma, chronic leukemia, dendritic / histiocytic neoplasm, or lymphoproliferative disorder.

[0144] In some embodiments of an above-described method, the hematologic cancer is acutemyeloid leukemia (AML).

[0145] In some embodiments of an above-described method, the method further comprisesadministering to the subject one or more additional anti-cancer therapies. 27 314460903v1Attorney Docket No: 243735.000428

[0146] In some embodiments of an above-described method, the one or more additional anti-cancer therapies comprise a chemotherapy, a targeted therapy, a radiotherapy, or a combination thereof.

[0147] In some embodiments of an above-described method, the immunotherapy is selectedfrom interferon beta-1a (IFN beta-1a), interferon beta-1b (IFN beta-1b), glatiramer acetate, mitoxantrone, natalizumab, teriflunomide, fingolimod, dimethyl fumarate, and alemtuzumab, or a variant or combination thereof.

[0148] In some embodiments of an above-described method, the autoimmune / autoinflammatorydisease is multiple sclerosis (MS) or Bechet’s disease.

[0149] In some embodiments of an above-described method, the method further comprisesadministering to the subject one or more additional anti-autoimmune / autoinflammatory therapies.

[0150] In some embodiments of an above-described method, the one or more additional anti-autoimmune / autoinflammatory therapies comprise an immunotherapy or a targeted therapy, or a combination thereof.

[0151] In some embodiments of an above-described method comprising administering to thesubject one or more additional anti-autoimmune / autoinflammatory therapies, the immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL- 17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19- targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38- targeted T cell engager, a BCMA-targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A 28 314460903v1Attorney Docket No: 243735.000428 (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

[0152] In some embodiments of an above-described method, the administering is via intrathecal,intratumoral, intravenous, intradermal, intraperitoneal, subcutaneous, intramuscular, inhalation, oral delivery, lipid nanoparticle (LNP)-based delivery, cellular delivery, viral and / or non-viral delivery, or gene editing, or as a cargo in a cell, or any combination thereof.

[0153] In some embodiments of an above-described method, the subject is human or veterinaryanimal. BRIEF DESCRIPTION OF THE DRAWINGS

[0154] Figures 1A-1E show TTYH2 as a negative regulator of myeloid cell-associated Type IIFN responses. PD-1 or LAG-3 mainly controls exhausted T cells in the tumor-site. Recombinant Type I IFN has shown modest efficacy in cancers with high toxicity profile. (Figure 1A) In this scenario, TTYH2 was identified as a myeloid cell-associated Type I IFN (MIFN) negative feedback inhibitor for immunotherapy based on the study of TTYH2 knock out (KO) mice as illustrated in Example 1. Figures 1B-1C depict the results of quantitative PCR (qPCR) analysis. Figures 1D-1E depict the results of cytokine analysis.

[0155] Figure 2 shows the results of flow cytometry analysis, demonstrating that anti-TTYH2antibodies from T36, T51, and T52 hybridoma clones are cross-reactive to human and mouse TTYH2, but not human TTYH1 and TTYH3.

[0156] Figures 3A-3C show the schematic of human TTYH2 secondary structure and results offlow cytometry and structural analyses, demonstrating that the second ectodomain (ECD2) of TTYH2 is the major binding site for anti-TTYH2 monoclonal antibodies (mAbs). Figure 3A shows a schematic of human TTYH2 secondary structure involving three ectodomains (ECD), three intracellular domains (ICD), and five transmembrane domains. Figure 3B depicts the results of flow cytometry analysis, demonstrating that the second ectodomain (ECD2) is the major binding site for anti-TTYH2 antibodies from T36, T51, and T52 hybridoma clones. Figure 3C depicts the results of structural analysis, demonstrating that T36 binds to TTYH2-ECD2.

[0157] Figures 4 shows the results of the sequence alignment and structural studies,demonstrating that the smallest conserved RGD (Arg-Gly-Asp) motif (highlighted in a solid line rectangular box) is present in TTYH2 across species. Figure 4 shows the sequence alignment of 29 314460903v1Attorney Docket No: 243735.000428 TTYH2 protein sequence among species by Clustal Omega, demonstrating that FAARGDY (SEQ ID NO: 25) motif (highlighted in dashed line rectangular box) in ECD2 domain is very conserved among species. Figure 4 discloses SEQ ID NOS 73-80, respectively, in order of appearance.

[0158] Figures 5A-5C depict that the conserved RGD containing motif is the binding site forthe TTYH2 mAbs. Figure 5A depicts a schematic representation of a TTYH2 protein and a TTYH2 RGD mutant protein fused to Flag-tag. The TTYH2 RGD mutant comprises LAGRPEF(SEQ ID NO: 49) motif from TTYH3. Figure 5A also discloses “FAARGDY” as SEQ ID NO:25. Figure 5B depicts the results of flow cytometry analysis, demonstrating that the RGD containing motif is the binding site of anti-TTYH2 antibodies from T36, T51, and T52 hybridoma clones. Figure 5C depicts an illustration of the TTYH2 (comprises FAARGDY (SEQ ID NO: 25) motif) and TTYH2 mutant (LAGRPEF (SEQ ID NO: 49) motif from TTYH3). The TTYH2 amino acids (aa) 161-167 were replaced with TTYH3 aa 158-164, as indicated in Figure 5C. The mutant comprises LAGRPEF (SEQ ID NO: 49) motif.

[0159] Figures 6A-6B demonstrate myeloid-dominant expression of TTYH2 on human (Figure6A) or mouse (Figure 6B) immune cells by flow cytometry with anti-TTYH2 clone T36. Figure6A shows the TTYH2 protein expression in human peripheral immune cells. Figure 6B shows the TTYH2 expression in mouse immune cells from peripheral blood (Figure 6B, left panel), as well as in mouse bone marrow-derived dendritic cells (BMDC) and bone marrow-derived macrophages (BMDM) (Figure 6B, right panel).

[0160] Figures 7A-7C show STING agonist (DMXAA) or toll-like receptor (TLR) agonists-associated feedback regulation of TTYH2. Figure 7A depicts the results of quantitative PCR (qPCR) analysis of STING in WT or TTYH2 KO myeloid cells, with or without STING agonist DMXAA stimulation. Figure 7B depicts the results of flow cytometry analysis of TTYH2 expression with clone T36 by STING (2’3’-cGAMP) or Toll-like receptor agonists stimulation. Figure 7C depicts a schematic of TTYH2 as a myeloid negative feedback modulator for Type-I IFN responses.

[0161] Figures 8A-8B show results demonstrating that TTYH2 deficient peritonealmacrophages have enhanced STING-TBK1-IRF3 (stimulator of interferon genes - TANK (TRAF Family Member Associated NF-κB Activator)-binding kinase 1) - interferon regulatory factor 3) pathway signaling. Figures 8A shows the results of western blots, demonstrating that the TTYH2 deficiency promotes STING / IRF3 / NF-κB responses and the inhibitory role of TTYH2. Figure 30 314460903v1Attorney Docket No: 243735.000428 8B shows the results demonstrating that the human TTYH2 overexpression (indicated with ‘+’ sign) suppresses IFN-β (IFNB1) and TNF-α measured by quantitative PCR (qPCR) analysis in human myeloid T cell leukemia patient-derived cell line 1 (THP-1) cells.

[0162] Figures 9A-9B show the results of TTYH2 surface staining and phorbol myristate acetate(PMA) stimulation, demonstrating that the TTYH2 overexpression suppresses NF-κB activation in Jurkat human T cell line and the reversion by an anti-TTYH2 antibody.

[0163] Figures 10A-10B show the tumor single cell RNA-seq data from melanoma patientsdemonstrating representative expression of TTYH2 on both tumor cells and M2 macrophages.Figure 10A depicts the data of Uniform Manifold Approximation and Projection (UMAP) analysisof cell clusters. Figure 10B depicts the expression of TTYH2, TREM1 (M1 macrophage marker; Figure 10B), and M2 macrophage markers (e.g., TREM2, MRC1). Figures were adapted from the (Tumor Immune Single Cell Hub 2) TISCH2 database.

[0164] Figures 11A-11H show the data, demonstrating an abundant expression of TTYH2 inhuman cancers and the results of survival analysis demonstrating that high TTYH2 expressionassociates with cancer survival and immunotherapy resistance. Figure 11A shows a TTYH2 RNAexpression profile across major human cancer types from the GEPIA2 which analyzed the TCGA data. Dominant expression of TTYH2 in multiple cancer types, such as in melanoma (SKCM, skin cutaneous melanoma), THYME (Thymic carcinoma), LGG (Low-Grade Glioma), LAML (Acute Myeloid Leukemia) compared to corresponding normal tissues, was shown. Figure 11B depicts an association of the high expression of TTYH2 with patient’s worse survival in melanoma cohorts. Figure 11C depicts the TTYH2 RNA expression across human cancer cell lines from the Human Protein Atlas database. Figures 11D-11G depict the results of survival analysis of TTYH2 high versus TTYH2 low patients in cancers from the TCGA OncoLnc database. Figure 11H depicts the results of survival analysis of TTYH2 high versus TTYH2 low patient survival upon of anti-PD-L1 treatment analyzed by the Tumor Immune Dysfunction and Exclusion (TIDE) database.

[0165] Figures 12A-12B depict representative flow cytometry analysis (Figure 12A) andkinetics of OT-I (also referred to herein as OT-1 or OT1) T cells among total CD8+ T cells at day 5 (Figure 12B) upon OVA257-264 peptide and TLR agonist (Poly i:c) stimulation in TTYH2 KO or wildtype (WT) mice, with or without Type-I IFN receptor antibody (anti-IFNAR1 antibody, 31 314460903v1Attorney Docket No: 243735.000428 MAR1-5A3) blockade, demonstrating that TTYH2 deficiency promotes antigen-specific T cell responses in a Type I IFN dependent manner.

[0166] Figures 13A-13B depict representative flow cytometry analysis (Figure 13A) andquantification of OT-I T cells among total CD8+ T cells at day 5 (Figure 13B) upon OVA257-264peptide and STING agonist (2’3’-cGAMP) stimulation in TTYH2 KO or wildtype (WT) mice, demonstrating that TTYH2 deficiency amplifies antigen-specific T cell responses with STING agonist.

[0167] Figures 14A-14B show quantification of tumor volumes and Kaplan-Meier survivalcurves of WT or TTYH2 KO mice transplanted with Yummer1.7 mouse melanoma cancer cells (Figure 14A) or MC38 mouse colon cancer cells (Figure 14B), with or without anti-PD-1 antibody (RMP1-14) blockade. These data show that TTYH2 deficiency in combination with anti- PD-1 therapy controls both tumor growth, while TTYH2 itself has effects in Yummer1.7 tumor.

[0168] Figures 15A-15D show quantification of the tumor volumes over time as individualmouse (Figure 15A) or grouped together (Figure 15B), Kaplan-Meier survival curves (Figure 15C), and comparison with or without CD8+T cell deletion of mice transplanted with Yummer1.7 mouse melanoma cancer cells and treated with control mouse IgG or an anti-TTYH2 T36 antibody. Figure 15D shows the comparison of mice with or without CD8+T cell deletion antibody i.p. for a total of 3 doses. These data show that anti-TTYH2 as monotherapy controls Yummer1.7 melanoma tumor growth.

[0169] Figures 16A-16B show quantification of the tumor volumes and Kaplan-Meier survivalcurves of mice transplanted with Yummer1.7 cells (Figure 16A) or MC38 colon tumor cells (Figure 16B). From day 5, mice were treated twice weekly with control mouse IgG or 200 µg of anti-TTYH2 T36 antibody, for a total of 5 doses, with or without 100 µg of anti-mouse PD-1RMP1-14 antibody, intraperitoneally (i.p.), for a total of 3 doses. These data show that TTYH2antibody has single agent effects in Yummer 1.7 tumor, and are very efficacious in combination with anti-PD-1 therapy to markedly control both Yummer 1.7 and MC38 tumor growth.

[0170] Figures 17A-17B show quantification of the tumor volumes (Figure 17A) and Kaplan-Meier survival curves (Figure 17B) of mice transplanted with MC38 cells. From day 5 after tumorinoculation, mice were treated twice weekly with control mouse IgG antibody or 200 µg anti- TTYH2 T36 antibody intraperitoneally (i.p.), for a total of 5 doses, with or without intra-tumoral 32 314460903v1Attorney Docket No: 243735.000428 injection of 5 µg STING agonist 2’3’-cGAMP on day 5 and day10. These data show that anti- TTYH2 controls MC38 tumor growth in the presence of STING agonist.

[0171] Figures 18A-18B show quantification of the tumor volumes (Figure 18A) and Kaplan-Meier survival curves (Figure 18B) of C57BL / 6 mice transplanted with KrasG12D, Trp53R172H, Pdx-1-Cre (KPC) pancreatic cancer cells. From day 5 after tumor inoculation, mice were intraperitoneally (i.p.) treated with control mouse IgG, or 200 µg anti-TTYH2 T36 antibody, or a combination of 200 µg anti-PD1 RMP1-14 in combination with 200 µg anti-LAG3 antibody, twice weekly, for a total of 4 doses. These data show that anti-TTYH2 controls tumor growth better than anti-PD-1 / LAG-3 combo in a pancreatic cancer model.

[0172] Figures 19A-19B show quantification of the tumor volumes in WT C57BL / 6 (Figure19A) and STING knockout (Figure 19B) mice transplanted with Yummer1.7 cells. From day 5, mice were treated twice weekly with 200 µg anti-TTYH2 antibody T36, or a combination of 200 µg anti-TTYH2 antibody T36 and anti-mouse IFNAR1 blocking antibody (MAR1-5A3), or mIgG2a control antibody intraperitoneally (i.p.) as indicated for a total of 4 doses (Figure 19A). These data demonstrate that the effect of anti-TTYH2 is dependent on STING / type-I IFN pathway.

[0173] Figures 20A-20C show quantification of the tumor volumes (Figures 20A-20B) andKaplan-Meier survival curves (Figure 20C) of mice transplanted with MC38 cells. From day 5, mice were treated twice weekly with 200 µg anti-TTYH2 (T36) / 100 µg PD-1(RMP1-14) combinational therapy, or anti-TTYH2(T36) / anti-PD-1(RMP1-14) combinational therapy plus 200 µg anti-mouse IFNAR1 blocking antibody (MAR1-5A3), or mIgG2a control antibody intraperitoneally (i.p.) for a total of 4 doses. These data demonstrate that the effect of anti- TTYH2 / PD-1 combinational therapy is dependent on type-I IFN.

[0174] Figures 21A-21C show quantification of the tumor volumes (Figures 21A-21B) andKaplan-Meier survival curves (Figure 21C) of mice transplanted with MC38 cells in STING knockout mice. From day 5, mice were treated twice weekly with 200 µg anti-TTYH2 T36, 100 µg anti-mouse PD-1 (RMP1-14), anti-TTYH2(T36) / anti-PD-1(RMP1-14) combinational therapy, or mIgG2a control antibody intraperitoneally (i.p.) for a total of 4 doses. These data demonstrate that the effect of anti-TTYH2 / PD-1 combinational therapy is dependent on STING.

[0175] Figures 22A-22C show results demonstrating that TTYH2 deficiency alleviatesExperimental autoimmune encephalomyelitis (EAE) symptoms and promotes Type-I IFN production in microglia (Figures 22A-22C). Figure 22A shows an EAE progression in WT or 33 314460903v1Attorney Docket No: 243735.000428 TTYH2 KO mice. Figure 22B shows an EAE progression in anti-TTYH2 or control antibody treated WT mice. Figures 22A-22B depict the data demonstrating the effect of TTYH2 in experimental autoimmune encephalomyelitis (EAE) by analyzing mean clinical score in TTYH2KO and WT mice, as well as anti-TTYH2 or control antibody treated WT mice. Figure 22Cdepicts the level of cytokines and chemokines in the TTYH2 deficient mouse microglial cells after stimulation with poly(dA:dT).

[0176] Figures 23A-23C depict a central concept of targeting immune feedback modulators forimmunotherapy. PD-1 / PD-L1 represents a negative feedback modulator in the disease tissue (Figure 23A), and the concept of TTYH2 as a TLR or STING induced myeloid negative feedback modulator (Figures 23B-23C).

[0177] Figures 24A-24B represent an approach involving anti-TTYH2-based feedbackimmunotherapies for cancer and autoimmunity versus recombinant Type-I IFN therapy.

[0178] Figures 25A-25C show the results of functional screening of myeloid-associated humanmembrane proteins, identifying TTYH2 as a negative regulator for IRF3 / NF-κB responses. cDNAs coding ~300 human myeloid-associated membrane proteins were individually overexpressed in human HEK-293T-based IRF3 / NF-κB reporter cells (HEK-Dual™ cells, invivogen). Figure 25A shows the data of IRF3 luciferase reporter signal upon Poly(dA:dT) stimulation. TTYH2 was among the top hits inhibiting IRF3 reporter signal. Figures 25B-25C show significant suppression of TTYH2 among the three TTYH Tweety family members, in the functional screening assay stimulated by Poly(dA:dT) (Figure 25B) or Poly(I:C) (Figure 25C).

[0179] Figures 26A-26B show evidence of TTYH2 as a homodimer in cis and in trans. Figure26A depicts a human TTYH2 structure that shows cis (protein data bank (PDB) ID: 7RTT) and trans (PDB ID: 7RTU) homodimers from the PDB database. Figure 26B depicts the results of a fluorescence resonance energy transfer (FRET) experiment demonstrating that TTYH2 can form cis-dimers.

[0180] Figure 27 shows RNA expression of TTYH2 in human peripheral immune cells (adaptedfrom the Human Protein Atlas (HPA) database). The dominant expression of TTYH2 was observed in human peripheral myeloid cells.

[0181] Figures 28A-28B show the dominant expression of TTYH2 in mouse myeloid cells. Datashown is the RNA expression of TTYH2 in mouse immune cells, with peritoneal macrophages (PMs) have the highest expression (Figure 28A), and induction by Toll-like receptor stimulation 34 314460903v1Attorney Docket No: 243735.000428 in bone-marrow-derived macrophages (e.g., LPS) (Figure 28B). (adapted from the Biology Gene Portal Services (BioGPS) database).

[0182] Figures 29A-29C show TTYH2 as a negative regulator for myeloid cell-associatedresponses. Mouse peritoneal macrophages (PMs) were isolated from WT or TTYH2 knockout mice, then stimulated with STING agonist DMXAA (5 µg / mL) for 3 hours, cytokines in the supernatant was analyzed (Figure 29A). STING agonist 2’3’-cGAMP (10 mg / kg) was intraperitoneal injected to TTYH2 knockout (KO) or wild-type (WT) littermates, serum was collected 3 hours later for cytokine / chemokine analysis (Figures 29B-29C).

[0183] Figure 30 shows data from RNAseq analysis demonstrating that TTYH2 KOmacrophages greatly shift to M1 phenotype in static condition. Mouse PMs were isolated from WT or TTYH2 KO mice. RNA sequencing data showed that TTYH2 KO PMs in static condition have higher expression of M1 macrophage signature genes (e.g., B2m, Lpl, Clec7a, and Siglec-1), and fewer M2 macrophage signature genes (e.g., Arg1, Fn1, Ncf1, and Atp2b4) compared to WT PMs.

[0184] Figures 31A-31C show that TTYH2 KO macrophages maintain M1 phenotype afterstimulation. Mouse PMs were isolated from WT or TTYH2 KO mice. RNA sequencing data showed that TTYH2 KO PMs expressed more Type-I IFN related genes (e.g., Irf8, Cmpk2, Cxcl9 and Gbp2) upon STING agonist DMXAA treatment (Figures 31A-31C).

[0185] Figures 32A-32B show suppression of NF-κB activation due to human TTYH2overexpression and reversion of NF-κB activation by anti-TTYH2 antibody. Figure 32B depicts a bar plot of fluorescent intensity reading data.

[0186] Figures 33A-33C show that TTYH2 deficiency affects polarization of tumor-associatedmacrophages. Figure 33A depicts an experiment setting for the single-cell RNA sequencing of tumors from both TTYH2 KO or WT mice. Figure 33B depicts UMAP of tumor infiltrating CD45+cells from 2 WT and 3 KO mice at day 10 after tumor injection. The frequencies of the identified cell types in WT and KO mice are indicated. Figure 33C depicts that the M2-like macrophages were significantly reduced, while M1-like macrophages were increased, in TTYH2 KO tumors.

[0187] Figures 34A-34D depict the results of UMAP and Gene Set Enrichment Analysis(GSEA) demonstrating that TTYH2 regulates tumor-associated macrophage signature genes expression. Figure 34A depicts expression levels of selected M1 and M2 marker genes. Figures 34B-34C depict top 10 upregulated pathways in GSEA of M1 and M2-like macrophage clusters. 35 314460903v1Attorney Docket No: 243735.000428 Pathway analysis was performed using the Hallmark and Gene Ontology Biological Process (GOBP) databases. Pathways with a p-value of p<0.05 were ranked by normalized enrichment score (NES) calculated from the comparison between Ttyh2 KO vs WT. Top 6 upregulated and downregulated regulators in IPA upstream analysis of M1 and M2-like macrophage clusters are indicated. Figure 34D depicts regulators with a p-value of p<0.05 were ranked by Z score calculated from the comparison between TTYH2 KO vs WT. The intensity of the shading in the Figure 34D reduces as the Z-score decreases.

[0188] Figures 35A-35B show the results of UMAP plot and frequencies of the lymphocyticsubsets demonstrating that TTYH2 deficiency affects intratumoral CD8+T cell / Treg balance. Figure 35A depicts UMAP plot of five tumor-associated lymphocytic subsets identified by sub- clustering cells from clusters 7, 9, and 10 (Figure 34). Figure 35B depicts frequencies of the identified lymphocytic subsets in TTYH2 WT and KO mice.

[0189] Figures 36A-36B show quantification of the tumor volumes (Figure 36A) and Kaplan-Meier survival curves (Figure 36B) of mice transplanted with A375 human melanoma cell line in NOD scid gamma mouse (NSG) mice. These data show that TTYH2 antibody therapy markedly controls human melanoma in NSG mice.

[0190] Figures 37A-37B depict the effect of anti-TTYH2 prophylactic or therapeutic treatmentfor experimental autoimmune encephalomyelitis (EAE). Figure 37A depicts the potential role of Type-I IFN in the control of multiple sclerosis (MS). Figure 37B depicts the effect of anti-TTYH2 in EAE, a mouse model of MS.

[0191] Figure 38 depicts the results of hematoxylin and eosin staining (H&E) andimmunohistochemistry (IHC) staining, demonstrating that TTYH2 deficiency reduces inflammation in the spinal cord. DETAILED DESCRIPTION

[0192] As described in Background section above, there is a need to understand how PD-L1 / PD-1 works to identify new targets that can overcome the limitations of the current therapies. The key principles of PD-1 / PD-L1 regulation are to: 1) target immune regulation in the tumor-site; 2) target key tumor-immune negative feedback modulators; and 3) recover or repair polyclonal T cell responses. In this setting, tumor cells express PD-L1 in response to IFN-γ from activated T cells. Subsequently, PD-L1 interacts with the PD-1 receptor on activated T cells, leading to the suppression of T cell activity, creating a negative feedback loop within the TME. Therefore, the 36 314460903v1Attorney Docket No: 243735.000428 inventors of the present disclosure aimed to identify key tumor-immune negative feedback modulators beyond the PD-1 / PD-L1 pathway that critically affect tumor immunity in the TME, and to develop novel targeted cancer immunotherapies (Figure 1A). The PD-1 / PD-L1 pathway represents immune checkpoints and key targets for both cancer immunotherapy (antagonists) and autoimmune diseases (agonists). However, therapies targeting many other immune checkpoints beyond PD-1 / PD-L1 do not work effectively to date (except anti-CTLA-4 and LAG-3 therapies). Mechanistically, PD-1 / PD-L1 is not a simple immune checkpoint, it forms a negative feedback loop in the disease microenvironment (Figure 23A). One of the aspects of the present disclosure is to focus on identifying PD-1 / PD-L1-like key immune feedback modulators in the disease tissue, rather than general immune checkpoints, for immunotherapy. In this manner, an immunity can be repaired in the disease lesion and achieve better efficacy with reasonable toxicity profile of an immunotherapy for a cancer, an autoimmunity, and other human diseases (Figure 23B). It is important to address these unmet clinical needs to overcome immunotherapy resistance by identifying novel immune checkpoints and / or immune feedback modulators and their associated molecular mechanisms.

[0193] Type I IFN is critical for tumor immunity; however, its use is also limited by highimmuno-toxicity, poor tolerability of regimes for patients, and modest efficacy through systemic administration. Type I IFNs play a crucial role in the immune system’s response to a broad spectrum of human diseases. Other than defense to pathogens, their major functions include: 1) Stimulation of immune responses: Type I IFNs enhance the immune response by promoting the maturation and activation of dendritic cells (DCs), monocytes, macrophages, natural killer (NK) cells, as well as B cell antibody responses. They also facilitate the differentiation of CD4+T cells into Th1 cells, the priming of CD8+cytotoxic T cells2, which are crucial for cellular immunity particularly against tumor cells. 2) Regulation of inflammation: Type I IFNs modulate inflammation by regulating the production of pro-inflammatory cytokines and chemokines, such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 13 (IL-13), interleukin 17A (IL-17A), interleukin 17F (IL-17F), chemokine (C- C motif) ligand 2 (CCL2), C-X-C motif chemokine ligand 10 (CXCL10),C-X-C motif chemokine ligand 10 (CXCL10), C-C motif chemokine ligand 19 (CCL19) etc., antigen presentation machinery, and costimulatory molecules. They can also paradoxically promote an anti- inflammatory environment by inducing immune suppressive molecules and immune cells, 37 314460903v1Attorney Docket No: 243735.000428 depending on the specific disease or conditions.3) Anti-proliferative function and the induction of Apoptosis: Type I IFNs can induce apoptosis (programmed cell death) in host cells including cancer cells, promoting the elimination of tissue cells and cancer cells by the immune system. Chronic IFN exposure may be also detrimental to immune responses, such as T cells due to enhanced apoptosis. Use of recombinant Type I Interferons in therapeutics

[0194] In cancers: From the 1980s onward, members of Type I IFN family have been thestandard care as immunotherapeutic agents in cancer therapy. In particular, IFNα has been approved by the US Food and Drug Administration (FDA) for cancer. To date, pharmaceutical companies produce several types of recombinant and pegylated IFNα for clinical use; e.g., IFNα2a (Roferon-A, Roche), IFNα2b (Intron-A, Schering-Plough) and pegylated IFNα2b (Sylatron, Schering Corporation) for treatment of hairy cell leukemia, melanoma, renal cell carcinoma, Kaposi’s sarcoma, multiple myeloma, follicular and non-Hodgkin lymphoma, and chronic myelogenous leukemia (Lazear et al.,Immunity.,2019 Apr 16;50(4):p907-923). Human IFNβ (Feron, Toray ltd.) has also been approved in Japan to treat glioblastoma, medulloblastoma, astrocytoma, and melanoma.

[0195] Combinational therapy with PD-1 / PD-L1 inhibitors: By combining PD-1 / PD-L1inhibitors with Type I interferons, researchers aim to tackle multiple resistance mechanisms and enhance the overall anti-tumor immune response. The approach is supported by preclinical and clinical studies that show promising synergistic effects, particularly in melanoma and renal carcinoma. These studies reveal increased infiltration and activation of T cells within the tumor microenvironment, the development of memory T cells, and prolonged patient survival. There is no combinational therapy approved yet possibly due to low efficacy and high immuno-toxicity.

[0196] In multiple sclerosis and other inflammatory diseases: Currently, there are four FDAapproved variants of IFN-β1 in use as a treatment for relapsing multiple sclerosis. IFN-β1 is not an appropriate treatment for patients with progressive, non-relapsing forms of multiple sclerosis. Whilst the mechanism of action is not completely understood, the use of IFN-β1 has been found to reduce brain lesions, increase the expression of anti-inflammatory cytokines and reduce T cell infiltration into the brain. Moreover, IFNα has been used in the treatment of Behcet’s uveitis or Behcet’s disease. Role of Type I IFN in protecting multiple sclerosis 38 314460903v1Attorney Docket No: 243735.000428

[0197] The mechanism of recombinant Type I IFN in protecting multiple sclerosis involves theirability to modulate inflammation and regulate immune responses, preserve Blood-Brain Barrier (BBB) integrity, as well as promote remyelination etc., making them an important therapeutic target in the management of MS. Specifically, 1) anti-Inflammatory Effects: Type I IFNs help to dampen the inflammatory response in the central nervous system (CNS) by reducing the activation and proliferation of pro-inflammatory immune cells such as T cells and macrophages. This modulation of inflammation helps to prevent damage to myelin and nerve fibers in MS.2) Immune Regulation: Type I IFNs promote the differentiation of regulatory T cells (Tregs), which are crucial for maintaining immune tolerance and preventing autoimmune attacks against self-antigens, including myelin proteins in MS.3) Reduction of Blood-Brain Barrier Permeability: Type I IFNs can enhance the integrity of the blood-brain barrier (BBB), limiting the infiltration of immune cells into the CNS and reducing the risk of inflammatory lesions and demyelination.4) Promotion of Remyelination: Type I IFNs have been shown to stimulate the production and differentiation of oligodendrocyte precursor cells (OPCs), which contribute to remyelination and repair of damaged myelin sheaths in MS lesions.

[0198] Compared to PD-1 immunotherapy approved for many cancer types, Type I IFN is onlyapproved for some cancers, such as hairy leukemia, kidney cancer, and melanoma, with weak efficacy. There are significant unmet clinical needs to identifying pathways that can be used to design more effective approaches to enhance Type I IFN activity preferentially at the tumor site.

[0199] The present disclosure is based on the central hypothesis that tumors may upregulatemembrane-associated immune regulators that form negative feedback loops to evade key pathways of tumor immunity, such as the production of Type I IFN essential for innate sensing, T cell priming2,9and targeting these pathways could recover endogenous immune responses with beneficial efficacy to side-effect ratio to facilitate the control of cancer and other human diseases (Figure 1A).

[0200] Recombinant Type-I IFNs (IFN-α / IFN-β) have been therapeutically approved for bothcancer (e.g., Melanoma, Renal-cell carcinoma, leukemia, lymphoma etc.) and autoimmune diseases (e.g., multiple sclerosis, and some off-label use for Behcet’s disease). However, systemic injection of Type-I IFNs broadly activates the immune system, leading to significant immuno- toxicities and limited efficacy (Figure 24A). Anti-TTYH2 antagonists represent a different approach through targeting disease-associated negative immune feedback modulators, which 39 314460903v1Attorney Docket No: 243735.000428 recovering endogenous myeloid cell responses (e.g., Type-I IFNs) (Figure 24B). As illustrated in the present disclosure, TTYH2 was identified as a negative feedback modulator for myeloid cell activities, particularly for Type-I IFN responses that are critical for innate sensing and the priming of adaptive immunity, such as T cells (Figure 23C).

[0201] Type-I interferons (IFNs, IFN-α / IFN-β) are key cytokine mediators bridging innate andadaptive immunity. While systemic administration of recombinant type-I IFNs and STING agonists directly amplifying Type-I IFNs has been extensively explored, their efficacy is often limited by dose-dependent immuno-toxicities. Restoring endogenous type-I IFNs in diseased tissues remains a major challenge for immunotherapy. As illustrated herein, TTYH2, a poorly characterized five-transmembrane protein was identified as a myeloid-associated inhibitor of type- I IFN responses. TTYH2 was weakly expressed in peripheral myeloid cells but was upregulated upon activation, where it potently suppressed the downstream production of type-I IFN and other proinflammatory cytokines. TTYH2 also controls other aspects of immune responses, such as TNF-α, IL-6, IL-10, CCL2, etc., as well as myeloid cell differentiation. Thus, the TTYH2 protein may function as myeloid inhibitor. Mechanistically, TTYH2 deficiency enhances both NF-κB and IRF3-mediated signaling and markedly promotes M1 macrophage polarization, and type-I IFN- driven antigen-specific T cell responses. As illustrated herein, TTYH2 antibody blockade targeting the FAARGDY (SEQ ID NO: 25) motif in the second ectodomain greatly promoted STING / type- I IFN-dependent tumor immunity, particularly in cold tumors, and provided potent synergy with immunotherapies. Furthermore, TTYH2 was overexpressed across multiple cancers and associated with patient survival and immunotherapy responses. Both TTYH2 deficiency and antibody blockade also significantly ameliorated disease symptoms and inflammation in mouse models of multiple sclerosis, in which Type-I IFN is therapeutic to the diseases. These findings of the present disclosure establish a myeloid-associated negative feedback modulator and a promising immunotherapeutic target for cancer and autoimmunity.

[0202] The studies described herein illustrate TTYH2 (Nalamalapu RR, Yue M, Stone AR,Murphy S, Saha MS. The tweety Gene Family: From Embryo to Disease. Front Mol Neurosci. 2021 Jun 28;14:672511) as a cell-surface negative feedback regulator for the STING / Type I IFN pathway. The present disclosure illustrates that TTYH2 is specifically expressed in myeloid cells over lymphoid immune subsets and most of the tissues. The studies described herein illustrate that TTYH2 is induced by STING or Toll-like receptor agonists, is mainly expressed on tumor- 40 314460903v1Attorney Docket No: 243735.000428 associated myeloid cells and some cancer cells, and suppresses Type I IFN production. Mechanistically, TTYH2 was identified as a receptor which plays a role in inhibiting NF-κB and TBK1 / IRF3 signaling critical for Type I IFN production. TTYH2 knockout mice or antibody blockade was also found to amplify T cell responses and tumor immunity in a STING / Type I IFN- dependent manner, as well as in a pancreatic cold tumor model.

[0203] The studies described herein illustrate the upregulation of TTYH2 in multiple humancancer types, such as melanoma, glioblastoma, acute myeloid leukemia, and Thymoma and Thymic Carcinoma, and that TTYH2 is mainly expressed on tumor-associated M2 macrophages and some cancer cells in the TME. Importantly, TTYH2 knock-out (KO) mice showed amplified T cell responses in a Type I IFN dependent manner. Moreover, TTYH2 KO demonstrated functional restoration of tumor immunity across multiple syngeneic tumor models. TTYH2 specific antibodies of the present disclosure that bind to unique epitopes of TTYH2, are cross- reactive to human and mouse antigens, and can antagonize the inhibitory function of TTYH2. TTYH2 antibody blockade strongly boosted tumor immunity in a STING / Type I IFN-dependent manner, and demonstrated therapeutic effects in a pancreatic cold tumor model having little response to PD-1 / LAG-3 checkpoint blockade immunotherapies. Anti-TTYH2 blockade was also found to provide synergistic effects with PD-1 checkpoint blockade and STING agonists. Moreover, TTYH2 antibody blockade was effective in a mouse model of multiple sclerosis (MS) where Type I IFNs can protect against disease. Definitions

[0204] The term “antibody” refers to an immunoglobulin molecule capable of specific bindingto a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site, located in the variable region(s) of the immunoglobulin molecule. As used herein, the term “antibody”, e.g., anti-TTYH2 antibody, encompasses not only intact (e.g., full- length) polyclonal or monoclonal antibodies, but also antigen-binding fragments thereof (such as Fab, Fab’, F(ab’)2, Fv), single chain (scFv), mutants thereof, fusion proteins comprising an antibody portion, humanized antibodies, chimeric antibodies, diabodies, nanobodies, linear antibodies, single chain antibodies, multi-specific antibodies (e.g., bispecific antibodies) and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site of the required specificity, including glycosylation variants of antibodies, amino 41 314460903v1Attorney Docket No: 243735.000428 acid sequence variants of antibodies, and covalently modified antibodies. An antibody, e.g., anti- TTYH2 antibody, includes an antibody of any class, such as IgD, IgE, IgG, IgA, or IgM (or sub- class thereof), and the antibody need not be of any particular class. Depending on the antibody amino acid sequence of the constant domain of its heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy-chain constant domains that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0205] A typical antibody molecule comprises a heavy chain variable region (HCVR or VH)and a light chain variable region (LCVR or VL), which are usually involved in antigen binding. The VH and VL regions can be further subdivided into regions of hypervariability, also known as “complementarity determining regions” (“CDR”), interspersed with regions that are more conserved, which are known as “framework regions” (“FR”). Each VH and VL is typically composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the framework region and CDRs can be precisely identified using methodology known in the art, for example, by the Kabat definition, the Chothia definition, the AbM definition, the EU definition, the “Contact” numbering scheme, the “IMGT” numbering scheme, the “Aho” numbering scheme, and / or the contact definition, all of which are well known in the art. (See, e.g., Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No.91-3242, Chothia et al., (1989) Nature 342:877; Chothia, C. et al. (1987) J. Mol. Biol.196:901-917, Al-lazikani et al. (1997) J. Molec. Biol.273:927-948; Edelman et al., Proc Natl Acad Sci U S A.1969 May;63(1):78-85; and Almagro, J. Mol. Recognit.17:132-143 (2004); MacCallum et al., J. Mol. Biol.262:732-745 (1996), Lefranc M P et al., Dev Comp Immunol, 2003 January; 27(1):55-77; and Honegger A and Pluckthun A, J Mol Biol, 2001 Jun.8; 309(3):657-70. See also hgmp.mrc.ac.uk and bioinf.org.uk / abs).

[0206] In some embodiments, the anti-TTYH2 antibody described herein is a full-lengthantibody, which contains two heavy chains and two light chains, each including a variable domain and a constant domain. Alternatively, the anti-TTYH2 antibody can be an antigen-binding 42 314460903v1Attorney Docket No: 243735.000428 fragment of a full-length antibody. Examples of binding fragments encompassed within the term “antigen-binding fragment” of a full length antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab’)2 fragment, a bivalent fragment including two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544- 546), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR) that retains functionality. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules known as single chain Fv (scFv). See e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883.

[0207] Any of the antibodies described herein, e.g., anti-TTYH2 antibody, can be eithermonoclonal or polyclonal. A “monoclonal antibody” refers to a homogenous antibody population and a “polyclonal antibody” refers to a heterogeneous antibody population. These two terms do not limit the source of an antibody or the manner in which it is made.

[0208] As used herein “specifically binds”, “specific binding”, “specifically recognizes” or“specifically recognition” refers to the ability of the antibodies or antigen-binding fragments of the disclosure to bind to a predetermined antigen (e.g., TTYH2 protein) with a dissociation constant (KD) of, for example, about 1×10−6M or less, for example about 1×10−7M or less, about 1×10−8M or less, about 1×10−9M or less, about 1×10−10M or less, about 1×10−11M or less, about 1×10−12M or less, or about 1×10−13M or less. In some embodiments, the KD may be, for example, about 1×10-8or less. As another example, a KD described herein may be about 2×10−7M or less, about 2×10−8M or less, about 2×10−9M or less, about 2×10−10M or less, about 2×10−11M or less, about 2×10−12M or less, or about 2×10−13M or less. In some embodiments, the KD may be, for example, about 2×10-9or less. Typically, the antibody or antigen-binding fragment binds to an antigen (e.g., TTYH2 protein) with a KDthat is at least ten-fold less than its KDfor a nonspecific antigen (for example BSA or casein) as measured by e.g., surface plasmon resonance using for example a Proteon Instrument (BioRad).

[0209] “Isolated” means a biological component (such as a nucleic acid, peptide or protein) hasbeen substantially separated, produced apart from, or purified away from other biological 43 314460903v1Attorney Docket No: 243735.000428 components of the organism in which the component naturally occurs, i.e., other chromosomal and extrachromosomal DNA and RNA, and proteins. Nucleic acids, peptides, and proteins that have been “isolated” thus include nucleic acids and proteins purified by standard purification methods. “Isolated” nucleic acids, peptides, and proteins can be part of a composition and still be isolated if such composition is not part of the native environment of the nucleic acid, peptide, or protein. The term also embraces nucleic acids, peptides, and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids in the polynucleotides that encode for an antibody or antigen-binding fragment described herein. An “isolated” antibody or antigen-binding fragment, as used herein, is intended to refer to an antibody or antigen-binding fragment which is substantially free of other antibodies or antigen-binding fragments having different antigenic specificities; for instance, an antibody that specifically binds to an intended antigen (e.g., TTYH2 protein) can be substantially free of other antibodies that specifically bind antigens other than the intended antigen (e.g., TTYH2 protein).

[0210] The term “polynucleotide” as referred to herein means a polymeric form of nucleotidesof at least 10 bases in length, either ribonucleotides or deoxyribonucleotides or a modified form of either type of nucleotide. The term includes single and double stranded forms.

[0211] The term “isolated polynucleotide” as used herein means a polynucleotide of genomic,cDNA, or synthetic origin or some combination thereof, which by virtue of its origin or source of derivation, the “isolated polynucleotide” has one to three of the following: (1) is not associated with all or a portion of a polynucleotides with which the “isolated polynucleotide” is found in nature, (2) is operably linked to a polynucleotide to which it is not linked in nature, or (3) does not occur in nature as part of a larger sequence. “Operably linked” sequences include both expression control sequences that are contiguous with the gene of interest and expression control sequences that act in trans or at a distance to control the gene of interest.

[0212] The term “expression control sequence” as used herein means polynucleotide sequencesthat are necessary to effect the expression and / or processing of coding sequences to which they are ligated. Expression control sequences include, without limitation, appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (e.g., Kozak consensus sequence); sequences that enhance protein stability; and when desired, sequences that enhance protein secretion. The nature of such 44 314460903v1Attorney Docket No: 243735.000428 control sequences differs depending upon the host organism; in prokaryotes, such control sequences generally, can include promoter, ribosomal binding site, and transcription termination sequences; in eukaryotes, generally, such control sequences can include promoters and transcription termination sequence.

[0213] The term “control sequences” is intended to include, at a minimum, all componentswhose presence is essential for expression and / or processing, and can also include additional components whose presence is advantageous, for example, leader sequences and fusion partner sequences.

[0214] The term “vector”, as used herein, means a vehicle capable of transporting a nucleic acidinto a host cell. In some embodiments, the vector is a plasmid, e.g., a circular double stranded DNA loop into which additional DNA segments may be ligated. In some embodiments, the vector is a viral vector, and additional DNA segments may be ligated into the viral genome. In some embodiments, the vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). In other embodiments, the vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “recombinant expression vectors” (or simply, “expression vectors”).

[0215] The term “promoter” as used herein is defined as a DNA sequence recognized by thesynthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence. As used herein, the term “regulatory sequence” means a nucleic acid sequence which can regulate expression of a gene product operably linked to the regulatory sequence. In some instances, this sequence may be the core promoter sequence and in other instances, this sequence may also include an enhancer sequence and other regulatory elements which are required for expression of the gene product. The promoter or regulatory sequence may, for example, be one which expresses the gene product in a tissue specific manner.

[0216] The term “recombinant host cell” (or simply “host cell”), as used herein, means a cellinto which an exogenous nucleic acid and / or recombinant vector has been introduced. It should be understood that “recombinant host cell” and “host cell” mean not only the particular subject cell but also the progeny of such a cell. Because certain modifications may occur in succeeding 45 314460903v1Attorney Docket No: 243735.000428 generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term “host cell” as used herein.

[0217] The term “percent sequence identity” means a ratio, expressed as a percent of the numberof identical residues over the total number of residues compared. Sequence identity for nucleic acid sequences may be analyzed over a stretch of at least about nine nucleotides, usually at least about 18 nucleotides, more usually at least about 24 nucleotides, typically at least about 28 nucleotides, more typically at least about 32 nucleotides, and preferably at least about 36, 48, or more nucleotides. There are a number of different algorithms known in the art which can be used to measure nucleotide sequence identity. For instance, polynucleotide sequences can be compared using FASTA, Gap or Bestfit, which are programs in Wisconsin Package Version 10.0, Genetics Computer Group (GCG), Madison, Wis. FASTA, which includes, e.g., the programs FASTA2 and FASTA3, provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson, Methods Enzymol.183:63-98 (1990); Pearson, Methods Mol. Biol. 132:185-219 (2000); Pearson, Methods Enzymol. 266:227-258 (1996); Pearson, J. Mol. Biol. 276:71-84 (1998); herein incorporated by reference). Unless otherwise specified, default parameters for a particular program or algorithm are used. For instance, percent sequence identity between nucleic acid sequences can be determined using FASTA with its default parameters (a word size of 6 and the NOPAM factor for the scoring matrix) or using Gap with its default parameters as provided in GCG Version 6.1, herein incorporated by reference. A reference to a nucleotide sequence encompasses its complement unless otherwise specified. Thus, a reference to a nucleic acid having a particular sequence should be understood to encompass its complementary strand, with its complementary sequence. Sequence identity for polypeptides, is typically measured using sequence analysis software. Protein analysis software matches sequences using measures of similarity assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For instance, GCG contains programs such as “Gap” and “Bestfit” which can be used with default parameters, as specified with the programs, to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild-type protein and a mutein thereof. See, e.g., GCG Version 6.1. Polypeptide sequences also can be compared using FASTA using default or recommended parameters, see GCG Version 6.1. (University of 46 314460903v1Attorney Docket No: 243735.000428 Wisconsin Wis.) FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson, Methods Enzymol.183:63-98 (1990); Pearson, Methods Mol. Biol.132:185-219 (2000)). Another preferred algorithm when comparing a sequence of the invention to a database containing a large number of sequences from different organisms is the computer program BLAST, especially blastp or tblastn, using default parameters, as supplied with the programs. See, e.g., Altschul et al., J. Mol. Biol.215:403-410 (1990); Altschul et al., Nucleic Acids Res.25:3389-402 (1997).

[0218] The length of polypeptide sequences compared for homology will generally be at leastabout 16 amino acid residues, usually at least about 20 residues, more usually at least about 24 residues, typically at least about 28 residues, and preferably more than about 35 residues. When searching a database containing sequences from a large number of different organisms, it is preferable to compare amino acid sequences.

[0219] The term “substantial similarity” or “substantial sequence similarity”, when referring toa nucleic acid or fragment thereof, means that when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 85%, preferably at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the nucleotide bases, as measured by any well-known algorithm of sequence identity, such as FASTA, BLAST, or Gap, as discussed above. As applied to polypeptides, the term “substantial identity” means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, as supplied with the programs, share at least 70%, 75%, 80% or 85% sequence identity, preferably at least 90%, 91%, 92%, 93%, 94% 95%, 96%, 97%, 98% or 99% sequence identity. In certain embodiments, residue positions that are not identical differ by conservative amino acid substitutions.

[0220] A “conservative amino acid substitution” is one in which an amino acid residue issubstituted by another amino acid residue having a side chain R group with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well-known to those of skill in the art. See, e.g., Pearson, Methods Mol. Biol. 243:307-31 (1994). Examples of groups of amino acids that have side chains with 47 314460903v1Attorney Docket No: 243735.000428 similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic-hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine and methionine. Conservative amino acids substitution groups include: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine- arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine. Alternatively, a conservative substitution or replacement, as the terms are used interchangeably herein, is any change having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al., Science 256:1443-45 (1992), herein incorporated by reference. A “moderately conservative” replacement is any change having a nonnegative value in the PAM250 log-likelihood matrix.

[0221] The terms “treat” or “treatment” of a state, disease, disorder, or condition, or the like,include: (1) preventing, delaying, or reducing the incidence and / or likelihood of the appearance of at least one clinical or sub-clinical symptom of the state, disease, disorder or condition developing in a subject that may be afflicted with or predisposed to the state, disease, disorder, or condition but does not yet experience or display clinical or subclinical symptoms of the state, disease, disorder, or condition; or (2) inhibiting the state, disease, disorder, or condition, e.g., arresting, reducing, or delaying the development of the state, disease, disorder, or condition; or a relapse thereof (in case of maintenance treatment) or at least one clinical or sub-clinical symptom thereof; or (3) relieving the state, disease, disorder, or condition, e.g., by causing regression of the state, disease, disorder, or condition or at least one of its clinical or sub-clinical symptoms. The benefit to a subject to be treated is either statistically significant or at least perceptible to the patient or to the physician.

[0222] As used herein, the term “therapeutically effective” applied to dose or amount refers tothat quantity of a compound or pharmaceutical composition that is sufficient to result in a desired activity upon administration to a subject in need thereof. Within the context of the present invention, the term “therapeutically effective” refers to that quantity of a compound or pharmaceutical composition that is sufficient to reduce or eliminate at least one symptom of a cancer, inflammation, autoimmune, or related disorder. Note that when a combination of active ingredients is administered the effective amount of the combination may or may not include amounts of each ingredient that would have been effective if administered individually. 48 314460903v1Attorney Docket No: 243735.000428

[0223] As used herein, the phrase “pharmaceutically acceptable” refers to molecular entities andcompositions that are generally regarded as physiologically tolerable.

[0224] As used herein, the term “combination” of a compound or a composition, and at least asecond pharmaceutically active ingredient means at least two, but any desired combination of compound or composition can be delivered simultaneously or sequentially.

[0225] The terms “patient”, “individual”, “subject”, and “animal” are used interchangeablyherein and refer to mammals, including, without limitation, human and veterinary animals (e.g., cats, dogs, cows, horses, goats, sheep, pigs, etc.) and experimental animal models (e.g., rodents such as mice and rats, rabbits, and non-human primates). In a preferred embodiment, the subject is a human.

[0226] As used herein, the term “healthy subject” refers to a subject that is without knowninfections or autoimmune disorders or cancer by using conventional diagnostic methods. In certain embodiments, a healthy subject is a subject without a known first degree relative with an autoimmune disorder. In certain embodiments, a healthy subject is a subject without a known first degree relative with a cancer. In certain embodiments, a matched healthy subject is matched by age, gender, and / or ethnicity.

[0227] The term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which thecompound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water or aqueous solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions. Alternatively, the carrier can be a solid dosage form carrier, including but not limited to one or more of a binder (for compressed pills), a glidant, an encapsulating agent, a flavorant, and a colorant. Suitable pharmaceutical carriers are described in “Remington’s Pharmaceutical Sciences” by E.W. Martin.

[0228] The term “about” or “approximately” means within a statistically meaningful range of avalue. Such a range can be within an order of magnitude, preferably within 50%, more preferably within 20%, still more preferably within 10%, and even more preferably within 5% of a given value or range. The allowable variation encompassed by the term “about” or “approximately” depends on the particular system under study, and can be readily appreciated by one of ordinary skill in the art. 49 314460903v1Attorney Docket No: 243735.000428

[0229] The terms “a,” “an,” and “the” do not denote a limitation of quantity, but rather denotethe presence of “at least one” of the referenced item.

[0230] The practice of the present invention employs, unless otherwise indicated, conventionaltechniques of statistical analysis, molecular biology (including recombinant techniques), microbiology, cell biology, and biochemistry, which are within the skill of the art. Such tools and techniques are described in detail in e.g., Sambrook et al. (2001) Molecular Cloning: A Laboratory Manual.3rded. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York; Ausubel et al. eds. (2005) Current Protocols in Molecular Biology. John Wiley and Sons, Inc.: Hoboken, NJ; Bonifacino et al. eds. (2005) Current Protocols in Cell Biology. John Wiley and Sons, Inc.: Hoboken, NJ; Coligan et al. eds. (2005) Current Protocols in Immunology, John Wiley and Sons, Inc.: Hoboken, NJ; Coico et al. eds. (2005) Current Protocols in Microbiology, John Wiley and Sons, Inc.: Hoboken, NJ; Coligan et al. eds. (2005) Current Protocols in Protein Science, John Wiley and Sons, Inc.: Hoboken, NJ; and Enna et al. eds. (2005) Current Protocols in Pharmacology, John Wiley and Sons, Inc.: Hoboken, NJ. Additional techniques are explained, e.g., in U.S. Patent No.7,912,698 and U.S. Patent Appl. Pub. Nos.2011 / 0202322 and 2011 / 0307437. Antibodies and Antigen-binding Fragments

[0231] The present disclosure provides antibodies, or antigen-binding fragments thereof, thatspecifically bind to tweety family member 2 (TTYH2) protein. In some embodiments, antibodies or antigen-binding fragments thereof may bind to a human TTYH2 protein which has the amino acid sequence MQAARVDYIAPWWVVWLHSVPHVGLRLQPVNSTFSPGDESYQESLLFLGLVAAVCLGLNLIFLVAYLVCA CHCRRDDAVQTKQHHSCCITWTAVVAGLICCAAVGVGFYGNSETNDGAYQLMYSLDDANHTFSGIDALVS GTTQKMKVDLEQHLARLSEIFAARGDYLQTLKFIQQMAGSVVVQLSGLPVWREVTMELTKLSDQTGYVEY YRWLSYLLLFILDLVICLIACLGLAKRSKCLLASMLCCGALSLLLSWASLAADGSAAVATSDFCVAPDTF ILNVTEGQISTEVTRYYLYCSQSGSSPFQQTLTTFQRALTTMQIQVAGLLQFAVPLFSTAEEDLLAIQLL LNSSESSLHQLTAMVDCRGLHKDYLDALAGICYDGLQGLLYLGLFSFLAALAFSTMICAGPRAWKHFTTR NRDYDDIDDDDPFNPQAWRMAAHSPPRGQLHSFCSYSSGLGSQTSLQPPAQTISNAPVSEYMNQAMLFGR NPRYENVPLIGRASPPPTYSPSMRATYLSVADEHLRHYGNQFPA (Uniport KB Accession No. Q9BSA4-1) (SEQ ID NO: 26)

[0232] In some embodiments, antibodies, or antigen-binding fragments thereof may bind to amouse TTYH2 protein which has the amino acid sequence. 50 314460903v1Attorney Docket No: 243735.000428 MPAARVEYIAPWWVVWLHSVPHLGLRLQRVDSTFSPGDETYQESLLFLGVLAAIGLGLNLIFLTVYLVCT CCCRRDHTVQTKQQESCCVTWTAVVAGLLCCAAVGVGFYGNSETNDGMHQLIYSLDNANHTFSGMDELVS ANTQRMKVDLEQHLARLSEIIAARGDYIQTLKFMQQMAGNVVSQLSGLPVWREVTTQLTKLSHQTAYVEY YRWLSYLLLFILDLVICLVTCLGLARRSKCLLASMLCCGILTLILSWASLAADAAAAVGTSDFCMAPDIY ILNNTGSQINSEVTRYYLHCSQSLISPFQQSLTTFQRSLTTMQIQVGGLLQFAVPLFPTAEKDLLGIQLL LNNSEISLHQLTAMLDCRGLHKDYLDALTGICYDGIEGLLFLGLFSLLAALAFSTLTCAGPRAWKYFINR DRDYDDIDDDDPFNPQARRIAAHNPTRGQLHSFCSYSSGLGSQCSLQPPSQTISNAPVSEYMNQAILFGG NPRYENVPLIGRGSPPPTYSPSMRPTYMSVADEHLRHYEFPS (Uniport KB Accession No. (Q3TH73- 1) (SEQ ID NO: 27)

[0233] The CDR and variable region amino acid sequence identifiers of exemplary anti- TTYH2antibodies of the present disclosure are shown in Table 1. Table 1. Amino Acid Sequence Identifiers for CDRs and Variable Regions of Exemplary Anti- TTYH2 Antibodies Antibody VH HCDR1 HCDR2 HCDR3 VL LCDR1 LCDR2 LCDR3 l n (SEQ ID (SEQ ID (SEQ ID (SEQ ID (SEQ (SEQ (SEQ (SEQ .)

[0234] Amino acid sequences of CDRs contained within the amino acid sequences of HCVRsand LCVRs of the present disclosure are presented in bold text below. T36 Heavy Chain Variable Domain QVQLKESGPVLVAPSQSLSVTCSVSGFALTSYGVHWVRQPPGKGLEWLGVIWAGGSTNYNSALM SRLSISKDNSKRQLFLKMNSLQTDDTAIYYCAREGPYYAMDYWGQGTSVTVSS (SEQ ID NO.: 1) HCDR1: SYGVH (SEQ ID NO.: 2) HCDR2: VIWAGGSTNYNSALMS (SEQ ID NO.: 3) HCDR3: EGPYYAMDY (SEQ ID NO.: 4) T36 Light Chain Variable Domain (κ) NIVLTQSPASLAVSPGQRATISCRASESVDRYGSSFIHWYQQKPGQPPKLLIYLASTLDSGVPA RFSGSGSRTDFTLTIDPVEADDATTYYCQQCYEDPSTFGGGTKLEIK (SEQ ID NO.: 5) 51 314460903v1Attorney Docket No: 243735.000428 LCDR1: RASESVDRYGSSFIH (SEQ ID NO.: 6) LCDR2: LASTLDS (SEQ ID NO.: 7) LCDR3: QQCYEDPST (SEQ ID NO.: 8) T51 Heavy Chain Variable Domain QVQLKESGPGLVAPSQSLSITCTVSGFSLTTYGVSWVRQPPGKGLEWLGVIWTGGGTNYNSALK SRLSISKDNSKSQVFLRMSSLQIDDTARYYCTRDGVYMSMDYWGHGTSVTVSS (SEQ ID NO.: 9) HCDR1: TYGVS (SEQ ID NO.: 10) HCDR2: VIWTGGGTNYNSALKS (SEQ ID NO.: 11) HCDR3: DGVYMSMDY (SEQ ID NO.: 12) T51 Light Chain Variable Domain (κ) NIVLTQSPASLAVSLGQRATISCRASENVDTLGNSFMHWYQQKPGQPPKLLIYSASTLEYGVPA RFSGSGSRTDFTLTIDPVEADDTATYYCQQSNEDPRTFGGGTKLEIK (SEQ ID NO.: 13) LCDR1: RASENVDTLGNSFMH (SEQ ID NO.: 14) LCDR2: SASTLEY (SEQ ID NO.: 15) LCDR3: QQSNEDPRT (SEQ ID NO.: 16) T52 Heavy Chain Variable Domain QVQLKESGPVLVAPSQSLSVTCSVSGFSLTSYGVHWVRQPPGKGLEWLGVIWAGGNTNYNSALM SRLSISKDNSKRQLFLKINSLQTDDTAIYYCAREGPYYAMDYWGQGTSVTVSS (SEQ ID NO.: 17) HCDR1: SYGVH (SEQ ID NO.: 18) HCDR2: VIWAGGNTNYNSALMS (SEQ ID NO.: 19) HCDR3: EGPYYAMDY (SEQ ID NO.: 20) T52 Light Chain Variable Domain (κ) 52 314460903v1Attorney Docket No: 243735.000428 NIVVTQSPASLTVSPGQRATISCRASESVDRYGSSFIHWYQQKPGQPPKLLIYLASALDSGVPA RFSGSGSKTDFTLIIDPVEADDAAIYYCQQCYEDPSTFGGGTKLEIK (SEQ ID NO: 21) LCDR1: RASESVDRYGSSFIH (SEQ ID NO: 22) LCDR2: LASALDS (SEQ ID NO: 23) LCDR3: QQCYEDPST (SEQ ID NO: 24)

[0235] In some embodiments, an antibody or antigen binding fragment described hereincomprises a heavy chain complementarity determining region 1 (CDR1), a heavy chain CDR2, a heavy chain CDR3 contained within a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 1, 9, or 17. In some embodiments, an antibody or antigen binding fragment described herein comprises a VH comprising an amino acid sequence of SEQ ID NO: 1, 9 or 17, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95.5%, 96%, 97%, 98%, 99%, 99.5%, or 99.9% sequence identity to the amino acid sequence of SEQ ID NO: 1, 9, or 17.

[0236] In some embodiments, an antibody or antigen binding fragment described hereincomprises a light chain complementarity determining region 1 (CDR1), a light chain CDR2, a light chain CDR3 contained within a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 5, 13, or 21. In some embodiments, an antibody or antigen binding fragment described herein comprises a VL comprising an amino acid sequence of SEQ ID NO: 5, 13, or 21, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95.5%, 96%, 97%, 98%, 99%, 99.5%, or 99.9% sequence identity to the amino acid sequence of SEQ ID NO: 5, 13, or 21.

[0237] In some embodiments, the antibody, or antigen-binding fragment thereof, comprises aheavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3 of a heavy chain variable region (VH) comprising an amino acid sequence of SEQ ID NO: 1, 9, or 17, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; and a light chain CDR1, a light chain CDR2, and a light chain CDR3 of a light chain variable region (VL) comprising an amino acid sequence of SEQ ID NO: 5, 13, or 21, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto. 53 314460903v1Attorney Docket No: 243735.000428

[0238] In some embodiments, the antibody, or antigen-binding fragment thereof, comprises aheavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3 of a heavy chain variable region (VH) comprising an amino acid sequence of SEQ ID NO: 1, 9, or 17; and a light chain CDR1, a light chain CDR2, and a light chain CDR3 of a light chain variable region (VL) comprising an amino acid sequence of SEQ ID NO: 5, 13, or 21.

[0239] In some embodiments, the heavy chain CDR1 comprises the amino acid sequence setforth in SEQ ID NO: 2, the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 3, and the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 4.

[0240] In some embodiments, the heavy chain CDR1 comprises the amino acid sequence setforth in SEQ ID NO: 10, the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 11, and the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 12.

[0241] In some embodiments, the heavy chain CDR1 comprises the amino acid sequence setforth in SEQ ID NO: 18, the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20.

[0242] In some embodiments, the light chain CDR1 comprises the amino acid sequence set forthin SEQ ID NO: 6, the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, and the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8.

[0243] In some embodiments, the light chain CDR1 comprises the amino acid sequence set forthin SEQ ID NO: 14, the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15, and the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 16.

[0244] In some embodiments, the light chain CDR1 comprises the amino acid sequence set forthin SEQ ID NO: 22, the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 23, and the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 24.

[0245] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising an amino acid sequence of SEQ ID NO: 1, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; 54 314460903v1Attorney Docket No: 243735.000428 and / or a VL comprising an amino acid sequence of SEQ ID NO: 5, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto.

[0246] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising an amino acid sequence of SEQ ID NO: 9, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; and / or a VL comprising an amino acid sequence of SEQ ID NO: 13, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto.

[0247] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising an amino acid sequence of SEQ ID NO: 17, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; and / or a VL comprising an amino acid sequence of SEQ ID NO: 21, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto.

[0248] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising an amino acid sequence of SEQ ID NO: 1; and / or a VL comprising an amino acid sequence of SEQ ID NO: 5.

[0249] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising an amino acid sequence of SEQ ID NO: 9; and / or a VL comprising an amino acid sequence of SEQ ID NO: 13.

[0250] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising an amino acid sequence of SEQ ID NO: 17; and / or a VL comprising an amino acid sequence of SEQ ID NO: 21.

[0251] In some embodiments, an antibody or antigen-binding fragment disclosed hereincomprises a heavy chain CDR1, a heavy chain CDR2, a heavy chain CDR3 of a VH comprising the amino acid sequence of SEQ ID NO: 1, and a light chain CDR1, a light chain CDR2, and a light chain CDR3 of a VL comprising the amino acid sequence of SEQ ID NO: 5.

[0252] In some embodiments of the above-described antibody or antigen-binding fragment, theantibody or antigen-binding fragment comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 2, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID 55 314460903v1Attorney Docket No: 243735.000428 NO: 3, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 4, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 6, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8.

[0253] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising the amino acid sequence of SEQ ID NO: 1, and a VL comprising the amino acid sequence of SEQ ID NO: 5.

[0254] In some embodiments, an antibody or antigen-binding fragment disclosed hereincomprises a heavy chain CDR1, a heavy chain CDR2, a heavy chain CDR3 of a VH comprising the amino acid sequence of SEQ ID NO: 9, and a light chain CDR1, a light chain CDR2, and a light chain CDR3 of a VL comprising the amino acid sequence of SEQ ID NO: 13.

[0255] In some embodiments of the above-described antibody or antigen-binding fragment, theantibody or antigen-binding fragment comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 12, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 14, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 15, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 16.

[0256] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising the amino acid sequence of SEQ ID NO: 9, and a VL comprising the amino acid sequence of SEQ ID NO: 13.

[0257] In some embodiments, an antibody or antigen-binding fragment disclosed hereincomprises a heavy chain CDR1, a heavy chain CDR2, a heavy chain CDR3 of a VH comprising the amino acid sequence of SEQ ID NO: 17, and a light chain CDR1, a light chain CDR2, and a light chain CDR3 of a VL comprising the amino acid sequence of SEQ ID NO: 21.

[0258] In some embodiments of the above-described antibody or antigen-binding fragment, theantibody or antigen-binding fragment comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 18, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 19, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 20, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 22, a light chain CDR2 56 314460903v1Attorney Docket No: 243735.000428 comprising the amino acid sequence of SEQ ID NO: 23, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 24.

[0259] In some embodiments, the antibody or antigen-binding fragment comprises a VHcomprising the amino acid sequence of SEQ ID NO: 17, and a VL comprising the amino acid sequence of SEQ ID NO: 21.

[0260] In some embodiments, an antibody or antigen-binding fragment thereof described hereincan compete for binding to TTYH2 protein with an antibody or antigen-binding fragment of the present disclosure.

[0261] In some embodiments, an antibody or antigen-binding fragment thereof described hereincan bind to the same epitope of TTYH2 protein as an antibody or antigen-binding fragment of the present disclosure.

[0262] In some embodiments, an anti-TTYH2 antibody, or antigen-binding fragment thereof,may bind to the second ectodomain (ECD2) of TTYH2 protein. In some embodiments, the epitope on TTYH2 protein comprises, consists essentially of, or consists of the sequence FAARGDY (SEQ ID NO: 25).

[0263] In certain embodiments, the antibody or antigen-binding fragment comprises one or moreamino acid substitutions. In certain embodiments, amino acid substitutions of an antibody or portion thereof are those which: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity for forming protein complexes, or (4) confer or modify other physicochemical or functional properties. For example, single or multiple amino acid substitutions (preferably conservative amino acid substitutions) may be made in the naturally occurring sequence.

[0264] A conservative amino acid substitution should not substantially change the structuralcharacteristics of the parent sequence. Examples of art-recognized polypeptide secondary and tertiary structures are described in Proteins, Structures and Molecular Principles (Creighton, Ed., W. H. Freeman and Company, New York (1984)); Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); and Thornton et al., Nature 354:105 (1991), which are each incorporated herein by reference.

[0265] As used herein, the twenty naturally occurring amino acids and their abbreviations followconventional usage. See Immunology—A Synthesis (2ndEdition, E. S. Golub and D. R. Green, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. 57 314460903v1Attorney Docket No: 243735.000428

[0266] In some embodiments, the antibody or antigen-binding fragment is recombinant. Incertain embodiments, the antibody or antigen-binding fragment is a human antibody, a humanized antibody, a chimeric antibody, a murine antibody, a monoclonal antibody, a single chain antibody, a bispecific antibody or antigen-binding fragment thereof, a bi-epitopic antibody or antigen- binding fragment thereof, a Fab, a Fab’, a F(ab’)2, a Fv, an scFv, a VH domain, or a nanobody.

[0267] In some embodiments, the antibodies or antigen-binding fragments described herein arehumanized antibodies.

[0268] In some embodiments, the antibodies or antigen-binding fragments described herein arechimeric antibodies.

[0269] In some embodiments, the antigen-binding fragment is a Fab.

[0270] In some embodiments, the antigen-binding fragment is an scFv.

[0271] In some embodiments, an antibody or antigen-binding fragment thereof can be abispecific antibody or antigen-binding fragment thereof. In some embodiments, the bispecific antibody or antigen-binding fragment thereof can comprise at least one antigen-binding domain. An antigen-binding domain can include any portion of an antigen-binding molecule (i.e., a molecule that is capable of specific binding to an antigen) which can specifically bind to an antigen. As a non-limiting example, an antigen-binding domain may be a portion of an antigen- binding fragment such as, without limitation, a Fab, an scFv, or a VH domain. In some embodiments, the bispecific antibody of antigen-binding fragment can comprise a first antigen- binding domain and a second antigen-binding domain.

[0272] In some embodiments, a bispecific antibody or antigen-binding fragment thereof maycomprise a bi-epitopic antibody or antigen-binding fragment. For example, without limitation, in such embodiments a bi-epitopic antibody may comprise a first antigen-binding domain which may bind to a first epitope that is the same as an antibody or antigen-binding fragment thereof disclosed herein (e.g., any of various antibodies or antigen-binding fragments thereof set forth in Table 1 or a substantially similar sequence thereof having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto), and a second antigen-binding domain which may bind to a second epitope that is the same as an antibody or antigen-binding disclosed herein (e.g., any of various antibodies or antigen-binding fragments thereof set forth in Table 1 or a substantially similar sequence thereof having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto) but is different from the first epitope. 58 314460903v1Attorney Docket No: 243735.000428

[0273] In some embodiments, a bispecific antibody or bispecific antigen-binding fragment ofthe present disclosure is a bispecific T-cell engager (BiTE), bispecific NK-cell engager (BiKE), or a bi-specific macrophage engager (BiME).

[0274] In some embodiments, the bispecific NK-cell engager (BiKE) antibody comprises anantigen-binding domain that specifically binds to CD3, alpha beta TCR, or gamma delta TCR. In some embodiments, the bispecific NK-cell engager (BiKE) antibody comprises an antigen-binding domain that specifically binds to Fc gamma receptor III (FcγRIII or CD16) , NKG2D, or NCR3.

[0275] In some embodiments, the BiME comprises an antigen-binding domain that specificallybinds to Fc gamma receptor III (FcγRIII or CD16), dendritic cell-associated C-type lectin 1 (Dectin-1), cluster of differentiation (CD47), signal regulatory protein alpha (SIRPA), or Tyro3, Axl, and MerTK (TAM) receptor(s).

[0276] In some embodiments, a bispecific antibody or antigen-binding fragment thereof of thepresent disclosure may comprise a first antigen-binding domain which binds to a first epitope that is the same as an antibody or antigen-binding fragment of the present disclosure (e.g., any of various antibodies or antigen-binding fragments thereof set forth in Table 1 or a substantially similar sequence thereof having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto), and a second antigen-binding domain which binds to a molecule on the cell surface. In some embodiments, the second antigen-binding domain which binds to a molecule on the cell surface is an immune checkpoint, a cytokine or a receptor thereof, a tumor-associated antigen (TAA), or an immune stimulatory receptor. In some embodiments, the second antigen-binding domain is an immune checkpoint, a cytokine or a receptor thereof, a tumor- associated antigen (TAA), or an immune stimulatory receptor.

[0277] In some embodiments, the second antigen-binding domain is an antigen-binding domainthat specifically binds to tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 13 (IL-13), interleukin 17A (IL-17A), interleukin 17F (IL-17F), chemokine (C-C motif) ligand 2 (CCL2), C-X-C motif chemokine ligand 10 (CXCL10), IFN-α, IFN-β, very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)- integrin or CD49d / CD29), cluster of differentiation 20 (CD20), B activating factor (BAFF), transmembrane activator and CAML interactor (TACI), cluster of differentiation 19 (CD19), interleukin-23 receptor (IL-23R), cluster of differentiation 19 (CD22), cluster of differentiation 38 (CD38), B-cell maturation antigen (BCMA), PD-1, BTLA, CD40, CD40 ligand (CD40L), OX40, 59 314460903v1Attorney Docket No: 243735.000428 OX40L, B7, CD28, CTLA-4, tumor necrosis factor-like cytokine 1A (TL1A), DR3, or thymic stromal lymphopoietin (TSLP), or a variant thereof.

[0278] In some embodiments, the second antigen-binding domain is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20- targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA- targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

[0279] Exemplary cytokines include, but are not limited to, interleukin-2 (IL-2), interleukin-6(IL-6), interleukin-10 (IL-10), interleukin-15 (IL-15), interleukin-21 (IL-21), IFN-α, IFN-β, IFN- γ, CCL19, CCL21, interleukin-18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), a receptor thereof, and any combinations thereof.

[0280] In some embodiments, the immune checkpoint is programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), lymphocyte activation gene 3 (LAG-3 or CD223), cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), cluster of differentiation 47 (CD47), T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3), B7 homolog 3 protein (B7- H3 or CD276), B7-H4, V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)), T cell immunoreceptor with Ig and ITIM Domains (TIGIT), signal regulatory protein alpha (SIRPA), signaling lymphocytic activation molecule family members (SLAMF), poliovirus receptor-related immunoglobulin domain-containing 60 314460903v1Attorney Docket No: 243735.000428 protein (PVRIG or CD112R), adenosine A2A receptor (A2aR), adenosine A2b receptor (A2bR), G protein-coupled receptor 171 (GPR171), insulin like growth factor binding protein 7 (IGFBP7), cluster of differentiation 93 (CD93), CD96, CD226, natural killer group protein 2A (NKG2A), natural killer group protein 2D (NKG2D), a killer cell lectin like receptor G1 (KLRG1), a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2), a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3), sialic acid-binding immunoglobulin-like lectin (Siglec)-15, cluster of differentiation 24 (CD24), sialic acid-binding immunoglobulin-like lectin (Siglec)-10, P-selectin glycoprotein ligand-1 (PSGL-1), V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11), leucine rich repeats and immunoglobulin like domains 1 (LRIG1), fibrinogen-like protein 1 (FGL1), B and T lymphocyte attenuator (BTLA), leukocyte associated immunoglobulin like receptor 1 (LAIR-1), cluster of differentiation 160 (CD160), leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2), leukocyte immunoglobulin-like receptor (LILRB4), angiopoietin 2 (ang2), or vascular endothelial growth factor (VEGF). In some embodiments, the immune checkpoint is PD-1, PD-L1, LAG-3, or CTLA-4, and any combination thereof.

[0281] In some embodiments, the second antigen-binding domain which binds to an immunecheckpoint is an immune checkpoint inhibitor. Exemplary immune checkpoint inhibitors include, but are not limited to, a programmed cell death protein 1 (PD-1) inhibitor, a programmed death- ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a B7-H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 61 314460903v1Attorney Docket No: 243735.000428 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)- 15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin- like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, a vascular endothelial growth factor (VEGF) inhibitor, and any combinations thereof. In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, or a CTLA-4 inhibitor, or a combination thereof.

[0282] In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor. ExemplaryPD-1 inhibitors include, but are not limited to, nivolumab, pembrolizumab, BAT1308, acrixolimab, balstilimab, budigalimab, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, ociperlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tiragolumab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, zimberelimab, and a variant or any combinations thereof. In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor. Exemplary PD-L1 inhibitors include, but are not limited to, durvalumab, avelumab, atezolizumab, bintrafusp alfa, and a variant or any combinations thereof.

[0283] In some embodiments, the immune checkpoint inhibitor is a LAG-3 inhibitor. ExemplaryLAG-3 inhibitors include, but are not limited to, relatlimab (BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), 62 314460903v1Attorney Docket No: 243735.000428 pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO- 7247669), TSR-033, tuparstobart (INCAGN02385), BGA-1953, and a variant or any combinations thereof. In certain specific embodiments, the LAG-3 inhibitor is relatlimab (BMS-986016), or a variant thereof.

[0284] In some embodiments, the immune checkpoint inhibitor is a CTLA-4 inhibitor.Exemplary CTLA-4 inhibitors include, but are not limited to, ipilimumab, tremelimumab, XmAb20717, ONC-392, XmAb22841, BMS-986249, ADG116, ATOR-1015, ADG126, YH001, botensilimab, HBM4003, lorigerlimab, SI-B003, AK104, KN046, quavonlimab, BNT316 / ONC- 392 (gotistobart), porustobart (HBM4003), and a variant or a combination thereof. In some embodiments, the CTLA-4 inhibitor is ipilimumab, tremelimumab, quavonlimab, BNT316 / ONC- 392 (gotistobart), or porustobart (HBM4003), or a variant or a combination thereof.

[0285] In some embodiments, the immune checkpoint inhibitor is a TIGIT inhibitor. ExemplaryTIGIT inhibitors include, but are not limited to, BMS‐986207, ociperlimab, BGB‐A1217, tiragolumab, domvanalimab, ASP8374, vibostolimab, IBI‐939, etigilimab, COM902, M6223, EOS884448, BAT6021, HLX301, and a variant or any combinations thereof.

[0286] In some embodiments, the immune checkpoint inhibitor is a SLAMF inhibitor. In someembodiments, the SLAMF inhibitor is elotuzumab or a variant thereof. In some embodiments, the immune checkpoint inhibitor is a PVRIG inhibitor. In some embodiments, the PVRIG inhibitor is COM701, JS009, or a variant thereof. In some embodiments, the immune checkpoint inhibitor is a CD96 inhibitor. In some embodiments, the CD96 inhibitor is GSK6097608 or a variant thereof.

[0287] In some embodiments, the immune stimulatory receptor is the immune stimulatoryreceptor is CD28, ICOS, CD28H / TMIGD2, NCR3, NCR1, NCR2, 4-1BB, OX40, CD30, CD40, DR3, CD226, CRTAM, CD27, HVEM, TNFR1, TNFR2, CD2, CD7, TLR4, TLR7, TLR9, or GITR, or a combination thereof.

[0288] In some embodiments, the immune stimulatory receptor is CD28, ICOS, TMIGD2,NCR3, NCR1, NCR2, 4-1BB, OX40, CD30, CD40, DR3, CD226, CRTAM, CD27, HVEM, TNFR1, TNFR2, CD2, CD7 , TLR4, TLR7, or TLR9.

[0289] In some embodiments, the tumor-associated antigen is 5T4, Ang2, BCMA, CD123,CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY-ESO-1, P-cadherin, PRAME , PSCA, 63 314460903v1Attorney Docket No: 243735.000428 PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, or RAF, or a combination thereof.

[0290] However, disclosure of immune checkpoints as cell surface on a cell surface is notintended to be limiting. Thus, it is well within the knowledge of one skilled in the art that where anti-cell surface molecule antibodies or antigen-binding fragments thereof may be used, such anti- cell surface antibodies or antigen-binding fragments may target any of various cell surface molecules known to a skilled artisan. In some embodiments, TAA targeting agent is a VEGF inhibitor, a VEGFR inhibitor, a CD20 inhibitor, a CD38 inhibitor, a TROP2 inhibitor, a KRAS inhibitor, or a RAF inhibitor.

[0291] In some embodiments, the CD20 inhibitor is rituximab. In some embodiments, the VEGFinhibitor is bevacizumab. In some embodiments, the CD38 inhibitor is daratumumab. In some embodiments, the TROP2 inhibitor is sacituzumab govitecan.

[0292] In some embodiments, an antibody or antigen-binding fragment thereof may comprise aKineTAC such as, e.g., a KineTAC similar to, or the same as, those which are described by Pance, K. et. Al., Nature Biotechnology, 41, 2023, 273-281 (PMID 36138170), the content of which is incorporated herein by reference in its entirety for all purposes. Without wishing to be bound by theory, KineTACs are a modular cytokine receptor-targeting chimera. KineTACs are genetically encoded bispecific antibodies that are used for targeted degradation of both cell surface and extracellular proteins via lysosomal delivery. KineTACs are made of a target-binding arm (e.g., a TTYH2-binding Fab), which can be bound to a protein of interest (e.g., TTYH2 protein), and a cytokine arm, which can bind its cognate cytokine receptor. The cytokine arm may be, but is not limited to, C-X-C motif chemokine ligand 12 (CXCL12), C-X-C motif chemokine ligand 11 (CXCL11), viral macrophage inflammatory protein-II (vMIPII), and / or IL-2 for lysosomal degradation applications. The KineTAC cytokine arm may bind C-X-C chemokine receptor type 7 (CXCR7) or Interleukin-2 receptor (IL-2R).

[0293] In some embodiments, an antibody or antigen-binding fragment described herein, cancomprise a first antigen-binding domain and a cytokine receptor-targeting molecule (e.g., a cytokine).

[0294] In some embodiments, the first antigen-binding domain can bind to a first epitope that isthe same as an antibody or antigen-binding fragment of the present disclosure (e.g., any of various antibodies or antigen-binding fragments thereof set forth in Table 1 or a substantially similar 64 314460903v1Attorney Docket No: 243735.000428 sequence thereof having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto), and the cytokine receptor-targeting molecule (e.g., a cytokine) may bind to its cognate cytokine receptor.

[0295] In some embodiments, the anti-TTYH2 antibodies disclosed herein, having the heavychain CDRs disclosed herein, contains framework regions derived from a subclass of germline VH fragment. Such germline VH regions are well known in the art. See, e.g., the IMGT database (imgt.org) or at vbase2.org / vbstat.php. Examples include the IGHV1 subfamily (e.g., IGHV1-2, IGHV1-3, IGHV1-8, IGHV1-18, IGHV1-24, IGHV1-45, IGHV1-46, IGHV1-58, and IGHV1-69), the IGHV2 subfamily (e.g., IGHV2-5, IGHV2-26, and IGHV2-70), the IGHV3 subfamily (e.g., IGHV3-7, IGHV3-9, IGHV3-11, IGHV3-13, IGHV3-15, IGHV3-20, IGHV3-21, IGHV3-23, IGHV3-30, IGHV3-33, IGHV3-43, IGHV3- 48, IGHV3-49, IGHV3-53, IGHV3-64, IGHV3-66, IGHV3-72, and IGHV3-73, IGHV3-74), the IGHV4 subfamily (e.g., IGHV4-4, IGHV4-28, IGHV4-31, IGHV4-34, IGHV4-39, IGHV4-59, IGHV4-61, and IGHV4-B), the IGHV subfamily (e.g., IGHV5-51, or IGHV6-1), and the IGHV7 subfamily (e.g., IGHV7-4-1).

[0296] Alternatively, or in addition, in some embodiments, the anti-TTYH2 antibody, havingthe light chain CDRs disclosed herein, contains framework regions derived from a germline Vκ fragment. Examples include an IGKV1 framework (e.g., IGKV1-05, IGKV1-12, IGKV1-27, IGKV1-33, or IGKV1-39), an IGKV2 framework (e.g., IGKV2-28), an IGKV3 framework (e.g., IGKV3-11, IGKV3-15, or IGKV3-20), and an IGKV4 framework (e.g., IGKV4-1). In other instances, the anti-TTYH2 antibody comprises a light chain variable region that contains a framework derived from a germline Vλ fragment. Examples include an IGλ1 framework (e.g., IgλV1-36, IgλV1-40, IgλV1-44, IgλV1-47, IgλV1-51), an Igλ2 framework (e.g., IgλV2-8, IgλV2- 11, IgλV2-14, IgλV2-18, IgλV2-23), an Igλ3 framework (e.g., IgλV3-1, IgλV3-9, IgλV3-10, IgλV3-12, IgλV3-16, IgλV3-19, IgλV3-21, IgλV3-25, IgλV3-27), an Igλ4 framework (e.g., IgλV4- 3, IgλV4-60, IgλV4-69), an Igλ5 framework (e.g., IgλV5-39, IgλV5-45), an Igλ6 framework (e.g., IgλV6-57), an Igλ7 framework (e.g., IgλV7-43, IgλV7-46), an Igλ8 framework (e.g., IgλV8-61), an Igλ9 framework (e.g., IgλV9-49), or an Igλ10 framework (e.g., IgλV10-54).

[0297] In some embodiments, an anti-TTYH2 antibody, or antigen-binding fragment thereof,may bind to human TTYH2 protein. In some embodiments, the antibody or antigen-binding fragment thereof may bind to mouse TTYH2 protein. In some embodiments, an anti-TTYH2 antibody, or antigen-binding fragment thereof, may bind to both human TTYH2 protein and mouse 65 314460903v1Attorney Docket No: 243735.000428 TTYH2 protein. In some embodiments, an anti-TTYH2 antibody, or antigen-binding fragment thereof, may bind to either human TTYH2 protein or mouse TTYH2 protein.

[0298] In some embodiments, an anti-TTYH2 antibody, or antigen-binding fragment thereofmay enhance T cell and macrophage immune response upon binding to TTYH2. In some embodiments, an anti-TTYH2 antibody, or antigen-binding fragment thereof may promote production of Type I interferons such as IFNα and IFNβ upon binding to TTYH2.

[0299] In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof asdescribed herein has a suitable binding affinity for the target antigen (e.g., TTYH2 protein) or antigenic epitopes thereof. As used herein, “binding affinity” refers to the apparent association constant or KA. The KAis the reciprocal of the dissociation constant (KD). In various embodiments, the anti-TTYH2 antibody, or antigen-binding fragment thereof described herein, may have a binding affinity (KD) of at least about 1x10-6M or less, about 1x10-7M or less, about 1x10-8M or less, about 1x10-9M or less, about 1x10-10M or less, about 1x10-11M or less, about 1x10-12M or less, or about 1x10-13M or less for the target antigen or antigenic epitope. An increased binding affinity corresponds to a decreased KD.

[0300] In various embodiments, the antibodies of the disclosure have the ability to bind to apredetermined antigen (e.g., TTYH2 protein) with a dissociation constant (KD) of 1x10-6M or less, 5x10-7M or less, 1x10-7M or less, 5x10-8M or less, 4x10-8M or less, 3x10-8M or less, 2x10-8M or less, about 1x10-8M or less, 9x10-9M or less, 8x10-9M or less, 7x10-9M or less, 6x10-9M or less, 5x10-9M or less, 4x10-9M or less, 3x10-9M or less, 2x10-9M or less, about 1x10-9M or less, 9x10-10M or less, 8x10-10M or less, 7x10-10M or less, 6x10-10M or less, 5x10-10M or less, 4x10-10M or less, 3x10-10M or less, 2x10-10M or less, 1x10-10M or less, 5x10-11M or less, about 1x10-11M or less, 5x10-12M or less, about 1x10-12M or less, 5x10-13M or less, or about 1x10-13M or less.

[0301] In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof,described herein may bind to human TTYH2 protein at a dissociation constant (KD) of about 1x10-6M or less, 5x10-7M or less, 1x10-7M or less, 5x10-8M or less, 4x10-8M or less, 3x10-8M or less, 2x10-8M or less, about 1x10-8M or less, 9x10-9M or less, 8x10-9M or less, 7x10-9M or less, 6x10-9M or less, 5x10-9M or less, 4x10-9M or less, 3x10-9M or less, 2x10-9M or less, about 1x10-9M or less, 9x10-10M or less, 8x10-10M or less, 7x10-10M or less, 6x10-10M or less, 5x10-10M or less, 4x10-10M or less, 3x10-10M or less, 2x10-10M or less, 1x10-10M or less, 5x10-11M or less, 66 314460903v1Attorney Docket No: 243735.000428 about 1x10-11M or less, 5x10-13M or less, about 1x10-12M or less, 5x10-12M or less, or about 1x10-13M or less. As a non-limiting example, the anti-TTYH2 antibody, or antigen-binding fragment thereof, may bind to human TTYH2 protein at a dissociation constant (KD) of about 1x10-8M or less. In some embodiments, the anti-TTYH2 antibody, or antigen-binding fragment thereof, may bind to human TTYH2 protein at a dissociation constant (KD) of about 1x10-7M or less. In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof described herein may bind human TTYH2 protein at a KDof about 2x10-9M or less. In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof described herein may bind human TTYH2 protein at a KD of about 5x10-9M or less.

[0302] In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereofdescribed herein may bind to mouse TTYH2 protein at a dissociation constant (KD) of about 1x10-6M or less, 5x10-7M or less, 1x10-7M or less, 5x10-8M or less, 4x10-8M or less, 3x10-8M or less, 2x10-8M or less, about 1x10-8M or less, 9x10-9M or less, 8x10-9M or less, 7x10-9M or less, 6x10-9M or less, 5x10-9M or less, 4x10-9M or less, 3x10-9M or less, 2x10-9M or less, about 1x10-9M or less, 9x10-10M or less, 8x10-10M or less, 7x10-10M or less, 6x10-10M or less, 5x10-10M or less, 4x10-10M or less, 3x10-10M or less, 2x10-10M or less, 1x10-10M or less, 5x10-11M or less, about 1x10-11M or less, 5x10-12M or less, about 1x10-12M or less, 5x10-13M or less, or about 1x10-13M or less. As a non-limiting example, the anti-TTYH2 antibody, or antigen-binding fragment thereof, may bind mouse TTYH2 protein at a dissociation constant (KD) of about 1x10-8M or less. In some embodiments, the anti-TTYH2 antibody, or antigen-binding fragment thereof, may bind to human TTYH2 protein at a dissociation constant (KD) of about 1x10-7M or less. In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof described herein may bind mouse TTYH2 protein at a KD of about 2x10-9M or less. In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof described herein may bind to mouse TTYH2 protein at a KD of about 5x10-9M or less.

[0303] Binding affinity (or binding specificity) can be determined by a variety of methodsincluding equilibrium dialysis, equilibrium binding, gel filtration, ELISA, biolayer interferometry (BLI), surface plasmon resonance (SPR), bead-based assay, radioimmunoassay, or spectroscopy (e.g., using a fluorescence assay). In certain embodiments, a TTYH2 antigen, or derivative thereof, is coated onto a bead or onto the surface of an ELISA plate or other solid phase used for measurement. Examples of bead-based binding assays which may be used for measuring KDof the 67 314460903v1Attorney Docket No: 243735.000428 anti-TTYH2 antibodies and / or antigen-binding fragments herein are described in, e.g., Nishikori et al., J Mol Biol. 2012 Dec 14;424(5):391-9 (PMID 23041298) and Hattori et al., J Immunol Methods. 2021 Mar;490:112952 (PMID 33358997), each of which is incorporated herein by reference in its entirety and for all purposes as if fully set forth herein.

[0304] These techniques can be used to measure the concentration of bound antibody or antigen-binding fragment as a function of target antigen concentration. Under certain conditions, the fractional concentration of bound antibody or antigen-binding fragment ([Bound] / [Total]) is generally related to the concentration of total target antigen ([Target]) by the following equation: [Bound] / [Total] = [Target] / (KD+[Target])

[0305] It is not always necessary to make an exact determination of KD, though, since sometimesit is sufficient to obtain a quantitative measurement of affinity, e.g., determined using a method such as enzyme-linked immunosorbent assay (ELISA) or fluorescence-activated cell sorting (FACS) analysis, is proportional to KD, and thus can be used for comparisons, such as determining whether a higher affinity is, e.g., 2-fold higher, to obtain a qualitative measurement of affinity, or to obtain an inference of affinity, e.g., by activity in a functional assay, e.g., an in vitro or in vivo assay. In some cases, the in vitro binding assay is indicative of in vivo activity. In other cases, the in vitro binding assay is not necessarily indicative of in vivo activity. In some cases, tight binding is beneficial, but in other cases tight binding is not as desirable in the in vivo setting, and an antibody with lower binding affinity is more desirable.

[0306] In some embodiments, the heavy chain of any of any of the anti-TTYH2 antibodies asdescribed herein further comprise a heavy chain constant region (CH) or a portion thereof (e.g., CH1, CH2, CH3, or a combination thereof). The heavy chain constant region can be of any suitable origin, e.g., human, mouse, rat, or rabbit. In some embodiments, the heavy chain constant region is from an IgD, IgE, IgG, IgA, or IgM class, or sub-class thereof. In some embodiments, the heavy chain constant region is from a human IgG (a gamma heavy chain) or any IgG subfamily as described herein. In some embodiments, the heavy chain constant region is from an IgG1, IgG2, IgG3, or IgG4 subclass.

[0307] In some embodiments, the heavy chain constant region of the antibodies described hereincomprise a single domain (e.g., CH1, CH2, or CH3) or a combination of any of the single domains, of a constant region. In some embodiments, the light chain constant region of the antibodies described herein comprise a single domain (e.g., CL), of a constant region. 68 314460903v1Attorney Docket No: 243735.000428

[0308] In some embodiments, the anti-TTYH2 antibody or antigen-binding fragment describedherein is of an IgG1 subclass.

[0309] In some embodiments, the anti-TTYH2 antibody or antigen-binding fragment describedherein is of an IgG2 subclass.

[0310] In some embodiments, the anti-TTYH2 antibody or antigen-binding fragment describedherein is of an IgG4 subclass.

[0311] In some embodiments, anti-TTYH2 antibodies or antigen-binding fragments describedherein comprises a fragment crystallizable (Fc) region. In some embodiments, anti-TTYH2 antibodies or antigen-binding fragment described herein comprises a modified Fc region.

[0312] In some embodiments, an Fc region described herein is altered by replacing at least oneamino acid residue with a different amino acid residue to alter the effector function(s) of the antibody. For example, one or more amino acids selected from the following positions: 234, 235, 236, 237, 297, 318, 320, 322, 330, and / or 331 may be substituted with a different amino acid residue such that the antibody has an altered affinity for an effector ligand but retains the antigen- binding ability of the parent antibody. The effector ligand to which affinity is altered can be, for example, an Fc receptor or the C1 component of complement. See, e.g., U.S. Pat. Nos.5,624,821 and 5,648,260, both of which are incorporated herein by reference in their entirety.

[0313] In one embodiment, an Fc region described herein comprises one or more amino acidssubstitutions at amino acid residues 329, 331, and 322 such that the antibody has altered C1q binding and / or reduced or abolished complement dependent cytotoxicity (CDC). See, e.g., U.S. Pat. No.6,194,551, which is incorporated herein by reference in its entirety.

[0314] In some examples, an Fc region described herein comprises one or more amino acidresidues at amino acid positions 231 and / or 239 to alter the ability of the antibody to fix complement. See, e.g., PCT Publication WO 94 / 29351, which is incorporated herein by reference in its entirety.

[0315] In some examples, an Fc region described herein can be modified to decrease antibodydependent cellular cytotoxicity (ADCC) and / or to decrease the affinity for an Fcγ receptor by modifying one or more amino acids at the following positions: 234, 235, 236, 238, 239, 240, 241, 243, 244, 245, 247, 248, 249, 252, 254, 255, 256, 258, 262, 263, 264, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 299, 301, 303, 305, 307, 309, 312, 313, 315, 320, 322, 324, 325, 326, 327, 329, 330, 331, 332, 333, 334, 335, 337, 69 314460903v1Attorney Docket No: 243735.000428 338, 340, 360, 373, 376, 378, 382, 388, 389, 398, 414, 416, 419, 430, 433, 434, 435, 436, 437, 438, or 439. Exemplary substitutions include but are not limited to 236A, 239D, 239E, 268D, 267E, 268E, 268F, 324T, 332D, and 332E. Exemplary variants include but are not limited to 239D / 332E, 236A / 332E, 236A / 239D / 332E, 268F / 324T, 267E / 268F, 267E / 324T, and 267E / 268F / 324T. In some embodiments, an exemplary variant can comprise L234A / L235A / P329G (LALA-PG), e.g., as described in Lo et al., J Biol Chem. 2017 Mar 3;292(9):3900-3908 (PMID 28077575). In some embodiments, the anti-TTYH2 antibody or antigen-binding fragment described herein comprises an Fc region comprising L234A / L235A / P329G (LALA-PG) amino acid substitutions, wherein the residues are numbered according to the EU index of Kabat.

[0316] In one embodiment, the anti-TTYH2 antibody or antigen-binding fragment describedherein comprises a variant of human IgG1 Fc region, having an amino acid sequence: IgG1 L234A / L235A / P329G (mutations are indicated in bold letters in the sequence below.) DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPG (SEQ ID NO: 50)

[0317] In some embodiments, an Fc region described herein include modifications that reduceor ablate binding to FcγR and / or complement proteins, thereby reducing or ablating Fc-mediated effector functions such as ADCC, ADCP, and CDC. Exemplary modifications include but are not limited to substitutions, insertions, and deletions at amino acid residues (EU numbering) 234, 235, 236, 237, 267, 269, 325, 328, 330, and / or 331. Exemplary substitutions include but are not limited to 234A, 235E, 236R, 237A, 267R, 269R, 325L, 328R, 330S, and 331S (e.g., 330S, and 331S). An exemplary Fc variant can comprise 236R / 328R. Other modifications for reducing FcγR and complement interactions include substitutions 297A, 234A, 235A, 237A, 318A, 228P, 236E, 268Q, 309L, 330S, 331S, 220S, 226S, 229S, 238S, 233P, and 234V, as well as removal of the glycosylation at position 297 by e.g., mutational or enzymatic means or by production in organisms such as bacteria that do not glycosylate proteins.

[0318] In one embodiment, an Fc region described herein is a human IgG1.3 Fc constant regioncomprising L234A, L235E, and G237A substitutions. In one embodiment, an Fc region described herein is a IgG1fa.P238K (or IgG1.P238K) comprising a P238K substitution. In one embodiment, 70 314460903v1Attorney Docket No: 243735.000428 an Fc region described herein is a IgG1.1f variant comprising L234A, L235E, G237A, A330S, and P331S substitutions.

[0319] In some embodiments, an Fc region described herein is modified to enhance affinity foran inhibitory receptor FcγRIIb. Such modification can provide an Fc fusion protein with immunomodulatory activities related to FcγRIIb cells, including for example B cells and monocytes. For example, the Fc variants may provide selectively enhanced affinity to FcγRIIb relative to one or more activating receptors. Modifications that alter binding to FcγRIIb include one or more modifications at a position (EU numbering) selected from the group consisting of 234, 235, 236, 237, 239, 266, 267, 268, 325, 326, 327, 328, 330, 331, and 332. Exemplary substitutions for enhancing FcγRIIb affinity include but are not limited to 234A, 234D, 234E, 234F, 234W, 235D, 235E, 235F, 235R, 235Y, 236D, 236N, 237A, 237D, 237N, 239D, 239E, 266M, 267D, 267E, 268D, 268E, 327D, 327E, 328F, 328W, 328Y, 330S, 331S, and 332E. Exemplary substitutions include 235Y, 236D, 239D, 266M, 267E, 268D, 268E, 328F, 328W, and 328Y. Other Fc variants for enhancing binding to FcγRIIb include 235Y / 267E, 236D / 267E, 239D / 268D, 239D / 267E, 267E / 268D, 267E / 268E, and 267E / 328F.

[0320] Other modifications for enhancing FcγR and complement interactions include but are notlimited to substitutions 298A, 333A, 334A, 326A, 2471, 339D, 339Q, 280H, 290S, 298D, 298V, 243L, 292P, 300L, 396L, 3051, and 396L. These and other modifications are reviewed in Strohl, 2009, Current Opinion in Biotechnology 20:685-691. Fc modifications that increase binding to an Fcγ receptor include amino acid modifications at any one or more of amino acid positions (EU numbering) 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 279, 280, 283, 285, 298, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 312, 315, 324, 327, 329, 330, 335, 337, 338, 340, 360, 373, 376, 379, 382, 388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438 or 439 of the Fc region (see., Patent Publication No. WO 00 / 42072).

[0321] The affinities and binding properties of an Fc region for its ligand can be determined bya variety of in vitro methods (e.g., biochemical or immunological based assays) known in the art including but not limited to, equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA), or radioimmunoassay (RIA)), or kinetics (e.g., BIACORE analysis), and other methods such as indirect binding assays, competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis and chromatography (e.g., gel filtration). These and other methods can utilize a label on one or more of the components being examined and / or employ a 71 314460903v1Attorney Docket No: 243735.000428 variety of detection methods including but not limited to fluorescent, luminescent, chromogenic, or isotopic labels.

[0322] In some embodiments, the glycosylation of an antibody is altered. Glycosylation can bealtered to, for example, increase the affinity of the antibody for antigen. Such carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the antibody sequence. For example, glycosylation of the constant region on N297 can be prevented or reduced by mutating the N297 residue to another residue, e.g., N297A, or N297D and / or by mutating an adjacent amino acid, e.g., 298. Additionally, one or more amino acid substitutions can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site. Such aglycosylation can increase the affinity of the antibody for antigen. See, e.g., U.S. Pat. Nos.5,714,350 and 6,350,861, both of which are incorporated herein by reference in their entirety.

[0323] In some embodiments, the antibody can be modified to increase its biological half-life.For example, the Fc region can be modified to increase its binding affinity for FcRn, by mutating one or more of following residues: 252, 254, 256, 433, 435, 436 (see, e.g., U.S. Pat. No.6,277,375, which is incorporated herein by reference in its entirety). Specific exemplary mutations include one or more of the following: T252L, T254S, and / or T256F. In some embodiments, the antibody can be modified within the CH1 or CL region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG, as described in U.S. Pat. Nos. 5,869,046 and 6,121,022, each of which is incorporated herein by reference in its entirety.

[0324] Other exemplary variants that increase Fc binding to FcRn include: 250E, 250Q, 428L,428F, 250Q / 428L (Hinton et al. 2004, J. Biol. Chem. 279(8): 6213-6216; Hinton et al. 2006 Journal of Immunology 176:346-356, each of which is incorporated herein by reference in its entirety), 256A, 272A, 286A, 305A, 307A, 307Q, 311A, 312A, 376A, 378Q, 380A, 382A, 434A (Shields et al., Journal of Biological Chemistry, 2001, 276(9):6591-6604, which is incorporated herein by reference in its entirety), 252F, 252T, 252Y, 252W, 254T, 256S, 256R, 256Q, 256E, 256D, 256T, 309P, 311S, 433R, 433S, 4331, 433P, 433Q, 434H, 434F, 434Y, 252Y / 254T / 256E, 433K / 434F / 436H, 308T / 309P / 311S (Dall Acqua et al. Journal of Immunology, 2002, 169:5171- 5180, Dall’Acqua et al., 2006, Journal of Biological Chemistry 281:23514-23524, each of which is incorporated herein by reference in its entirety). Additional exemplary variants that increase binding to FcRn and / or improve pharmacokinetic properties include mutations at positions 259, 72 314460903v1Attorney Docket No: 243735.000428 308, 428, and 434, including for example 259I, 308F, 428L, 428M, 434S, 434H.434F, 434Y, and 434M. Further modifications for modulating FcRn binding include those described in Yeung et al., 2010, J Immunol, 182:7663-7671, which is incorporated herein by reference in its entirety.

[0325] In some embodiments, antibodies described herein are of hybrid IgG isotypes. In someembodiments, an IgG1 / IgG2 hybrid variant can be constructed by substituting IgG2 positions in the CH2 and / or CH3 region with amino acids from IgG1 at positions where the two isotypes differ. In some embodiments, a hybrid variant IgG antibody can be constructed that comprises one or more substitutions, e.g., one or more of the following amino acid substitutions: 233E, 234L, 235L, - 236G (referring to an insertion of a glycine at position 236), and 327A. In some embodiments, an IgG1 / IgG3 hybrid variant can be constructed by substituting IgG1 positions in the CH2 and / or CH3 region with the amino acids from IgG3 at positions where the two isotypes differ. In some embodiments, a hybrid variant IgG antibody can be constructed that comprises one or more substitutions, e.g., 274Q, 276K, 300F, 339T, 356E, 358M, 384S, 392N, 397M, 4221, 435R, and / or 436F.

[0326] Moreover, the binding sites on human IgG1 for FcγRI, FcγRII, FcγRIII and FcRn havebeen mapped and variants with improved binding have been described (see Shields, R. L. et al. (2001) J. Biol. Chem. 276:6591-6604). Specific mutations at positions 256, 290, 298, 333, 334 and 339 were shown to improve binding to FcγRIII. Additionally, the following combination mutants were shown to improve FcγRIII binding: T256A / S298A, S298A / E333A, S298A / K224A and S298A / E333A / K334A, which has been shown to exhibit enhanced FcγRIIIa binding and ADCC activity (Shields et al., 2001). Other IgG1 variants with strongly enhanced binding to FcγRIIIa have been identified, including variants with S239D / I332E and S239D / I332E / A330L mutations Ih showed the greatest Increase in affinity for FcγRIIIa, a decrease in FcγRIIb binding, and strong cytotoxic activity in cynomolgus monkeys (Lazar et al., 2006). Introduction of the triple mutations into antibodies such as alemtuzumab (CD52-specific), trastuzumab (HER2 / neu- specific), rituximab (CD20-specific), and cetuximab (EGFR-specific) translated into greatly enhanced ADCC activity in vitro, and the S239D / I332E variant showed an enhanced capacity to deplete B cells in monkeys (Lazar et al., 2006).

[0327] In addition, IgG1 mutants containing L235V, F243L, R292P, Y300L, and P396Lmutations which exhibited enhanced binding to FcγRIIIa and concomitantly enhanced ADCC activity in transgenic mice expressing human FcγRIIIa in models of B cell malignancies and breast 73 314460903v1Attorney Docket No: 243735.000428 cancer have been identified (Stavenhagen et al., 2007; Nordstrom et al., 2011). Other Fc mutants that can be used include: S298A / E333A / L334A, S239D / I332E, S239D / I332E / A330L, L235V / F243L / R292P / Y300L / P396L, and M428L / N434S.

[0328] In some embodiments, an Fc region described herein may have reduced binding to FcγRs.For example, the Fc region (e.g., IgG1 Fc) with reduced FcγR binding may comprise the following three amino acid substitutions: L234A, L235E and G237A.

[0329] In some embodiments, an Fc region described herein may have reduced complementfixation. For example, the Fc region (e.g., IgG1 Fc) with reduced complement fixation may have the following two amino acid substitutions: A330S and P331S.

[0330] In some embodiments, an Fc region described herein may have essentially no effectorfunction, i.e., it has reduced binding to FcγRs and reduced complement fixation. For example, an “effectorless” Fc region (e.g., IgG1 Fc) may comprise the following five mutations: L234A, L235E, G237A, A330S and P331S.

[0331] In some embodiments, an Fc region described herein may include mutations L234Aand / or L235A (EU numbering), which can suppress FcγR binding, and / or a P329G mutation (EU numbering) to abolish complement C1q binding, e.g., to abolish all immune effector functions.

[0332] In some embodiments, an IgG4 Fc region described herein may include a S228Pmutation, e.g., to stabilize stabilizes IgG4 molecules and suppress formation of half-antibodies. In some embodiments, an hIgG4 Fab described herein may comprise an exchange mutant sequence which may include a S228P mutation which can suppress Fab arm exchange.

[0333] In some embodiments, the Fc region may comprise a non-naturally occurring amino acidresidue at additional and / or alternative positions as described in, e.g., PCT Patent Publications WO 00 / 42072; WO 01 / 58957; WO 02 / 06919; WO 04 / 016750; WO 04 / 029207; WO 04 / 035752; WO 04 / 063351; WO 04 / 074455; WO 04 / 099249; WO 05 / 040217; WO 05 / 070963; WO 05 / 092925 and WO 06 / 020114; and U.S. Pat. Nos. 5,624,821; 6,277,375; 6,737,056; 6,194,551; 7,317,091; 8,101,720; each of which is incorporated herein by reference in its entirety). Other suitable Fc modifications are described in WO 2016 / 081746 or WO 2017 / 087678, both of which are incorporated herein by reference in their entirety.

[0334] In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereofdescribed herein is capable of depleting TTYH2 protein from a tumor microenvironment of a subject. In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof 74 314460903v1Attorney Docket No: 243735.000428 described herein antagonizes the inhibitory function of TTYH2 on a myeloid cell or cancer cell. In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof described herein stimulates Type I IFN response upon binding to TTYH2 protein expressed on a myeloid cell or cancer cell. In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereof described herein activates STING / Type I IFN pathway signaling, NF-κB pathway signaling, or TANK-binding kinase 1 / interferon regulatory factor 3 (TBK1 / IRF3) pathway signaling, or a combination thereof, upon binding to TTYH2 protein expressed on a myeloid cell or cancer cell.

[0335] In some embodiments, anti-TTYH2 antibody or antigen-binding fragment thereofdescribed herein (i) antagonizes the inhibitory function of TTYH2 on a myeloid cell or cancer cell; (ii) stimulates Type I IFN response upon binding to TTYH2 protein expressed on a myeloid cell or cancer cell; (iii) activates STING / Type I IFN pathway signaling, NF-κB pathway signaling, or TBK1 / IRF3 pathway signaling, or a combination thereof, upon binding to TTYH2 protein expressed on a myeloid cell or cancer cell; and / or (iv) controls TNF-α, IL-6, CCL2, IL-10, and / or CXCL10, or myeloid cell differentiation.

[0336] In some embodiments, the myeloid cell is a macrophage or a dendritic cell. In someembodiments, the macrophage is an M2 macrophage and / or tumor-associated macrophage.

[0337] In some embodiments, an anti-TTYH2 antibody or antigen-binding fragment thereofdescribed herein is capable of selectively recognizing TTYH2 in the tumor microenvironment (TME), that is, an anti-TTYH2 antibody or antigen-binding fragment thereof described herein has a higher affinity to TTYH2 that is present in the TME than TTYH2 in healthy tissues. Such anti- TTYH2 antibodies or antigen-binding fragments thereof described herein may exhibit higher safety. Reported antibody engineering technologies may be exploited for developing such anti- TTYH2 antibodies or antigen-binding fragments thereof described herein. Antibody-drug Conjugates

[0338] In some embodiments, the present disclosure also provides an antibody-drug conjugatecomprising the antibody or antigen-binding fragment described herein conjugated to a heterologous moiety. Non-limiting examples of a heterologous moiety include a cytotoxic agent, 75 314460903v1Attorney Docket No: 243735.000428 siRNA, an antisense oligonucleotide, a radionucleotide, a lysosome-targeting chimera (LYTAC), an immune checkpoint inhibitor, a cytokine, a tumor-associated antigen (TAA)- targeting agent, TLR agonist, STING agonist, and an immune agonist. In some embodiments, an antibody- drug conjugate is an immune-stimulating antibody conjugate (ISAC) or an antibody-oligonucleotide conjugate (AOC).

[0339] In some embodiments, an antibody-drug conjugate comprising the antibody orantigen-binding fragment described is conjugated to a heterologous moiety selected from an immune checkpoint inhibitor, a cytokine, a tumor-associated antigen (TAA) targeting agent, an immune agonist, a cytotoxic agent, an siRNA, and an antisense oligonucleotide.

[0340] In some embodiments, a heterologous moiety of the present disclosure is an immunecheckpoint inhibitor. Exemplary immune checkpoint inhibitors include, but are not limited to, a programmed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a B7-H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich 76 314460903v1Attorney Docket No: 243735.000428 repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, a vascular endothelial growth factor (VEGF) inhibitor, and any combinations thereof. In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, or a CTLA-4 inhibitor, or a combination thereof.

[0341] In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor. ExemplaryPD-1 inhibitors include, but are not limited to, nivolumab, pembrolizumab, BAT1308, acrixolimab, balstilimab, budigalimab, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, ociperlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tiragolumab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, zimberelimab, and a variant or any combinations thereof. In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor. Exemplary PD-L1 inhibitors include, but are not limited to, durvalumab, avelumab, atezolizumab, bintrafusp alfa, and a variant or any combinations thereof.

[0342] In some embodiments, the immune checkpoint inhibitor is a LAG-3 inhibitor. ExemplaryLAG-3 inhibitors include, but are not limited to, relatlimab (BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO- 7247669), TSR-033, tuparstobart (INCAGN02385), BGA-1953, and a variant or any combinations thereof. In certain specific embodiments, the LAG-3 inhibitor is relatlimab (BMS-986016), or a variant thereof.

[0343] In some embodiments, the immune checkpoint inhibitor is a CTLA-4 inhibitor.Exemplary CTLA-4 inhibitors include, but are not limited to, ipilimumab, tremelimumab, 77 314460903v1Attorney Docket No: 243735.000428 XmAb20717, ONC-392, XmAb22841, BMS-986249, ADG116, ATOR-1015, ADG126, YH001, botensilimab, HBM4003, lorigerlimab, SI-B003, AK104, KN046, quavonlimab, BNT316 / ONC- 392 (gotistobart), porustobart (HBM4003), or a variant or combination thereof. In some embodiments, the CTLA-4 inhibitor is ipilimumab, tremelimumab, quavonlimab, BNT316 / ONC- 392 (gotistobart), porustobart (HBM4003), and a variant or a combination thereof.

[0344] In some embodiments, the immune checkpoint inhibitor is a TIGIT inhibitor. ExemplaryTIGIT inhibitors include, but are not limited to, BMS‐986207, ociperlimab, BGB‐A1217, tiragolumab, domvanalimab, ASP8374, vibostolimab, IBI‐939, etigilimab, COM902, M6223, EOS884448, BAT6021, HLX301, and a variant or any combinations thereof.

[0345] In some embodiments, the immune checkpoint inhibitor is a SLAMF inhibitor. In someembodiments, the SLAMF inhibitor is elotuzumab or a variant thereof. In some embodiments, the immune checkpoint inhibitor is a PVRIG inhibitor. In some embodiments, the PVRIG inhibitor is COM701, JS009, or a variant thereof. In some embodiments, the immune checkpoint inhibitor is a CD96 inhibitor. In some embodiments, the CD96 inhibitor is GSK6097608 or a variant thereof.

[0346] In some embodiments, a heterologous moiety of the present disclosure is an immuneagonist. Exemplary immune agonists include, but are not limited to, a 4-1BB agonist, an OX40 agonist, a CD40 agonist, a CD30 agonist, a GITR agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a CD28H / TMIGD2 agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a 4-1BB agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, and any combinations thereof.

[0347] In some embodiments, a heterologous moiety of the present disclosure is a cytokine.Exemplary cytokines include, but are not limited to, interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-15 (IL-15), interleukin-21 (IL-21), IFN-α, IFN-β, IFN-γ, CCL19, CCL21, interleukin-18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), and a receptor thereof, or any combinations thereof.

[0348] In some embodiments, a heterologous moiety of the present disclosure is a tumor-associated antigen (TAA)- targeting agent. Exemplary TAA-targeting agents include, but are not limited to, 5T4, Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, 78 314460903v1Attorney Docket No: 243735.000428 FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY-ESO-1, P-cadherin, PRAME , PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, RAF, and any combinations thereof. In some embodiments, the TAA targeting agent is a VEGF inhibitor, a VEGFR inhibitor, a KRAS inhibitor, a CD20 inhibitor, a CD38 inhibitor, a TROP2 inhibitor, or a RAF inhibitor.

[0349] In some embodiments, the CD20 inhibitor is rituximab. In some embodiments, the VEGFinhibitor is bevacizumab. In some embodiments, the CD38 inhibitor is daratumumab. In some embodiments, the TROP2 inhibitor is sacituzumab govitecan.

[0350] In some embodiments, the heterologous moiety is a tumor necrosis factor-alpha (TNF-α)inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL- 4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA-targeted T- cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

[0001] In some embodiments, a cytotoxic agent can include, e.g., an anti-tubulin agent, a tubulinpolymerization inhibitor, a DNA synthesis inhibitor, a DNA intercalating agent, a DNA alkylating agent, a DNA cross-linking agent, a DNA cleaving agent, a platinum-based agent, topoisomerase I inhibitor, topoisomerase II inhibitor, an anti-microtubule agent, an anti-mitotic agent, a taxane- related anti-neoplastic agent, an anti-metabolite, an anti-tumor plant alkaloid, and an RNA polymerase II inhibitor. 79 314460903v1Attorney Docket No: 243735.000428

[0351] Non-limiting examples of cytotoxic agents are doxorubicin, nemorubicin, PNU- 159682,paclitaxel, docetaxel, auristatin E, auristatin F, dolastatin 10, dolastatin 15, monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), monomethyl auristatin D (MMAD), maytansine, mertansine (DM1), maytansinoid DM4, calicheamicin, N-acetyl-calicheamicin, vinblastine, vincristine, vindesine, vinorelbine, camptothecin, topotecan, irinotecan, SN-38, duocarmycin A, duocarmycin Bl, duocarmycin B2, duocarmycin CI, duocarmycin C2, duocarmycin D, duocarmycin SA, duocarmycin TM, duocarmycin MB, duocarmycin DM, mitomycin C, rachelmycin, epothilone A, epothilone B, epothilone C, tubulysin B, tubulysin M, pyrrolobenzodiazepine (PBD), bortezomib, ranpimase, hTNF, IL-12, ranpimase, human ribonuclease (RNAse), Bovine pancreatic Rnase, pokeweed antiviral protein, Pseudomonas exotoxin A, gelonin, ricin-A, interferon-alpha, interferon-lambda, urease, amatoxin, alpha- amanitin, beta-amanitin, gamma-amanitin, epsilon- amanitin, bouganin, and staphylococcal enterotoxin.

[0352] Further non-limiting examples of cytotoxic agents are antiviral drugs (e.g., abacavir,acyclovir, ampligen, cidofovir, delavirdine, didanosine, efavirenz, entecavir, fosfonet, ganciclovir, ibacitabine, immunovir, idoxuridine, inosine, lopinavir, methisazone, nexavir, nevirapine, oseltamivir, penciclovir, stavudine, trifluridine, 54PG EMM, valaciclovir, and zanamivir), daunorubicin hydrochloride, daunoriycin, rubidomycin, cerubidine, idarubicin, doxorubicin, epirubicin and morpholino derivatives, phenoxizone biscyclopeptides (e.g., dactinomycin), basic glycopeptides (e.g., bleomycin), anthraquinone glycosides (e.g., plicamycin and mithramycin), anthracenediones (e.g., mitoxantrone), azirinopyrrolo indolediones (e.g., mitomycin), macrocyclic immunosuppressants (e.g., cyclosporine, FK-506, tacrolimus, prograf, and rapamycin), navelbene, CPT-11, anastrazole, letrazole, capecitabine, reloxafine, droloxafine, allocolchicine, Halichondrin B, colchicine and colchicine derivatives, rhizoxin, thiocolchicine, trityl cysterin, vinblastine sulfate, hydroxyurea, N-methylhydrazine, epidophyllotoxin, procarbazine, mitoxantrone, leucovorin, and tegafur, combretatstatin, chalicheamicin, maytansine, DM-I, netropsin, podophyllotoxin (e.g., etoposide and teniposide), baccatin and its derivatives, anti-tubulin agents, cryptophysin, combretastatin, vincristine, vincristine sulfate, vinblastine, vindesine, vinorelbine, VP-16, camptothecin, epothilone A, epothilone B, nocodazole, colchicines, colcimid, estramustine, cemadotin, discodermolide, eleutherobin, mechlorethamine, cyclophosphamide, melphalan, carmustine, lomustine, semustine, streptozocin, chlorozotocin, uracil mustard, 80 314460903v1Attorney Docket No: 243735.000428 chlormethine, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, dacarbazine, temozolomide, ytarabine, cytosine arabinoside, fluorouracil, 5-fluorouracil (5-FU), floxuridine, 6-thioguanine, 6-mercaptopurine, pentostatin, methotrexate, 10-propargyl- 5,8-dideazafolate, 5,8-dideazatetrahydrofolic acid, leucovorin, NCA1, auristatin, auristatin E, DNA minor groove binding agents, DNA minor groove alkylating agents, enediyne, lexitropsin, duocarmycin, taxane, puromycin, dolastatin, maytansinoid, vinca alkaloid, AFP, MMAF, MMAE, AEB, AEVB, 54PG EMM (e.g., paclitaxel and paclitaxel derivatives (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE®(American Pharmaceutical Partners, Schaumberg, Ill.), as well as docetaxel and docetaxel derivatives), CC-1065, SN-38, topotecan, morpholino-doxorubicin, rhizoxin, cyanomorpholino-doxorubicin, dolastatin-10, echinomycin, fludarabine phosphate, pentostatine, gemcitabine, Ara-C, deoxycoformycin, mitomycins such as mitomycin-C, L-asparaginase, azathioprine, brequinar, antibiotics (e.g., anthracycline, gentamicin, cefalotin, vancomycin, telavancin, daptomycin, azithromycin, erythromycin, rocithromycin, furazolidone, amoxicillin, ampicillin, carbenicillin, flucloxacillin, methicillin, penicillin, ciprofloxacin, moxifloxacin, ofloxacin, doxycycline, minocycline, oxytetracycline, tetracycline, streptomycin, rifabutin, ethambutol, and rifaximin), enediyne antibiotics (e.g., calicheamicin, calicheamicin gamma1I and calicheamicin omegaI1, and dynemicin, including dynemicin A),. “Taxanes” include paclitaxel, as well as any active taxane derivative or pro-drug. Chemotherapeutic agents such as erlotinib (TARCEVA®, Genentech / OSI Pharm.), bortezomib (VELCADE®, Millenium Pharm.), fulvestrant (FASLODEX®, AstraZeneca), sunitinib (Sutent®, Pfizer), letrozole (FEMARA®, Novartis), imatinib mesylate (GLEEVEC®, Novartis), PTK787 / ZK 222584 (Novartis), oxaliplatin (Eloxatin®, Sanofi), leucovorin, lapatinib (TYKERB®, GSK572016, GlaxoSmithKline), lonafarnib (SCH 66336), sorafenib (BAY43-9006, Bayer Labs.), and gefitinib (IRESSA®, AstraZeneca), AG1478, AG1571 (SU 5271; Sugen), alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; antifolate antineoplastic such as pemetrexed (ALIMTA®Eli Lilly); aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylomelamine; acetogenins (such as bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogues adozelesin, carzelesin and bizelesin); 81 314460903v1Attorney Docket No: 243735.000428 cryptophycins (such as cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including its synthetic analogues KW-2189 and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, and uracil mustard; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; 54PG EMM55zu; defofamine; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; 55PG EMM, e.g., paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE™ Cremophor-free, albumin, nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE®doxetaxel (Rhone- Poulenc Rorer, Antony, France); 55PG EMM55zuma; GEMZAR®gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); mitoxantrone; NAVELBINE®vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores, aclacinomysins, actinomycin, anthramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (ADRIAMYCIN®) (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2- pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-FU; folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, 56PG EMM56zum, 6-azauridine, 82 314460903v1Attorney Docket No: 243735.000428 carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; losoxantrone; podophyllinic acid; 2- ethylhydrazide; procarbazine; PSK®polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2’,2’’- trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; and pharmaceutically acceptable salts, esters, acids, prodrugs, or derivatives of any of the above.

[0353] In some embodiments, a heterologous moiety of the present disclosure is a lysosome-targeting chimera (LYTAC). In some embodiments, a LYTAC described herein may be used, e.g., to target TTYH2 protein for degradation (e.g., via a lysosomal-based degradation pathway). A LYTAC can target membrane proteins and extracellular proteins for degradation, such as is described by Ahn, G. et. Al., Nat Chem Biol, 17(9), 2021, 937-946 (PMID 33767387), Banik, S. M. et. Al., Nature, 584(7820), 2020, 291-297 (PMID 32728216) and Caianiello et al., Nat Chem Biol. 2021 Sep;17(9):947-953 (PMID 34413525), each of which is incorporated herein by reference in its entirety and for all purposes as if fully set forth herein. Without wishing to be bound by theory, LYTACs can bridge the extracellular domain of a target protein (e.g., TTYH2 protein) to a cell-surface lysosomal targeting receptor. The motif that binds the lysosomal targeting receptor may be, e.g., without limitation, N-acetylgalactosamine (GalNAc), galactose (Gal), or mannose-6-phosphonate (M6Pn). The lysosomal targeting receptor be, e.g., without limitation, a cation-independent mannose-6-phosphate receptor (CI-M6PR) or asialoglycoprotein receptor (ASGPR). The lysosomal targeting receptor can allow for degradation of a protein (e.g., TTYH2 protein) in a cell type-specific manner. In some embodiments, a heterologous moiety of the present disclosure is ZNFR3 (Marei, H., Tsai, WT.K., Kee, YS. et al. Antibody targeting of E3 ubiquitin ligases for receptor degradation. Nature 610, 182–189 (2022)

[0354] In some embodiments, a heterologous moiety of the present disclosure is anoligonucleotide (e.g., siRNA or antisense oligonucleotide). In some embodiments, an oligonucleotide is described herein may be used, e.g., to silence TTYH2.

[0355] Methods for making the antibody-drug conjugates disclosed herein is also contemplatedas within the scope of this disclosure. In one aspect, provided herein is a method for making an antibody-drug conjugate described herein comprising 83 314460903v1Attorney Docket No: 243735.000428 (a) contacting the antibody or antigen-binding fragment with the heterologous moiety under the conditions favorable for conjugation of the antibody or antigen-binding fragment to the heterologous moiety; and (b) optionally, isolating the antibody-drug conjugate produced in step (a).

[0356] In order to facilitate the coupling between the heterologous moiety and the antibody, itis possible to directly conjugate the two agents, or to introduce a spacer molecule between them. Suitable spacers include poly(alkylene) glycols such as polyethylene glycol, and peptide linkers. Many suitable coupling techniques are well known in the art. Suitable agents allowing covalent, electrostatic or noncovalent binding of the moiety to the antibody include benzoquinone, carbodiimide and more particularly EDC (1-ethyl-3-[3-dimethyl-aminopropyl]-carbodiimide hydrochloride), dimaleimide, dithiobis-nitrobenzoic acid (DTNB), N-succinimidyl S-acetyl thio- acetate (SATA), the bridging agents having one or more phenylazide groups reacting with the ultraviolets (U.V.) and preferably N-[-4-(azidosalicylamino)butyl]-3’-(2’-pyridyldithio)- propionamide (APDP), N-succinimid-yl 3-(2-25 pyridyldithio)propionate (SPDP), 6-hydrazino- nicotinamide (HYNIC). Another form of coupling, especially for the radioelements, includes the use of a bifunctional ion chelator. For example, chelates derived from EDTA or DTPA which have been developed for binding metals, especially radioactive metals, and immunoglobulins. Thus, DTPA and its derivatives can be substituted by different groups on the carbon chain in order to increase the stability and the rigidity of the ligand-metal complex, as is well known in the art.

[0357] In some embodiments, conjugation of a heterologous moiety to an antibody or antigen-binding fragment described herein is carried out via the covalent-nature binding, or binding by the use of adapter molecules or linkers.

[0358] Covalent binding requires prior activation of the heterologous moieties. In someembodiments, covalent strategies occur via carbodiimide chemistry, maleimide chemistry or “click chemistry”, as discussed in detail below.

[0359] In some embodiments, a heterologous moiety described herein is conjugated to an anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, a heterologous moiety described herein is conjugated to an anti-TTYH2 antibody or antigen-binding fragment directly. In some embodiments, a heterologous moiety described herein is conjugated to an anti-TTYH2 antibody or antigen-binding fragment via a linker covalently connecting the anti-TTYH2 antibody or antigen-binding fragment with the heterologous moiety. 84 314460903v1Attorney Docket No: 243735.000428

[0360] In some embodiments, the heterologous moiety described herein is conjugated to the anti-TTYH2 antibody or antigen-binding fragment by a chemical ligation process. In some embodiments, the heterologous moiety described herein is conjugated to the anti-TTYH2 antibody or antigen-binding fragment by a native ligation. In some embodiments, the conjugation is as described in e.g., Dawson, et al. Science 1994, 266, 776-779; Dawson, et al. J. Am. Chem. Soc. 1997, 119, 4325-4329; Hackeng, et al. Proc. Natl. Acad. Sci. USA 1999, 96, 10068-10073; or Wu, et al. Angew. Chem. Int. Ed. 2006, 45, 4116-4125, each of which is herein incorporated by reference in its entirety. In some embodiments, the conjugation is as described in U.S. Patent No. 8,936,910, which is herein incorporated by reference in its entirety. In some embodiments, the heterologous moiety described herein is conjugated to the anti-TTYH2 antibody or antigen- binding fragment either site-specifically or non-specifically via native ligation chemistry.

[0361] Sortase (SrtA), a transpeptidase from Staphylococcus aureus, catalyzes a cell-wallsorting reaction at an LPXTG (SEQ ID NO: 28) motif by cleaving between threonine and glycine and subsequently joining the carboxyl group of threonine to an amino group of pentaglycine on the cell wall peptidoglycan. In some embodiments, sortase-mediated ligation may be utilized to covalently link an antibody or an antigen-binding fragment thereof according to the present disclosure comprising a sortase recognition sequence (e.g., a LPXT sequence) to a heterologous moiety comprising a (G), sequence (see, e.g., Proft T. Sortase-mediated protein ligation: an emerging biotechnology tool for protein modification and immobilization. Biotechnol Lett.2010, 32(1):1-10). In one embodiment, an antibody or an antigen-binding fragment thereof according to the present disclosure comprising an LPXT motif can be specifically ligated to a linker or linker- payload comprising an aminoglycine via an amide bond in the presence of sortase.

[0362] Subtiligase is a variant of the serine protease subtilisin BPN’ that has been engineered tocatalyze a ligation reaction between a peptide ester donor substrate and the N-terminal α-amine of a peptide or protein (Abrahmsen et al., 1991). Based on its ability to catalyze peptide bond formation with absolute chemoselectivity for N-terminal α-amines over lysine α-amines, subtiligase has been utilized for both site-specific protein bioconjugation and for cellular N- terminomics studies (see, e.g., Weeks, A. M. Subtiligase-Catalyzed Peptide Ligation. Chemical Reviews 2020, 120(6): 3127-3160). In some embodiments, subtiligase-mediated ligation may be utilized to covalently link an antibody or an antigen-binding fragment thereof according to the present disclosure to a linker or linker-payload comprising an ester moiety. In one embodiment, 85 314460903v1Attorney Docket No: 243735.000428 an antibody or an antigen-binding fragment thereof according to the present disclosure can be specifically ligated to a linker or a linker-payload comprising an ester moiety via an amide bond in the presence of subtiligase.

[0363] In some embodiments, the heterologous moiety described herein is conjugated to the anti-TTYH2 antibody or antigen-binding fragment by a site-directed method utilizing an enzyme- catalyzed process. In some embodiments, the site-directed method utilizes SMARTagTM technology (Catalent, Inc.). In some embodiments, the SMARTagTM technology comprises generation of a formylglycine (Fgly) residue from cysteine by formylglycine-generating enzyme (FGE) through an oxidation process under the presence of an aldehyde tag and the subsequent conjugation of Fgly to an alkylhydraine-functionalized heterologous moiety described herein (e.g., a cytotoxic agent) via 59PG EMM59z-Pictet-Spengler (HIPS) ligation. (See Wu et al., PNAS 106(9): 3000-3005 (2009); Agarwal, et al., PNAS 110(1): 46-51 (2013), each of which is herein incorporated by reference in its entirety).

[0364] In some embodiments, the heterologous moiety described herein is conjugated to the anti-TTYH2 antibody or antigen-binding fragment by a site-directed method utilizing a “traceless” coupling technology (Philochem). In some embodiments, the “traceless” coupling technology utilizes an N-terminal 1,2-aminothiol group on the anti-TTYH2 antibody or antigen-binding fragment which is then conjugated with a heterologous moiety described herein (e.g., a cytotoxic agent) containing an aldehyde group. (See Casi et al., JACS 134(13): 5887-5892 (2012), which is herein incorporated by reference in its entirety).

[0365] In some embodiments, the heterologous moiety described herein is conjugated to the anti-TTYH2 antibody or antigen-binding fragment by a site-directed method utilizing an unnatural amino acid incorporated into the anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, the unnatural amino acid comprises p-acetylphenylalanine (pAcPhe). In some embodiments, the keto group of pAcPhe is selectively coupled to an alkoxy-amine derivatived conjugating moiety to form an oxime bond. (See Axup et al., PNAS 109(40): 16101-16106 (2012), which is herein incorporated by reference in its entirety).

[0366] In some embodiments, the enzyme-catalyzed process comprises transglutaminase (TG),e.g., microbial transglutaminase (mTG). In some cases, the heterologous moiety described herein (e.g., a cytotoxic agent) is conjugated to the anti-TTYH2 antibody or antigen-binding fragment utilizing a microbial transglutaminase-catalyzed process. In some embodiments, mTG catalyzes 86 314460903v1Attorney Docket No: 243735.000428 the formation of a covalent bond between the amide side chain of a glutamine within the recognition sequence and a primary amine of a functionalized heterologous moiety described herein (In some embodiments, mTG is produced from Streptomyces mobarensis. (See Strop et al., Chemistry and Biology 20(2) 161-167 (2013), which is herein incorporated by reference in its entirety).

[0367] In some embodiments, a sequence of amino acids comprising an acceptor glutamineresidue are incorporated into (e.g., appended to) a polypeptide sequence, under suitable conditions, for recognition by a TG. This sequence leads to cross-linking by the TG through a reaction between an amino acid side chain within the sequence of amino acids and a reaction partner. The recognition tag may be a peptide sequence that is not naturally present in the polypeptide comprising the TG recognition tag. In some embodiments, the TG recognition tag comprises at least one Gln.

[0368] In some embodiments, the TGase recognition tag comprises an amino acid sequenceXXQX, wherein X is any amino acid (e.g., conventional amino acid Leu, Ala, Gly, Ser, Val, Phe, Tyr, His, Arg, Asn, Glu, Asp, Cys, Gln, Ile, Met, Pro, Thr, Lys, or Trp or nonconventional amino acid). In some embodiments, the acyl donor glutamine-containing tag comprises an amino acid sequence selected from the group consisting of LLQ, LLQG (SEQ ID NO: 29), LLQGG (SEQ ID NO: 30), LSLSQG (SEQ ID NO: 31), GGGLLQGG (SEQ ID NO: 32), GLLQ (SEQ ID NO: 33), GLLQG (SEQ ID NO: 34), GLLQGGG (SEQ ID NO: 35), GLLQGG (SEQ ID NO: 36), GSPLAQSHGG (SEQ ID NO: 37), LLQLLQGA (SEQ ID NO: 38), LLQGA (SEQ ID NO: 39), LLQYQGA (SEQ ID NO: 40), LLQGSG (SEQ ID NO: 41), LLQYQG (SEQ ID NO: 42), LLQLLQG (SEQ ID NO: 43), SLLQG (SEQ ID NO: 44), LLQLQ (SEQ ID NO: 45), LLQLLQ (SEQ ID NO: 46), and LLQGR (SEQ ID NO: 47). See, e.g., PCT Publication No. WO2012 / 059882, which is herein incorporated by reference in its entirety. In some embodiments, the acyl donor glutamine-containing tag is present at the N-terminus of the antibody or antigen- binding fragment. In some embodiments, the acyl donor glutamine-containing tag is present at the C-terminus of the antibody or antigen-binding fragment. In some embodiments, the acyl donor glutamine-containing tag is present both at the N-terminus and the C-terminus of the antibody or antigen-binding fragment.

[0369] In some embodiments, the heterologous moiety described herein (e.g., a cytotoxic agent)is conjugated to the anti-TTYH2 antibody or antigen-binding fragment by a method which utilizes 87 314460903v1Attorney Docket No: 243735.000428 a sequence-specific transpeptidase (see, e.g., PCT Publication No. WO2014 / 140317, which is herein incorporated by reference in its entirety). Other conjugation methods include those described in U.S. Patent Publication Nos. 2015 / 0105539 and 2015 / 0105540, each of which is herein incorporated by reference in its entirety.

[0370] In some embodiments, the heterologous moiety described herein (e.g., a cytotoxic agent)is conjugated to the anti-TTYH2 antibody or antigen-binding fragment utilizing Azide-Alkyne Cycloaddition (CuAAC) click chemistry. Azides and alkynes can undergo catalyst free [3+2] cycloaddition by a using the reaction of activated alkynes with azides. Such catalyst-free [3+2] cycloaddition can be used in the methods described herein to conjugate an anti-TTYH2 antibody or antigen-binding fragment and the heterologous moiety described herein (e.g., a cytotoxic agent). Alkynes can be activated by ring strain such as, by way of example only, eight-membered ring structures, or nine-membered, appending electron-withdrawing groups to such alkyne rings, or alkynes can be activated by the addition of a Lewis acid such as, by way of example only, Au(I) or Au(III).

[0371] Alkynes activated by ring strain have been described and used in “copperless” [3+2]cycloaddition. Non-limiting examples include cyclooctynes and difluorocyclooctynes (Agard et al., J. Am. Chem. Soc., 126 (46):15046-15047 (2004)), dibenzocyclooctynes (PCT International Publication No. WO 2009 / 067663 Al (2009)), aza-dibenzocyclooctynes (Debets et al., Chem. Comm., 46:97-99 (2010)), and cyclononynes (Dommerholt et al., Angew. Chem.122:9612-9615 (2010)). In some embodiments, a tetrazine (Tzn)-activated anti-TTYH2 antibody or antigen- binding fragment may be cross-linked to a trans-cyclooctene (TCO)-activated heterologous moiety described herein (e.g., a cytotoxic agent). In some embodiments, a TCO-activated anti-TTYH2 antibody or antigen-binding fragment may be crosslinked to a Tzn-activated heterologous moiety described herein (e.g., a cytotoxic agent).

[0372] Complexes described herein may comprise a linker that connects an antibody or antigen-binding fragment to a heterologous moiety (e.g., a cytotoxic agent). A linker comprises at least one covalent bond. In some embodiments, a linker may be a single bond, e.g., a disulfide bond or disulfide bridge, that connects an antibody or antigen-binding fragment to a heterologous moiety (e.g., a cytotoxic agent). However, in some embodiments, a linker may connect an antibody or antigen-binding fragment to a heterologous moiety (e.g., a cytotoxic agent) through multiple covalent bonds. A linker is generally stable in vitro and in vivo, and may be stable in certain cellular 88 314460903v1Attorney Docket No: 243735.000428 environments. Additionally, generally a linker does not negatively impact the functional properties of either the antibody or antigen-binding fragment or the heterologous moiety (e.g., a cytotoxic agent).

[0373] A precursor to a linker typically will contain two different reactive species that allow forattachment to both the antibody or antigen-binding fragment and a heterologous moiety (e.g., a cytotoxic agent. In some embodiments, the two different reactive species may be a nucleophile and / or an electrophile. In some embodiments, a linker is connected to an antibody or antigen- binding fragment via conjugation to a lysine residue or a cysteine residue of the antibody or antigen-binding fragment. In some embodiments, a linker is connected to a cysteine residue of an antibody or antigen-binding fragment via a maleimide-containing linker, wherein optionally the maleimide-containing linker comprises a maleimidocaproyl or maleimidomethyl cyclohexane- 1 - carboxylate group. In some embodiments, a linker is connected to a cysteine residue of an antibody or antigen-binding fragment or thiol functionalized heterologous moiety via a 3-arylpropionitrile functional group. In some embodiments, a linker is connected to an antibody or antigen-binding fragment and / or a heterologous moiety (e.g., a cytotoxic agent) via an amide bond, a hydrazide, a triazole, a thioether or a disulfide bond.

[0374] In some embodiments, a linker described herein is a cleavable linker or a non-cleavablelinker. In some embodiments, the linker is a cleavable linker. In other embodiments, the linker is a non-cleavable linker.

[0375] A cleavable linker may be a protease-sensitive linker, a pH-sensitive linker, or aglutathione-sensitive linker. These linkers are generally cleavable only intracellularly and are preferably stable in extracellular environments.

[0376] Protease-sensitive linkers are cleavable by protease enzymatic activity. These linkerstypically comprise peptide sequences and may be 2-10 amino acids, about 2-5 amino acids, about 5-10 amino acids, about 10 amino acids, about 5 amino acids, about 3 amino acids, or about 2 amino acids in length. In some embodiments, a peptide sequence may comprise naturally- occurring amino acids, e.g. cysteine, alanine, or non-naturally-occurring or modified amino acids. Non-naturally occurring amino acids include 3-amino acids, homo-amino acids, proline derivatives, 3-substituted alanine derivatives, linear core amino acids, N-methyl amino acids, and others known in the art. In some embodiments, a protease-sensitive linker comprises a valine- 89 314460903v1Attorney Docket No: 243735.000428 citrulline or alanine-citrulline dipeptide sequence. In some embodiments, a protease-sensitive linker can be cleaved by a lysosomal protease, e.g. cathepsin B, and / or an endosomal protease.

[0377] A pH-sensitive linker is a covalent linkage that readily degrades in high or low pHenvironments. In some embodiments, a pH-sensitive linker may be cleaved at a pH in a range of 4 to 6. In some embodiments, a pH-sensitive linker comprises a hydrazone or cyclic acetal. In some embodiments, a pH-sensitive linker is cleaved within an endosome or a lysosome.

[0378] In some embodiments, a glutathione-sensitive linker comprises a disulfide moiety. Insome embodiments, a glutathione-sensitive linker is cleaved by a disulfide exchange reaction with a glutathione species inside a cell. In some embodiments, the disulfide moiety further comprises at least one amino acid, e.g. a cysteine residue.

[0379] In some embodiments, non-cleavable linkers may be used. Generally, a non-cleavablelinker cannot be readily degraded in a cellular or physiological environment. In some embodiments, a non-cleavable linker comprises an optionally substituted alkyl group, wherein the substitutions may include halogens, hydroxyl groups, oxygen species, and other common substitutions. In some embodiments, a linker may comprise an optionally substituted alkyl, an optionally substituted alkylene, an optionally substituted arylene, a heteroarylene, a peptide sequence comprising at least one non-natural amino acid, a truncated glycan, a sugar or sugars that cannot be enzymatically degraded, an azide, an alkyneazide, a peptide sequence comprising a LPXT sequence, a thioether, a biotin, a biphenyl, repeating units of polyethylene glycol or equivalent compounds, acid esters, acid amides, sulfamides, and / or an alkoxy-amine linker.

[0380] In some embodiments, a linker may comprise a substituted alkylene, an optionallysubstituted alkenylene, an optionally substituted alkynylene, an optionally substituted cycloalkylene, an optionally substituted cycloalkenylene, an optionally substituted arylene, an optionally substituted heteroarylene further comprising at least one heteroatom selected from N, O, and S; an optionally substituted heterocyclylene further comprising at least one heteroatom selected from N, O, and S; an imino, an optionally substituted nitrogen species, an optionally substituted oxygen species, an optionally substituted sulfur species, or a poly(alkylene oxide), e.g. polyethylene oxide or polypropylene oxide.

[0381] In some cases, the linker is a non-polymeric linker. A non-polymeric linker refers to alinker that does not contain a repeating unit of monomers generated by a polymerization process. Exemplary non-polymeric linkers include, but are not limited to, C1-C30 alkyl group (e.g., a C5, 90 314460903v1Attorney Docket No: 243735.000428 C4, C3, C2, or C1 alkyl group), homobifunctional cross linkers, heterobifunctional cross linkers, peptide linkers, traceless linkers, self-immolative linkers, maleimide-based linkers, or combinations thereof. In some cases, the non-polymeric linker comprises a C1-C30 alkyl group (e.g., a C5, C4, C3, C2, or C1 alkyl group), a homobifunctional cross linker, a heterobifunctional cross linker, a peptide linker, a traceless linker, a self-immolative linker, a maleimide-based linker, or a combination thereof. In additional cases, the non-polymeric linker does not comprise more than two of the same type of linkers, e.g., more than two homobifunctional cross linkers, or more than two peptide linkers. In further cases, the non-polymeric linker optionally comprises one or more reactive functional groups. In some cases, the non-polymeric linker does not encompass a polyalkylene oxide (e.g., PEG). In some cases, the non-polymeric linker does not encompass a PEG.

[0382] In some embodiments, the linker comprises a homobifunctional linker. Exemplaryhomobifunctional linkers include, but are not limited to, bis(sulfosuccinimidyl)suberate (BS), disuccinimidyl tartrate (DST), disulfosuccinimidyl tartrate (sulfo DST), ethylene glycobis(succinimidylsuccinate) (EGS), disuccinimidyl glutarate (DSG), N,N’-disuccinimidyl carbonate (DSC), dimethyl adipimidate (DMA), dimethyl pimelimidate (DMP), dimethyl suberimidate (DMS), dimethyl-3,3’-dithiobispropionimidate (DTBP), 1,4-di-3’-(2’- pyridyldithio)propionamido) butane (DPDPB), bismaleimidohexane (BMH), aryl halide- containing compound (DFDNB), such as e.g.1,5-difluoro-2,4-dinitrobenzene or 1,3-difluoro-4,6- dinitrobenzene, 4,4’-difluoro-3,3’-dinitrophenylsulfone (DFDNPS), bis-113-(4- azidosalicylamido)ethyl]disulfide (BASED), organoazide, organoalkyne, Lomant’s reagent dithiobis (succinimidylpropionate) DSP, 3’3’-dithiobis(sulfosuccinimidyl proprionate (DTSSP), disuccinimidyl suberate (DSS), formaldehyde, glutaraldehyde, 1,4-butanediol diglycidyl ether, adipic acid dihydrazide, carbohydrazide, o-toluidine, 3,3’-dimethylbenzidine, benzidine, a,a’-p- diaminodiphenyl, diiodo-p-xylene sulfonic acid, N,N’-ethylene-bis(iodoacetamide), or N,N’- hexamethylene-bis(iodoacetamide).

[0383] In some embodiments, the linker comprises a heterobifunctional linker. Non-limitingexamples of heterobifunctional linker include carbonyl-reactive and sulfhydrylreactive cross- linkers such as 4-(4-N-maleimidophenyl) butyric acid hydrazide (MPBH), 4-(N- maleimidomethyl) cyclohexane-l-carboxyl-hydrazide-8 (M2C2H), 3-(2- pyridyldithio)propionyl hydrazide (PDPH), amine-reactive and sulfhydryl cross-linkers such as N-succinimidyl 3-(2- 91 314460903v1Attorney Docket No: 243735.000428 pyridyldithio) propionate (sPDP), long-chain N-succinimidyl 3-(2-pyridyldithio) propionate (LC- sPDP), water-soluble-long-chain N-succinimidyl 3-(2-pyridyldithio) propionate (sulfo-LCsPDP), succinimidyloxycarbonyl-a-methyl-a-(2-pyridyldithio) toluene (sMPT), sulfosuccinimidy1-6-[a- methyl-a-(2-pyridyldithio)toluamido]hexanoate (sulfo-LC-sMPT), succinimidy1-4-(N- maleimidomethyl) cyclohexane-1-car-boxylate (sMCC), sulfosuccinimidyl-4-(N- maleimidomethyl) cyclohexane- 1-carboxylate (sulfo-sMCC), m-maleimidobenzoyl-N- hydroxysuccinimide ester (MBs), m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester (sulfo- MB s), N-succinimidyl (4-iodoacteyl)aminobenzoate (sIAB), sulfosuccinimidyl (4- iodoacteyl)aminobenzoate (sulfo-sIAB), succinimidyl-4-(p-maleimidophenyl)butyrate (sMPB), sulfosuccinimidyl-4-(p-maleimidophenyl)butyrate (sulfo-sMPB), N-(y- maleimidobutyryloxy)succinimide ester (GMBs), N-(y-maleimidobutyryloxy)sulfosuccinimide ester (sulfo-GMBs), succinimidyl 6-((iodoacetyl)amino)hexanoate (sIAX), succinimidyl 6-[6- (((iodoacetyl)amino) hexanoyl)amino]hexanoate (sIAXX), succinimidyl 4-(((iodoacetyl) amino)methyl)cyclohexane-l-carboxylate (sIAC), succinimidyl 6-((((4- iodoacetyl)amino)methyl)cyclohexane-1 -carbonyl)amino) hexanoate (sIACX), p-nitrophenyl iodoacetate (NPIA), amine-reactive and photoreactive cross-linkers such as N- hydroxysuccinimidyl-4-azidosalicylic acid (NHs-AsA), N-hydroxysulfosuccinimidyl-4- azidosalicylic acid (sulfo-NHs-AsA), sulfosuccinimidyl-(4-azidosalicylamido)hexanoate (sulfo- NHs-LC-AsA), sulfosuccinimidy1-2-(p-azidosalicylamido)ethyl1,3’-dithiopropionate (sAsD), N- hydroxysuccinimidyl-4-azidobenzoate (HsAB), N-hydroxysulfosuccinimidyl-4-azidobenzoate (sulfo-HsAB), N-succinimidyl-6-(4’-azido2’-nitrophenylamino)hexanoate (sANPAH), sulfo succinimidyl- 6- (4’ -azido-2’-nitrophenylamino)hexanoate (sulfo-sANPAH), N-5-azido-2- nitrobenzoyloxysuccinimide (ANB-NOs), sulfosuccinimidy1-2-(m-azido-o-nitrobenzamido)- ethyl-1,3’-dithiopropionate (sAND), N-succinimidyl-4(4-azidopheny1)1,3’-dithiopropionate (sADP), N-sulfosuccinimidyl(4-azidophenyl)-1,3’-dithiopropionate (sulfo-sADP), sulfosuccinimidyl 4-(p-azidophenyl) butyrate (sulfo-sAPB), sulfosuccinimidyl 2-(7-azido-4- methylcoumarin-3-acetamide)ethy1-1,3’-dithiopropionate (sAED), sulfosuccinimidyl 7-azido-4- methylcoumain-3-acetate (sulfo-sAMCA), p-nitrophenyl diazopyruvate (pNPDP), p-nitrophenyl- 2-diazo-3,3,3-trifluoropropionate (PNP-DTP), carboxylate-reactive and photoreactive cross- linkers such as 4-(p-azidosalicylamido) butylamine (AsBA), and arginine-reactive and photoreactive cross-linkers such as p-azidophenyl glyoxal (APG); sulfhydryl-reactive and 92 314460903v1Attorney Docket No: 243735.000428 photoreactive crosslinkers such as 1-(p-Azidosalicylamido)-4-(iodoacetamido) butane (AsIB), N- [4-(p-azidosalicylamido)buty1]-3’-(2’-pyridyldithio)propionamide (APDP), benzophenone-4- iodoacetamide, benzophenone-4-maleimide carbonylreactive and photoreactive cross-linkers such as p-azidobenzoyl hydrazide (ABH).

[0384] In some embodiments, the linker comprises a reactive functional group. In some cases,the reactive functional group comprises a nucleophilic group that is reactive to an electrophilic group present on an anti-TTYH2 antibody or antigen-binding fragment. Exemplary electrophilic groups include carbonyl groups such as aldehyde, ketone, carboxylic acid, ester, amide, enone, acyl halide or acid anhydride. In some embodiments, the reactive functional group is aldehyde. Exemplary nucleophilic groups include hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate, and arylhydrazide.

[0385] In some embodiments, the linker comprises a maleimide group. In some embodiments,the maleimide group is also referred to as a maleimide spacer. In some embodiments, the maleimide group further encompasses a caproic acid, forming maleimidocaproyl (mc). In some cases, the linker comprises maleimidocaproyl (mc). In some cases, the linker is maleimidocaproyl (mc). In other embodiments, the maleimide group comprises a maleimidomethyl group, such as succinimidy1-4-(N-maleimidomethyl)cyclohexane-l-carboxylate (sMCC) or sulfosuccinimidy1- 4-(N-maleimidomethyl)cyclohexane-1 -carboxylate (sulfo-sMCC) described above.

[0386] In some embodiments, the maleimide group is a self-stabilizing maleimide. In someembodiments, the self-stabilizing maleimide utilizes diaminopropionic acid (DPR) to incorporate a basic amino group adjacent to the maleimide to provide intramolecular catalysis of tiosuccinimide ring hydrolysis, thereby eliminating maleimide from undergoing an elimination reaction through a retro-Michael reaction. In some embodiments, the self-stabilizing maleimide is a maleimide group described in Lyon, et al., Nat. Biotechnol.32(10):1059-1062 (2014). In some embodiments, the linker comprises a self-stabilizing maleimide. In some embodiments, the linker is a self-stabilizing maleimide.

[0387] In some embodiments, the linker is a traceless linker or a linker in which after cleavagedoes not leave behind a linker moiety (e.g., an atom or a linker group) to an anti-TTYH2 antibody or antigen-binding fragment or a polynucleotide B. Exemplary traceless linkers include, but are not limited to, aryl-triazene linkers, germanium linkers, silicium linkers, sulfur linkers, selenium linkers, nitrogen linkers, phosphorus linkers, boron linkers, chromium linkers, or phenylhydrazide 93 314460903v1Attorney Docket No: 243735.000428 linker. In some embodiments, the linker is a traceless linker described in Blaney, et al., Chem. Rev.102: 2607-2024 (2002) or U.S. Patent No.6,821,783.

[0388] In some embodiments, the linker is a linker described in U.S. Pat. Nos. 6,884,869;7,498,298; 8,288,352; 8,609,105; or 8,697,688; U.S. Patent Publication Nos. US2014 / 0127239; US2013 / 028919; US2014 / 286970; US2013 / 0309256; US2015 / 037360; and US2014 / 0294851; or International Application Publication Nos. WO2015 / 057699; WO2014 / 080251; WO2014 / 197854; WO2014 / 145090; WO2014 / 177042, WO2022 / 015656, each of which is herein incorporated by reference in its entirety.

[0389] In some embodiments, a linker is absent. In some cases, a linker is a non-polymericlinker. In some cases, a linker is a polymeric linker.

[0390] In some embodiments, the linker comprises an alkyl group. In some embodiments, thelinker comprises a C1-C30 alkyl group, or a C1-C24 alkyl group, or a C1-C20 alkyl group, or a C1-C16 alkyl group, or a C1-C12 alkyl group, or a C1-C10 alkyl group, or a C1-C8 alkyl group, or a C1-C6 alkyl group, or a C1-C4 alkyl group. In some cases, a linker is a C1-C6 alkyl group, such as for example, a C3, C4, C3, C2, or C1 alkyl group. In some cases, the C1-C6 alkyl group is an unsubstituted C1-C6 alkyl group. As used in the context of a linker, alkyl means a saturated straight or branched hydrocarbon radical containing up to six carbon atoms. In some embodiments, the linker comprises a homobifunctional linker or a heterobifunctional linker described herein.

[0391] In some cases, a linker is an oligomeric or a polymeric linker. In some embodiments, alinker is a natural or synthetic oligomer or polymer, consisting of branched or unbranched monomers, and / or cross-linked network of monomers in two or three dimensions. In some embodiments, the linker comprises a polysaccharide, lignin, rubber, or polyalkylen oxide (e.g., polyethylene glycol).

[0392] In some embodiments, polymeric linker includes, but is not limited to, alpha-, omega-dihydroxylpolyethyleneglycol, biodegradable lactone-based polymer, e.g. polyacrylic acid, polylactide acid (PLA), poly(glycolic acid) (PGA), polypropylene, polystyrene, polyolefin, polyamide, polycyanoacrylate, polyimide, polyethylene terephthalate (also known as poly(ethylene terephthalate), PET, PETG, or PETE), polytetramethylene glycol (PTG), or polyurethane as well as mixtures thereof. In some embodiments, the linker comprises polyalkylene oxide. In some embodiments, the linker comprises PEG. In some embodiments, the linker comprises polyethylene imide (PEI) or hydroxy ethyl starch (HES). 94 314460903v1Attorney Docket No: 243735.000428

[0393] In some embodiments, the linker comprises a polyalkylene oxide (e.g., PEG) comprisingdiscrete ethylene oxide units. In some cases, the linker comprises between about 2 and about 48 ethylene oxide units. In some cases, the polymer linker comprises about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 24, about 30, about 36, about 42, or about 48 ethylene oxide units.

[0394] In some embodiments, the anti-TTYH2 antibody or antigen-binding fragment isconjugated to the heterologous moiety described herein (e.g., a cytotoxic agent) using a protamine linker (see, e.g., U.S. Patent Application Publication Nos. US2002 / 0132990, US2004 / 0023902, US2007 / 012152, and US2010 / 0209440, each of which is herein incorporated by reference in its entirety). In some embodiments, a protamine linker encompassed for use in the antibody-drug conjugates described herein comprises a sequence disclosed in US 2010 / 0209440, which is herein incorporated by reference in its entirety.

[0395] Acid cleavable linkers can also be used in the antibody-drug conjugates described hereinand include, but are not limited to, bismaleimideothoxy propane, adipic acid dihydrazide linkers and acid labile transferrin conjugates that contain a sufficient portion of transferrin to permit entry into the intracellular transferrin cycling pathway. Conjugates linked via acid cleavable linkers should be preferentially cleaved in acidic intracellular compartments, such as the endosome.

[0396] Photocleavable linkers can also be used with in the antibody-drug conjugates describedherein. Photocleavable linkers are cleaved upon exposure to light, thereby releasing the targeted agent upon exposure to light. Example photocleavable linkers may comprise nitrobenzyl group as a photocleavable protective group for cysteine; water soluble photocleavable copolymers, including hydroxypropylmethacrylamide copolymer, glycine copolymer, fluorescein copolymer and methylrhodamine copolymer; and nitrobenzyloxy carbonyl chloride cross linking reagents that produce photocleavable linkages. Such linkers are particularly useful in treating dermatological or ophthalmic conditions. In addition, other tissues, such as blood vessels that can be exposed to light using fiber-optics during angioplasty in the prevention or treatment of restenosis may benefit from the use of photocleavable linkers. After administration of the conjugate, the body part is exposed to light, resulting in release of the targeted moiety from the conjugate. Heat sensitive linkers would also have similar applicability. 95 314460903v1Attorney Docket No: 243735.000428

[0397] In some embodiments, antibodies can be conjugated or recombinantly fused to adiagnostic, detectable or therapeutic agent or any other molecule. The conjugated or recombinantly fused antibodies can be useful, e.g., for monitoring or prognosing the onset, development, progression and / or severity of a disease as part of a clinical testing procedure, such as determining the efficacy of a particular therapy. Such diagnosis and detection can be accomplished by coupling the antibody to detectable substances including, but not limited to, various enzymes, such as, but not limited to, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; prosthetic groups, such as, but not limited to, streptavidin / biotin and avidin / biotin; fluorescent materials, such as, but not limited to, umbelliferone, fluorescein, fluorescein isothiocynate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; luminescent materials, such as, but not limited to, luminol; bioluminescent materials, such as but not limited to, luciferase, luciferin, and aequorin; radioactive materials, such as, but not limited to, iodine (131I, 125I, 123I, and 121I,), carbon (14C), sulfur (35S), tritium (3H), indium (115In, 113In, 112In, and 111In,), technetium (99Tc), thallium (201Ti), gallium (68Ga, 67Ga), palladium (103Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), 153Sm, 177Lu, 159Gd, 149Pm, 140La, 175Yb, 166Ho, 90Y, 47Sc, 186Re, 187Re, 142Pr, 105Rh, 97Ru, 68Ge, 57Co, 65Zn, 85Sr, 32P, 153Gd, 169Yb, 51Cr, 54Mn, 75Se, 113Sn, and 117Sn; and positron emitting metals using various positron emission tomographies, and non-radioactive paramagnetic metal ions.

[0398] Encompassed herein are antibodies recombinantly fused or chemically conjugated(including both covalent and non-covalent conjugations) to a heterologous protein or polypeptide (or fragment thereof, preferably to a polypeptide of about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 100 amino acids) to generate fusion proteins. In particular, provided herein are fusion proteins comprising an antigen-binding fragment of a monoclonal antibody (e.g., a Fab fragment, an Fd fragment, an Fv fragment, a F(ab)2 fragment, a VH domain, a 68PG EM, a VL domain or a VL CDR) and a heterologous protein, polypeptide, or peptide. In a specific embodiment, the heterologous protein, polypeptide, or peptide that the antibody is fused to is useful for targeting the antibody to a particular cell type.

[0399] In one embodiment, a fusion protein provided herein comprises an anti-TTYH2 antibodydescribed herein, or an antigen-binding fragment thereof, and a heterologous polypeptide. In another embodiment, a fusion protein provided herein comprises one, two, or more VH domains 96 314460903v1Attorney Docket No: 243735.000428 having the amino acid sequence of any one of the VH domains of an anti-TTYH2 antibody described herein or one or more VL domains having the amino acid sequence of any one of the VL domains of an anti-TTYH2 antibody described herein and a heterologous polypeptide. In another embodiment, a fusion protein provided herein comprises one, two, or more VH CDRs having the amino acid sequence of any one of the VH CDRs of an anti-TTYH2 antibody described herein and a heterologous polypeptide. In another embodiment, a fusion protein comprises one, two, or more 68PG EMM having the amino acid sequence of any one of the 68PG EMM of an anti-TTYH2 antibody described herein and a heterologous polypeptide. In another embodiment, a fusion protein provided herein comprises at least one VH domain and at least one VL domain of an anti-TTYH2 antibody described herein and a heterologous polypeptide. In yet another embodiment, a fusion protein provided herein comprises at least one 68PG EM and at least one VL CDR of an anti-TTYH2 antibody described herein and a heterologous polypeptide. In certain embodiments, the above-referenced antibodies comprise a modified IgG (e.g., IgG1) constant domain, or FcRn binding fragment thereof (e.g., the Fc domain or hinge-Fc domain), described herein.

[0400] Moreover, antibodies can be fused to marker sequences, such as a peptide to facilitatepurification. In preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide HHHHHH (SEQ ID NO: 48) (i.e., His-tag), such as the tag provided in a pQE vector (QIAGEN, Inc.), among others, many of which are commercially available. As described in Gentz et al., 1989, Proc. Natl. Acad. Sci. USA 86:821-824, for instance, hexa-histidine (SEQ ID NO: 48) provides for convenient purification of the fusion protein. Other peptide tags useful for purification include, but are not limited to, the hemagglutinin (“HA”) tag, which corresponds to an epitope derived from the Influenza hemagglutinin protein (Wilson et al., 1984, Cell 37:767), and the “flag” tag.

[0401] Methods for fusing or conjugating therapeutic moieties (including polypeptides) toantibodies are well known, see, e.g., Amon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp.243-56 (Alan R. Liss, Inc.1985); Hellstrom et al., “Antibodies For Drug Delivery”, in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp.623-53 (Marcel Dekker, Inc.1987); Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in Monoclonal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); “Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In 97 314460903v1Attorney Docket No: 243735.000428 Cancer Therapy”, in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp.303-16 (Academic Press 1985), Thorpe et al., 1982, Immunol. Rev.62:119-58; —C— U.S. Pat. Nos. 5,336,603, 5,622,929, 5,359,046, 5,349,053, 5,447,851, 5,723,125, 5,783,181, 5,908,626, 5,844,095, and 5,112,946; EP 307,434; EP 367,166; EP 394,827; PCT publications WO 91 / 06570, WO 96 / 04388, WO 96 / 22024, WO 97 / 34631, and WO 99 / 04813; Ashkenazi et al., Proc. Natl. Acad. Sci. USA, 88: 10535-10539, 1991; Traunecker et al., Nature, 331:84-86, 1988; Zheng et al., J. Immunol., 154:5590-5600, 1995; Vil et al., Proc. Natl. Acad. Sci. USA, 89:11337- 11341, 1992; which are incorporated herein by reference in their entireties.

[0402] In particular, fusion proteins may be generated, for example, through the techniques ofgene-shuffling, motif-shuffling, exon-shuffling, and / or codon-shuffling (collectively referred to as “DNA shuffling”). DNA shuffling may be employed to alter the activities of the monoclonal antibodies described herein or generated in accordance with the methods provided herein (e.g., antibodies with higher affinities and lower dissociation rates). See, e.g., U.S. Pat. Nos.5,605,793, 5,811,238, 5,830,721, 5,834,252, and 5,837,458; Patten et al., 1997, Curr. Opinion Biotechnol. 8:724-33; Harayama, 1998, Trends Biotechnol. 16(2):76-82; Hansson, et al., 1999, J. Mol. Biol. 287:265-76; and Lorenzo and Blasco, 1998, Biotechniques 24(2):308-313 (each of these patents and publications are hereby incorporated by reference in its entirety). Antibodies, or the encoded antibodies, may be altered by being subjected to random mutagenesis by error-prone PCR, random nucleotide insertion, or other methods prior to recombination. A polynucleotide encoding a monoclonal antibody described herein or generated in accordance with the methods provided herein may be recombined with one or more components, motifs, sections, parts, domains, fragments, etc. of one or more heterologous molecules.

[0403] An antibody can also be conjugated to a second antibody to form an antibody hetero-conjugate as described in U.S. Pat. No.4,676,980, which is incorporated herein by reference in its entirety.

[0404] An antibody can also be linked directly or indirectly to one or more antibodies to producebispecific / multispecific antibodies.

[0405] An antibody can also be attached to solid supports, which are particularly useful forimmunoassays or purification of an antigen. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene. 98 314460903v1Attorney Docket No: 243735.000428

[0406] In some embodiments, one or more heterologous moieties are conjugated to an anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 20, 24, 30, 36, or more heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 1 heterologous moiety is conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 2 heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 3 heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 4 heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 5 heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 6 heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen- binding fragment. In some embodiments, 7 heterologous moieties are conjugated to one anti- TTYH2 antibody or antigen-binding fragment. In some embodiments, 8 heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 9 heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. In some embodiments, 10 or more heterologous moieties are conjugated to one anti-TTYH2 antibody or antigen-binding fragment. Polynucleotides and Vectors

[0407] In one aspect, the present disclosure provides one or more polynucleotides encoding anantibody or antigen-binding fragment as described herein. The isolated polynucleotides capable of encoding the variable domain segments provided herein as well as constant domains segments as described herein may be included on the same, or different, vectors to produce antibodies or antigen-binding fragments.

[0408] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 1, 9, or 17, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; and / or a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 5, 13, or 21, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto. 99 314460903v1Attorney Docket No: 243735.000428

[0409] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 1, 9, or 17; and / or a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 5, 13, or 21.

[0410] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 1, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; and / or a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 5, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto.

[0411] In some embodiments, an isolated polynucleotide encoding an anti- TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 1; and a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 5.

[0412] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 9, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; and / or a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 13, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto.

[0413] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 9; and a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 13.

[0414] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 17, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto; and / or a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 21, or a sequence having at least 70%, 75%, 80%, 85%, 90%, preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identity thereto. 100 314460903v1Attorney Docket No: 243735.000428

[0415] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibody orantigen-binding fragment comprises a nucleotide sequence encoding a VH amino acid sequence of SEQ ID NO: 17; and a nucleotide sequence encoding a VL amino acid sequence of SEQ ID NO: 21.

[0416] In some embodiments, an isolated polynucleotide encoding an anti-TTYH2 antibodycomprises a nucleotide sequence encoding an Fc region comprising L234A / L235A / P329G (LALA-PG) amino acid substitutions, wherein the residues are numbered according to the EU index of Kabat.

[0417] In some embodiments, the polynucleotide is a DNA molecule or a derivative thereof.

[0418] In some embodiments, the polynucleotide is an RNA molecule (e.g., mRNA) or aderivative thereof.

[0419] Also provided are vectors comprising the polynucleotides described herein. The vectorscan be expression vectors. Recombinant expression vectors containing a sequence encoding a polypeptide of interest are thus contemplated as within the scope of this disclosure. The expression vector may contain one or more additional sequences such as but not limited to regulatory sequences (e.g., promoter, enhancer), a selection marker, and a polyadenylation signal. Vectors for transforming a wide variety of host cells are well known and include, but are not limited to, plasmids, phagemids, cosmids, baculoviruses, bacmids, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), as well as other bacterial, yeast and viral vectors. In some embodiments, the vector is a viral vector.

[0420] Recombinant expression vectors within the scope of the description include synthetic,genomic, or cDNA-derived nucleic acid fragments that encode at least one recombinant protein which may be operably linked to suitable regulatory elements. Such regulatory elements may include a transcriptional promoter, sequences encoding suitable mRNA ribosomal binding sites, and sequences that control the termination of transcription and translation. Expression vectors, especially mammalian expression vectors, may also include one or more nontranscribed elements such as an origin of replication, a suitable promoter and enhancer linked to the gene to be expressed, other 5’ or 3’ flanking nontranscribed sequences, 5’ or 3’ nontranslated sequences (such as necessary ribosome binding sites), a polyadenylation site, splice donor and acceptor sites, or transcriptional termination sequences. An origin of replication that confers the ability to replicate in a host may also be incorporated. 101 314460903v1Attorney Docket No: 243735.000428

[0421] Vectors described herein may contain one or more Internal Ribosome Entry Site(s)(IRES). Inclusion of an IRES sequence into fusion vectors may be beneficial for enhancing expression of some proteins. In some embodiments the vector system will include one or more polyadenylation sites (e.g., SV40), which may be upstream or downstream of any of the aforementioned nucleic acid sequences. Vector components may be contiguously linked, or arranged in a manner that provides optimal spacing for expressing the gene products (i.e., by the introduction of “spacer” nucleotides between the ORFs), or positioned in another way. Regulatory elements, such as the IRES motif, may also be arranged to provide optimal spacing for expression.

[0422] The vectors may comprise selection markers, which are well known in the art. Selectionmarkers include positive and negative selection markers, for example, antibiotic resistance genes (e.g., neomycin resistance gene, a hygromycin resistance gene, a kanamycin resistance gene, a tetracycline resistance gene, a penicillin resistance gene, a puromycin resistance gene, a blasticidin resistance gene), glutamate synthase genes, HSV-TK, HSV-TK derivatives for ganciclovir selection, or bacterial purine nucleoside phosphorylase gene for 6-methylpurine selection (Gadi et al., 7 Gene Ther.1738-1743 (2000)). A nucleic acid sequence encoding a selection marker or the cloning site may be upstream or downstream of a nucleic acid sequence encoding a polypeptide of interest or cloning site.

[0423] Non-limiting examples of additional vectors that can be used in accordance with thepresent disclosure include Moloney murine leukemia viruses (MLV), Moloney murine leukemia viruses pseudotyped with vesicular stomatitis virus G protein (MLV-VSV-G), murine stem cell viruses (MSCV), lentiviruses, lentiviruses pseudotyped with vesicular stomatitis virus G protein (LV-VSV-G), adenoviruses (e.g., Ad5, Ad41), adeno-associated viruses (such as AAV1, AAV2, AAV5, AAV8, AAV9, AAV10, AAV6), and variants and derivatives thereof. Additional suitable vectors include those described in Buckinx and Timmermans, Histochem Cell Biol (2016) 146:709–720, which is incorporated herein by reference in its entirety.

[0424] In some embodiments, bacteriophages may be engineered to incorporate a nucleotidesequence encoding an antibody described herein into the genetic material of the phage so that the antibody may be exposed on the surface of the phage.

[0425] The vectors described herein may be used to transform various cells with the genesencoding the described antibodies or antigen-binding fragments. For example, the vectors may be used to generate TTYH2-specific antibody or antigen-binding fragment-producing cells. Thus, in 102 314460903v1Attorney Docket No: 243735.000428 another aspect is provided host cells transformed with vectors comprising a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof that specifically binds a TTYH2 protein, such as the antibodies or antigen-binding fragments described and exemplified herein.

[0426] In some embodiments, a host cell comprising a polynucleotide described herein or avector described herein can express an antibody or an antigen-binding fragment of the present disclosure. Any of a wide variety of host cells within the knowledge of one of skill in the art may be used in the practice of the present disclosure. A non-limiting example of a host cell is a hybridoma. In some embodiments, the antibody or antigen-binding fragment is recombinantly produced.

[0427] Numerous techniques are known in the art for the introduction of foreign genes into cellsand may be used to construct the recombinant cells for purposes of carrying out the described methods, in accordance with the various embodiments described and exemplified herein. The technique used should provide for the stable transfer of the heterologous gene sequence to the host cell, such that the heterologous gene sequence is heritable and expressible by the cell progeny, and so that the necessary development and physiological functions of the recipient cells are not disrupted. Techniques which may be used include but are not limited to chromosome transfer (e.g., cell fusion, chromosome mediated gene transfer, micro cell mediated gene transfer), physical methods (e.g., transfection, spheroplast fusion, microinjection, electroporation, liposome carrier), viral vector transfer (e.g., recombinant DNA viruses, recombinant RNA viruses) and the like (described in Cline, 29 Pharmac. Ther. 69-92 (1985)). Calcium phosphate precipitation and polyethylene glycol (PEG)-induced fusion of bacterial protoplasts with mammalian cells may also be used to transform cells. Preparation of Antibodies or Antigen-Binding Fragments

[0428] Antibodies or antigen-binding fragments capable of binding TTYH2 protein as describedherein can be made by any method known in the art, including but not limited to, recombinant technology.

[0429] For example, nucleic acids encoding the heavy and light chain of an anti-TTYH2antibody as described herein can be cloned into one expression vector, each nucleotide sequence being in operable linkage to a suitable promoter. In one example, each of the nucleotide sequences encoding the heavy chain and light chain is in operable linkage to a distinct promoter. 103 314460903v1Attorney Docket No: 243735.000428

[0430] Alternatively, the nucleotide sequences encoding the heavy chain and the light chain canbe in operable linkage with a single promoter, such that both heavy and light chains are expressed from the same promoter. When necessary, an internal ribosomal entry site (IRES) can be inserted between the heavy chain and light chain encoding sequences.

[0431] In some examples, the nucleotide sequences encoding the two chains of the antibody arecloned into two vectors, which can be introduced into the same or different cells. When the two chains are expressed in different cells, each of them can be isolated from the host cells expressing such and the isolated heavy chains and light chains can be mixed and incubated under suitable conditions allowing for the formation of the antibody.

[0432] Generally, a nucleic acid sequence encoding one or all chains of an antibody can becloned into a suitable expression vector in operable linkage with a suitable promoter using methods known in the art. For example, the nucleotide sequence and vector can be contacted, under suitable conditions, with a restriction enzyme to create complementary ends on each molecule that can pair with each other and be joined together with a ligase. Alternatively, synthetic nucleic acid linkers can be ligated to the termini of a gene. These synthetic linkers contain nucleic acid sequences that correspond to a particular restriction site in the vector. The selection of expression vectors / promoter would depend on the type of host cells for use in producing the antibodies.

[0433] A variety of promoters can be used for expression of the antibodies described herein,including, but not limited to, cytomegalovirus (CMV) intermediate early promoter, a viral LTR such as the Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR, the simian virus 40 (SV40) early promoter, E. coli lac UV5 promoter, and the herpes simplex TK virus promoter.

[0434] Regulatable promoters can also be used. Such regulatable promoters include those usingthe lac repressor from E. coli as a transcription modulator to regulate transcription from lac operator-bearing mammalian cell promoters [Brown, M. et al., Cell, 49:603-612 (1987)], those using the tetracycline repressor (tetR) [Gossen, M., and Bujard, H., Proc. Natl. Acad. Sci. USA 89:5547-5551 (1992); Yao, F. et al., Human Gene Therapy, 9:1939-1950 (1998); Shockelt, P., et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)]. Other systems include FK506 dimer, VP16 or p65 using astradiol, RU486, diphenol murislerone, or rapamycin. Inducible systems are available from Invitrogen, Clontech and Ariad.

[0435] Regulatable promoters that include a repressor with the operon can be used. In oneembodiment, the lac repressor from E. coli can function as a transcriptional modulator to regulate 104 314460903v1Attorney Docket No: 243735.000428 transcription from lac operator-bearing mammalian cell promoters (M. Brown et al., Cell, 49:603- 612 (1987); Gossen and Bujard (1992); M. Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992)) combined the tetracycline repressor (tetR) with the transcription activator (VP 16) to create a tetR-mammalian cell transcription activator fusion protein, tTa (tetR-VP 16), with the tetO- bearing minimal promoter derived from the human cytomegalovirus (hCMV) major immediate- early promoter to create a tetR-tet operator system to control gene expression in mammalian cells. In one embodiment, a tetracycline inducible switch is used. The tetracycline repressor (tetR) alone, rather than the tetR-mammalian cell transcription factor fusion derivatives can function as potent trans-modulator to regulate gene expression in mammalian cells when the tetracycline operator is properly positioned downstream for the TATA element of the CMVIE promoter (Yao et al., Human Gene Therapy, 10(16):1392-1399 (2003)). One particular advantage of this tetracycline inducible switch is that it does not require the use of a tetracycline repressor-mammalian cells transactivator or repressor fusion protein, which in some instances can be toxic to cells (Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992); Shockett et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)), to achieve its regulatable effects.

[0436] Additionally, the vector can contain, for example, some or all of the following: aselectable marker gene, such as the neomycin gene for selection of stable or transient transfectants in mammalian cells; enhancer / promoter sequences from the immediate early gene of human CMV for high levels of transcription; transcription termination and RNA processing signals from SV40 for mRNA stability; SV40 polyoma origins of replication and ColE1 for proper episomal replication; internal ribosome binding sites (IRESes), versatile multiple cloning sites; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNA. Suitable vectors and methods for producing vectors containing transgenes are well known and available in the art.

[0437] Examples of polyadenylation signals useful to practice the methods described hereininclude, but are not limited to, human collagen I polyadenylation signal, human collagen II polyadenylation signal, and SV40 polyadenylation signal.

[0438] One or more vectors (e.g., expression vectors) comprising nucleic acids encoding any ofthe antibodies may be introduced into suitable host cells for producing the antibodies. The host cells can be cultured under suitable conditions for expression of the antibody or any polypeptide chain thereof. Such antibodies or polypeptide chains thereof can be recovered by the cultured cells (e.g., from the cells or the culture supernatant) via a conventional method, e.g., affinity 105 314460903v1Attorney Docket No: 243735.000428 purification. If necessary, polypeptide chains of the antibody can be incubated under suitable conditions for a suitable period of time allowing for production of the antibody.

[0439] In some embodiments, methods for preparing an antibody described herein involve arecombinant expression vector that encodes both the heavy chain and the light chain of an anti- TTYH2 antibody, as also described herein. The recombinant expression vector can be introduced into a suitable host cell by a conventional method, e.g., calcium phosphate-mediated transfection. Positive transformant host cells can be selected and cultured under suitable conditions allowing for the expression of the two polypeptide chains that form the antibody, which can be recovered from the cells or from the culture medium. When necessary, the two chains recovered from the host cells can be co-incubated under suitable conditions allowing for the formation of the antibody.

[0440] In one example, two recombinant expression vectors are provided, one encoding theheavy chain of the anti-TTYH2 antibody and the other encoding the light chain of the anti-TTYH2 antibody. Both of the two recombinant expression vectors can be introduced into a suitable host cell by a conventional method, e.g., calcium phosphate-mediated transfection. Alternatively, each of the expression vectors can be introduced into suitable host cells. Positive transformants can be selected and cultured under suitable conditions allowing for the expression of the polypeptide chains of the antibody. When the two expression vectors are introduced into the same host cells, the antibody produced therein can be recovered from the host cells or from the culture medium. If necessary, the polypeptide chains can be recovered from the host cells or from the culture medium and then incubated under suitable conditions allowing for formation of the antibody. When the two expression vectors are introduced into different host cells, each of them can be recovered from the corresponding host cells or from the corresponding culture media. The two polypeptide chains can then be incubated under suitable conditions for formation of the antibody.

[0441] In some embodiments, the antibody or antigen-binding fragment described herein areproduced by a method comprising culturing a host cell described herein (e.g., a host cell expressing an antibody or antigen binding fragment described herein comprising a polynucleotide described herein or a vector described herein) and isolated the antibody or antigen-binding fragment.

[0442] Standard molecular biology techniques are used to prepare the recombinant expressionvector, transfect the host cells, select for transformants, culture the host cells and recovery of the antibodies from the culture medium. For example, some antibodies can be isolated by affinity chromatography with a Protein A or Protein G coupled matrix. 106 314460903v1Attorney Docket No: 243735.000428

[0443] Any of the nucleic acids encoding the heavy chain, the light chain, or both of an anti-TTYH2 antibody as described herein, vectors (e.g., expression vectors) containing such; and host cells comprising the vectors are within the scope of the present disclosure.

[0444] In some embodiments, the antibody or antigen-binding fragment described herein areisolated from an animal immunized with a TTYH2-containing antigen, or genetically modified to produce the antibody or antigen-binding fragment. The animal may be genetically modified with the polynucleotide or the vector described herein.

[0445] In some embodiments, the animal is a dairy animal, such as, but not limited to, a goat, acow, a buffalo, a sheep, or a camel. In such cases, the antibody or antigen-binding fragment may be isolated from milk produced by the dairy animal.

[0446] Anti-TTYH2 antibodies prepared as described herein can be characterized using methodsknown in the art, whereby reduction, amelioration, or neutralization of biological activity associated with the target TTYH2 protein is detected and / or measured. For example, in some embodiments, an ELISA-type assay is suitable for qualitative or quantitative measurement of binding of the antibody to the target TTYH2 protein. Pharmaceutical Compositions

[0447] Any of the antibodies or antigen-binding fragments, antibody-drug conjugates,polynucleotides, or vectors described herein can be present in a pharmaceutical composition (such as a formulation) that can includes other agents, excipients, or stabilizers. In various embodiments, a pharmaceutical composition described herein may comprise (i) an antibody or antigen-binding fragment described herein, (ii) an antibody-drug conjugate described herein, (iii) a polynucleotide described herein, and / or (iv) a recombinant vector described herein, and a pharmaceutically acceptable carrier or adjuvant.

[0448] It is understood that the compounds of the present disclosure can use amino acidsindependently selected from L and D forms (e.g., the peptide may contain two serine residues, each serine residue having the same or opposite absolute stereochemistry), etc., are intended for the use of both L- and D-form amino acids.

[0449] Accordingly, the compounds of the present disclosure also include substantially purestereoisomeric form of the specific compound with respect to the asymmetric center of the amino acid residue, for example about 90% de, such as greater than about 95% to 97% de, or 99% de. 107 314460903v1Attorney Docket No: 243735.000428 For larger compounds, as well as mixtures thereof (such as racemic mixtures). Such diastereomers may be prepared, for example, by asymmetric synthesis using chiral intermediates, or the mixture may be divided by conventional methods, such as chromatography or the use of dividing agents.

[0450] If the compounds of the disclosure require purification, chromatographic techniques suchas high-performance liquid chromatography (HPLC) and reverse phase HPLC can be used. Peptides may be characterized by mass spectrometry and / or other suitable methods.

[0451] If the compound contains one or more functional groups that can be protonated ordeprotonated (e.g., at physiological pH), the compound can be prepared and / or isolated as a pharmaceutically acceptable salt. It will be appreciated that the compound can be zwitterion at a given pH. As used herein, the expression “pharmaceutically acceptable salt” refers to a salt of a given compound, which salt is suitable for pharmaceutical administration. Such salts can be formed, for example, by reacting an acid or base with an amine or carboxylic acid group, respectively.

[0452] Pharmaceutically acceptable acid addition salts can be prepared from inorganic andorganic acids. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Examples of organic acids include acetic acid, propionic acid, glycolic acid, pyruvate, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartrate acid, citrate, benzoic acid, cinnamic acid, mandelic acid, Examples thereof include methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid.

[0453] Pharmaceutically acceptable base addition salts can be prepared from inorganic andorganic bases. Corresponding counterions derived from inorganic bases include salts of sodium, potassium, lithium, ammonium, calcium and magnesium. Organic bases include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2- dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, prokine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, Substituted amines such as primary, secondary and tertiary amines such as N-alkylglucamine, theobromine, purines, piperazine, piperazine and N-ethylpiperidine, substituted amines such as natural substituted amines and cyclic amines can be mentioned.

[0454] Acid / base addition salts tend to be more soluble in aqueous solvents than thecorresponding free acid / base forms. 108 314460903v1Attorney Docket No: 243735.000428

[0455] In some embodiments, it is envisioned that two or more combinations of the compoundsof the disclosure will be administered to the subject. It is believed that the compound (s) may also be administered in combination with one or more additional therapeutic agents. This combination can allow separate, continuous or simultaneous administration with the other active ingredients of the above compounds. This combination may be provided in the form of a pharmaceutical composition.

[0456] Combination agents can be administered, for example, simultaneously or staggered intime (i.e., at different times and at equal or different time intervals for any part of a kit). The ratio of the total amount of combination agents administered in a combination can vary, e.g., to address the needs of a subpopulation of patients to be treated or the needs of a single patient, and different needs are the age of the patient, it can be due to gender, weight, etc.

[0457] In some embodiments, the pharmaceutical composition further comprises one or moreimmune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include, but are not limited to, a programmed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD- L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T- lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a B7-H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin-like lectin 109 314460903v1Attorney Docket No: 243735.000428 (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, a vascular endothelial growth factor (VEGF) inhibitor, and any combinations thereof. In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, or a CTLA-4 inhibitor, or any combination thereof.

[0458] In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor. ExemplaryPD-1 inhibitors include, but are not limited to, nivolumab, pembrolizumab, BAT1308, acrixolimab, balstilimab, budigalimab, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, ociperlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tiragolumab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, zimberelimab, and a variant or any combinations thereof. In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor. Exemplary PD-L1 inhibitors include, but are not limited to, durvalumab, avelumab, atezolizumab, bintrafusp alfa, and a variant or any combinations thereof.

[0459] In some embodiments, the immune checkpoint inhibitor is a LAG-3 inhibitor. ExemplaryLAG-3 inhibitors include, but are not limited to, relatlimab (BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO- 7247669), TSR-033, tuparstobart (INCAGN02385), BGA-1953, and a variant or any combinations thereof. In certain specific embodiments, the LAG-3 inhibitor is relatlimab (BMS-986016), or a variant thereof. 110 314460903v1Attorney Docket No: 243735.000428

[0460] In some embodiments, the immune checkpoint inhibitor is a CTLA-4 inhibitor.Exemplary CTLA-4 inhibitors include, but are not limited to, ipilimumab, tremelimumab, XmAb20717, ONC-392, XmAb22841, BMS-986249, ADG116, ATOR-1015, ADG126, YH001, botensilimab, HBM4003, lorigerlimab, SI-B003, AK104, KN046, quavonlimab, BNT316 / ONC- 392 (gotistobart), porustobart (HBM4003), or a variant or combination thereof. In some embodiments, the CTLA-4 inhibitor is ipilimumab, tremelimumab, quavonlimab, BNT316 / ONC- 392 (gotistobart), porustobart (HBM4003), and a variant or a combination thereof.

[0461] In some embodiments, the immune checkpoint inhibitor is a TIGIT inhibitor. ExemplaryTIGIT inhibitors include, but are not limited to, BMS‐986207, ociperlimab, BGB‐A1217, tiragolumab, domvanalimab, ASP8374, vibostolimab, IBI‐939, etigilimab, COM902, M6223, EOS884448, BAT6021, HLX301, and a variant or any combinations thereof.

[0462] In some embodiments, the SLAMF inhibitor is elotuzumab or a variant thereof. In someembodiments, the PVRIG inhibitor is COM701, JS009, or a variant thereof. In some embodiments, the CD96 inhibitor is GSK6097608 or a variant thereof.

[0463] In some embodiments, the pharmaceutical composition further comprises one or morecytokines, tumor-associated antigen (TAA)-targeting agents, immune agonists, cytotoxic agents, siRNAs, or antisense oligonucleotides.

[0464] Exemplary cytokines include, but are not limited to, interleukin-2 (IL-2), interleukin-6(IL-6), interleukin-10 (IL-10), interleukin-15 (IL-15), interleukin-21 (IL-21), IFN-α, IFN-β, IFN- γ, CCL19, CCL21, interleukin-18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), and a receptor thereof, or any combinations thereof.

[0465] Exemplary tumor-associated antigen -targeting agents include, but are not limited to,5T4, Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY-ESO-1, P- cadherin, PRAME , PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, RAF, and any combinations thereof. In some embodiments, TAA targeting agent is a VEGF inhibitor, a VEGFR inhibitor, a CD20 inhibitor, a CD38 inhibitor, a TROP2 inhibitor, a KRAS inhibitor, or a RAF inhibitor. 111 314460903v1Attorney Docket No: 243735.000428

[0466] Exemplary immune agonists include, but are not limited to, a 4-1BB agonist, an OX40agonist, a CD40 agonist, a CD30 agonist, a GITR agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a CD28H / TMIGD2 agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, and any combinations thereof.

[0467] In some embodiments, the pharmaceutical composition further comprises stimulator ofinterferon genes (STING) agonist or toll-like receptor (TLR) agonist or combination thereof. Exemplary STING agonists include, but are not limited to, DMXAA, 2’3’-cGAMP, ADU- S100 / MIW815, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING (BI 1387446), E7766, TAK-676, SNX281, SYNB1891, and a variant or a combination thereof. Exemplary TLR agonists include, but are not limited to, a TLR1 / TLR2 agonist, a TLR2 / TLR6 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, and a combination thereof.

[0468] In some embodiments, the pharmaceutical composition further comprises one or more ofa tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL- 17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19- targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38- targeted T cell engager, a BCMA-targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof. 112 314460903v1Attorney Docket No: 243735.000428

[0469] The route of administration and the type of pharmaceutically acceptable carrier willdepend on the condition being treated and the type of mammal. Formulations containing the active compound may be prepared such that the activity of the compound is not disrupted during the process and the compound can reach its site of action without disruption. In some cases, it may be necessary to protect the compound by means known in the art, such as microencapsulation. Similarly, the route of dosing selected should be such that the compound reaches its site of action.

[0470] In some embodiments, the composition further comprises a targeting agent or a carrierthat promotes the delivery of the inhibitors of endocytosis to an area affected by the chronic pain. Exemplary carriers include, but are not limited to, liposomes, micelles, nanodisperse albumin and its modifications, polymer nanoparticles, dendrimers, and inorganic nanoparticles of different compositions.

[0471] The appropriate formulation for the compound of the disclosure can be adjusted for pH.Buffer systems are routinely used to provide pH values in the desired range and include carboxylic acid buffers such as acetates, citrates, lactates, and succinates. In some embodiments, the composition is formulated to have a pH range of about 4.5 to about 9.0, including for example pH ranges of about any of 5.0 to about 8.0, about 6.5 to about 7.5, and about 6.5 to about 7.0. In some embodiments, the pH of the composition is formulated to no less than about 6, including for example no less than about any of 6.5, 7, or 8 (such as about 8). The composition can also be made to be isotonic with blood by the addition of a suitable tonicity modifier, such as glycerol.

[0472] The formulation may also include suitable excipients, such as antioxidants. Non-limitingexamples of antioxidants include phenolic compounds such as BHT or Vitamin E, reducing agents such as methionine or sulfites, and metal chelating agents such as EDTA.

[0473] The compounds or pharmaceutically acceptable salts thereof described herein can beprepared in parenteral dosage forms such as those suitable for, e.g., intravascular (intravenous or intraarterial), subcutaneous, intraperitoneal, intratumoral, intraventricular, intrapleural or intramuscular administration delivery. Suitable pharmaceutical forms for injectable use include sterile injectable or dispersions and sterile powders for the immediate preparation of sterile injectable solutions. They must be stable under manufacturing and storage conditions and protected from reduction or oxidation and the contaminating effects of microorganisms such as bacteria or fungi. 113 314460903v1Attorney Docket No: 243735.000428

[0474] The solvent or dispersion medium for the injectable solution or dispersion may includeeither conventional solvents or carrier systems for the active compound, e.g., water, ethanol, polyols (e.g., glycerol, propylene glycol and). Liquid polyethylene glycol, etc., suitable mixtures thereof, and vegetable oils may be included. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, the maintenance of the required particle size in the case of dispersions, and the use of surfactants. Prevention of the action of microorganisms can be performed as needed by incorporating various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it may be preferable to include agents that regulate osmotic pressure, such as sugar o...

Claims

Attorney Docket No: 243735.000428 Claims 1. An antibody, or antigen-binding fragment thereof, that specifically binds to tweety familymember 2 (TTYH2) protein, comprising: (i) a heavy chain variable region (HCVR) that comprises the heavy chain complementarity- determining region 1 (HCDR1), HCDR2, and HCDR3 of an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1, 9, or 17, or a variant thereof; and / or (ii) a light chain variable region (LCVR) comprising the light chain complementarity- determining region 1 (LCDR1), LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, 13, or 21, or a variant thereof.

2. The antibody or antigen-binding fragment of claim 1, comprising(a) an HCVR that comprises the HCDR1, HCDR2, and HCDR3 of an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1, or a variant thereof, and an LCVR that comprises the LCDR1, LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, or a variant thereof; (b) an HCVR that comprises the HCDR1, HCDR2, and HCDR3 of an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 9, or a variant thereof, and an LCVR that comprises the LCDR1, LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 13, or a variant thereof; and / or (c) an HCVR that comprises the HCDR1, HCDR2, and HCDR3 of an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 17, or a variant thereof, and an LCVR that comprises the LCDR1, LCDR2, and LCDR3 of an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 21, or a variant thereof.

3. The antibody or antigen-binding fragment of claim 1 or claim 2, comprising(a) an HCVR that comprises an HCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 2, an HCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 3, and an HCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 4, and an LCVR that comprises an LCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 6, an LCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 7, and an LCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 8; 188 314460903v1Attorney Docket No: 243735.000428 (b) an HCVR that comprises an HCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 10, an HCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 11, and an HCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 12, and a LCVR that comprises an LCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 14, an LCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 15, and an LCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 16; and / or (c) an HCVR that comprises an HCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 18, an HCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 19, and an HCDR3 that comprises the amino acid sequence set forth in SEQ ID NO: 20, and an LCVR that comprises an LCDR1 that comprises the amino acid sequence set forth in SEQ ID NO: 22, an LCDR2 that comprises the amino acid sequence set forth in SEQ ID NO: 23, and an LCDR3 that comprises the amino acid sequence set forth in SEQ ID NO:

24.

4. The antibody or antigen-binding fragment of any one of claims 1-3, comprising:(a) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1, 9, or 17, or a variant thereof, and / or (b) an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, 13, or 21, or a variant thereof.

5. The antibody or antigen-binding fragment of any one of claims 1-4, comprising:(a) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 5, or a variant thereof; (b) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 9, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 13, or a variant thereof; and / or (c) an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 17, or a variant thereof, and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 21, or a variant thereof. 189 314460903v1Attorney Docket No: 243735.000428 6. An antibody, or antigen-binding fragment thereof, that competes for binding to TTYH2protein with the antibody or antigen-binding fragment of any one of claims 1-5.

7. An antibody, or antigen-binding fragment thereof, that binds to the same epitope of TTYH2protein as the antibody or antigen-binding fragment of any one of claims 1-6.

8. An antibody, or antigen-binding fragment thereof, that specifically binds to TTYH2 protein,wherein the antibody or antigen-binding fragment thereof binds to the second ectodomain (ECD2) of TTYH2 protein.

9. The antibody or antigen-binding fragment of claim 8, wherein the antibody or antigen-binding fragment thereof binds to an epitope on TTYH2 protein comprising, consisting essentially of, or consisting of the sequence FAARGDY (SEQ ID NO: 25).

10. The antibody or antigen-binding fragment of any one of claims 1-9, wherein the antibodyor antigen-binding fragment binds to human TTYH2 protein and / or mouse TTYH2 protein.

11. The antibody or antigen-binding fragment of any one of claims 1-10, wherein the antibodyor antigen-binding fragment is recombinant.

12. The antibody or antigen-binding fragment of any one of claims 1-11, wherein the antibodyor antigen-binding fragment is a human antibody, a humanized antibody, a chimeric antibody, a murine antibody, a monoclonal antibody, a single chain antibody, a bispecific antibody or antigen-binding fragment thereof, a bi-epitopic antibody or antigen-binding fragment thereof, a Fab, a Fab’, a F(ab’)2, a Fv, an scFv, a VH domain, or a nanobody.

13. The antibody or antigen-binding fragment of claim 12, wherein the bispecific antibody orantigen-binding fragment thereof is a bispecific T-cell engager (BiTE), bispecific NK-cell engager (BiKE), or a bi-specific macrophage engager (BiME). 190 314460903v1Attorney Docket No: 243735.000428 14. The antibody or antigen-binding fragment of claim 13, wherein the BiTE comprises anantigen-binding domain that specifically binds to cluster of differentiation (CD)3 (CD3), alpha beta T cell receptor (TCR), or gamma delta TCR.

15. The antibody or antigen-binding fragment of claim 13, wherein the BiKE comprises anantigen-binding domain that specifically binds to Fc gamma receptor III (FcγRIII or CD16), natural killer group 2, member D (NKG2D), or natural cytotoxicity triggering receptor 3 (NCR3).

16. The antibody or antigen-binding fragment of claim 13, wherein the BiME comprises anantigen-binding domain that specifically binds to Fc gamma receptor III (FcγRIII or CD16), dendritic cell-associated C-type lectin 1 (Dectin-1), cluster of differentiation (CD47), signal regulatory protein alpha (SIRPA), or Tyro3, Axl, and MerTK (TAM) receptor(s).

17. The antibody or antigen-binding fragment of claim 12, wherein the bispecific antibody orantigen-binding fragment thereof comprises an antigen-binding domain that specifically binds to an immune checkpoint, a cytokine or a receptor thereof, a tumor-associated antigen (TAA), or an immune stimulatory receptor.

18. The antibody or antigen-binding fragment of claim 17, wherein the immune checkpoint isprogrammed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), lymphocyte activation gene 3 (LAG-3 or CD223), cytotoxic T-lymphocyte-associated antigen 4 (CTLA- 4), cluster of differentiation 47 (CD47), T cell immunoglobulin and mucin domain- containing protein 3 (TIM-3), B7 homolog 3 protein (B7-H3 or CD276), B7-H4, V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)), T cell immunoreceptor with Ig and ITIM Domains (TIGIT), signal regulatory protein alpha (SIRPA), signaling lymphocytic activation molecule family members (SLAMF), poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R), adenosine A2A receptor (A2aR), adenosine A2b receptor (A2bR), G protein- coupled receptor 171 (GPR171), insulin like growth factor binding protein 7 (IGFBP7), cluster of differentiation 93 (CD93), CD96, CD226, natural killer group protein 2A 191 314460903v1Attorney Docket No: 243735.000428 (NKG2A), natural killer group protein 2D (NKG2D), a killer cell lectin like receptor G1 (KLRG1), a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2), a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3), sialic acid-binding immunoglobulin-like lectin (Siglec)-15, cluster of differentiation 24 (CD24), sialic acid-binding immunoglobulin-like lectin (Siglec)-10, P-selectin glycoprotein ligand-1 (PSGL-1), V-set and Ig domain-containing protein 3 (VSIG3, also known as B7 and T cell immunoglobulin domain-containing superfamily member (BT-IgSF) or immunoglobulin superfamily member 11 (IGSF11)), leucine rich repeats and immunoglobulin like domains 1 (LRIG1), fibrinogen-like protein 1 (FGL1), B and T lymphocyte attenuator (BTLA), leukocyte associated immunoglobulin like receptor 1 (LAIR-1), cluster of differentiation 160 (CD160), leukocyte immunoglobulin- like receptor subfamily B member 2 (LILRB2), leukocyte immunoglobulin-like receptor (LILRB4), angiopoietin 2 (Ang2), or vascular endothelial growth factor (VEGF).

19. The antibody or antigen-binding fragment of claim 18, wherein the immune checkpoint isPD-1, PD-L1, LAG-3, and / or CTLA-4.

20. The antibody or antigen-binding fragment of claim 17, wherein the cytokine is interleukin-2 (IL-2), IL-6, IL-10, IL-15, IL-21, IFN-α, IFN-β, IFN-γ, chemokine (C-C motif) ligand 19 (CCL19), chemokine (C-C motif) ligand 21 (CCL21), interleukin-18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF), or a receptor thereof.

21. The antibody or antigen-binding fragment of claim 17, wherein the tumor-associatedantigen (TAA) is oncofetal antigen 5T4 (5T4), angiopoietin 2 (Ang2), B-cell maturation antigen (BCMA), cluster of differentiation 123 (CD123), cluster of differentiation 19 (CD19), cluster of differentiation 20 (CD20), cluster of differentiation 22 (CD22), cluster of differentiation 33 (CD33), cluster of differentiation 38 (CD38), cluster of differentiation 47 (CD47), carcinoembryonic antigen (CEA), carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), carcinoembryonic antigen-related cell adhesion 192 314460903v1Attorney Docket No: 243735.000428 molecule 6 (CEACAM6), Claudin 6, Claudin 18.2, c-type lectin domain family 12 member A (CLEC12A), delta-like ligand 3 (DLL3), epidermal growth factor receptor (EGFR), epithelial cell adhesion molecule (EpCAM), Fc receptor homolog 5 (FcRH5), fms-like tyrosine kinase 3 (FLT3), disialoganglioside (GD2), Glypican-3, glycoprotein A33 (gpA33), G protein-coupled receptor class C group 5 member D (GPRC5D), human epidermal growth factor receptor 2 (Her2), human epidermal growth factor receptor 3 (Her3), melanoma antigen family A4 (MAGE-A4), mesenchymal-epithelial transition factor (MET), mucin 16 (MUC16), mucin 17 (MUC17), New York esophageal squamous cell carcinoma 1 (NY-ESO-1), placental cadherin (P-cadherin), preferentially expressed antigen in melanoma (PRAME), prostate stem cell antigen (PSCA), prostate-specific membrane antigen (PSMA), somatostatin receptor 2 (SSTR2), six transmembrane epithelial antigen of the prostate 1 (STEAP1), tumor-associated calcium signal transducer 2 (TROP2, also known as tumor-associated calcium signal transducer 2 (TACSTD2)), angiopoietin 2 (Ang2), TEK tyrosine kinase (Tie2), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR), gamma delta T-cell receptor (γδTCR), Kirsten rat sarcoma viral oncogene homolog (KRAS), or rapidly accelerated fibrosarcoma (RAF).

22. The antibody or antigen-binding fragment of claim 17, wherein the immune stimulatoryreceptor is cluster of differentiation 28 (CD28), inducible T-cell costimulatory (ICOS), transmembrane and immunoglobulin domain containing 2 (TMIGD2), natural cytotoxicity receptor 3 (NCR3), natural cytotoxicity receptor 1 (NCR1), natural cytotoxicity receptor 2 (NCR2), tumor necrosis factor receptor superfamily member 9 (4-1BB, also known as cluster of differentiation 137 (CD137)), tumor necrosis factor receptor superfamily member 4 (OX40), cluster of differentiation 30 (CD30), cluster of differentiation 40 (CD40), death receptor 3 (DR3), cluster of differentiation 226 (CD226, also known as DNAX accessory molecule 1 (DNAM-1)), class I restricted T cell-associated molecule (CRTAM), cluster of differentiation 27 (CD27), herpes virus entry mediator (HVEM), tumor necrosis factor receptor 1 (TNFR1), tumor necrosis factor receptor 2 (TNFR2), cluster of differentiation 2 (CD2), cluster of differentiation 7 (CD7), toll-like receptor 4 (TLR4), toll-like receptor 7 (TLR7), or toll-like receptor 9 (TLR9). 193 314460903v1Attorney Docket No: 243735.000428 23. The antibody or antigen-binding fragment of claim 12, wherein the bispecific antibody orantigen-binding fragment thereof comprises an antigen-binding domain that specifically binds to tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 13 (IL-13), interleukin 17A (IL-17A), interleukin 17F (IL-17F), chemokine (C-C motif) ligand 2 (CCL2), C-X-C motif chemokine ligand 10 (CXCL10), IFN-α, IFN-β, very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29), cluster of differentiation 20 (CD20), B activating factor (BAFF), transmembrane activator and CAML interactor (TACI), cluster of differentiation 19 (CD19), interleukin-23 receptor (IL-23R), cluster of differentiation 19 (CD22), cluster of differentiation 38 (CD38), B-cell maturation antigen (BCMA), PD-1, BTLA, CD40, CD40 ligand (CD40L), OX40, OX40L, B7, CD28, CTLA-4, tumor necrosis factor-like cytokine 1A (TL1A), DR3, or thymic stromal lymphopoietin (TSLP), or a variant thereof.

24. The antibody or antigen-binding fragment of any one of claims 1-23, wherein the antibodyor antigen-binding fragment is an IgG antibody.

25. The antibody or antigen-binding fragment of any one of claims 1-24, wherein the antibodyor antigen-binding fragment is of IgG1, IgG2, IgG3, or IgG4 subclass.

26. The antibody or antigen-binding fragment of any one of claim 1-25, wherein the antibodyor antigen-binding fragment is of IgG1 or IgG4 subclass.

27. The antibody or antigen-binding fragment of any one of claim 1-25, wherein the antibodycomprises an Fc region comprising L234A / L235A / P329G (LALA-PG) amino acid substitutions, wherein the residues are numbered according to the EU index of Kabat.

28. The antibody or antigen-binding fragment of any one of claims 1-27, wherein the antibodyor antigen-binding fragment (i) antagonizes the inhibitory function of TTYH2 on a myeloid cell or cancer cell; (ii) stimulates Type I interferon (IFN) response or myeloid function upon binding to TTYH2 protein expressed on a myeloid cell or cancer cell; 194 314460903v1Attorney Docket No: 243735.000428 (iii) activates stimulator of interferon genes (STING) / Type I IFN pathway signaling, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway signaling, or TANK-binding kinase 1 / interferon regulatory factor 3 (TBK1 / IRF3) pathway signaling, or a combination thereof, upon binding to TTYH2 protein expressed on a myeloid cell or cancer cell; and / or (iv) controls TNF-α, IL-6, CCL2, IL-10, and / or CXCL10, or myeloid cell differentiation.

29. The antibody of antigen-binding fragment thereof of claim 28, wherein the myeloid cell isa macrophage or a dendritic cell.

30. The antibody of antigen-binding fragment thereof of claim 29, wherein the macrophage isan M2 macrophage and / or tumor-associated macrophage.

31. An antibody-drug conjugate comprising the antibody or antigen-binding fragment of anyone of claims 1-30, conjugated to a heterologous moiety.

32. The antibody-drug conjugate of claim 31, wherein the heterologous moiety is an immunecheckpoint inhibitor, a cytokine, a tumor-associated antigen (TAA)-targeting agent, an immune agonist, a cytotoxic agent, an siRNA, or an antisense oligonucleotide.

33. The antibody-drug conjugate of claim 32, wherein the immune checkpoint inhibitor is aprogrammed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3, also known as cluster of differentiation 223 (CD223)) inhibitor, a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain- containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3, also known as cluster of differentiation 276 (CD276)) inhibitor, a B7-H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as PD-1H) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing 195 314460903v1Attorney Docket No: 243735.000428 protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein-coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a cluster of differentiation 96 (CD96) inhibitor, a cluster of differentiation 226 (CD226) inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat- associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL-1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as B7 and T cell immunoglobulin domain-containing superfamily member (BT-IgSF) or immunoglobulin superfamily member 11 (IGSF11)) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen- like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin-like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, or a vascular endothelial growth factor (VEGF) inhibitor, or a combination thereof.

34. The antibody-drug conjugate of claim 33, wherein the immune checkpoint inhibitor is a PD-1 inhibitor or a PD-L1 inhibitor.

35. The antibody-drug conjugate of claim 34, wherein the PD-1 inhibitor or PD-L1 inhibitor isnivolumab, pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, 196 314460903v1Attorney Docket No: 243735.000428 penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof.

36. The antibody-drug conjugate of claim 33, wherein the immune checkpoint inhibitor is aLAG-3 inhibitor.

37. The antibody-drug conjugate of claim 36, wherein the LAG-3 inhibitor is relatlimab (BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA-1953, or a variant or a combination thereof.

38. The antibody-drug conjugate of claim 37, wherein the LAG-3 inhibitor is relatlimab (BMS-986016), or a variant thereof.

39. The antibody-drug conjugate of claim 33, wherein the immune checkpoint inhibitor is aCTLA-4 inhibitor.

40. The antibody-drug conjugate of claim 39, wherein the CTLA-4 inhibitor is ipilimumab,tremelimumab, quavonlimab, BNT316 / ONC-392 (gotistobart), or porustobart (HBM4003), or a variant or a combination thereof.

41. The antibody-drug conjugate of claim 32, wherein the cytokine is interleukin (IL)-2 (IL-2),IL-6, IL-10, IL-15, IL-21, IFN-α, IFN-β, IFN-γ, chemokine (C-C motif) ligand 19 (CCL19), chemokine (C-C motif) ligand 21 (CCL21), interleukin-18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage 197 314460903v1Attorney Docket No: 243735.000428 colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF) or a receptor thereof.

42. The antibody-drug conjugate of claim 32, wherein the TAA-targeting agent targets 5T4,Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY- ESO-1, P-cadherin, PRAME , PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, or RAF, or a combination thereof.

43. The antibody-drug conjugate of claim 32, wherein the immune agonist is a 4-1BB agonist,an OX40 agonist, a CD40 agonist, a CD30 agonist, a glucocorticoid-induced tumor necrosis factor receptor-related protein (GITR) agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a transmembrane and immunoglobulin domain-containing protein 2 (TMIGD2, also known as CD28H) agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

44. The antibody-drug conjugate of claim 31, wherein the heterologous moiety is a tumornecrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B- cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted 198 314460903v1Attorney Docket No: 243735.000428 T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA- targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

45. A polynucleotide encoding the antibody or antigen-binding fragment of any one of claims1-30.

46. A vector comprising the polynucleotide of claim 45.

47. A host cell expressing the antibody or antigen-binding fragment of any one of claims 1-30,or comprising the polynucleotide of claim 45 or the vector of claim 46.

48. The host cell of claim 47, wherein the antibody or antigen-binding fragment isrecombinantly produced.

49. A method of producing the antibody or antigen-binding fragment of any one of claims 1-30, wherein said method comprises culturing the host cell of claim 47 or 48, and isolating said antibody or antigen-binding fragment.

50. A pharmaceutical composition comprising the antibody or antigen-binding fragment of anyone of claims 1-30, the antibody-drug conjugate of any one of claims 31-44, the polynucleotide of claim 45, or the vector of claim 46, and a pharmaceutically acceptable carrier or diluent.

51. The pharmaceutical composition of claim 50, further comprising one or more immunecheckpoint inhibitors, cytokines, tumor-associated antigen (TAA)-targeting agents, immune agonists, cytotoxic agents, siRNAs, or antisense oligonucleotides. 199 314460903v1Attorney Docket No: 243735.000428 52. The pharmaceutical composition of claim 51, wherein the immune checkpoint inhibitor is aprogrammed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T- lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a B7-H4 inhibitor, a V- domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD-1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein- coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL- 1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin- like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, or a vascular endothelial growth factor (VEGF) inhibitor, or a combination thereof. 200 314460903v1Attorney Docket No: 243735.000428 53. The pharmaceutical composition of claim 52, wherein the immune checkpoint inhibitor is aPD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a CTLA-4 inhibitor, or a combination thereof.

54. The pharmaceutical composition of claim 53, wherein the PD-1 inhibitor or PD-L1 inhibitoris nivolumab, pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof.

55. The pharmaceutical composition of claim 53, wherein the LAG-3 inhibitor is relatlimab(BMS-986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA-1953, or a variant or a combination thereof.

56. The pharmaceutical composition of claim 55, wherein the LAG-3 inhibitor is relatlimab(BMS-986016), or a variant thereof.

57. The pharmaceutical composition of claim 53, wherein the immune checkpoint inhibitor is aCTLA-4 inhibitor. 201 314460903v1Attorney Docket No: 243735.000428 58. The pharmaceutical composition of claim 53, wherein the CTLA-4 inhibitor is ipilimumab,tremelimumab, quavonlimab, BNT316 / ONC-392 (gotistobart), or porustobart (HBM4003), or a variant or a combination thereof.

59. The pharmaceutical composition of claim 51, wherein the cytokine is interleukin-2 (IL-2),IL-6, IL-10, IL-15, IL-21, IFN-α, IFN-β, IFN-γ, IL-18, CCL19, CCL21, monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF), or a receptor thereof.

60. The pharmaceutical composition of claim 51, wherein the TAA-targeting agent targets 5T4,Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY- ESO-1, P-cadherin, PRAME, PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, or RAF, or a combination thereof.

61. The pharmaceutical composition of claim 51, wherein the immune agonist is a 4-1BBagonist, an OX40 agonist, a CD40 agonist, a CD30 agonist, a GITR agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a CD28H / TMIGD2 agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

62. The pharmaceutical composition of any one of claims 50-61, further comprising a stimulatorof interferon genes (STING) agonist or toll-like receptor (TLR) agonist, or a combination thereof.

63. The pharmaceutical composition of claim 62, wherein the STING agonist is DMXAA, 2’3’-cGAMP, ADU-S100 / MIW815, MK-1454, MK-2118, SB11285, GSK3745417, BMS- 202 314460903v1Attorney Docket No: 243735.000428 986301, BI-STING (BI 1387446), E7766, TAK-676, SNX281, or SYNB1891, or a variant or a combination thereof.

64. The pharmaceutical composition of claim 62, wherein the TLR agonist is a TLR1 / TLR2agonist, a TLR2 / TLR6 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof.

65. The pharmaceutical composition of claim 50, further comprising one or more of a tumornecrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B- cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA- targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

66. A kit comprising (i) the antibody or antigen-binding fragment of any one of claims 1-30,the antibody-drug conjugate of any one of claims 31-44, the polynucleotide of claim 45, or the vector of claim 46, and (ii) optionally, packaging for the same and / or instructions for use. 203 314460903v1Attorney Docket No: 243735.000428 67. A method for stimulating Type I IFN response of a myeloid cell or cancer cell or enhancingmyeloid function in a subject in need thereof, the method comprising administering to the subject an effective amount of an agent that targets TTYH2 such that the Type I IFN response of the myeloid cells or cancer cell is stimulated.

68. The method of claim 67, wherein the myeloid cell is a macrophage or a dendritic cell.

69. The method of claim 68, wherein the macrophage is an M2 macrophage and / or tumor-associated macrophage.

70. A method for depleting a cancer cell that expresses TTYH2 protein in a subject in needthereof, the method comprising administering to the subject an effective amount of an agent that targets TTYH2 such that the cancer cell is depleted.

71. A method of treating or preventing a cancer in a subject in need thereof, the methodcomprising administering to the subject an effective amount of an agent that targets TTYH2.

72. The method of any one of claims 67-71, wherein the agent that targets TTYH2 is selectedfrom a small molecule, a protein, a protein-drug conjugate, a nucleotide molecule, a gene editing system, an engineered cell system, and any combination thereof.

73. The method of claim 72, wherein the protein is selected from a peptide; an antibody orantigen-binding fragment thereof; a monobody; engineered, low-density-lipoprotein- receptor-derived, A domain (LDLR-A) (e.g., Avimers™); a designed ankyrin repeat protein (DARPin) lipocalin (e.g., anticalins); an affibody; engineered, Protein-A-derived, Z domain (Affibodies™) CTLD3 (e.g., Tetranectin); C-type lectin-like domain scaffolds; Sac7d- derived polypeptides (e.g., Nanoffitins®or affitins); engineered, tenascin-derived, tenascin type III domain (e.g., Centyrin™), thioredoxin (e.g., peptide aptamer); KALBITOR®; the β-sandwich (e.g., iMab); gamma-B crystallin-derived scaffold or engineered, ubiquitin- derived scaffold (e.g., Affilins); engineered, protease inhibitor-derived, Kunitz domain (e.g., EETI-II / AGRP, BPTI / LACI-D1 / ITI-D2); engineered antibody mimics; miniproteins; engineered, Fyn-derived, SH2 domain (e.g., Fynomers®); genetically manipulated 204 314460903v1Attorney Docket No: 243735.000428 counterparts of the foregoing that retains its binding functionality, and any combination thereof.

74. The method of claim 72, wherein the antibody or antigen-binding fragment thereof isselected from an intact antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a single-chain variable fragment (scFv), a VH domain, a nanobody, a Bi-specific T-cell engager (BiTE), a bispecific killer cell engager (BiKE), a bi-specific macrophage engager (BiME), a CrossMab, a tri-specific binding partner, and any combination thereof.

75. The method of claim 72, wherein the nucleotide molecule is selected from an antisenseoligonucleotide, a micro RNA (miRNA), a small interfering RNA (siRNA), a short hairpin RNA (shRNA), a single guide RNA (sgRNA), and any combination thereof.

76. The method of claim 72, wherein the gene editing system comprises a CRISPR-associatedprotein (Cas) nuclease, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN), a meganuclease, any endo- or exo-nuclease, variants thereof, fragments thereof, or any combination thereof.

77. The method of claim 72, wherein the engineered cell system comprises a chimeric antigenreceptor (CAR) modified cell comprising a fragment that targets TTYH2.

78. The method of claim 77, wherein the CAR modified cell is include a CAR-modified T cell(CAR-T cell), a CAR-modified natural killer (NK) cell (CAR-NK cell), or a CAR- macrophage (CAR-M).

79. The method of any one of claims 67-78, wherein the agent is conjugated to a detectablelabel, a chemotherapeutic agent, a radioisotope, an affinity tag, or a toxin.

80. The method of any one of claims 67-74, wherein the agent that targets TTYH2 comprisesthe antibody or antigen-binding fragment of any one of claims 1-30, the antibody-drug 205 314460903v1Attorney Docket No: 243735.000428 conjugate of any one of claims 31-44, the polynucleotide of claim 45, the vector of claim 46, or the pharmaceutical composition of any one of claims 50-65.

81. The method of any one of claim 71-80, wherein the cancer is resistant to an immunotherapycomprising one or more immune checkpoint inhibitors.

82. The method of claim 81, wherein the immune checkpoint inhibitor is a programmed celldeath protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T-lymphocyte- associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a (B7-H4) inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD- 1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein- coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL- 1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) 206 314460903v1Attorney Docket No: 243735.000428 inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin- like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, or a vascular endothelialgrowth factor (VEGF) inhibitor, or a combination thereof.

83. The method of claim 82, wherein the immune checkpoint inhibitor comprises a PD-1inhibitor or PD-L1 inhibitor.

84. The method of claim 83, wherein the PD-1 inhibitor or PD-L1 inhibitor is nivolumab,pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof.

85. The method of claim 82, wherein the immune checkpoint inhibitor is a LAG-3 inhibitor.

86. The method of claim 85, wherein the LAG-3 inhibitor is relatlimab (BMS-986016),ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA- 1953, or a variant or a combination thereof.

87. The method of claim 86, wherein the LAG-3 inhibitor is relatlimab (BMS-986016), or avariant thereof. 207 314460903v1Attorney Docket No: 243735.000428 88. The method of claim 82, wherein the immune checkpoint inhibitor is a CTLA-4 inhibitor.

89. The method of claim 88, wherein the CTLA-4 inhibitor is ipilimumab, tremelimumab,quavonlimab, BNT316 / ONC-392 (gotistobart), or porustobart (HBM4003), or a variant or combination thereof.

90. The method of any one of claims 71-89, wherein the method further comprisesadministering to the subject one or more additional anti-cancer therapies.

91. The method of claim 90, wherein the one or more additional anti-cancer therapies comprisean immunotherapy, a chemotherapy, a targeted therapy, a radiotherapy, or a combination thereof.

92. The method of claim 91, wherein the immunotherapy is interferon beta-1a (IFN beta-1a),interferon beta-1b (IFN beta-1b), glatiramer acetate, mitoxantrone, natalizumab, teriflunomide, fingolimod, dimethyl fumarate, or alemtuzumab, or a variant or combination thereof.

93. A method of treating or preventing an autoimmune / autoinflammatory disease in a subjectin need thereof, the method comprising administering to the subject an effective amount of an agent that targets TTYH2.

94. The method of claim 93, wherein the method further comprises administering to the subjectone or more additional anti-autoimmune / autoinflammatory therapies.

95. The method of claim 94, wherein the one or more additional anti-autoimmune / autoinflammatory therapies comprise an immunotherapy or a targeted therapy, or a combination thereof.

96. The method of claim 94 or claim 95, wherein the immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, 208 314460903v1Attorney Docket No: 243735.000428 an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA- targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof.

97. The method of claim 95, wherein the immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, or a combination thereof.

98. A method of increasing effectiveness of an immunotherapy in a subject in need thereof, themethod comprising co-administering to the subject an immunotherapy with an effective amount of an agent that targets TTYH2.

99. The method of any one of claims 93-98, wherein the agent that targets TTYH2 is selectedfrom a small molecule, a protein, a protein-drug conjugate, a nucleotide molecule, a gene editing system, an engineered cell system, and any combination thereof. 100.The method of claim 99, wherein the protein is selected from a peptide; an antibody or antigen-binding fragment thereof; a monobody; engineered, low-density-lipoprotein- 209 314460903v1Attorney Docket No: 243735.000428 receptor-derived, A domain (LDLR-A) (e.g., Avimers™); a designed ankyrin repeat protein (DARPin) lipocalin (e.g., anticalins); an affibody; engineered, Protein-A-derived, Z domain (Affibodies™) CTLD3 (e.g., Tetranectin); C-type lectin-like domain scaffolds; Sac7d- derived polypeptides (e.g., Nanoffitins®or affitins); engineered, tenascin-derived, tenascin type III domain (e.g., Centyrin™), thioredoxin (e.g., peptide aptamer); KALBITOR®; the β-sandwich (e.g., iMab); gamma-B crystallin-derived scaffold or engineered, ubiquitin- derived scaffold (e.g., Affilins); engineered, protease inhibitor-derived, Kunitz domain (e.g., EETI-II / AGRP, BPTI / LACI-D1 / ITI-D2); engineered antibody mimics; miniproteins; engineered, Fyn-derived, SH2 domain (e.g., Fynomers®); genetically manipulated counterparts of the foregoing that retains its binding functionality, and any combination thereof. 101.The method of claim 99, wherein the antibody or antigen-binding fragment thereof is selected from an intact antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a single-chain variable fragment (scFv), a VH domain, a nanobody, a Bi-specific T-cell engager (BiTE), a bispecific killer cell engager (BiKE), a bi-specific macrophage nano-engager (BiME), a CrossMab, a tri- specific binding partner, and any combination thereof. 102.The method of claim 99, wherein the nucleotide molecule is selected from an antisense oligonucleotide, a micro RNA (miRNA), a small interfering RNA (siRNA), a short hairpin RNA (shRNA), a single guide RNA (sgRNA), and any combination thereof. 103.The method of claim 99, wherein the gene editing system comprises a CRISPR-associated protein (Cas) nuclease, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN), a meganuclease, any endo- or exo-nuclease, variants thereof, fragments thereof, or any combination thereof. 104.The method of claim 99, wherein the engineered cell system comprises a chimeric antigen receptor (CAR) modified cell comprising a fragment that targets TTYH2. 210 314460903v1Attorney Docket No: 243735.000428 105.The method of claim 104, wherein the CAR modified cell is include a CAR-modified T cell (CAR-T cell), a CAR-modified natural killer (NK) cell (CAR-NK cell), or a CAR- macrophage (CAR-M). 106.The method of any one of claims 93-105, wherein the agent is conjugated to a detectable label, a chemotherapeutic agent, a radioisotope, an affinity tag, or a toxin. 107.The method of any one of claims 93-106, wherein the agent that targets TTYH2 comprises the antibody or antigen-binding fragment of any one of claims 1-30, the antibody-drug conjugate of any one of claims 31-44, the polynucleotide of claim 45, the vector of claim 46, or the pharmaceutical composition of any one of claims 50-65. 108.The method of any one of claims 91-92, and 95-107, wherein the immunotherapy and the agent that targets TTYH2 are administered sequentially. 109.The method of any one of claims 91-92, and 95-107, wherein the immunotherapy and the agent that targets TTYH2 are administered simultaneously in one composition or in separate compositions. 110.The method of any one of claims 91-92, and 98-109, wherein the immunotherapy comprises one or more immune checkpoint inhibitors, cytokines, tumor-associated antigen (TAA)-targeting agents, or immune agonists. 111.The method of claim 110, wherein the immune checkpoint inhibitor is a programmed cell death protein 1 (PD-1) inhibitor, a programmed death-ligand 1 (PD-L1) inhibitor, a lymphocyte activation gene 3 (LAG-3 or CD223) inhibitor, a cytotoxic T-lymphocyte- associated antigen 4 (CTLA-4) inhibitor, a cluster of differentiation 47 (CD47) inhibitor, a T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) inhibitor, a B7 homolog 3 protein (B7-H3 or CD276) inhibitor, a B7-H4 inhibitor, a V-domain Ig suppressor of T cell activation (VISTA, also known as programmed death-1 homolog (PD- 1H)) inhibitor, a T cell immunoreceptor with Ig and ITIM Domains (TIGIT) inhibitor, a signal regulatory protein alpha (SIRPA) inhibitor, a signaling lymphocytic activation 211 314460903v1Attorney Docket No: 243735.000428 molecule family members (SLAMF) inhibitor, a poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG or CD112R) inhibitor, an adenosine A2A receptor (A2aR) inhibitor, an adenosine A2b receptor (A2bR) inhibitor, a G protein- coupled receptor 171 (GPR171) inhibitor, an insulin like growth factor binding protein 7 (IGFBP7) inhibitor, a cluster of differentiation 93 (CD93) inhibitor, a CD96 inhibitor, a CD226 inhibitor, a natural killer group protein 2A (NKG2A) inhibitor, a natural killer group protein 2D (NKG2D) inhibitor, a killer cell lectin like receptor G1 (KLRG1) inhibitor, a human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) inhibitor, a killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-15 inhibitor, a cluster of differentiation 24 (CD24) inhibitor, a sialic acid-binding immunoglobulin-like lectin (Siglec)-10 inhibitor, a P-selectin glycoprotein ligand-1 (PSGL- 1) inhibitor, a V-set and Ig domain-containing protein 3 (VSIG3, also known as BT-IgSF or IGSF11) inhibitor, a leucine rich repeats and immunoglobulin like domains 1 (LRIG1) inhibitor, a fibrinogen-like protein 1 (FGL1) inhibitor, a B and T lymphocyte attenuator (BTLA) inhibitor, a leukocyte associated immunoglobulin like receptor 1 (LAIR-1) inhibitor, a cluster of differentiation 160 (CD160) inhibitor, a leukocyte immunoglobulin- like receptor subfamily B member 2 (LILRB2) inhibitor, a leukocyte immunoglobulin-like receptor (LILRB4) inhibitor, an angiopoietin 2 (Ang2) inhibitor, a vascular endothelial growth factor (VEGF) inhibitor, or a combination thereof. 112.The method of claim 111, wherein the immune checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a LAG3 inhibitor, a CTLA-4 inhibitor, or a combination thereof. 113.The method of claim 111 or claim 112, wherein the PD-1 inhibitor or PD-L1 inhibitor is nivolumab, pembrolizumab, BAT1308, durvalumab, avelumab, atezolizumab, acrixolimab, balstilimab, budigalimab, bintrafusp alfa, cadonilimab, camrelizumab, cemiplimab, cetrelimab, danvilostomig, dostarlimab, eciskafusp alfa, enlonstobart, ezabenlimab, fanastomig, fidasimtamab, finotonlimab, geptanolimab, iparomlimab, ivonescimab, izuralimab, lipustobart, lodapolimab, lomvastomig, lorigerlimab, nofazinlimab, penpulimab, peresolimab, pidilizumab, pimivalimab, pradusinstobart, prolgolimab, 212 314460903v1Attorney Docket No: 243735.000428 pucotenlimab, reozalimab, retifanlimab, rilvegostomig, rosnilimab, rulonilimab, sabestomig, sasanlimab, serplulimab, sintilimab, spartalizumab, tebotelimab, tislelizumab, tobemstomig, toripalimab, volrustomig, vudalimab, zeluvalimab, or zimberelimab, or a variant or a combination thereof. 114.The method of claim 111 or claim 112, wherein the LAG-3 inhibitor is relatlimab (BMS- 986016), ABL501, CB213, EMB-02, favezelimab (MK-420 / 22D2), fianlimab (REGN3767), FS118, GSK2831781 (IMP731), IBI323, ieramilimab (LAG525 / IMP701 / BAP050), miptenalimab (BI-754111 / 496G6), pavunalimab (XmAb841), Sym022, tebotelimab (MGD013), tobemstomig (RG-6139 / RO-7247669), TSR-033, tuparstobart (INCAGN02385), or BGA-1953, or a variant or a combination thereof. 115.The method of claim 114, wherein the LAG-3 inhibitor is relatlimab (BMS-986016), or a variant thereof 116.The method of claim 111 or claim 112, wherein the immune checkpoint inhibitor is a CTLA-4 inhibitor. 117.The method of claim 116, wherein the CTLA-4 inhibitor is ipilimumab, tremelimumab, or quavonlimab, BNT316 / ONC-392 (gotistobart), or porustobart (HBM4003), or a variant or a combination thereof. 118.The method of claim 110, wherein the cytokine is interleukin-2 (IL-2), IL-6, IL-10, IL-15, IL-21, IFN-α, IFN-β, IFN-γ, IL-18, CCL19, CCL21, monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF), or a receptor thereof. 119.The method of claim 110, wherein the TAA targeting agent targets 5T4, Ang2, BCMA, CD123, CD19, CD20, CD22, CD33, CD38, CD47, CEA, CEACAM5, CEACAM6, Claudin 6, Claudin 18.2, CLEC12A, DLL3, EGFR, EpCAM, FcRH5, FLT3, GD2, Glypican-3, 213 314460903v1Attorney Docket No: 243735.000428 gpA33, GPRC5D, Her2, Her3, MAGE-A4, MET, MUC16, MUC17, NY-ESO-1, P- cadherin, PRAME, PSCA, PSMA, SSTR2, STEAP1, TROP2 (TACSTD2), Ang2, Tie2, VEGF, VEGFR, γδTCR, KRAS, or RAF, or a combination thereof. 120.The method of claim 110, wherein the immune agonist is a 4-1BB agonist, an OX40 agonist, a CD40 agonist, a CD30 agonist, a GITR agonist, an ICOS agonist, a CD27 agonist, a CD28 agonist, a CD28H / TMIGD2 agonist, an NCR3 agonist, an NCR1 agonist, an NCR2 agonist, a DR3 agonist, a CD226 agonist, a CRTAM agonist, a HVEM agonist, a TNFR1 agonist, a TNFR2 agonist, a CD2 agonist, a CD7 agonist, a TLR4 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof. 121.The method of any one of claims 98-120, wherein the immunotherapy comprises a stimulator of interferon genes (STING) agonist or toll-like receptor (TLR) agonist or a combination thereof. 122.The method of claim 121, wherein the STING agonist is DMXAA, 2’3’-cGAMP, ADU- S100 / MIW815, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING (BI 1387446), E7766, TAK-676, SNX281, or SYNB1891, or a variant or a combination thereof. 123.The method of claim 121, wherein the TLR agonist is a TLR1 / TLR2 agonist, TLR2 / TLR6 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist, or a combination thereof. 124.The method of any one of claims 98-109, wherein the immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster 214 314460903v1Attorney Docket No: 243735.000428 of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA- targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof. 125.The method of any one of claims 67-92, and 98-123, wherein the subject has a solid cancer. 126.The method of claim 125, wherein the solid cancer develops as a hot tumor with abundant anti-tumor immune responses particularly T cell activities. 127.The method of claim 125, wherein the solid cancer develops as a cold tumor with weak anti-tumor immune responses particularly T cell activities. 128.The method of any one of claims 125-127, wherein the solid cancer is a lung cancer, a glioma, a medulloblastoma, a thyroid cancer, a colorectal cancer, a head and neck cancer, a gastric cancer, a stomach cancer, a liver cancer, a pancreatic cancer, a renal cancer, a urothelial cancer, a prostate cancer, a testis cancer, a breast cancer, a cervical cancer, an endometrial cancer, an ovarian cancer, a gallbladder cancer, a sarcoma, or a melanoma. 129.The method of claim 128, wherein the glioma is a glioblastoma or an astrocytoma. 130.The method of claim 128, wherein the solid cancer is a colorectal cancer or a melanoma. 215 314460903v1Attorney Docket No: 243735.000428 131.The method of claim 128 or 129, wherein the solid cancer is a pancreatic cancer or a glioblastoma. 132.The method of any one of claims 67-92, and 98-123, wherein the subject has a hematologic cancer. 133.The method of claim 132, wherein the hematologic cancer is acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), acute leukemia of ambiguous lineage, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, chronic myeloid neoplasm, non-Hodgkin lymphoma (e.g., follicular non-Hodgkin lymphoma), Hodgkin lymphoma, chronic leukemia, dendritic / histiocytic neoplasm, or lymphoproliferative disorder. 134.The method of claim 133, wherein the hematologic cancer is acute myeloid leukemia (AML). 135.The method of any one of claims 67-92, 98-123, and 125-134, wherein the method further comprises administering to the subject one or more additional anti-cancer therapies. 136.The method of claim 135, wherein the one or more additional anti-cancer therapies comprise an immunotherapy, a chemotherapy, a targeted therapy, a radiotherapy, or a combination thereof. 137.The method of claim 136, wherein the immunotherapy is selected from interferon beta-1a (IFN beta-1a), interferon beta-1b (IFN beta-1b), glatiramer acetate, mitoxantrone, natalizumab, teriflunomide, fingolimod, dimethyl fumarate, and alemtuzumab, or a variant or combination thereof. 138.The method of any one of claims 93-109 and 124, wherein the autoimmune / autoinflammatory disease is multiple sclerosis (MS) or Bechet’s disease. 216 314460903v1Attorney Docket No: 243735.000428 139.The method of any one of claims 93-109, 124, and 138, wherein the method further comprises administering to the subject one or more additional anti- autoimmune / autoinflammatory therapies. 140.The method of claim 139, wherein the one or more additional anti- autoimmune / autoinflammatory therapies comprise an immunotherapy or a targeted therapy, or a combination thereof. 141.The method of claim 140, wherein the immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, an interleukin 4 (IL-4) inhibitor, an interleukin 5 (IL-5) inhibitor, an interleukin 13 (IL-13) inhibitor, an interleukin 17A (IL-17A) inhibitor, an interleukin 17F (IL-17F) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, an IFN-α inhibitor, an IFN-β inhibitor, a very late antigen 4 (VLA4, also known as alpha 4 beta 1 (α4β1)-integrin or CD49d / CD29) inhibitor, a cluster of differentiation 20 (CD20) inhibitor, a B activating factor (BAFF) inhibitor, a transmembrane activator and CAML interactor (TACI) inhibitor, a cluster of differentiation 19 (CD19) inhibitor, an interleukin-23 receptor (IL-23R) inhibitor, a cluster of differentiation 22 (CD22) inhibitor, a cluster of differentiation 38 (CD38) inhibitor, a B-cell maturation antigen (BCMA) inhibitor, a CD19-targeted T cell engager, a CD20-targeted T cell engager, a CD22-targeted T cell engager, a CD38-targeted T cell engager, a BCMA- targeted T-cell engager, a PD-1 agonist, a BTLA agonist, a CD40 inhibitor, a CD40 ligand (CD40L) inhibitor, an OX40 inhibitor, an OX40L inhibitor, a B7 inhibitor, a CD28 inhibitor, a CTLA-4-Fc fusion protein, a tumor necrosis factor-like cytokine 1A (TL1A) inhibitor, a DR3 inhibitor, or a thymic stromal lymphopoietin (TSLP) inhibitor, or a combination thereof. 142.The method of claim 142, wherein the immunotherapy is a tumor necrosis factor-alpha (TNF-α) inhibitor, an interleukin 6 (IL-6) inhibitor, an interleukin 10 (IL-10) inhibitor, a chemokine (C-C motif) ligand 2 (CCL2) inhibitor, a C-X-C motif chemokine ligand 10 (CXCL10) inhibitor, or a combination thereof. 217 314460903v1Attorney Docket No: 243735.000428 143.The method of any one of claims 67-142, wherein the administering is via intrathecal, intratumoral, intravenous, intradermal, intraperitoneal, subcutaneous, intramuscular, inhalation, oral delivery, lipid nanoparticle (LNP)-based delivery, cellular delivery, viral and / or non-viral delivery, or gene editing, or as a cargo in a cell, or any combination thereof. 144.The method of any one of claims 67-143, wherein the subject is human or veterinary animal. 218 314460903v1

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