Chimeric shuttling receptor

Abstract

A chimeric shuttling receptor, comprising: (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a polypeptide derived from a Golgi-related shuttling receptor. The chimeric shuttling receptor is used for reducing the expression of the target protein. Also provided are methods for producing and using the chimeric shuttling receptor.

Description

Chimeric shuttle receptor Technical Field

[0001] This application relates to a chimeric shuttle receptor comprising: (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a polypeptide derived from a Golgi-associated shuttle receptor; this application also relates to methods for preparing and using these constructs. Background Technology

[0002] Protein expression and localization play crucial roles in many aspects of cellular function, including in disease contexts and in engineered cells. The need for regulation of target protein expression in cells, including for disease treatment, remains unmet. This application addresses this and other needs.

[0003] All publications, patents, patent publications, and patent application publications mentioned in this article are incorporated herein by reference in their entirety. Summary of the Invention

[0004] In one aspect, this application provides a chimeric shuttle receptor comprising a polypeptide derived from a Golgi-associated shuttle receptor. In some embodiments, according to any of the above-described chimeric shuttle receptors, it comprises (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a polypeptide derived from a Golgi-associated shuttle receptor. In some embodiments, the polypeptide derived from a Golgi-associated shuttle receptor comprises a transmembrane domain or a portion thereof. In some embodiments, the polypeptide derived from a Golgi-associated shuttle receptor comprises a cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor further comprises a transmembrane domain or a portion thereof located between the heterologous binding portion and the cytoplasmic tail region or a portion thereof.

[0005] In one aspect, this application provides a chimeric shuttle receptor comprising a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, according to any of the above chimeric shuttle receptors, it comprises (a) a heterologous binding portion capable of specifically binding a target protein, and (b) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, according to any of the above chimeric shuttle receptors, it optionally comprises a cytoplasmic tail region or a portion thereof, said cytoplasmic tail region or portion thereof may or may not be derived from a Golgi-associated shuttle receptor.

[0006] In one aspect, this application provides a chimeric shuttle receptor comprising a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, according to any of the chimeric shuttle receptors described above, it comprises (a) a heterologous binding portion capable of specifically binding a target protein, and (b) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, according to any of the chimeric shuttle receptors described above, it further comprises a transmembrane domain or a portion thereof located between the heterologous binding portion and the cytoplasmic tail region or a portion thereof, said transmembrane domain or a portion thereof may or may not be derived from a Golgi-associated shuttle receptor.

[0007] In some embodiments, according to any of the chimeric shuttle receptors described above, the polypeptide derived from the Golgi-associated shuttle receptor includes a transmembrane domain or a portion thereof and a cytoplasmic tail region or a portion thereof. In some embodiments, the transmembrane domain or a portion thereof may be derived from the same Golgi-associated shuttle receptor as the cytoplasmic tail region or a portion thereof.

[0008] In some embodiments, according to any of the chimeric shuttle receptors described above, the Golgi-associated shuttle receptor is a sorting protein (SORT), a Sortilin-related receptor (SORL), a central nervous system-expressed Sortilin-related receptor 1 (SORCS1), a central nervous system-expressed Sortilin-related receptor 2 (SORCS2), a central nervous system-expressed Sortilin-related receptor 3 (SORCS3), or a lysosomal membrane protein 2 (SCARB2). In some embodiments, according to any of the chimeric shuttle receptors described above, the Golgi-associated shuttle receptor is a VPS10P domain receptor.

[0009] In some embodiments, according to any of the chimeric shuttle receptors described above, the polypeptide derived from a Golgi-associated shuttle receptor includes a transmembrane domain or a portion thereof. In some embodiments, the transmembrane domain or a portion thereof is derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some embodiments, according to any of the chimeric shuttle receptors described above, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-36, or an amino acid sequence having at least 75% sequence identity with it. In some embodiments, the transmembrane domain or a portion thereof is not derived from a Golgi-associated shuttle receptor; for example, the transmembrane domain or a portion thereof is derived from CD8α, programmed death receptor 1 (PD1), or low-density lipoprotein receptor-associated protein (LRP). In some embodiments, according to any of the chimeric shuttle receptors described above, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:99, SEQ ID NO:101, or SEQ ID NO:114, or an amino acid sequence having at least 75% sequence identity with it.

[0010] In some embodiments, according to any of the chimeric shuttle receptors described above, the polypeptide derived from a Golgi-associated shuttle receptor includes a cytoplasmic tail region or a portion thereof. In some embodiments, the cytoplasmic tail region or a portion thereof is derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some embodiments, according to any of the chimeric shuttle receptors described above, the cytoplasmic tail region or a portion thereof contains the amino acid sequence shown in any of SEQ ID NO:37-65, or an amino acid sequence having at least 75% sequence identity with it. In some embodiments, according to any of the chimeric shuttle receptors described above, the cytoplasmic tail region or a portion thereof contains the amino acid sequence shown in any of SEQ ID NO:37-59, or an amino acid sequence having at least 75% sequence identity with it. In some embodiments, according to any of the chimeric shuttle receptors described above, the cytoplasmic tail region or a portion thereof is derived from a Golgi-associated shuttle receptor that is the same as or a portion thereof as the transmembrane domain. In some embodiments, according to any of the chimeric shuttle receptors described above, the cytoplasmic tail region or a portion thereof and the transmembrane domain or a portion thereof are derived from different molecules. In some embodiments, the cytoplasmic tail region or a portion thereof is not derived from a Golgi-associated shuttle receptor; for example, the cytoplasmic tail region or a portion thereof is derived from low-density lipoprotein receptor-associated protein (LRP). In some embodiments, according to any of the chimeric shuttle receptors described above, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:115, or an amino acid sequence having at least 75% sequence identity with it.

[0011] In some embodiments, based on any of the chimeric shuttle receptors described above, the heterologous binding portion comprises an antibody or an antigen-binding fragment thereof. In some embodiments, the heterologous binding portion is an antibody.

[0012] In some implementations, the antibody or its antigen-binding fragment, depending on any of the chimeric shuttle receptors described above, is IgG, a single-chain variable fragment (scFv), a single-domain antibody (sdAb), VHH, Fab, (Fab)2, (scFv')2, or a nanobody.

[0013] In some implementations, the heterologous binding moiety, depending on any of the chimeric shuttle receptors described above, includes a peptide, ligand, aptamer, designed ankyrin repeat protein (DARPin), artificially designed binding protein, or D-Domain.

[0014] In some embodiments, the heterologous binding portion binds to extracellular protein molecules, membrane protein molecules, or intracellular molecules, depending on any of the chimeric shuttle receptors described above. In some embodiments, the heterologous binding portion binds to inhibitory immune receptors, ligands of inhibitory immune receptors, or immune checkpoint molecules.

[0015] In some embodiments, the heterologous binding portion can bind to any protein that is desired to be downregulated, depending on any of the chimeric shuttle receptors described above. In some embodiments, the heterologous binding portion binds to one or more components of the major histocompatibility complex (MHC) or the CD3 / T cell receptor (TCR) complex. In some embodiments, the heterologous binding portion binds to MHC class I molecules (HLA-A, HLA-B, HLA-C, HLA-G, or HLA-E), MHC class II molecules (HLA-DP, HLA-DQ, HLA-DR), or MHC class III molecules. In some embodiments, the heterologous binding portion binds to the T-cell receptor (TCR) or its subunits, the CD3 / TCR complex or its subunits (CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, and CD3ζ), programmed cell death protein 1 (PD-1), programmed cell death ligand 1 (PD-L1), CTLA-4, CTLA-1, TIM3, LAG3, TIGIT, epidermal growth factor receptor (EGFR), glucocorticoid receptor (GR), CD38, CD7, CD25, CD69, CD70, CD56, CD22, FcγRII-B, 4-1BB, 4-BBL, β2-microglobulin (B 2M), CIITA, NKG2A, NKG2D, CD94, TGF-β receptor, cytotoxic immunoglobulin-like receptor (KIR)2DL1, KIR2DL2, KIR2DL3, HLA, BCMA, CFH, ARMS2, MYOC, OPTN, VEGF, HIF1A, RB1, MUC16, TGFBR1, TGFBR2, OPN1SW, OPN1MW, MTTP, FOXC2, MSX1, KERA, SLC4A11, PAX6, OTX2, RHO, P23H, USH2A, MYO7A, LXS, CRYAA, CRYAB, ABCA4, CEP290 or MHCI. In some implementations, the heterogeneous binding portion is bound to NR3C1, Perforin1, GUCY2C, CLEC12A, PRLR, ERBB2, IMPDH, ITK, TMEM30A, Regnase-1, IFNGR, CD58, HSP90B1, or PIM3.

[0016] On the other hand, this application provides peptides or combinations of peptides that comprise any chimeric shuttle receptor described herein. In some embodiments, the peptides or combinations of peptides also comprise any antigen recognition receptor described herein. In some embodiments, the antigen recognition receptor is an engineered T-cell receptor (TCR), a chimeric antigen receptor (CAR), a T-cell antigen conjugate (TAC), a chimeric TCR (cTCR), a T-cell antigen conjugate-like (TAC) chimeric receptor, a chimeric switch receptor, a signal transduction receptor, an inducible-regulated dimerization-activated receptor (DARIC), a chimeric cytokine receptor, a co-stimulatory receptor, a dominant-negative receptor, or a portion thereof.

[0017] In some embodiments, the chimeric shuttle receptor and antigen recognition receptor in the polypeptide combination may be unconnected. In other embodiments, the chimeric shuttle receptor and antigen recognition receptor in the polypeptide combination may be connected or fused, i.e., as a single polypeptide containing both a chimeric shuttle receptor region and an antigen recognition receptor region.

[0018] In some embodiments, according to the above-described polypeptides or combinations of polypeptides, the chimeric shuttle receptor and the antigen recognition receptor are separated by a self-cleaving linker. In some embodiments, the self-cleaving linker is a 2A peptide, such as P2A, T2A, E2A, or F2A peptide. In some embodiments, according to the above-described polypeptides or combinations of polypeptides, the chimeric shuttle receptor and the antigen recognition receptor are separated by an internal ribosome entry site (IRES). In some embodiments, according to the above-described polypeptides or combinations of polypeptides, the chimeric shuttle receptor and the antigen recognition receptor are separated by an enzyme cleavage site. In some embodiments, the enzyme cleavage site includes, for example, a trypsin cleavage site or a thrombin cleavage site.

[0019] In some implementations, the chimeric shuttle receptor is located upstream of the antigen recognition receptor, based on the aforementioned polypeptide or combination of polypeptides.

[0020] In some implementations, the chimeric shuttle receptor is located downstream of the antigen recognition receptor, based on the aforementioned polypeptide or combination of polypeptides.

[0021] In some embodiments, the polypeptide or combination of polypeptides described above further includes a detection marker. In some embodiments, the detection marker includes, but is not limited to, antibodies or ligands, resistance genes (e.g., antibiotic resistance genes such as puromycin resistance gene (Puro), G418 resistance gene (Neo), blast fungicide resistance gene (BSD)) or selection markers (e.g., affinity tags such as histidine tags), and fluorescent proteins.

[0022] In some embodiments, the polypeptide or polypeptide combination further comprises exogenous inosine monophosphate dehydrogenase (IMPDH). In some embodiments, according to the above-described polypeptide or polypeptide combination, the chimeric shuttle receptor, the antigen recognition receptor, and the exogenous IMPDH are separated by a self-cleaving linker, including a 2A peptide linker, such as P2A, T2A, E2A, or F2A peptides. In some embodiments, according to the above-described polypeptide or polypeptide combination, the chimeric shuttle receptor is separated from the antigen recognition receptor and the exogenous IMPDH by an internal ribosome entry site (IRES). In some embodiments, according to the above-described polypeptide or polypeptide combination, the chimeric shuttle receptor is separated from the antigen recognition receptor and the exogenous IMPDH by an enzyme cleavage site. In some embodiments, the enzyme cleavage site comprises, for example, a trypsin cleavage site or a thrombin cleavage site.

[0023] In some embodiments, the exogenous inosine monophosphate dehydrogenase (IMPDH) is resistant to purine biosynthesis inhibitors. In some embodiments, the exogenous IMPDH is a non-human IMPDH or a variant of human IMPDH. In some embodiments, the exogenous IMPDH is derived from prokaryotic IMPDH, such as bacterial IMPDH or a variant thereof. In some embodiments, the bacterial IMPDH is Gram-positive bacterial IMPDH or a variant thereof. In some embodiments, the Gram-positive bacterial IMPDH is derived from Bacillus subtilis, Lactobacillus plantarum, or Staphylococcus aureus. In some embodiments, the bacterial IMPDH is Gram-negative bacterial IMPDH or a variant thereof. In some embodiments, the Gram-negative bacterial IMPDH is derived from Escherichia coli or Mesoplasma florum. In some specific embodiments, the exogenous IMPDH is Lactobacillus plantarum IMPDH, Escherichia coli IMPDH, Bacillus subtilis IMPDH, Staphylococcus aureus IMPDH, or Mesoplasma florum IMPDH, or a variant thereof.

[0024] On the other hand, this application provides isolated nucleic acids encoding any chimeric shuttle receptor described herein. In some embodiments, the isolated nucleic acid further comprises a second nucleic acid encoding an antigen recognition receptor. In some embodiments, the isolated nucleic acid further comprises a third nucleic acid encoding an exogenous IMPDH.

[0025] In some embodiments, based on the isolated nucleic acids described above, the nucleic acid encoding the chimeric shuttle receptor is located upstream of the nucleic acid encoding the antigen recognition receptor. In some embodiments, based on the isolated nucleic acids described above, the nucleic acid encoding the chimeric shuttle receptor is located downstream of the nucleic acid encoding the antigen recognition receptor. In some embodiments, based on the isolated nucleic acids described above, the order of the nucleic acids encoding the chimeric shuttle receptor, the antigen recognition receptor, and / or the exogenous IMPDH is not limited.

[0026] In some implementations, the isolated nucleic acid described above contains a nucleic acid encoding a polypeptide or combination of polypeptides as described herein.

[0027] In some implementations, the isolated nucleic acid also includes a nucleic acid encoding a detection marker.

[0028] On the other hand, this application provides a vector containing any isolated nucleic acid described herein.

[0029] On the other hand, this application provides engineered cells comprising any chimeric shuttle receptor described herein, any antigen recognition receptor described herein, any polypeptide or combination of polypeptides described herein, any isolated nucleic acid described herein, or any vector described herein.

[0030] In some implementations, the engineered cells described above contain nucleic acids encoding any of the chimeric shuttle receptors described herein.

[0031] In some embodiments, the engineered cells further comprise nucleic acids encoding antigen recognition receptors, optionally wherein the antigen recognition receptors are selected from engineered T-cell receptors (TCRs), chimeric antigen receptors (CARs), T-cell antigen conjugates (TACs), chimeric TCRs (cTCRs), T-cell antigen conjugates (TACs), chimeric switch receptors, signal transduction receptors, induced regulation dimerization activation receptors (DARICs), chimeric cytokine receptors, co-stimulatory receptors, dominant and negative receptors, or portions thereof.

[0032] In some embodiments, the antigen recognition receptor described herein comprises: (a) an extracellular antigen-binding domain comprising a single-domain antibody (sdAb) domain that binds to GUCY2C; (b) a transmembrane domain; and (c) an intracellular signal transduction domain.

[0033] In some embodiments, the engineered cells described above further comprise detection markers or nucleic acids encoding detection markers, wherein the detection markers optionally include, but are not limited to, antibodies or ligands, resistance genes (e.g., antibiotic resistance genes, such as puromycin resistance gene (Puro), G418 resistance gene (Neo), blast fungicide resistance gene (BSD)) or selection markers (e.g., affinity tags, such as histidine tags), and fluorescent proteins.

[0034] In some embodiments, the engineered cells, based on the aforementioned engineered cells, are immune cells. In some embodiments, the immune cells are T cells, natural killer (NK) cells, γδT cells, αβT cells, cytotoxic T cells (CTLs), regulatory T (Treg) cells, natural killer T (NKT) cells, peripheral blood mononuclear cells (PBMCs), hematopoietic stem cells, pluripotent stem cells, embryonic stem cells, memory T cells, T memory stem cells (TSCMs), naive T cells, effector T cells, T helper cells, stem cells, induced pluripotent stem (iPSC) cells, ocular tissue cells, hepatocytes, kidney cells, cardiac cells, or bone marrow cells, or tumor-infiltrating lymphocytes (TILs).

[0035] In some implementations, the engineered cells are allogeneic cells or autologous cells.

[0036] On the other hand, this application provides a method for producing any engineered cells described herein, the method comprising introducing any isolated nucleic acid provided herein or any vector described herein into engineered cells.

[0037] On the other hand, this application provides a method for modulating target proteins in cells, comprising introducing a nucleic acid encoding any of the chimeric shuttle receptors described herein into cells, wherein the heterologous binding portion is capable of specifically binding to the target protein. In some embodiments, modulating the target protein involves downregulating the expression level of the target protein. In some embodiments, a viral vector is used to introduce the nucleic acid encoding the chimeric shuttle receptor into cells, optionally wherein the viral vector is a retroviral (e.g., lentiviral) vector, an adenovirus vector, or an adeno-associated virus vector.

[0038] On the other hand, this application provides pharmaceutical compositions comprising any chimeric shuttle receptor described herein, any antigen recognition receptor described herein, any polypeptide or combination of polypeptides described herein, any isolated nucleic acid described herein, or any carrier described herein.

[0039] On the other hand, this application provides pharmaceutical compositions comprising any of the engineered cells described herein.

[0040] In some embodiments, according to any of the above-described pharmaceutical compositions, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient. In some embodiments, according to any of the above-described pharmaceutical compositions, the pharmaceutical composition further comprises an additional therapeutic agent. In some embodiments, according to any of the above-described pharmaceutical compositions, the pharmaceutical composition further comprises an agent that inhibits the function of antigen-presenting cells (APCs). In some embodiments, the agent is abatacept, tofacitinib, secukinumab, ipilimumab, cyclosporine A, fingolimod, methotrexate, or equivalents thereof. In some embodiments, according to any of the above-described pharmaceutical compositions, the pharmaceutical composition further comprises a purine biosynthesis inhibitor. In some embodiments, the purine biosynthesis inhibitor is mycophenolic acid (MPA), mycophenolate ester, mycophenolate mofetil (MMF), sodium mycophenolate (MPS), calcium mycophenolate, potassium mycophenolate, or derivatives thereof. In some embodiments, the purine biosynthesis inhibitor is ribavirin, mizoribine, tiazofurin, AVN-944 (VX-944), FF-10501, AS2643361, or BMS-986126, or their pharmaceutically acceptable esters, salts, or prodrugs.

[0041] On the other hand, this application provides a method for treating an individual's disease or condition, comprising administering to the individual an effective amount of any chimeric shuttle receptor, any antigen recognition receptor, any polypeptide or combination of polypeptides described herein, any isolated nucleic acid described herein, any carrier described herein, any engineered cell described herein, or any pharmaceutical composition described herein. In some embodiments, the engineered cells in the pharmaceutical composition are allogeneic to the individual. In some embodiments, the treatment method is allogeneic cell therapy or autologous cell therapy.

[0042] On the other hand, this application provides a method for treating graft-versus-host disease (GVHD) or host-versus-graft reaction (HVGR) in a subject, comprising administering to an individual an effective amount of any chimeric shuttle receptor, any antigen recognition receptor, any polypeptide or combination of polypeptides described herein, any isolated nucleic acid described herein, any vector described herein, any engineered cell described herein, or any pharmaceutical composition described herein. In some embodiments, treating GVHD or HVGR is to prevent GVHD or HVGR, or to reduce the risk of developing GVHD or HVGR. In some embodiments, host-versus-graft reaction (HVGR) includes acute HVGR and chronic HVGR. In some embodiments, graft-versus-host disease (GVHD) includes acute GVHD and chronic GVHD. In some embodiments, graft-versus-host disease (GVHD) is steroid-refractory acute GVHD.

[0043] On the other hand, this application provides for the use of any chimeric shuttle receptor, any antigen recognition receptor, any polypeptide or combination of polypeptides, any isolated nucleic acid, or any carrier, any engineered cell, or any pharmaceutical composition described herein as a drug or for the preparation of a drug. In some embodiments, the drug is used to treat a disease or condition. In some embodiments, the treatment is allogeneic cell therapy or autologous cell therapy. In some embodiments, the drug is used to treat graft-versus-host disease (GVHD) or host-versus-graft reaction (HVGR). Attached Figure Description

[0044] Figure 1 shows the TCR degradation efficiency in T cells expressing chimeric shuttle receptor (SR)-CAR constructs or control CAR constructs; where WD103 (SORT), WD105 (SORL), WD106 (SORCS1), WD107 (SORCS2), WD108 (SORCS3), and WD110 (SCARB2) represent CAR-T cells transduced based on SR-CAR constructs constructed from Golgi-associated shuttle receptors SORT, SORL, SORCS1, SORCS2, SORCS3, and SCARB2, respectively; the control group (WD115 ER) refers to CAR-T cells containing endoplasmic reticulum (ER) retention signals but not Golgi-associated shuttle receptors; and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0045] Figure 2 shows the change in the percentage of TCR+ cells in T cells expressing the chimeric shuttle receptor (SR)-CAR construct or the control CAR construct; WD103 (SORT) refers to CAR-T cells transduced with the SR-CAR construct based on the Golgi-associated shuttle receptor SORT, and the control group (WD115 ER) refers to CAR-T cells in which the Golgi-associated shuttle receptor was replaced with endoplasmic reticulum (ER) retention signals.

[0046] Figure 3 shows the changes in the percentage of TCR+ cells in T cells expressing chimeric shuttle receptor (SR)-CAR constructs or control CAR constructs. WD103(SORT) refers to CAR-T cells transduced with an SR-CAR construct based on the Golgi-associated shuttle receptor SORT, and WD103(SORT)+T84 refers to CAR-T cells expressing the SR-CAR construct co-cultured with T84 tumor cells. The control group (WD117LRP) refers to CAR-T cells with the Golgi-associated shuttle receptor replaced by LRP molecules, and the control group (WD117 LRP)+T84 refers to CAR-T cells expressing the LRP-CAR construct co-cultured with T84 tumor cells.

[0047] Figure 4 shows fluorescence images obtained by laser scanning confocal microscopy, where red fluorescence signals represent the localization of CD3ε protein molecules, green fluorescence signals represent the localization of the endoplasmic reticulum (ER), yellow fluorescence signals represent the localization of the Golgi apparatus, blue fluorescence signals represent the localization of the nucleus, and merged images represent a combined image of the fluorescence signals of CD3ε, ER, Golgi apparatus, and nucleus.

[0048] Figure 5 shows the TCR degradation efficiency in T cells expressing chimeric shuttle receptor (SR)-CAR constructs or control CAR constructs with different hinge region lengths; where WD103 (H13) refers to CAR-T cells transduced with the WD103 SR-CAR construct (hinge region length of 13 amino acids); WD103-2 (H12) refers to CAR-T cells with the hinge region length of the WD103 SR-CAR construct shortened to 12 amino acids; WD103-3 (H11) refers to CAR-T cells with the hinge region length of the WD103 SR-CAR construct shortened to 11 amino acids; and so on, WD104 (H4) refers to CAR-T cells with the hinge region length of the WD103 SR-CAR construct shortened to 4 amino acids; WD103-10 (H3) refers to CAR-T cells with the hinge region length of the WD103 SR-CAR construct shortened to 3 amino acids; WD103-11 (H2) refers to WD103... CAR-T cells with a hinge region length shortened to 2 amino acids in the SR-CAR construct; WD103-12(H1) refers to CAR-T cells with a hinge region length shortened to 1 amino acid in the WD103 SR-CAR construct; WD103-13(H0) refers to CAR-T cells with no hinge region at all; the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0049] Figure 6 shows the TCR degradation efficiency in T cells expressing SR-CAR constructs or control CAR constructs with different hinge region lengths; where WD105 (SORL H18) refers to CAR-T cells transduced with the WD105 SR-CAR construct (hinge region length of 18 amino acids); WD105-2 (SORL H4) refers to CAR-T cells with the hinge region length of the WD105 SR-CAR construct shortened to 4 amino acids; and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0050] Figure 7 shows the TCR degradation efficiency in T cells expressing SR-CAR constructs or control CAR constructs with different hinge region lengths; where WD106 (SORCS1 H11) refers to CAR-T cells transduced with the WD106 SR-CAR construct (hinge region length of 11 amino acids); WD106-2 (SORCS1 H4) refers to CAR-T cells with the hinge region length of the WD106 SR-CAR construct shortened to 4 amino acids; and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0051] Figure 8 shows the TCR degradation efficiency in T cells expressing SR-CAR constructs or control CAR constructs with different hinge region lengths; where WD107 (SORCS2 H11) refers to CAR-T cells transduced with the WD107 SR-CAR construct (hinge region length of 11 amino acids); WD107-2 (SORCS2 H4) refers to CAR-T cells with the hinge region length of the WD107 SR-CAR construct shortened to 4 amino acids; and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0052] Figure 9 shows the TCR degradation efficiency in T cells expressing SR-CAR constructs or control CAR constructs with different hinge region lengths; where WD108 (SORCS3 H9) refers to CAR-T cells transduced with the WD108 SR-CAR construct (hinge region length of 9 amino acids); WD108-2 (SORCS3 H4) refers to CAR-T cells with the hinge region length of the WD108 SR-CAR construct shortened to 4 amino acids; and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0053] Figure 10 shows the TCR degradation efficiency in T cells expressing SR-CAR constructs or control CAR constructs with different hinge region lengths; where WD110 (SCARB2 H13) refers to CAR-T cells transduced with the WD110 SR-CAR construct (hinge region length of 13 amino acids); WD110-2 (SCARB2 H4) refers to CAR-T cells with the hinge region length of the WD110 SR-CAR construct shortened to 4 amino acids; and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0054] Figure 11 shows the changes in the percentage of TCR+ cells in CAR-T cells expressing SR-CAR constructs or control CAR constructs with different hinge region lengths. WD103, WD104, WD103-2 to WD103-13 refer to CAR-T cells with hinge region lengths of 13 to 0 amino acids in the SR-CAR constructs. The control group (WD115 ER) refers to CAR-T cells in which the Golgi-associated shuttle receptor was replaced with endoplasmic reticulum (ER) retention signals.

[0055] Figure 12 shows the change in the percentage of TCR+ cells in CAR-T cells co-cultured with T84 tumor cells.

[0056] Figure 13 shows the TCR degradation efficiency in CAR-T cells expressing SR-CAR constructs with cytoplasmic tail regions or control CAR constructs. WD104 refers to CAR-T cells with SR-CAR constructs without cytoplasmic tail regions, WD132 to WD146 refer to CAR-T cells with SR-CAR constructs with cytoplasmic tail regions, and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0057] Figure 14 shows the TCR degradation efficiency in CAR-T cells expressing SR-CAR constructs with cytoplasmic tail regions or control CAR constructs. WD106-2 refers to CAR-T cells with SR-CAR constructs without cytoplasmic tail regions, WD150 to WD152 refer to CAR-T cells with SR-CAR constructs having cytoplasmic tail regions, and the control group (WD116 CAR) refers to CAR-T cells without Golgi-associated shuttle receptors.

[0058] Figures 15A-15B show the changes in the percentage of TCR+ cells in CAR-T cells co-cultured with (Figure 15A) or (Figure 15B) T84 tumor cells. WD104 refers to CAR-T cells in SR-CAR constructs without cytoplasmic tail region modification, while WD132 to WD146 refer to CAR-T cells in SR-CAR constructs with cytoplasmic tail region modification.

[0059] Figure 16 shows the TCR degradation efficiency in CAR-T cells expressing SR-CAR constructs or control CAR constructs. WD146, WD165 to WD177 refer to CAR-T cells whose SR-CAR constructs contain wild-type or variant transmembrane regions and have no cytoplasmic tail regions. The control group (WD116 CAR) refers to CAR-T cells that do not contain Golgi-associated shuttle receptors.

[0060] Figure 17 shows the TCR degradation efficiency in CAR-T cells expressing SR-CAR constructs with different transmembrane lengths or control CAR constructs with chimeric shuttle receptors. WD146(TM23) refers to CAR-T cells transduced with the WD146 SR-CAR construct (transmembrane length of 23 amino acids); WD146-2(TM22) refers to CAR-T cells with the transmembrane length of the WD146 SR-CAR construct shortened to 22 amino acids; WD146-3(TM21) refers to CAR-T cells with the transmembrane length of the WD146 SR-CAR construct shortened to 21 amino acids; and so on, with WD146-16(TM8) referring to CAR-T cells with the transmembrane length of the WD146 SR-CAR construct shortened to 8 amino acids.

[0061] Figure 18 shows the TCR degradation efficiency in CAR-T cells expressing the heterologous transmembrane region (TM) plus the Golgi-associated shuttle receptor cytoplasmic tail region (CT) or the control CAR construct, where WD178 (CD8a TM+SORT CT), WD179 (CD8a TM+SORT CT), WD180 (CD8a TM+SORCS1 CT), WD181 (CD8a TM+SORCS2 CT), WD182 (CD8a TM+SORCS3 CT), and WD183 (CD8a TM+SCARB2 CT) are listed. CT) refers to CAR-T cells whose chimeric shuttle receptor transmembrane region originates from a heterologous transmembrane region (the transmembrane region of a non-Golgi shuttle receptor molecule CD8α) and whose cytoplasmic tail region originates from Golgi-associated shuttle receptors SORT, SORL, SORCS2, SORCS3, and SCARB2, respectively; the control group WD184 (CD8aTM) refers to CAR-T cells that only have a heterologous transmembrane region (the transmembrane region of a non-Golgi shuttle receptor molecule CD8α) and no cytoplasmic tail region; the control group (WD116 CAR) refers to CAR-T cells that do not contain Golgi-associated shuttle receptors.

[0062] Figure 19 shows the TCR degradation efficiency in CAR-T cells of SR-CAR constructs or control CAR constructs expressing a heterologous transmembrane region (TM) plus a Golgi-associated shuttle receptor cytoplasmic tail region (CT). WD185 (PD1 TM+SORT CT) refers to CAR-T cells where the transmembrane region of the chimeric shuttle receptor comes from a heterologous transmembrane region (the transmembrane region of a non-Golgi shuttle receptor PD1) and the cytoplasmic tail region comes from the Golgi shuttle receptor SORT. The control group WD186 (PD1 TM) refers to CAR-T cells with only a heterologous transmembrane region (the transmembrane region of a non-Golgi shuttle receptor molecule PD1) and no cytoplasmic tail region. The control group (WD116 CAR) refers to CAR-T cells without a Golgi-associated shuttle receptor.

[0063] Figure 20 shows the changes in the percentage of TCR+ cells in CAR-T cells co-cultured with or without T84 tumor cells. WD104 (SORT™+SORT CT) refers to CAR-T cells in which both the transmembrane region and cytoplasmic tail region of the chimeric shuttle receptor are derived from the Golgi shuttle receptor molecule SORT. WD178 (CD8a™+SORT CT) refers to CAR-T cells in which the transmembrane region of the chimeric shuttle receptor is derived from the heterologous transmembrane region (the transmembrane region of the non-Golgi shuttle receptor molecule CD8α), and the cytoplasmic tail region is derived from the Golgi shuttle receptor molecule SORT.

[0064] Figure 21 shows the killing efficiency of CAR-T cells expressing chimeric shuttle receptors at various effector-to-target ratios. WD104 (SORT) refers to CAR-T cells transduced with an SR-CAR construct based on the Golgi-associated shuttle receptor SORT; the control group (WD115 ER) refers to CAR-T cells in which the Golgi-associated shuttle receptor was replaced with endoplasmic reticulum (ER) retention signals; and Mock-T refers to T cells that were not transduced with viral vectors.

[0065] Figure 22 shows the expansion of CAR-T cells expressing chimeric shuttle receptors under multiple rounds of target cell stimulation. WD104 (SORT) refers to CAR-T cells transduced based on the SR-CAR construct constructed from the Golgi-associated shuttle receptor SORT; the control group (WD115 ER) refers to CAR-T cells in which the Golgi-associated shuttle receptor was replaced with endoplasmic reticulum (ER) retention signals; and the control group (WD116 CAR) refers to CAR-T cells without the Golgi-associated shuttle receptor.

[0066] Figures 23A-23B show the in vitro inhibitory effect of CAR-T cells expressing chimeric shuttle receptors on graft-versus-host disease (GVHD) in mixed cell experiments. Figure 23A shows the change in the proportion of recipient cells, and Figure 23B shows the change in the number of recipient cells. WD104 refers to CAR-T cells transduced with the SR-CAR construct; Mock-T refers to T cells that have not been transduced with the viral vector.

[0067] Figures 24A-24F illustrate the degradation function of chimeric shuttle receptors for different or dual targets. WD187, WD188, WD189, WD190, and WD191 refer to cells that target and degrade chimeric shuttle receptors targeting CD38, NKG2A, BCMA, CD7, and 4-1BB, respectively. WD192 and WD193 refer to cells that express chimeric shuttle receptors targeting and degrading CD25 and 4-1BB. The control group WD116 refers to cells that do not contain Golgi-associated shuttle receptors.

[0068] Figure 25 shows the TCR degradation efficiency in Jurkat cells expressing chimeric shuttle receptors. WD103(SORT), WD104(SORT), WD105(SORL), WD106(SORCS1), WD107(SORCS2), WD108(SORCS3), and WD110(SCARB2) represent CAR-Jurkat cells transduced with SR-CAR constructs based on Golgi-associated shuttle receptors SORT, SORL, SORCS1, SORCS2, SORCS3, and SCARB2, respectively. WD116(CAR-Jurkat) refers to CAR-Jurkat cells without Golgi-associated shuttle receptors.

[0069] Figures 26A-26B illustrate the CD7 degradation function of the chimeric shuttle receptor in different T cell types (Figure 26A: γδT, Figure 26B: αβT), where WD190 refers to cells expressing the chimeric shuttle receptor that targets CD7 degradation; and the control group WD116 refers to cells that do not contain the Golgi-associated shuttle receptor.

[0070] Figure 27 shows the HLA degradation efficiency of the chimeric shuttle receptor in iPSC cells, where WD194 refers to cells expressing the chimeric shuttle receptor that targets HLA degradation; and the control group WD116 refers to cells that do not contain the Golgi-associated shuttle receptor.

[0071] Figures 28A-28D show the in vivo inhibitory effect of CAR-T cells expressing chimeric shuttle receptors on graft-versus-host disease (GVHD). Figure 28A shows the GVHD scoring curve region, Figure 28B shows the trend of mouse body weight change, Figure 28C shows the immunohistochemical detection of target organs (liver, intestine, and skin tissue), and Figure 28D shows the mouse survival curve analysis. WD104 refers to CAR-T cells expressing chimeric shuttle receptors; Mock-T refers to T cells without viral vector transduction; and HBSS refers to the solution control group.

[0072] Figure 29 shows the proportion of TCR-CAR+ cells in CAR-T cells that co-express the chimeric shuttle receptor and IMPDH genes; where WDTD80 refers to CAR-T cells that co-express the chimeric shuttle receptor and IMPDH genes; WDTD80+MMF refers to CAR-T cell culture medium supplemented with MPA drug (MMF). Detailed Implementation

[0073] In one aspect, this application provides a chimeric shuttle receptor comprising: a polypeptide derived from a Golgi-associated shuttle receptor. In some embodiments, according to any of the above-described chimeric shuttle receptors, it comprises (a) a heterologous binding portion capable of specifically binding a target protein, and (b) a polypeptide derived from a Golgi-associated shuttle receptor. In some aspects, the polypeptide derived from a Golgi-associated shuttle receptor comprises a transmembrane domain or a portion thereof. In some aspects, the polypeptide derived from a Golgi-associated shuttle receptor comprises a cytoplasmic tail region or a portion thereof. In some aspects, the chimeric shuttle receptor further comprises a transmembrane domain or a portion thereof located between the heterologous binding portion and the cytoplasmic tail region or a portion thereof.

[0074] In some embodiments, the Golgi-associated shuttle receptor is a sorting protein (SORT), a Sortilin-related receptor (SORL), a centrally expressed Sortilin-related receptor 1 (SORCS1), a centrally expressed Sortilin-related receptor 2 (SORCS2), a centrally expressed Sortilin-related receptor 3 (SORCS3), or a lysosomal membrane protein 2 (SCARB2). In some embodiments, the Golgi-associated shuttle receptor is a VPS10P domain receptor.

[0075] In some aspects, the chimeric shuttle receptor optionally further comprises a hinge domain or a portion thereof. In some embodiments, the chimeric shuttle receptor further comprises a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof may be derived from or not derived from a Golgi-associated shuttle receptor. In some embodiments, the hinge domain or a portion thereof may be derived from a Golgi-associated shuttle receptor that is the same as or different from the transmembrane domain or a portion thereof and / or the cytoplasmic tail region or a portion thereof.

[0076] In some embodiments, according to any of the chimeric shuttle receptors described above, the hinge domain or a portion thereof is derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:66-88, or an amino acid sequence having at least 75% identity with it. In some embodiments, according to any of the chimeric shuttle receptors described above, the hinge domain or a portion thereof is derived from a Golgi-associated shuttle receptor that is identical to the transmembrane domain or a portion thereof and / or the cytoplasmic tail region or a portion thereof. In some embodiments, according to any of the chimeric shuttle receptors described above, the hinge domain or a portion thereof is derived from a different molecule than the transmembrane domain or a portion thereof and / or the cytoplasmic tail region or a portion thereof.

[0077] In some embodiments, according to any of the chimeric shuttle receptors described above, the hinge domain or a portion thereof is not derived from a Golgi-associated shuttle receptor, for example, the hinge domain or a portion thereof is derived from CD8α or PD1. In some embodiments, according to any of the chimeric shuttle receptors described above, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 100 or 239, or SEQ ID NO: 102, or an amino acid sequence having at least 75% sequence identity with it. In some embodiments, according to any of the chimeric shuttle receptors described above, the hinge domain or a portion thereof comprises an artificial linker. In some embodiments, according to any of the chimeric shuttle receptors described above, the hinge domain or a portion thereof comprises GG, GS, SG, SS, GSS, SSG, GSG, SGS, SGG, G4S, (G4S)2, (G4S)3, or (G4S)4.

[0078] In some aspects, the chimeric shuttle receptor also comprises a signal peptide. In some embodiments, the signal peptide may or may not be derived from a Golgi-associated shuttle receptor. In some embodiments, the signal peptide may be derived from a Golgi-associated shuttle receptor that is the same as or different from a portion thereof of the transmembrane domain and / or a portion thereof of the cytoplasmic tail region. In some embodiments, according to any of the chimeric shuttle receptors described above, the signal peptide is derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some embodiments, the signal peptide comprises the amino acid sequence shown in any of SEQ ID NO:89-94, or an amino acid sequence having at least 75% identity with it. In some embodiments, according to any of the chimeric shuttle receptors described above, the signal peptide is derived from the same Golgi-associated shuttle receptor as the hinge structure or a portion thereof, the transmembrane domain or a portion thereof, and / or the cytoplasmic tail region or a portion thereof. In some embodiments, according to any of the chimeric shuttle receptors described above, the signal peptide is derived from a molecule that is different from the hinge structure or a portion thereof, the transmembrane domain or a portion thereof, and / or the cytoplasmic tail region or a portion thereof. In some embodiments, the signal peptide is not derived from a Golgi-associated shuttle receptor; for example, the signal peptide is derived from mouse immunoglobulin κ light chain (IgGκ), low-density lipoprotein receptor-associated protein (LRP), or CD8α. In some embodiments, according to any of the chimeric shuttle receptors described above, the signal peptide comprises the amino acid sequence shown in SEQ ID NO: 97, 113, and 240, or an amino acid sequence having at least 75% sequence identity with it.

[0079] The need to control and regulate the expression of specific proteins (e.g., disease-associated proteins) in cells remains unmet. The chimeric shuttle receptors described herein offer a novel approach to specifically downregulate unwanted intracellular target proteins. Without being bound by theory, it is believed that upon binding to a specific target protein, a chimeric shuttle receptor mediates the delivery of the target protein into the endosomal system, leading to its translocation to and degradation within the lysosome. Therefore, chimeric shuttle receptor expression results in the downregulation or knockout of a specific target protein. Chimeric shuttle receptors have broad applications in situations requiring target protein downregulation, such as modifying cellular phenotypes induced by the target protein, treating diseases or conditions associated with the expression or overexpression of a specific target protein, or reducing or eliminating adverse effects caused by the target protein.

[0080] In some embodiments of this disclosure, cells containing chimeric shuttle receptors optionally, but not necessarily, also contain or express antigen recognition receptors. In some embodiments, the chimeric shuttle receptor in the cell can exert its biological activity in the absence of antigen recognition receptors. In some embodiments, the antigen recognition receptor has immune-effecting biological activity. In some embodiments, the antigen recognition receptor and the chimeric shuttle receptor produce an additive or synergistic stimulatory effect. In some embodiments, to facilitate the detection of chimeric shuttle receptors in transduced cells, the chimeric shuttle receptor may be transduced into the cells or co-expressed with additional detection markers. In some embodiments, detection markers include, but are not limited to, antibodies or ligands, resistance genes (e.g., antibiotic resistance genes such as puromycin resistance gene (Puro), G418 resistance gene (Neo), blast fungicide resistance gene (BSD)) or selection markers (e.g., affinity tags such as histidine tags), and fluorescent proteins. Detection markers that are co-expressed with the protein being detected without affecting its expression and biological activity are well known in the art. In some embodiments, antigen recognition receptors are introduced for the detection of chimeric shuttle receptor expression.

[0081] For example, in some embodiments, cells containing chimeric shuttle receptors also express antigen recognition receptors (e.g., chimeric antigen receptors, CARs), wherein the chimeric shuttle receptors improve the cytotoxicity, proliferation, and / or persistence of cells expressing antigen recognition receptors (e.g., CARs). In some embodiments, overexpression of a specific target protein (e.g., an inhibitory immune receptor) leads to undesirable effects, such as reduced cell viability, proliferation, killing function, and / or increased undesirable host immune responses. Without being bound by theory, expression of chimeric shuttle receptors targeting target proteins associated with undesirable side effects can downregulate the expression of the target protein, thereby reducing the likelihood of host rejection and / or increasing the cytotoxicity, proliferation, and / or persistence of cells expressing antigen recognition receptors (e.g., CARs).

[0082] In some embodiments, the antigen recognition receptor (e.g., a chimeric antigen receptor, CAR) described herein comprises: (a) an extracellular antigen-binding domain comprising a single-domain antibody (sdAb) domain binding to GUCY2C, wherein the sdAb domain comprises the amino acid sequence of any one of SEQ ID NO:215-235; (b) a transmembrane domain; and (c) an intracellular signaling domain. In some embodiments, the transmembrane domain is derived from molecules selected from CD8α, CD4, CD28, CD137, CD80, CD86, CD152, and PD1. In some embodiments, the transmembrane domain is derived from CD8α, and optionally comprises the amino acid sequence of SEQ ID NO:99. In some embodiments, the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell. In some embodiments, the primary intracellular signaling domain is derived from CD3ζ, and optionally, the primary intracellular signaling domain comprises the amino acid sequence of SEQ ID NO:236. In some embodiments, the intracellular signal transduction domain includes a co-stimulatory signal transduction domain. In some embodiments, the co-stimulatory signal transduction domain is derived from a co-stimulatory molecule selected from ligands of CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, CD83, and combinations thereof. In some embodiments, the co-stimulatory signal transduction domain includes a cytoplasmic domain of CD28, and optionally, the co-stimulatory signal transduction domain includes the amino acid sequence of SEQ ID NO:237. In some embodiments, the co-stimulatory signal transduction domain includes a cytoplasmic domain of CD137 (4-1BB), and optionally, the co-stimulatory signal transduction domain includes the amino acid sequence of SEQ ID NO:238. In some embodiments, the CAR of this disclosure further includes a hinge domain located between the C-terminus of the extracellular antigen-binding domain and the N-terminus of the transmembrane domain. In some embodiments, the hinge domain is derived from CD8α, and optionally, the hinge domain comprises the amino acid sequence of SEQ ID NO:239. In some embodiments, the CAR also comprises a signal peptide located at the N-terminus of the polypeptide. In some embodiments, the signal peptide is derived from CD8α, and optionally, the signal peptide comprises the amino acid sequence of SEQ ID NO:240.In some embodiments, the CAR provided herein comprises or consists of an amino acid sequence shown in any of SEQ ID NO:95, 241-261, or an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the sequence of any of SEQ ID NO:95, 241-261.

[0083] In some embodiments, the target protein can be any protein that is desired to be downregulated. In some embodiments, the target protein is one or more components of the major histocompatibility complex (MHC) or the CD3 / T cell receptor (TCR) complex. In some embodiments, the chimeric shuttle receptor targets MHC class I molecules (HLA-A, HLA-B, HLA-C, HLA-G, or HLA-E), MHC class II molecules (HLA-DP, HLA-DQ, HLA-DR), or MHC class III molecules. In some implementations, the chimeric shuttle receptor targets the T-cell receptor (TCR) or its subunits, the CD3 / TCR complex or its subunits (CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, and CD3ζ), programmed cell death protein 1 (PD-1), programmed cell death ligand 1 (PD-L1), CTLA-4, CTLA-1, TIM3, LAG3, TIGIT, epidermal growth factor receptor (EGFR), glucocorticoid receptor (GR), CD38, CD7, CD25, CD69, CD70, CD56, CD22, FcγRII-B, 4-1BB, 4-BBL, and β2-microglobulin (B 2M), CIITA, NKG2A, NKG2D, CD94, TGF-β receptor, cytotoxic immunoglobulin-like receptor (KIR)2DL1, KIR2DL2, KIR2DL3, HLA, BCMA, CFH, ARMS2, MYOC, OPTN, VEGF, HIF1A, RB1, MUC16, TGFBR1, TGFBR2, OPN1SW, OPN1MW, MTTP, FOXC2, MSX1, KERA, SLC4A11, PAX6, OTX2, RHO, P23H, USH2A, MYO7A, LXS, CRYAA, CRYAB, ABCA4, CEP290 or MHCI. In some implementations, the heterogeneous binding portion is bound to NR3C1, Perforin1, GUCY2C, CLEC12A, PRLR, ERBB2, IMPDH, ITK, TMEM30A, Regnase-1, IFNGR, CD58, HSP90B1, or PIM3.

[0084] This application also provides a polypeptide or combination of polypeptides comprising the chimeric shuttle receptor described herein. This application further provides a nucleic acid encoding the chimeric shuttle receptor (or a polypeptide portion thereof) described herein, or a vector comprising such nucleic acid.

[0085] This application also provides compositions (e.g., pharmaceutical compositions or formulations) comprising the chimeric shuttle receptor described herein, or effector cells (e.g., T cells expressing the chimeric shuttle receptor) expressed herein or associated with the chimeric shuttle receptor described herein.

[0086] This application also provides methods for preparing and using chimeric shuttle receptors (or effector cells expressing chimeric shuttle receptors or associated with chimeric shuttle receptors), for example, for regulating (e.g., downregulating) intracellular target proteins and / or for treating diseases, and for inclusion in kits and articles for treating diseases or conditions.

[0087] I. Definition

[0088] Unless otherwise defined, the terminology used herein is the same as that commonly used in the art.

[0089] The singular forms “a,” “an,” and “the” used in this specification and the appended claims include the plural forms unless otherwise expressly specified. Thus, for example, reference to “a molecule” optionally includes combinations of two or more such molecules, etc.

[0090] As used herein, “treatment” is a method of achieving a beneficial or desired outcome (including clinical outcomes). For the purposes of this application, a beneficial or desired clinical outcome includes, but is not limited to, one or more of the following: reducing one or more symptoms caused by a disease, reducing the severity of the disease, stabilizing the disease (e.g., preventing or delaying the worsening of the disease), preventing or delaying the spread of the disease, preventing or delaying the onset or recurrence of the disease, delaying or slowing the progression of the disease, improving the disease state, providing remission of the disease (whether partial or complete remission), reducing the dosage of one or more other medications required to treat the disease, delaying disease progression, improving quality of life, and / or prolonging survival. “Treatment” also includes reducing the pathological consequences of the disease. The methods of this application contemplate any one or more of these aspects of treatment.

[0091] The terms “individual,” “subject,” and “patient” are used interchangeably herein to describe mammals, including humans. In some embodiments, the individual is a person. In some embodiments, the individual has cancer. In some embodiments, the individual requires treatment.

[0092] As understood in the art, an "effective amount" means an amount of composition sufficient to produce the desired therapeutic outcome (e.g., reducing the severity or duration of cancer, stabilizing the severity of cancer, or eliminating one or more symptoms of cancer). For therapeutic applications, beneficial or desired outcomes include, for example, reducing one or more symptoms (biochemical, histological, and / or behavioral) caused by the disease, including complications and intermediate pathological phenotypes that occur during disease development, improving the patient's quality of life, reducing the dosage of other medications required to treat the disease, enhancing the effect of another medication, delaying disease progression, and / or prolonging patient survival. In some embodiments, an effective amount of the therapeutic agent may prolong survival (including overall survival and progression-free survival); result in an objective response rate (including complete or partial remission); alleviate one or more signs or symptoms of the disease or condition to some extent; and / or improve the subject's quality of life.

[0093] As used herein, the term "wild type" is a term understood by those skilled in the art as referring to the typical form of an organism, strain, gene, or trait found in nature, as distinct from mutated or variant forms.

[0094] The terms “non-naturally occurring” and “engineered” are used interchangeably and indicate human intervention. When referring to nucleic acid molecules or peptides, the term means that the nucleic acid molecule or peptide is at least substantially free of at least one other component that is naturally bound to it in nature and found in nature.

[0095] As used herein, “expression” refers to the process by which polynucleotides are transcribed from a DNA template (e.g., into mRNA or other RNA transcripts) and / or the transcribed mRNA is subsequently translated into peptides, polypeptides, or proteins. Transcripts and encoded polypeptides are collectively referred to as “gene products.” If the polynucleotides are derived from genomic DNA, expression may also include mRNA splicing in eukaryotic cells.

[0096] The term "antibody" is used in its broadest sense to encompass a wide range of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies, trispecific antibodies, etc.), humanized antibodies, chimeric antibodies, full-length antibodies and antigen-binding fragments, single-chain Fv, nanobodies, and Fc fusion proteins, as long as they exhibit the desired antigen-binding activity. Antibodies and / or antibody fragments can be derived from mouse antibodies, rabbit antibodies, chicken antibodies, human antibodies, fully humanized antibodies, camel antibody variable domains and humanized forms, shark antibody variable domains and humanized forms, and camel-derived antibody variable domains.

[0097] The term "chimeric," when used for example, cells, nucleic acids, proteins, or carriers, indicates that the cell, nucleic acid, protein, or carrier (or a derivative of the cell, nucleic acid, protein, or carrier) has been modified by introducing a heterologous nucleic acid or protein or by altering the native nucleic acid or protein, or that the cell is derived from such a modified cell. A "heterologous" nucleic acid is a non-natural nucleic acid relative to the native form of the cell, nucleic acid, or carrier. A "heterologous" protein is a non-natural protein, peptide, or polypeptide relative to the native form of the cell, protein, or carrier.

[0098] The term “virus” or “viral particle” is used in its usual sense in virology to refer to a virion that includes the viral genome (e.g., DNA, RNA, single-stranded, double-stranded), the viral capsid, and associated proteins, and in the case of enveloped viruses (e.g., herpesviruses, poxviruses), also includes the envelope (containing lipids and optional host cell membrane components) and / or viral proteins.

[0099] The “percentage of amino acid sequence identity (%)” for the peptide and antibody sequences specified herein is defined as the percentage of amino acid residues in the candidate sequence that are identical to amino acid residues in the peptide being compared, after sequence alignment with any conserved substitutions as part of sequence identity. Alignment used to determine the percentage of amino acid sequence identity can be performed in various ways within the scope of the art, such as using publicly available computer software, such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm required to achieve maximum alignment across the full length of the compared sequences. However, for the purposes of this document, the sequence comparison computer program MUSCLE (Edgar, RC, Nucleic Acids Research 32(5):1792-1797, 2004; Edgar, RC, BMC Bioinformatics 5(1):113, 2004), the entire contents of which are incorporated herein by reference for all purposes.

[0100] As used in this article, the term "epitope" refers to a specific atom or amino acid group on an antigen that an antibody or bivalent antibody binds to. If two antibodies or antibody portions exhibit competitive binding to an antigen, they can bind to the same epitope within the antigen.

[0101] The term “polypeptide” or “peptide” as used herein includes all kinds of naturally occurring and synthetic proteins, including protein fragments of all lengths, fusion proteins, and modified proteins, including but not limited to glycoproteins, as well as all other types of modified proteins (e.g., proteins produced by phosphorylation, acetylation, myristylation, palmitoylation, glycosylation, oxidation, formylation, amidation, polyglutamylation, ADP-ribosylation, PEGylation, biotinylation, etc.).

[0102] As used herein, the terms “specific binding,” “specific recognition,” and “specific targeting” refer to measurable and reproducible interactions, such as the binding between a target and an antibody (e.g., a bivalent antibody). In some embodiments, specific binding determines the presence of a target in the presence of a heterogeneous group of molecules, including biomolecules such as cell surface receptors. For example, an antibody that specifically recognizes a target (which may be an epitope) is an antibody (e.g., a bivalent antibody) that has greater affinity, cohesion, ease of binding, and / or longer duration of binding to that target compared to its binding to other molecules. In some embodiments, as measured by, for example, radioimmunoassay (RIA), the degree of binding of an antibody to unrelated molecules is less than about 10% of the antibody binding to the target. In some embodiments, the antibody that specifically binds to the target has ≤10 -5 M, ≤10 -6 M, ≤10 -7 M, ≤10 -8 M, ≤10 -9 M, ≤10 -10 M, ≤10 -11 M or ≤10 -12 The dissociation constant (KD) of M. In some embodiments, the antibody specifically binds to conserved protein epitopes in proteins from different species. In some embodiments, specific binding may include, but is not required to be, exclusive binding. The binding specificity of the antibody or antigen-binding domain can be experimentally determined by methods known in the art. These methods include, but are not limited to, Western blot, ELISA, RIA, ECL, IRMA, EIA, BIACORE™, and peptide scanning.

[0103] "Pharmaceutically acceptable carriers" refer to one or more components in a pharmaceutical preparation, other than the active ingredient, that are non-toxic to the subjects. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, cryoprotectants, tension agents, preservatives, and combinations thereof. Preferably, pharmaceutically acceptable carriers or excipients meet toxicological and manufacturing testing requirements and / or are approved by the relevant regulatory agency or listed in the Chinese Pharmacopoeia, United States Pharmacopoeia, European Pharmacopoeia, or other generally recognized pharmacopoeias for use in mammals, and more specifically, in humans.

[0104] It should be understood that the embodiments of the invention described herein include embodiments that are "composed of" and / or "substantially composed of".

[0105] The “approximate” values ​​or parameters mentioned in this article include (and describe) variations with respect to the value or parameter itself. For example, a description involving “approximately X” includes a description of “X”.

[0106] As used in this article, references to "not" values ​​or parameters generally mean and describe something "different from" that value or parameter. For example, "This method is not used to treat type X disease" means that this method is used to treat diseases other than type X.

[0107] The term “about XY” used in this article has the same meaning as “about X to about Y”.

[0108] Unless the context clearly specifies otherwise, the singular forms “a,” “an,” or “the” used herein and in the appended claims include the plural forms.

[0109] As used herein, the term "and / or" (e.g., "A and / or B") is intended to include both A and B; A or B; A (alone); and B (alone). Similarly, as used herein, the term "and / or" (e.g., the phrase "A, B and / or C") is intended to cover each of the following embodiments: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0110] The term "shuttling receptor" (SR) used in this article refers to a class of dynamically distributed proteins that recognize signals and transport molecules between the cell nucleus, cytoplasm, and / or organelles, thereby regulating biological processes. Specifically, the term "Golgi-associated shuttle receptor" refers to a class of membrane-integrated proteins located in the Golgi apparatus that specifically recognize and bind to target proteins during intracellular protein transport. These proteins transport the target proteins from the Golgi apparatus to specific destinations (e.g., lysosomes, endosomes) or reverse them, allowing the transported target proteins to be stored or degraded within the targeted organelles.

[0111] II. Chimeric shuttle receptors containing cytoplasmic tail regions derived from Golgi-associated shuttle receptors.

[0112] This application provides a chimeric shuttle receptor comprising a polypeptide derived from a Golgi-associated shuttle receptor. In some embodiments, the polypeptide derived from the Golgi-associated shuttle receptor comprises a cytoplasmic tail region or a portion thereof. In some embodiments, according to the chimeric shuttle receptor described above, it comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a polypeptide derived from the Golgi-associated shuttle receptor. In some embodiments, the polypeptide comprises a cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor further comprises a transmembrane domain or a portion thereof located between the heterologous binding portion and the cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, (b) a transmembrane domain or a portion thereof, and (c) a cytoplasmic tail region or a portion thereof derived from the Golgi-associated shuttle receptor. In some embodiments, according to the chimeric shuttle receptor described above, the transmembrane domain or a portion thereof may or may not be derived from the Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor further comprises a hinge domain or a portion thereof. In some embodiments, the chimeric shuttle receptor also includes a signal peptide. Exemplary components and characteristics of chimeric shuttle receptors are provided herein.

[0113] A. Cytoplasmic tail region

[0114] The chimeric shuttle receptors described herein include the cytoplasmic tail region or a portion thereof derived from the Golgi-associated shuttle receptor.

[0115] In some embodiments, the cytoplasmic tail region or a portion thereof comprises a truncated or elongated cytoplasmic tail region derived from a Golgi-associated shuttle receptor. In some embodiments, the cytoplasmic tail region or a portion thereof is truncated from the N-terminus. In some embodiments, the cytoplasmic tail region or a portion thereof is truncated from the C-terminus.

[0116] In some embodiments, the Golgi-related shuttle receptor is a sorting protein (SORT), a Sortilin-related receptor (SORL), a centrally expressed Sortilin-related receptor 1 (SORCS1), a centrally expressed Sortilin-related receptor 2 (SORCS2), a centrally expressed Sortilin-related receptor 3 (SORCS3), or a lysosomal membrane protein 2 (SCARB2). In some embodiments, the Golgi-related shuttle receptor is a VPS10P domain receptor. In some embodiments, the Golgi-related shuttle receptor includes Sortilin (SORT). In some embodiments, the Golgi-related shuttle receptor is a Sortilin-related receptor (SORL). In some embodiments, the Golgi-related shuttle receptor is a centrally expressed Sortilin-related receptor 1 (SORCS1). In some embodiments, the Golgi-associated shuttle receptor is Sortilin-associated receptor 2 (SORCS2) expressed in the central nervous system. In some embodiments, the Golgi-associated shuttle receptor is Sortilin-associated receptor 1 (SORCS3) expressed in the central nervous system. In some embodiments, the Golgi-associated shuttle receptor is lysosomal membrane protein 2 (SCARB2).

[0117] In some embodiments, the length of the cytoplasmic tail region or a portion thereof is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 5 2, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acids, or within any of the quantities defined above. In some embodiments, the length of the cytoplasmic tail region is between 1 and 50, 1 and 40, 1 and 30, 1 and 20, 1 and 10, 5 and 50, 5 and 40, 5 and 30, 5 and 20, 5 and 10, 10 and 50, 10 and 40, 10 and 30, 10 and 20, 15 and 50, 15 and 40, 15 and 30, 15 and 20, 20 and 50, 20 and 40, 20 and 30, 25 and 50, 25 and 40, 25 and 30, 30 and 50, 30 and 40, 35 and 50, 35 and 40, 40 and 50, 30 and 60, 40 and 70, 50 and 80, 35 and 90, and 45 and 100 amino acids.

[0118] In some embodiments, the cytoplasmic tail region or a portion thereof comprises a cytoplasmic tail region, a variant thereof, or a portion thereof from a Golgi-associated shuttle receptor. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a cytoplasmic tail region or a portion thereof from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 4 2, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 consecutive amino acids, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a portion of the cytoplasmic tail region from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, or 41. A continuous sequence of 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acids. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a variant of the cytoplasmic tail region from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of its cytoplasmic tail regions or a portion thereof.In some implementations, the cytoplasmic tail region or a portion thereof comprises a cytoplasmic tail region from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2.

[0119] In some embodiments, the cytoplasmic tail region or a portion thereof comprises the cytoplasmic tail region or a portion thereof of any of the Golgi-associated shuttle receptors shown in Table 1, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45. A continuous sequence of 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acids, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a portion of the cytoplasmic tail region of any of the Golgi-associated shuttle receptors shown in Table 1, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44. A continuous sequence of 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acids. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a variant of the cytoplasmic tail region of any of the Golgi-associated shuttle receptors shown in Table 1, for example, an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a cytoplasmic tail region from any of the Golgi-associated shuttle receptors shown in Table 1.

[0120] Table 1: Components of naturally derived shuttle receptors

[0121] In some embodiments, the cytoplasmic tail region or a portion thereof originates from a Golgi-associated shuttle receptor and contains a localization signal, such as one or more localization signals present in SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. The localization signal can guide a protein, such as a chimeric shuttle receptor, to a specific subcellular structure, such as an endosome or lysosome. In some embodiments, the localization signal transports a protein (e.g., a chimeric shuttle receptor) to an endosome or lysosome. In some embodiments, the localization signal recycles or directs a protein (e.g., a chimeric shuttle receptor) from an endosome or lysosome to another subcellular organelle (e.g., the Golgi apparatus). In some embodiments, the localization signal includes or is selected from Golgi retention sequences, endoplasmic reticulum sequences, or lysosomal localization sequences. In some embodiments, the localization signal includes a lysosomal localization sequence. Exemplary lysosomal localization sequences include adaptor protein (AP) binding motifs and Golgi-localized, γEAR-containing ARF-binding protein (GGA) motifs (see, for example, Robinson MS. Trends Cell Biol. 2004).

[0122] In some embodiments, the cytoplasmic tail region or a portion thereof contains an AP-binding motif. AP-binding motifs are present in subsets of VPS10P domain receptors, such as Sortilin, SORL, SORCS2, and SORCS3 (see, for example, Robinson MS. Trends Cell Biol. 2004; Dittie AS et al., EMBO, 1997; Borgne RL et al., J. Bio Chem, 1993; Bonifacino JS et al., Nat. Review Molec. Cell. Bio., 2006; Rogaeva E et al., Nat. Genet., 2007; Lu et al., Nature, 2004). In some embodiments, the cytoplasmic tail region contains at least one AP-binding motif. In some embodiments, the cytoplasmic tail region contains two or more AP-binding motifs. In some embodiments, the AP-binding motif contains a sequence recognized by any of the following APs: AP-1, AP-2, AP-3, AP-4, and AP-5. In some embodiments, the AP binding motif comprises a sequence recognized by AP-1 and / or AP-2. Exemplary AP binding motifs recognized by AP-1 and / or AP-2 include tyrosine-based motifs, casein kinase 2 (CK-2) sites, and dileucine-like sites.

[0123] In some embodiments, the cytoplasmic tail region or a portion thereof contains a tyrosine-based motif. Exemplary tyrosine-based motifs include... Where X is any amino acid. It is L, I, M, V, or F. In some embodiments, the cytoplasmic tail region or a portion thereof contains an amino acid sequence. Where X is any amino acid. It is L, I, M, V, or F. In some embodiments, the cytoplasmic tail region or a portion thereof contains the amino acid sequence YSVL (SEQ ID NO: 60). In some embodiments, the cytoplasmic tail region or a portion thereof contains the amino acid sequence YAQM (SEQ ID NO: 64). In some embodiments, the cytoplasmic tail region or a portion thereof contains the amino acid sequence YAQV (SEQ ID NO: 65).

[0124] In some embodiments, the AP binding motif includes a CK-2 site. In some embodiments, the cytoplasmic tail region or a portion thereof includes at least one CK-2 site. In some embodiments, the cytoplasmic tail region or a portion thereof includes two or more CK-2 sites. In some embodiments, the cytoplasmic tail region or a portion thereof includes the amino acid sequence DDSDED (SEQ ID NO:61) and / or the amino acid sequence DEDLL (SEQ ID NO:62). In some embodiments, the cytoplasmic tail region or a portion thereof includes the amino acid sequence DDLGEDDEDAPMI (SEQ ID NO:63).

[0125] In some embodiments, the AP-binding motif includes a bis-leucine-like site. In some embodiments, the cytoplasmic tail region or a portion thereof includes a bis-leucine-like site. In some embodiments, the cytoplasmic tail region or a portion thereof includes the amino acid sequence LL, DDLGEDDEDAPMI (SEQ ID NO:63), or DEDLL (SEQ ID NO:62). In some embodiments, the cytoplasmic tail region or a portion thereof includes the sequence shown in LL. In some embodiments, the cytoplasmic tail region or a portion thereof includes the sequence shown in DEDLL (SEQ ID NO:62). In some embodiments, the cytoplasmic tail region or a portion thereof includes the sequence shown in DDLGEDDEDAPMI (SEQ ID NO:63).

[0126] In some embodiments, the cytoplasmic tail region or a portion thereof contains a GGA motif (see, for example, Robinson MS. Trends Cell Biol. 2004; Dittie AS et al., EMBO, 1997; Borgne RL et al., J. Bio Chem, 1993; Bonifacino JS et al., Nat. Review Molec. Cell. Bio., 2006). On the other hand, the GGA motif is present in subsets of VPS10P domain receptors, such as Sortilin, SORLA, SORCS2, and SORCS3. In some embodiments, the cytoplasmic tail region or a portion thereof contains the amino acid sequence LL. In some embodiments, the cytoplasmic tail region or a portion thereof contains the amino acid sequence (D / E)XXLL, where X is any amino acid and (D / E) is D or E. In some embodiments, the cytoplasmic tail region or a portion thereof contains the amino acid sequence DEDLL (SEQ ID NO: 62).

[0127] In some embodiments, the cytoplasmic tail region or a portion thereof is derived from the VPS10P domain receptor. Exemplary VPS10P domain receptors include SORT, SORL, SORCS1, SORCS2, and SORCS3. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a cytoplasmic tail region, or a portion thereof, or a variant thereof, derived from SORT, SORL, SORCS1, SORCS2, or SORCS3.

[0128] In some embodiments, the cytoplasmic tail region or a portion thereof is derived from SORT. SORT refers to Sortilin, such as Sortilin 1. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORT cytoplasmic tail region, its continuous portions, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of the SORT cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of the SORT cytoplasmic tail region containing a localization signal. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORT cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof contains a localization signal. In some embodiments, the localization signal comprises an AP-binding motif. In some embodiments, the localization signal comprises a dileucine-like site. In some embodiments, the localization signal comprises the sequence shown in DEDLL (SEQ ID NO: 62). In some embodiments, the localization signal comprises a CK-2 site. In some embodiments, the localization signal comprises the sequence shown in DDSDED (SEQ ID NO: 61). In some embodiments, the localization signal comprises a tyrosine-based motif. In some embodiments, the localization signal comprises the sequence shown in YSVL (SEQ ID NO: 60). In some embodiments, the location signal includes a GGA motif. In some embodiments, the location signal includes the sequence shown in DEDLL (SEQ ID NO: 62). In some embodiments, the location signal comprises one or more (e.g., any 3, 4, 5, 6, 7, 8, or 9) of the motifs, sites, or sequences disclosed above. For example, in some embodiments, the location signal includes an AP-binding motif and a GGA motif. In some embodiments, the location signal includes the sequences shown in DDSDED (SEQ ID NO: 61) and DEDLL (SEQ ID NO: 62). In some embodiments, the location signal includes the sequences shown in YSVL (SEQ ID NO: 60) and DEDLL (SEQ ID NO: 62). In some embodiments, the location signal includes the sequences shown in DDSDED (SEQ ID NO: 61), YSVL (SEQ ID NO: 60), and DEDLL (SEQ ID NO: 62).

[0129] An exemplary SORT cytoplasmic tail region is shown in SEQ ID NO:37. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:37, a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:37, or a continuous portion thereof of at least 5 (e.g., at least 5, 10, 20, 30, 40, or 50) amino acids. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:37, or a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:37. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of at least five (e.g., at least 5, 10, 20, 30, 40, or 50) amino acids from SEQ ID NO:37. In some embodiments, the continuous portion of SEQ ID NO:37 includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof includes a positioning signal. In some embodiments, the positioning signal includes an AP-binding motif. In some embodiments, the positioning signal includes a dileucine-like site. In some embodiments, the localization signal includes the sequence shown in DEDLL (SEQ ID NO: 62). In some embodiments, the localization signal includes the CK-2 site. In some embodiments, the localization signal includes the sequence shown in DDSDED (SEQ ID NO: 61). In some embodiments, the localization signal includes a tyrosine-based motif. In some embodiments, the localization signal includes the sequence shown in YSVL (SEQ ID NO: 60). In some embodiments, the localization signal includes the GGA motif. In some embodiments, the localization signal includes the sequence shown in DEDLL (SEQ ID NO: 62). In some embodiments, the localization signal includes one or more (e.g., any 3, 4, 5, 6, 7, 8, or 9) of the motifs, sites, or sequences disclosed above. In some embodiments, the cytoplasmic tail region or a portion thereof includes the amino acid sequence shown in SEQ ID NO: 37. In some embodiments, the cytoplasmic tail region or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO: 37.In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORT cytoplasmic tail region, a portion thereof, or a variant thereof, which comprises, or is substantially composed of, the amino acid sequence shown in any of SEQ ID NO:37-51, for example, is composed of.

[0130] In some embodiments, the cytoplasmic tail region or a portion thereof is derived from SORL. SORL refers to a sorting protein-associated receptor with type A repeat sequences, also known as LR11, SORLA, and SORL1. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORL cytoplasmic tail region, its continuous portions, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of the SORL cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of the SORL cytoplasmic tail region containing a localization signal. In some embodiments, the cytoplasmic tail region or a portion thereof contains a localization signal. In some embodiments, the localization signal comprises an AP binding motif. In some embodiments, the localization signal comprises a dileucine-like site. In some embodiments, the localization signal comprises the sequence shown in DDLGEDDEDAPMI (SEQ ID NO: 63).

[0131] An exemplary SORL cytoplasmic tail region is shown in SEQ ID NO:52. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:52, a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:52, or a continuous portion thereof of at least 5 (e.g., at least 5, 10, 20, 30, 40, or 50) amino acids. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:52, or a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:52. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of at least five (e.g., at least 5, 10, 20, 30, 40, or 50) amino acids from SEQ ID NO:52. In some embodiments, the continuous portion of SEQ ID NO:52 includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof includes a positioning signal. In some embodiments, the positioning signal includes an AP-binding motif. In some embodiments, the positioning signal includes a dileucine-like site. In some embodiments, the localization signal comprises the sequence shown in DDLGEDDEDAPMI (SEQ ID NO: 63). In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 52. In some embodiments, the cytoplasmic tail region or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO: 52.

[0132] In some embodiments, the cytoplasmic tail region or a portion thereof is derived from SORCS1. SORCS1 refers to CNS-expressed Sortilin-associated receptor 1. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORCS1 cytoplasmic tail region, its continuous portions, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of the SORCS1 cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of the SORCS1 cytoplasmic tail region containing a localization signal. In some embodiments, the cytoplasmic tail region or a portion thereof contains a localization signal. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORCS1 cytoplasmic tail region.

[0133] An exemplary SORCS1 cytoplasmic tail region is shown in SEQ ID NO:53. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:53, a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:53, or a continuous portion thereof of at least 5 (e.g., at least 5, 10, 20, 30, or 40) amino acids. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:53, or a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:53. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of at least five (e.g., at least 5, 10, 20, 30, or 40) amino acids from SEQ ID NO:53. In some embodiments, the continuous portion of SEQ ID NO:53 includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:53. In some embodiments, the cytoplasmic tail region or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:53. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORL cytoplasmic tail region, a portion thereof, or a variant thereof, which comprises, or is substantially composed of, the amino acid sequence shown in any of SEQ ID NO:53-56.

[0134] In some embodiments, the cytoplasmic tail region or a portion thereof is derived from SORCS2. SORCS2 refers to CNS expression of Sortilin-associated receptor 2. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORCS2 cytoplasmic tail region, its continuous portions, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises continuous portions of the SORCS2 cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof comprises continuous portions of the SORCS2 cytoplasmic tail region containing a localization signal. In some embodiments, the cytoplasmic tail region or a portion thereof contains a localization signal. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORCS2 cytoplasmic tail region. In some embodiments, the localization signal comprises an AP-binding motif. In some embodiments, the localization signal comprises a tyrosine-based motif. In some embodiments, the localization signal comprises the sequence shown in YAQM (SEQ ID NO: 64).

[0135] An exemplary SORCS2 cytoplasmic tail region is shown in SEQ ID NO:57. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:57, a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:57, or a continuous portion of at least 5 (e.g., at least 5, 10, 20, 30, 40, 50, or 60) amino acids. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:57, or a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:57. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of at least five (e.g., at least 5, 10, 20, 30, 40, 50, or 60) amino acids from SEQ ID NO:57. In some embodiments, the continuous portion of SEQ ID NO:57 includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof includes a positioning signal. In some embodiments, the positioning signal includes an AP-binding motif. In some embodiments, the positioning signal includes a tyrosine-based motif. In some embodiments, the localization signal includes the sequence shown in YAQM (SEQ ID NO: 64). In some embodiments, the cytoplasmic tail region or a portion thereof contains the amino acid sequence shown in SEQ ID NO: 57. In some embodiments, the cytoplasmic tail region or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO: 57.

[0136] In some embodiments, the cytoplasmic tail region or a portion thereof is derived from SORCS3. In some aspects, SORCS3 refers to CNS expression of Sortilin-associated receptor 3. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORCS3 cytoplasmic tail region, its continuous portions, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises continuous portions of the SORCS3 cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof comprises continuous portions of the SORCS3 cytoplasmic tail region containing a localization signal. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SORCS3 cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof contains a localization signal. In some embodiments, the localization signal comprises an AP-binding motif. In some embodiments, the localization signal comprises a dileucine-like motif. In some embodiments, the localization signal comprises the sequence shown in LL. In some embodiments, the localization signal comprises a tyrosine-based motif. In some embodiments, the localization signal comprises the sequence shown in YAQV (SEQ ID NO: 65). In some embodiments, the localization signal comprises one or more (e.g., three) of the motifs, sites, or sequences disclosed above. For example, in some embodiments, the localization signal includes a bisleucine-like site and a tyrosine-based motif. In some embodiments, the localization signal includes the sequence shown in LL and the sequence shown in YAQV (SEQ ID NO: 65).

[0137] An exemplary SORCS3 cytoplasmic tail region is shown in SEQ ID NO:58. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:58, a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:58, or a continuous portion thereof of at least 5 (e.g., at least 5, 10, 20, 30, 40, 50, 60, or 70) amino acids. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:58, or a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:58. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of at least five (e.g., at least 5, 10, 20, 30, 40, 50, 60, or 70) amino acids of SEQ ID NO:58. In some embodiments, the continuous portion of SEQ ID NO:58 includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof includes a positioning signal. In some embodiments, the positioning signal includes an AP-binding motif. In some embodiments, the positioning signal includes a dileucine-like motif. In some embodiments, the positioning signal includes the sequence shown in LL. In some embodiments, the localization signal comprises a tyrosine-based motif. In some embodiments, the localization signal comprises the sequence shown in YAQV (SEQ ID NO: 65). In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 58. In some embodiments, the cytoplasmic tail region or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO: 58. In some embodiments, the localization signal comprises one or more (e.g., three) motifs, sites, or sequences disclosed above. For example, in some embodiments, the localization signal comprises a dileucine-like site and a tyrosine-based motif.

[0138] In some embodiments, the cytoplasmic tail region or a portion thereof is derived from SCARB2. SCARB2 refers to lysosomal membrane protein 2. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SCARB2 cytoplasmic tail region, its continuous portions, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of the SCARB2 cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the SCARB2 cytoplasmic tail region. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a positioning signal.

[0139] An exemplary SCARB2 cytoplasmic tail region is shown in SEQ ID NO:59. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:59, a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:59, or a continuous portion of at least five amino acids thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:59, or a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:59. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a continuous portion of at least five amino acids of SEQ ID NO:59. In some embodiments, the continuous portion of SEQ ID NO:59 includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof includes a positioning signal. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:59. In some embodiments, the cytoplasmic tail region or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:59.

[0140] In some embodiments, the cytoplasmic tail region or a portion thereof comprises any of the amino acid sequences shown in SEQ ID NO:37-59, a continuous portion of at least five amino acids thereof, or an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the cytoplasmic tail region or a portion thereof comprises any of the amino acid sequences shown in SEQ ID NO:37-59, or a continuous portion of at least five amino acids thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises any of the amino acid sequences shown in SEQ ID NO:37-59, or an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the cytoplasmic tail region or a portion thereof comprises any of the amino acid sequences shown in SEQ ID NO:37-59. In some embodiments, the cytoplasmic tail region or a portion thereof is substantially composed of (e.g., composed of) any of the amino acid sequences shown in SEQ ID NO:37-59.

[0141] The experimental results disclosed herein fully demonstrate that by adding the cytoplasmic tail region, a portion thereof, or a variant thereof derived from the Golgi-associated shuttle receptor, non-shuttle receptor molecules can be endowed with the ability to regulate target protein expression levels, achieving efficient degradation and maintaining good stability. Even with modifications to the cytoplasmic tail region of chimeric shuttle receptors through various structural modifications such as source substitution, length truncation, and sequence mutation, as long as the core region and / or localization signal sequence are retained, efficient target protein degradation function and long-term stability can be maintained. Non-homologous chimeric shuttle receptors containing the cytoplasmic tail region of the Golgi-associated shuttle receptor possess long-term functional stability comparable to homologous chimeric receptors.

[0142] On the one hand, this strongly demonstrates that when the cytoplasmic tail region of a chimeric shuttle receptor originates from the cytoplasmic tail region of a Golgi-associated shuttle receptor, the cytoplasmic tail region can serve as a core structural domain for protein degradation regulation. Its function is independent of the homologous transmembrane region, and even when chimeric with a non-homologous transmembrane region, it can still endow the chimeric receptor with highly efficient and stable degradation activity. This non-homologous chimerism strategy breaks the homology restriction between the transmembrane region and the cytoplasmic tail region, highlighting the structural independence and functional universality of the cytoplasmic tail region, and providing a more flexible design path and core functional module support for protein expression regulation technology. On the other hand, this indicates that the structural modification of the transmembrane region has broad flexibility. In the engineering optimization of shuttle receptors, the design of its transmembrane region is not limited by any form of structural modification, providing a solid theoretical and experimental basis for developing functionally stable shuttle receptors adapted to diverse application scenarios, and enabling unlimited innovative applications in cell therapy, protein regulation, and other fields.

[0143] B. Heterogeneous combination part

[0144] The chimeric shuttle receptor described herein includes a heterologous binding moiety. In some embodiments, the heterologous binding moiety specifically binds to target proteins in cells (e.g., the cells described in Section VI).

[0145] In some embodiments, the heterologous binding moiety includes a peptide, a ligand, an aptamer, a designed ankylosing protein repeat (DARPin), a man-made binding protein, or a D-Domain. In some embodiments, the heterologous binding moiety comprises a peptide. In some embodiments, the heterologous binding moiety comprises a ligand. In some embodiments, the heterologous binding moiety comprises an aptamer.

[0146] In some embodiments, the heterologous binding portion comprises an antibody or an antigen-binding fragment thereof. In some embodiments, the heterologous binding portion comprises a full-length antibody. In some embodiments, the heterologous binding portion comprises an antigen-binding fragment, such as those selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-chain variable fragments (scFv), single-domain antibodies (sdAb), and nanobodies. In some embodiments, the heterologous binding portion comprises scFv. In some embodiments, the heterologous binding portion is human, humanized, or semi-synthetic.

[0147] In some embodiments, the heterologous binding portion of the chimeric shuttle protein is a full-length antibody. In some embodiments, the heterologous binding portion of the chimeric shuttle receptor is an antigen-binding fragment of the target protein described herein (e.g., in Section IV), such antigen-binding fragments are selected, for example, from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAbs), and single-chain variable fragments (scFvs). In some embodiments, the heterologous binding portion of the chimeric shuttle receptor is scFv. In some embodiments, the heterologous binding portion of the chimeric shuttle receptor is sdAb. In some embodiments, the heterologous binding portion is human, humanized, or semi-synthetic.

[0148] In some embodiments, the chimeric shuttle receptor includes a heterologous binding portion that binds to human, mouse, rat, cynomolgus monkey, and / or rhesus monkey target proteins. In some embodiments, the chimeric shuttle receptor includes a heterologous binding portion that specifically binds to human target proteins. In some embodiments, the chimeric shuttle receptor includes a heterologous binding portion that specifically binds to target proteins present or expressed in the cell. In some embodiments, the cell is a eukaryotic cell. In some embodiments, the cell is a mammalian cell. In some embodiments, the cell is a human cell. In some embodiments, the cell is a T cell. In some embodiments, the cell expresses high levels of the target protein.

[0149] In some embodiments, the binding affinity of the heterologous binding portion (or the chimeric shuttle receptor containing the heterologous binding portion) to the target protein is at least about 10 times its binding affinity to non-target proteins (including, for example, at least about 10, 20, 30, 40, 50, 75, 100, 200, 300, 400, 500, 750, 1000 or more). In some embodiments, the Kd of the heterologous binding portion (or the chimeric shuttle receptor containing the heterologous binding portion) to the target protein is no more than about 1 / 10 times its Kd of binding to non-target proteins (e.g., no more than about 1 / 10, 1 / 20, 1 / 30, 1 / 40, 1 / 50, 1 / 75, 1 / 100, 1 / 200, 1 / 300, 1 / 400, 1 / 500, 1 / 750, 1 / 1000 or less).

[0150] In some embodiments, the Kd of the heterologous binding portion (or the chimeric shuttle receptor containing the heterologous binding portion) binding to the target protein is between about 0.1 μm and about 500 nM (e.g., any one of about 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any range between these values). In some embodiments, the Kd of the heterologous binding portion (or the chimeric shuttle receptor containing the heterologous binding portion) binding to the target protein is between about 1 pM and about 250 pM (e.g., any one of about 1, 10, 25, 50, 75, 100, 150, 200, or 250 pM, including any range between these values). In some implementations, the Kd of the heterologous binding portion (or the chimeric shuttle receptor containing the heterologous binding portion) binding to the target protein is between about 1 nM and about 500 nM (e.g., any one of about 1, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450 or 500 nM, including any range between these values).

[0151] In some embodiments, the heterologous binding moiety comprises a specific sequence or certain variants of such sequences. In some embodiments, amino acid substitutions in the variant sequence do not significantly reduce the ability of the heterologous binding moiety to bind to the target protein described herein (e.g., the target protein described in Section IV). For example, alterations can be made that substantially do not reduce the target protein binding affinity. Alterations that significantly increase the target protein binding affinity or affect some other property (e.g., specificity and / or cross-reactivity with related variants of the target protein) are also contemplated herein.

[0152] In some embodiments, the chimeric shuttle receptor includes a heterologous binding portion that binds to extracellular protein molecules, membrane protein molecules, or intracellular molecules. In some embodiments, the heterologous binding portion binds to extracellular protein molecules. In some embodiments, the heterologous binding portion binds to membrane protein molecules. In some embodiments, the heterologous binding portion binds to intracellular molecules. In some embodiments, the heterologous binding portion binds to a receptor or immune checkpoint molecule. In some embodiments, the heterologous binding portion binds to a receptor. In some embodiments, the receptor is an inhibitory immune receptor. In some embodiments, the heterologous binding portion binds to a ligand of an inhibitory immune receptor. In some embodiments, the heterologous binding portion binds to an immune checkpoint molecule. In some embodiments, the heterologous binding portion binds to protein molecules associated with ophthalmic diseases.

[0153] In some embodiments, the heterologous binding portion can bind to any protein that is desired to be downregulated, depending on any of the chimeric shuttle receptors described above. In some embodiments, the heterologous binding portion binds to one or more components of the major histocompatibility complex (MHC) or the CD3 / T cell receptor (TCR) complex. In some embodiments, the heterologous binding portion binds to MHC class I molecules (HLA-A, HLA-B, HLA-C, HLA-G, or HLA-E), MHC class II molecules (HLA-DP, HLA-DQ, HLA-DR), or MHC class III molecules. In some implementations, the heterologous binding portion binds to the T-cell receptor (TCR) or its subunits, the CD3 / TCR complex or its subunits (CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, and CD3ζ), programmed cell death protein 1 (PD-1), programmed cell death ligand 1 (PD-L1), CTLA-4, CTLA-1, TIM3, LAG3, TIGIT, epidermal growth factor receptor (EGFR), glucocorticoid receptor (GR), CD38, CD7, CD25, CD69, CD70, CD56, CD22, FcγRII-B, 4-1BB, 4-BBL, and β2-microglobulin (B2). M), CIITA, NKG2A, NKG2D, CD94, TGF-β receptor, cytotoxic immunoglobulin-like receptor (KIR)2DL1, KIR2DL2, KIR2DL3, HLA, BCMA, CFH, ARMS2, MYOC, OPTN, VEGF, HIF1A, RB1, MUC16, TGFBR1, TGFBR2, OPN1SW, OPN1MW, MTTP, FOXC2, MSX1, KERA, SLC4A11, PAX6, OTX2, RHO, P23H, USH2A, MYO7A, LXS, CRYAA, CRYAB, ABCA4, CEP290, or MHCI. In some implementations, the heterogeneous binding portion is bound to NR3C1, Perforin1, GUCY2C, CLEC12A, PRLR, ERBB2, IMPDH, ITK, TMEM30A, Regnase-1, IFNGR, CD58, HSP90B1, or PIM3.

[0154] In some embodiments, the heterologous binding portion binds to target proteins in the CD3 / T cell receptor (TCR) complex. Exemplary targets in the CD3 / TCR complex include CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, and CD3ζ. In some embodiments, the heterologous binding portion binds to targets selected from CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, and CD3ζ. In some embodiments, the heterologous binding portion binds to CD3ε. In some embodiments, the heterologous binding portion binds to target proteins in the major histocompatibility complex (MHC). In some embodiments, the heterologous binding portion binds to HLA-A. In some embodiments, the heterologous binding portion binds to B2M.

[0155] In some embodiments, the heterologous binding portion comprises the amino acid sequence shown in any of SEQ ID NO:103-111, or a variant having at least 75% (e.g., at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with any of the aforementioned sequences. In some embodiments, the heterologous binding portion comprises the amino acid sequence shown in any of SEQ ID NO:103-111. In some embodiments, the heterologous binding portion consists substantially of (e.g., composed of) the amino acid sequence shown in any of SEQ ID NO:103-111.

[0156] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to CD3ε. In some embodiments, the heterologous binding moiety is an anti-CD3ε antibody. In some embodiments, the heterologous binding moiety is a full-length anti-CD3ε antibody. In some embodiments, the binding moiety is an anti-CD3ε antigen-binding fragment, such as selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-CD3ε scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0157] In some embodiments, the heterologous binding moiety is an anti-CD3ε antibody or a fragment thereof. In some embodiments, the heterologous binding moiety comprising the anti-CD3ε antibody is human or mouse. In some embodiments, the heterologous binding moiety comprises the amino acid sequence shown in SEQ ID NO:103, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:103. In some embodiments, the heterologous binding moiety consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:103.

[0158] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to CD38. In some embodiments, the heterologous binding moiety is an anti-CD38 antibody. In some embodiments, the heterologous binding moiety is a full-length anti-CD38 antibody. In some embodiments, the binding moiety is an anti-CD38 antigen-binding fragment, such as selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-CD38 scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0159] In some embodiments, the heterologous binding moiety is an anti-CD38 antibody. In some embodiments, the heterologous binding moiety containing the anti-CD38 antibody is human or mouse. In some embodiments, the heterologous binding moiety comprises the amino acid sequence shown in SEQ ID NO:104, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:104. In some embodiments, the heterologous binding moiety consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:104.

[0160] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to NKG2A. In some embodiments, the heterologous binding moiety is an anti-NKG2A antibody. In some embodiments, the heterologous binding moiety is a full-length anti-NKG2A antibody. In some embodiments, the binding moiety is an anti-NKG2A antigen-binding fragment, such as selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-NKG2A scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0161] In some embodiments, the heterologous binding moiety is an anti-NKG2A antibody. In some embodiments, the heterologous binding moiety comprising an anti-NKG2A antibody is human or mouse. In some embodiments, the heterologous binding moiety comprises the amino acid sequence shown in SEQ ID NO:105, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:105. In some embodiments, the heterologous binding moiety consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:105.

[0162] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to BCMA. In some embodiments, the heterologous binding moiety is an anti-BCMA antibody. In some embodiments, the heterologous binding moiety is a full-length anti-BCMA antibody. In some embodiments, the binding moiety is an anti-BCMA antigen-binding fragment, such as selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-BCMA scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0163] In some embodiments, the heterologous binding moiety is an anti-BCMA antibody. In some embodiments, the heterologous binding moiety containing the anti-BCMA antibody is human or mouse. In some embodiments, the heterologous binding moiety comprises the amino acid sequence shown in SEQ ID NO:106, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:106. In some embodiments, the heterologous binding moiety consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:106.

[0164] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to CD7. In some embodiments, the heterologous binding moiety is an anti-CD7 antibody. In some embodiments, the heterologous binding moiety is a full-length anti-CD7 antibody. In some embodiments, the binding moiety is an anti-CD7 antigen-binding fragment, such as selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-CD7 scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0165] In some embodiments, the heterologous binding moiety is an anti-CD7 antibody. In some embodiments, the heterologous binding moiety containing the anti-CD7 antibody is human or mouse. In some embodiments, the heterologous binding moiety comprises the amino acid sequence shown in SEQ ID NO:107, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:107. In some embodiments, the heterologous binding moiety consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:107.

[0166] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to 41BB. In some embodiments, the heterologous binding moiety is an anti-41BB antibody. In some embodiments, the heterologous binding moiety is a full-length anti-41BB antibody. In some embodiments, the binding moiety is an anti-41BB antigen-binding fragment, such as selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-41BB scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0167] In some embodiments, the heterobinding moiety is an anti-41BB antibody. In some embodiments, the heterobinding moiety containing the anti-41BB antibody is human or mouse. In some embodiments, the heterobinding moiety comprises the amino acid sequence shown in SEQ ID NO:108, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:108. In some embodiments, the heterobinding moiety comprises substantially the amino acid sequence shown in SEQ ID NO:108 (e.g., composed of it).

[0168] In some embodiments, the heterobinding moiety (or a chimeric shuttle receptor containing the heterobinding moiety) specifically binds to 41BB and CD25. In some embodiments, the heterobinding moiety is a bivalent antibody against 41BB and anti-CD25. In some embodiments, the heterobinding moiety is a full-length bivalent antibody against 41BB and anti-CD25. In some embodiments, the binding moiety is an antigen-binding fragment against 41BB and anti-CD25, such as a fragment selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterobinding moiety is an anti-41BB and anti-CD25 scFv. In some embodiments, the heterobinding moiety is human, humanized, or semi-synthetic.

[0169] In some embodiments, the heterologous binding moiety is a bivalent antibody against 41BB and antiCD25. In some embodiments, the heterologous binding moiety comprising the bivalent antibody against 41BB and antiCD25 is human or mouse. In some embodiments, the heterologous binding moiety comprises the amino acid sequence shown in SEQ ID NO:109 or SEQ ID NO:110, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:109 or SEQ ID NO:110. In some embodiments, the heterologous binding moiety is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:109 or SEQ ID NO:110.

[0170] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to HLA-A (e.g., HLA-A2). In some embodiments, the heterologous binding moiety is an anti-HLA-A (e.g., HLA-A2) antibody. In some embodiments, the heterologous binding moiety is a full-length anti-HLA-A (e.g., HLA-A2) antibody. In some embodiments, the binding moiety is an anti-HLA-A (e.g., HLA-A2) antigen-binding fragment, such as those selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-HLA-A (e.g., HLA-A2) scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0171] In some embodiments, the heterologous binding moiety (or a chimeric shuttle receptor containing the heterologous binding moiety) specifically binds to B2M. In some embodiments, the heterologous binding moiety is an anti-B2M antibody. In some embodiments, the heterologous binding moiety is a full-length anti-B2M antibody. In some embodiments, the binding moiety is an anti-B2M antigen-binding fragment, such as selected from Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), single-domain antibodies (sdAb), and single-chain variable fragments (scFv). In some embodiments, the heterologous binding moiety is an anti-B2M scFv. In some embodiments, the heterologous binding moiety is human, humanized, or semi-synthetic.

[0172] In some embodiments, the heterologous binding portion is an anti-B2M antibody. In some embodiments, the heterologous binding portion containing the anti-B2M antibody is human or mouse. In some embodiments, the heterologous binding portion comprises the amino acid sequence shown in SEQ ID NO:110, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:110. In some embodiments, the heterologous binding portion consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:110.

[0173] C. Transmembrane domain

[0174] The chimeric shuttle receptor described herein includes a transmembrane domain or a portion thereof. In some embodiments, when the chimeric shuttle receptor includes a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor, the chimeric shuttle receptor includes a transmembrane domain or a portion thereof located between the heterologous binding portion and the cytoplasmic tail region or a portion thereof. In some embodiments, the transmembrane domain or a portion thereof may be derived from or not derived from a Golgi-associated shuttle receptor. In some embodiments, the transmembrane domain or a portion thereof may be derived from a Golgi-associated shuttle receptor that is the same as or different from the cytoplasmic tail region or a portion thereof.

[0175] In some embodiments, the transmembrane domain or a portion thereof comprises an amino acid sequence shown in any of SEQ ID NO:1-36, SEQ ID NO:99, SEQ ID NO:101, or SEQ ID NO:114, a continuous portion thereof, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the transmembrane domain or a portion thereof comprises an amino acid sequence shown in any of SEQ ID NO:1-36, SEQ ID NO:99, SEQ ID NO:101, or SEQ ID NO:114, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises an amino acid sequence shown in any of SEQ ID NO:1-36, SEQ ID NO:99, SEQ ID NO:101, or SEQ ID NO:114. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequences shown in any of SEQ ID NO:1-36, SEQ ID NO:99, SEQ ID NO:101 or SEQ ID NO:114.

[0176] In some embodiments, the transmembrane domain or a portion thereof comprises a truncated transmembrane domain. In some embodiments, the transmembrane domain or a portion thereof is truncated from the N-end. In some embodiments, the transmembrane domain or a portion thereof is truncated from the C-end.

[0177] In some embodiments, the transmembrane domain or a portion thereof is not derived from the Golgi-associated shuttle receptor. In some embodiments, the transmembrane domain or a portion thereof is derived from CD8α, 4-1BB, PD1, ERBB2, CD2, CD4, CD5, CD7, CD8b, CD25, CD28, or LRP.

[0178] In some embodiments, the transmembrane domain or a portion thereof is derived from CD8α. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:99, consecutive portions thereof, or an amino acid sequence having at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO:99. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:99 or consecutive portions thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:99. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:99.

[0179] In some embodiments, the transmembrane domain or a portion thereof is derived from PD1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:101, consecutive portions thereof, or an amino acid sequence having at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO:101. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:101 or consecutive portions thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:101. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:101.

[0180] In some embodiments, the transmembrane domain or a portion thereof is derived from LRP. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:114, consecutive portions thereof, or an amino acid sequence having at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO:114. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:114 or consecutive portions thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:114. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:114.

[0181] In some embodiments, the transmembrane domain or a portion thereof originates from a Golgi-associated shuttle receptor, such as any Golgi-associated shuttle receptor described herein. In some embodiments, the transmembrane domain or a portion thereof originates from the same Golgi-associated shuttle receptor as the cytoplasmic tail region or a portion thereof, such as any cytoplasmic tail region described in Section II.A. In some embodiments, the transmembrane domain or a portion thereof originates from a different Golgi-associated shuttle receptor than the cytoplasmic tail region.

[0182] In some embodiments, the Golgi-related shuttle receptor includes sorting protein (SORT), Sortilin-related receptor (SORL), Sortilin-related receptor central nervous system-expressed 1 (SORCS1), Sortilin-related receptor 2 (SORCS2), Sortilin-related receptor 3 (SORCS3), or lysosome membrane protein 2 (SCARB2). In some embodiments, the Golgi-related shuttle receptor includes a VPS10P domain receptor. In some embodiments, the Golgi-related shuttle receptor includes sorting protein (SORT). In some embodiments, the Golgi-related shuttle receptor is Sortilin-related receptor (SORL). In some embodiments, the Golgi-related shuttle receptor is Sortilin-related receptor 1 (SORCS1) expressed in the central nervous system. In some embodiments, the central nervous system expresses Sortilin-associated receptor 2 (SORCS2). In some embodiments, the Golgi-associated shuttle receptor is the central nervous system expressing Sortilin-associated receptor 3 (SORCS3). In some embodiments, the Golgi-associated shuttle receptor is lysosomal membrane protein 2 (SCARB2).

[0183] In some embodiments, the length of the transmembrane domain or a portion thereof is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids, or within any of the aforementioned quantities. In some embodiments, the length of the transmembrane domain or a portion thereof is between 1 and 35, 1 and 30, 1 and 25, 1 and 20, 1 and 15, 1 and 10, 5 and 35, 5 and 30, 5 and 25, 5 and 20, 5 and 10, 10 and 35, 10 and 30, 10 and 25, 10 and 20, 10 and 15, 20 and 35, 20 and 30, 20 and 25, 25 and 35, 25 and 30, 30 and 35 amino acids.

[0184] In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain or a portion thereof from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids, or a variant thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a portion of a transmembrane domain from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids. In some embodiments, the transmembrane domain or a portion thereof comprises a variant of a transmembrane domain from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of its transmembrane domains or a portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) transmembrane domains from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2.

[0185] In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain or a portion thereof of any of the Golgi-associated shuttle receptors shown in Table 1, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 amino acids, or a variant thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a portion of the transmembrane domain of any of the Golgi-associated shuttle receptors shown in Table 1, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 amino acids. In some embodiments, the transmembrane domain or a portion thereof comprises a variant of the transmembrane domain of any of the Golgi-associated shuttle receptors shown in Table 1, for example, an amino acid sequence having at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it. In some embodiments, the transmembrane domain or a portion thereof comprises the transmembrane domain of any of the Golgi-associated shuttle receptors shown in Table 1.

[0186] In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-36, its continuous portions, or an amino acid sequence having at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-36, or its continuous portions. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-36. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in any of SEQ ID NO:1-36.

[0187] In some embodiments, the transmembrane domain or a portion thereof is derived from the VPS10P domain receptor. Exemplary VPS10P domain receptors include SORT, SORL, SORCS1, SORCS2, and SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain, or a portion thereof, or a variant thereof, derived from SORT, SORL, SORCS1, SORCS2, or SORCS3.

[0188] In some embodiments, the transmembrane domain or a portion thereof is derived from a sorting protein (SORT). In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORT, a continuous portion thereof, or a variant thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORT. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORT. An exemplary SORT transmembrane domain is shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-26, or a variant thereof having at least 80% (e.g., at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-26.

[0189] In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1, or a variant having at least 80% (e.g., at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORT transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:1 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:2-4, or a variant having at least 80% (e.g., at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in any of SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:2). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:3). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:4).

[0190] In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of at least one amino acid (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23) of SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., of) the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of SEQ ID NO:1 with a length of 22 amino acids. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof comprises substantially composed of (e.g., of) the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of SEQ ID NO:1 with a length of 21 amino acids. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:6. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:6). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 20 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:7. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:7). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 19 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:8. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:8). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 18 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:9. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:9). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 17 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:10.In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO: 10). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 16 amino acids in length as shown in SEQ ID NO: 1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 11. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO: 11). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 15 amino acids in length as shown in SEQ ID NO: 11. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 12. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO: 12). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 14 amino acids in length as shown in SEQ ID NO: 11. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 13. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:13. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 13 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:14. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:14. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 12 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:15. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:15. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 11 amino acids in length as shown in SEQ ID NO:11. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:16. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:16. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 10 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:17.In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:17). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 9 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:18. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:18). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 8 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:19. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:19). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 7 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:20. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:20). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 6 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:21. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:21). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 5 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:22. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:22). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:23. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:23). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 3 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence (SVP) shown in SEQ ID NO:24. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:24.In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 2 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence (SV) shown in SEQ ID NO:25. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:25). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 1 amino acid in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence (S) shown in SEQ ID NO:26. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:26).

[0191] In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:2, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) with the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:3, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:4, a variant thereof, or a continuous portion thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:4.In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:5, a variant of, or a continuous portion thereof, having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:6, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:6. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:6. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:6.

[0192] In some implementations, the transmembrane domain or a portion thereof from SORT contains the sequence "LI".

[0193] In some embodiments, the transmembrane domain or a portion thereof is derived from SORL. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORL, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORL. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORL. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:27 or 28, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:27 or 28, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:27. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:27. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORL transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:27 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:28, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:28. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:28. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:28.

[0194] In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS1. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS1, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORCS1. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:29 or 30, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:29 or 30, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:29. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:29. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORCS1 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:29 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:30, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:30. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:30. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:30.

[0195] In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS2, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORCS2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:31 or 32, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:31 or 32, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:31. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:31. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORCS2 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:31 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:32, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:32. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:32. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:32.

[0196] In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS3, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:33 or 34, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:33 or 34, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:33. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:33. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORCS3 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:33 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:34, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:34. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:34. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:34.

[0197] In some embodiments, the transmembrane domain or a portion thereof is derived from SCARB2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SCARB2, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SCARB2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SCARB2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:35 or 36, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:35 or 36, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:35. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:35. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SCARB2 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:35 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:36, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:36. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:36. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:36.

[0198] D. Hinge domain

[0199] In some embodiments, the chimeric shuttle receptor described herein does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor further includes a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof may be derived from or not derived from a Golgi-associated shuttle receptor. In some embodiments, the hinge domain or a portion thereof may be derived from a Golgi-associated shuttle receptor that is the same as or different from the transmembrane domain or a portion thereof and / or the cytoplasmic tail region or a portion thereof.

[0200] In some embodiments, the hinge domain or a portion thereof comprises an amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102, a continuous portion of at least one amino acid, or an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the hinge domain or a portion thereof comprises an amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102, a continuous portion of at least one amino acid thereon. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102, or an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102. In some embodiments, the hinge domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in any of SEQ ID NO:100 or SEQ ID NO:102.

[0201] In some embodiments, the hinge domain or a portion thereof comprises an artificial connector. Exemplary artificial connectors include GS connectors, α-helical connectors, glycine-alanine polymer connectors, alanine-serine polymer connectors, and IgG4-Fc connectors. In some embodiments, the hinge domain or a portion thereof comprises a GS connector, α-helical connector, glycine-alanine polymer connector, alanine-serine polymer connector, or IgG4-Fc connector. In some embodiments, the hinge domain or a portion thereof is derived from a GS connector. In some embodiments, the hinge domain or a portion thereof comprises a GS connector. Exemplary GS connectors include GG, GS, SG, SS, GSS, SSG, GSG, SGS, SGG, G4S, (G4S)2, (G4S)3, or (G4S)4. In some embodiments, the hinge domain or a portion thereof comprises GG, GS, SG, SS, GSS, SSG, GSG, SGS, SGG, G4S, (G4S)2, (G4S)3, or (G4S)4. In some embodiments, the hinge domain or a portion thereof comprises GG. In some embodiments, the hinge domain or a portion thereof comprises GS. In some embodiments, the hinge domain or a portion thereof comprises SG. In some embodiments, the hinge domain or a portion thereof comprises SS. In some embodiments, the hinge domain or a portion thereof comprises GSS. In some embodiments, the hinge domain or a portion thereof comprises SSG. In some embodiments, the hinge domain or a portion thereof comprises GSG. In some embodiments, the hinge domain or a portion thereof comprises SGS. In some embodiments, the hinge domain or a portion thereof comprises SGG. In some embodiments, the hinge domain or a portion thereof comprises (G4S)n, where n = 1 to 14. In some embodiments, the hinge domain or a portion thereof comprises (G4S)2. In some embodiments, the hinge domain or a portion thereof comprises (G4S)3. In some embodiments, the hinge domain or a portion thereof comprises (G4S)4. In some embodiments, the hinge domain or a portion thereof comprises the sequence shown in SEQ ID NO:112.

[0202] In some embodiments, the hinge domain or a portion thereof is not derived from the Golgi-associated shuttle receptor. In some embodiments, the hinge domain or a portion thereof comprises a sequence derived from CD8α. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:100 or 239, consecutive portions thereof, or an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO:100 or 239. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:100 or consecutive portions thereof. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:100. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:100.

[0203] In some embodiments, the hinge domain or a portion thereof is not derived from the Golgi-associated shuttle receptor. In some embodiments, the hinge domain or a portion thereof comprises a sequence derived from PD1. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:102, consecutive portions thereof, or an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO:102. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:102 or consecutive portions thereof. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:102. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:102.

[0204] In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a Golgi-associated shuttle receptor, such as any Golgi-associated shuttle receptor described herein. In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a Golgi-associated shuttle receptor identical to or a portion of the cytoplasmic tail region, such as any cytoplasmic tail region or a portion thereof described in Section II.A. In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a Golgi-associated shuttle receptor identical to or a portion of the transmembrane domain, such as any transmembrane domain or a portion thereof described in Section II.C. In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a Golgi-associated shuttle receptor identical to both the cytoplasmic tail region or a portion thereof and the transmembrane domain or a portion thereof.

[0205] In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a Golgi-associated shuttle receptor, distinct from the cytoplasmic tail region or a portion thereof, such as any cytoplasmic tail region or a portion thereof described in Section II.A. In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a Golgi-associated shuttle receptor, distinct from the transmembrane domain or a portion thereof, such as any transmembrane domain or a portion thereof described in Section II.C.

[0206] In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a molecule different from the cytoplasmic tail region or a portion thereof. In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a molecule different from the transmembrane domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof contains a sequence derived from a molecule different from both the cytoplasmic tail region or a portion thereof and the transmembrane domain or a portion thereof.

[0207] In some embodiments, the hinge domain or a portion thereof is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids, or within any of the foregoing defined quantities. In some embodiments, the hinge domain or a portion thereof is an amino acid sequence shorter than 35, 30, 25, 20, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 amino acids. In some embodiments, the hinge domain or a portion thereof is an amino acid sequence shorter than 18 amino acids, shorter than 16 amino acids, shorter than 14 amino acids, shorter than 12 amino acids, shorter than 10 amino acids, shorter than 8 amino acids, shorter than 6 amino acids, or shorter than 4 amino acids. In some embodiments, the hinge domain or a portion thereof is an amino acid sequence longer than 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, or 35 amino acids. In some embodiments, the length of the hinge domain or a portion thereof is between 1 and 35, 1 and 30, 1 and 25, 1 and 20, 1 and 15, 1 and 10, 5 and 35, 5 and 30, 5 and 25, 5 and 20, 5 and 10, 10 and 35, 10 and 30, 10 and 25, 10 and 20, 10 and 15, 20 and 35, 20 and 30, 20 and 25, 25 and 35, 25 and 30, 30 and 35 amino acids.

[0208] In some embodiments, the hinge domain or a portion thereof is derived from a Golgi-associated shuttle receptor. In some embodiments, the Golgi-associated shuttle receptor includes Sortilin (SORT), Sortilin-related receptor (SORL), Sortilin-related receptor central nervous system-expressed 1 (SORCS1), Sortilin-related receptor 2 (SORCS2), Sortilin-related receptor 3 (SORCS3), or lysosome membrane protein 2 (SCARB2). In some embodiments, the Golgi-associated shuttle receptor is a VPS10P domain receptor. In some embodiments, the Golgi-associated shuttle receptor is Sortilin (SORT). In some embodiments, the Golgi-associated shuttle receptor is Sortilin-related receptor (SORL). In some embodiments, the Golgi-associated shuttle receptor is centrally expressed Sortilin-associated receptor 1 (SORCS1). In some embodiments, the Golgi-associated shuttle receptor is centrally expressed Sortilin-associated receptor 2 (SORCS2). In some embodiments, the Golgi-associated shuttle receptor is centrally expressed Sortilin-associated receptor 3 (SORCS3). In some embodiments, the Golgi-associated shuttle receptor is lysosomal membrane protein 2 (SCARB2).

[0209] In some embodiments, the hinge domain or a portion thereof comprises a hinge domain or a portion thereof derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids, or a variant thereof. In some embodiments, the hinge domain or a portion thereof comprises a portion of a hinge domain derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, a continuous portion having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids. In some embodiments, the hinge domain or a portion thereof comprises a variant of a hinge domain derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of its hinge domains or a portion thereof. In some embodiments, the hinge domain or a portion thereof comprises a hinge domain derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some implementations, the hinge domain or a portion thereof is substantially composed of (e.g., composed of) hinge domains from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2.

[0210] In some embodiments, the hinge domain or a portion thereof comprises the hinge domain or a portion thereof of any of the Golgi-associated shuttle receptors shown in Table 1, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 amino acids, or a variant thereof. In some embodiments, the hinge domain or a portion thereof comprises a portion of the hinge domain of any of the Golgi-associated shuttle receptors shown in Table 1, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids. In some embodiments, the hinge domain or a portion thereof comprises a variant of the hinge domain of any of the Golgi-associated shuttle receptors shown in Table 1, for example, an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it. In some embodiments, the hinge domain or a portion thereof comprises the hinge domain of any of the Golgi-associated shuttle receptors shown in Table 1.

[0211] In some embodiments, the hinge domain or a portion thereof comprises an amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102, a continuous portion of at least one amino acid, or an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the hinge domain or a portion thereof comprises an amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102, a continuous portion of at least one amino acid thereon. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102, or an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102. In some embodiments, the hinge domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequences shown in any of SEQ ID NO:66-88, SEQ ID NO:100 or 239, or SEQ ID NO:102.

[0212] In some embodiments, the hinge domain or a portion thereof is derived from the VPS10P domain receptor. Exemplary VPS10P domain receptors include SORT, SORL, SORCS1, SORCS2, and SORCS3. In some embodiments, the hinge domain or a portion thereof comprises a hinge domain, or a portion thereof, derived from SORT, SORL, SORCS1, SORCS2, or SORCS3.

[0213] In some embodiments, the hinge domain or a portion thereof is derived from the sorting protein (SORT). In some embodiments, the hinge domain or a portion thereof comprises a hinge domain derived from SORT, its continuous portions, or variations thereof. An exemplary sequence of the SORT hinge domain is shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:66, a variant thereof having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13) amino acids. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:66-78, or a variant thereof having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the hinge domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:66-78.

[0214] In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:66, or a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13) amino acids from SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 12 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:67. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:67. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 11 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:68. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:68. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 10 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:69. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:69. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 9 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:70. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:70. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 8 amino acids in length as shown in SEQ ID NO:66.In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:71. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:71). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 7 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:72. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:72). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 6 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:73. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:73). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 5 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:74. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:74. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:75. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:75. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 3 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence (KSN) shown in SEQ ID NO:76. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:76. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 2 amino acids in length as shown in SEQ ID NO:66. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence (SN) shown in SEQ ID NO:77. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:77. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of length one amino acid in SEQ ID NO:66.In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence (N) shown in SEQ ID NO:78. In some embodiments, the hinge domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:78.

[0215] In some embodiments, the hinge structure domain or a portion thereof is derived from SORL. In some embodiments, the hinge region or a portion thereof comprises a hinge region from SORL, a continuous portion thereof, or a variation thereof. An exemplary sequence of SORL hinge structure domains is shown in SEQ ID NO:79. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:79, a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:79, or a variant thereof, or at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18) consecutive amino acids. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:79 or SEQ ID NO:80, or a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:79 or SEQ ID NO:80. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:79 or SEQ ID NO:80.

[0216] In some embodiments, the hinge domain or a portion thereof comprises an amino acid sequence as shown in SEQ ID NO:79, a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:79. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18) amino acids as shown in SEQ ID NO:79. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:79. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:79 (e.g., constitutes thereof). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:79. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:80. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:80 (e.g., constitutes thereof).

[0217] In some embodiments, the hinge domain or a portion thereof is derived from SORCS1. In some embodiments, the hinge domain or a portion thereof comprises a hinge domain from SORCS1, its continuous portions, or variants thereof. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:81, a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:81, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) amino acids. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:81 or SEQ ID NO:82, or a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:81 or SEQ ID NO:82. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:81 or SEQ ID NO:82.

[0218] In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:81, a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:81. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) amino acids from SEQ ID NO:81. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:81. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:81 (e.g., constitutes thereof). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:81. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:82. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:82 (e.g., constitutes thereof).

[0219] In some embodiments, the hinge domain or a portion thereof is derived from SORCS2. In some embodiments, the hinge domain or a portion thereof comprises a hinge domain from SORCS2, its continuous portions, or variations thereof. An exemplary sequence of SORCS2 hinge domains is shown in SEQ ID NO:83. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:83, a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:83, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) amino acids. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:83 or SEQ ID NO:84, or a variant thereof having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the hinge domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:83 or SEQ ID NO:84.

[0220] In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:83, or a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:83. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) amino acids from SEQ ID NO:83. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:83. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:83 (e.g., constitutes thereof). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:83. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:84. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:84 (e.g., constitutes thereof).

[0221] In some embodiments, the hinge domain or a portion thereof is derived from SORCS3. In some embodiments, the hinge domain or a portion thereof comprises a hinge domain from SORCS3, its continuous portions, or variations thereof. An exemplary sequence of SORCS3 hinge domains is shown in SEQ ID NO:85. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:85, a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:85, or a continuous portion thereof containing at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, or 9) amino acids. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:85 or SEQ ID NO:86, or a variant thereof having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the hinge domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:85 or SEQ ID NO:86.

[0222] In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:85, or a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:85. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, or 9) amino acids from SEQ ID NO:85. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:85. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:85 (e.g., constitutes thereof). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:85. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:86. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:86 (e.g., constitutes thereof).

[0223] In some embodiments, the hinge domain or a portion thereof is derived from SCARB2. In some embodiments, the hinge domain or a portion thereof comprises a hinge domain from SCARB2, its continuous portions, or variations thereof. An exemplary sequence of SCARB2 hinge domains is shown in SEQ ID NO:87. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:87, a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:87, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13) amino acids. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:87 or SEQ ID NO:88, or a variant thereof having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the hinge domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:87 or SEQ ID NO:88.

[0224] In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:87 or a variant thereof, or a variant having at least 75% (e.g., at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:87. In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13) amino acids from SEQ ID NO:87. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:87. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:87 (e.g., constitutes thereof). In some embodiments, the hinge domain or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:87. In some embodiments, the hinge domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:88. In some embodiments, the hinge domain or a portion thereof consists substantially of the amino acid sequence shown in SEQ ID NO:88 (e.g., constitutes thereof).

[0225] E. signal peptide

[0226] In some embodiments, the chimeric shuttle receptor does not contain a signal peptide. In some embodiments, the chimeric shuttle receptor also contains a signal peptide, which can be subsequently cleaved upon expression. In some embodiments, the signal peptide may or may not be derived from a Golgi-associated shuttle receptor. In some embodiments, the signal peptide may be derived from a Golgi-associated shuttle receptor that is the same as or different from a transmembrane domain or a portion thereof and / or a cytoplasmic tail region or a portion thereof.

[0227] In some implementations, the signal peptide comprises a sequence derived from a Golgi-associated shuttle receptor, such as any Golgi-associated shuttle receptor described herein.

[0228] In some embodiments, the signal peptide comprises a sequence not derived from the Golgi-associated shuttle receptor. In some embodiments, the signal peptide comprises a sequence derived from CD8, mouse immunoglobulin κ light chain (IgGκ), low-density lipoprotein receptor-associated protein (LRP), or GM-CSF receptor.

[0229] In some embodiments, the signal peptide comprises a sequence derived from the same Golgi-associated shuttle receptor as the cytoplasmic tail region, such as any cytoplasmic tail region described in Section II.A. In some embodiments, the signal peptide comprises a sequence derived from the same Golgi-associated shuttle receptor as the transmembrane domain, such as any transmembrane domain described in Section II.C. In some embodiments, the signal peptide comprises a sequence derived from the same Golgi-associated shuttle receptor as both the cytoplasmic tail region and the transmembrane domain.

[0230] In some embodiments, the signal peptide comprises a sequence derived from a Golgi-associated shuttle receptor, distinct from the cytoplasmic tail region, such as any cytoplasmic tail region described in Section II.A. In some embodiments, the signal peptide comprises a sequence derived from a Golgi-associated shuttle receptor, distinct from the transmembrane domain, such as any transmembrane domain described in Section II.C.

[0231] In some embodiments, the signal peptide is derived from a Golgi-associated shuttle receptor. In some embodiments, the Golgi-associated shuttle receptor includes sortilin (SORT), sortilin-related receptor (SORL), central nervous system-expressed sortilin-related receptor 1 (SORCS1), central nervous system-expressed sortilin-related receptor 2 (SORCS2), central nervous system-expressed sortilin-related receptor 3 (SORCS3), or lysosomal membrane protein 2 (SCARB2). In some embodiments, the Golgi-associated shuttle receptor is a VPS10P domain receptor. In some embodiments, the Golgi-associated shuttle receptor includes sortilin (SORT). In some embodiments, the Golgi-associated shuttle receptor is a sortilin-related receptor (SORL). In some embodiments, the Golgi-associated shuttle receptor is centrally expressed Sortilin-associated receptor 1 (SORCS1). In some embodiments, the Golgi-associated shuttle receptor is centrally expressed Sortilin-associated receptor 2 (SORCS2). In some embodiments, the Golgi-associated shuttle receptor is centrally expressed Sortilin-associated receptor 3 (SORCS3). In some embodiments, the Golgi-associated shuttle receptor is lysosomal membrane protein 2 (SCARB2).

[0232] In some embodiments, the signal peptide comprises a signal peptide derived from a Golgi-associated shuttle receptor. In some embodiments, the signal peptide comprises a signal peptide or a portion thereof derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, a continuous portion of at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 amino acids, or a variant thereof. In some embodiments, the signal peptide comprises a portion of a signal peptide derived from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, a continuous portion having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 amino acids. In some embodiments, the signal peptide comprises a variant of the signal peptide from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of its signal peptides or portions thereof. In some embodiments, the signal peptide comprises a signal peptide from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some embodiments, the signal peptide is substantially composed of (e.g., composed of) the signal peptide from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2.

[0233] In some embodiments, the signal peptide comprises a signal peptide with an amino acid sequence shown in any of SEQ ID NO: 89-94, 97, 113, and 240, a continuous portion thereof, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the signal peptide comprises an amino acid sequence shown in any of SEQ ID NO: 89-94, 97, 113, and 240, or a continuous portion thereof. In some embodiments, the signal peptide comprises the amino acid sequence shown in any of SEQ ID NO:89-94, SEQ ID NO:97, or SEQ ID NO:113, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence shown in any of SEQ ID NO:89-94, 97, 113, and 240. In some embodiments, the signal peptide comprises the amino acid sequence shown in any of SEQ ID NO:89-94, 97, 113, and 240. In some embodiments, the signal peptide is substantially composed of (e.g., composed of) the amino acid sequence shown in any of SEQ ID NO:89-94, 97, 113, and 240.

[0234] In some embodiments, the signal peptide is derived from SORT. In some embodiments, the signal peptide comprises a signal peptide derived from SORT or a continuous portion thereof. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:89, a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:89, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) amino acids. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:89, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:89. In some embodiments, the signal peptide comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) amino acids from SEQ ID NO:89. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:89. In some embodiments, the signal peptide is essentially composed of the amino acid sequence shown in SEQ ID NO:89 (e.g., composed of it).

[0235] In some embodiments, the signal peptide is derived from SORL. In some embodiments, the signal peptide comprises a signal peptide derived from SORL or a continuous portion thereof. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:90, a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:90, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28) amino acids. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:90, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:90. In some embodiments, the signal peptide comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28) amino acids from SEQ ID NO:90. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:90. In some embodiments, the signal peptide is essentially composed of the amino acid sequence shown in SEQ ID NO:90 (e.g., composed of it).

[0236] In some embodiments, the signal peptide is derived from SORCS1. In some embodiments, the signal peptide comprises a signal peptide derived from SORCS1 or a continuous portion thereof. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:91, a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:91, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) amino acids. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:91, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:91. In some embodiments, the signal peptide comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) amino acids from SEQ ID NO:91. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:91. In some embodiments, the signal peptide is essentially composed of the amino acid sequence shown in SEQ ID NO:91 (e.g., composed of it).

[0237] In some embodiments, the signal peptide is derived from SORCS2. In some embodiments, the signal peptide comprises a signal peptide derived from SORCS2 or a continuous portion thereof. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:92, and... The amino acid sequence shown in NO:92 has a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity, or a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acids. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:92, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:92. In some embodiments, the signal peptide comprises a continuous portion of at least one amino acid (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50) from SEQ ID NO:92. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:92. In some embodiments, the signal peptide is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:92.

[0238] In some embodiments, the signal peptide is derived from SORCS3. In some embodiments, the signal peptide comprises a signal peptide derived from SORCS3 or a continuous portion thereof. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:93, a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:93, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) amino acids. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:93, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:93. In some embodiments, the signal peptide comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) amino acids from SEQ ID NO:93. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:93. In some embodiments, the signal peptide is essentially composed of the amino acid sequence shown in SEQ ID NO:93 (e.g., composed of it).

[0239] In some embodiments, the signal peptide is derived from SCARB2. In some embodiments, the signal peptide comprises a signal peptide derived from SCARB2 or a continuous portion thereof. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:94, a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:94, or a continuous portion thereof having at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26) amino acids. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:94, or a variant having at least 85% (e.g., at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:94. In some embodiments, the signal peptide comprises a continuous portion of at least one (e.g., at least 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26) amino acids from SEQ ID NO:94. In some embodiments, the signal peptide comprises the amino acid sequence shown in SEQ ID NO:94. In some embodiments, the signal peptide is essentially composed of the amino acid sequence shown in SEQ ID NO:94 (e.g., composed of it).

[0240] F. Exemplary Implementation

[0241] In some respects, this document provides exemplary chimeric shuttle receptors. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion and (b) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor, such as any cytoplasmic tail region or a portion thereof as described in Section II.A.

[0242] In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, such as any heterologous binding portion described in Section II.B; (b) a transmembrane domain or a portion thereof, such as any transmembrane domain or a portion thereof described in Section II.C; and (c) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor, such as any cytoplasmic tail region or a portion thereof described in Section II.A.

[0243] In some embodiments, the chimeric shuttle receptor further comprises a hinge domain or a portion thereof, such as any hinge domain or a portion thereof described in Section II.D. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, (b) a hinge domain or a portion thereof, (c) a transmembrane domain or a portion thereof, and (d) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor.

[0244] In some embodiments, the chimeric shuttle receptor further comprises a signal peptide, such as any signal peptide described in Section II.E. In some embodiments, the chimeric shuttle receptor comprises: (a) a signal peptide, (b) a heterologous binding portion capable of specifically binding to a target protein, (c) a transmembrane domain or a portion thereof, and (d) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises: (a) a signal peptide, (b) a heterologous binding portion capable of specifically binding to a target protein, (c) a hinge domain or a portion thereof, (d) a transmembrane domain or a portion thereof, and (e) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor.

[0245] In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, (a) a heterobinding portion, (b) a transmembrane domain or a portion thereof, and (c) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, (a) a signal peptide, (b) a heterobinding portion, (c) a transmembrane domain or a portion thereof, and (d) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, (a) a heterobinding portion, (b) a hinge domain or a portion thereof, (c) a transmembrane domain or a portion thereof, and (d) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, (a) a signal peptide, (b) a heterologous binding portion, (c) a hinge domain or a portion thereof, (d) a transmembrane domain or a portion thereof, and (e) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor.

[0246] Exemplary chimeric shuttle receptor constructs are shown in Tables E1, E3, E4, E7, E8, and E9. In some embodiments, the chimeric shuttle receptor comprises an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identical to the amino acid sequence shown in any of SEQ ID NO: 116-121, 125-142, 143-156, 158-160, 189-194, 196, 198-206, 207-214. In some embodiments, the chimeric shuttle receptor comprises the amino acid sequence shown in any one of SEQ ID NO: 116-121, 125-142, 143-156, 158-160, 189-194, 196, 198-206, 207-214. In some embodiments, the chimeric shuttle receptor is substantially composed of (e.g., composed of) the amino acid sequence shown in any one of SEQ ID NO: 116-121, 125-142, 143-156, 158-160, 189-194, 196, 198-206, 207-214.

[0247] In some aspects, this document provides a chimeric shuttle receptor comprising a cytoplasmic tail region or a portion thereof derived from SORT, such as a cytoplasmic tail region or a portion thereof comprising a cytoplasmic tail region or a portion thereof derived from SORT. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof derived from SORT (e.g., a cytoplasmic tail region or a portion thereof comprising a localization signal present in SORT). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof comprising the sequence shown in any of SEQ ID NO:37-51. In some embodiments, the heterologous binding portion comprises scFv. In some embodiments, the transmembrane domain or a portion thereof is derived from SORT. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or portion thereof located between the heterobinding portion and the transmembrane domain or portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or portion thereof located between the heterobinding portion and the transmembrane domain or portion thereof.

[0248] In some aspects, this document provides a chimeric shuttle receptor comprising a cytoplasmic tail region or a portion thereof derived from SORL, such as a cytoplasmic tail region or a portion thereof comprising a cytoplasmic tail region or a portion thereof derived from SORL. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof derived from SORL (e.g., a cytoplasmic tail region or a portion thereof containing a localization signal present in SORL). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof comprising the sequence shown in SEQ ID NO:52. In some embodiments, the heterologous binding portion comprises scFv. In some embodiments, the transmembrane domain or a portion thereof is derived from SORL. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof.

[0249] In some aspects, this document provides a chimeric shuttle receptor comprising a cytoplasmic tail region or a portion thereof derived from SORCS1, such as a cytoplasmic tail region or a portion thereof comprising a cytoplasmic tail region or a portion thereof derived from SORCS1. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof derived from SORCS1 (e.g., a cytoplasmic tail region or a portion thereof containing a localization signal present in SORCS1). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof comprising the sequence shown in any of SEQ ID NO:53-56. In some embodiments, the heterologous binding portion includes scFv. In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS1. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof.

[0250] In some aspects, this document provides a chimeric shuttle receptor comprising a cytoplasmic tail region or a portion thereof derived from SORCS2, such as a cytoplasmic tail region or a portion thereof comprising a cytoplasmic tail region or a portion thereof derived from SORCS2. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof derived from SORCS2 (e.g., a cytoplasmic tail region or a portion thereof containing a localization signal present in SORCS2). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof comprising the sequence shown in SEQ ID NO:57. In some embodiments, the heterologous binding portion comprises scFv. In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS2. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof.

[0251] In some aspects, this document provides a chimeric shuttle receptor comprising a cytoplasmic tail region or a portion thereof derived from SORCS3, such as a cytoplasmic tail region or a portion thereof comprising a cytoplasmic tail region or a portion thereof derived from SORCS3. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof derived from SORCS3 (e.g., a cytoplasmic tail region or a portion thereof containing a localization signal present in SORCS3). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof comprising the sequence shown in SEQ ID NO:58. In some embodiments, the heterologous binding portion comprises scFv. In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS3. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof.

[0252] In some aspects, this document provides a chimeric shuttle receptor comprising a cytoplasmic tail region or a portion thereof derived from SCARB2, such as a cytoplasmic tail region or a portion thereof comprising a cytoplasmic tail region or a portion thereof derived from SCARB2. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof derived from SCARB2 (e.g., a cytoplasmic tail region or a portion thereof containing a localization signal present in SCARB2). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section II.B herein), a transmembrane domain or a portion thereof (e.g., any transmembrane domain or a portion thereof described in Section II.C herein), and a cytoplasmic tail region or a portion thereof comprising the sequence shown in SEQ ID NO:59. In some embodiments, the heterologous binding portion comprises scFv. In some embodiments, the transmembrane domain or a portion thereof is derived from SCARB2. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterobinding portion and the transmembrane domain or a portion thereof.

[0253] III. Chimeric shuttle receptors containing transmembrane domains derived from Golgi-associated shuttle receptors

[0254] This application provides a chimeric shuttle receptor comprising a polypeptide derived from a Golgi-associated shuttle receptor. In some embodiments, the polypeptide derived from the Golgi-associated shuttle receptor comprises a transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor according to the above comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a polypeptide derived from the Golgi-associated shuttle receptor, wherein the polypeptide derived from the Golgi-associated shuttle receptor comprises a transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor does not comprise a cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor further comprises a cytoplasmic tail region or a portion thereof. In some embodiments, the cytoplasmic tail region or a portion thereof is derived from the Golgi-associated shuttle receptor. In some embodiments, the cytoplasmic tail region or a portion thereof is not derived from the Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a transmembrane domain or a portion thereof derived from the Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor further includes a cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor further includes a hinge domain or a portion thereof. In some embodiments, the chimeric shuttle receptor further includes a signal peptide. Exemplary components and characteristics of chimeric shuttle receptors are provided herein.

[0255] A. Transmembrane domain

[0256] In some implementations, the chimeric shuttle receptor includes a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor.

[0257] In some embodiments, the transmembrane domain or a portion thereof comprises a truncated or elongated transmembrane domain derived from a Golgi-associated shuttle receptor. In some embodiments, the transmembrane domain or a portion thereof is truncated from the N-terminus. In some embodiments, the transmembrane domain or a portion thereof is truncated from the C-terminus.

[0258] In some embodiments, the transmembrane domain or a portion thereof is derived from a Golgi-related shuttle receptor, such as any Golgi-related shuttle receptor described herein. In some embodiments, the Golgi-related shuttle receptor includes sorting protein (SORT), Sortilin-related receptor (SORL), Sortilin-related receptor central nervous system-expressed 1 (SORCS1), Sortilin-related receptor 2 (SORCS2), Sortilin-related receptor 3 (SORCS3), or lysosome membrane protein 2 (SCARB2). In some embodiments, the Golgi-related shuttle receptor is a VPS10P domain receptor.

[0259] In some embodiments, the length of the transmembrane domain or a portion thereof is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids, or within any of the aforementioned quantities. In some embodiments, the transmembrane domain or a portion thereof is between 1 to 35, 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, 5 to 35, 5 to 30, 5 to 25, 5 to 20, 5 to 10, 10 to 35, 10 to 30, 10 to 25, 10 to 20, 10 to 15, 20 to 35, 20 to 30, 20 to 25, 25 to 35, 25 to 30, 30 to 35 amino acids.

[0260] In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain or a portion thereof from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 amino acids in a continuous portion. In some embodiments, the transmembrane domain or a portion thereof comprises portions of a transmembrane domain from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, such as at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24. In some embodiments, the transmembrane domain or a portion thereof comprises a variant of a transmembrane domain from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, such as an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of its transmembrane domains or portions thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) transmembrane domains from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2.

[0261] In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain or a portion thereof of any of the Golgi-associated shuttle receptors shown in Table 1, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 amino acids, or a variant thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a portion of the transmembrane domain of any of the Golgi-associated shuttle receptors shown in Table 1, for example, a continuous portion of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 amino acids. In some embodiments, the transmembrane domain or a portion thereof comprises a variant of the transmembrane domain of any of the Golgi-associated shuttle receptors shown in Table 1, for example, an amino acid sequence having at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it. In some embodiments, the transmembrane domain or a portion thereof comprises the transmembrane domain of any of the Golgi-associated shuttle receptors shown in Table 1.

[0262] In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-36, its continuous portions, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of the aforementioned sequences. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-36, or its continuous portions. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-36. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in any of SEQ ID NO:1-36.

[0263] In some embodiments, the transmembrane domain or a portion thereof is derived from the VPS10P domain receptor. Exemplary VPS10P domain receptors include SORT, SORL, SORCS1, SORCS2, and SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain, or a portion thereof, or a variant thereof, derived from SORT, SORL, SORCS1, SORCS2, or SORCS3.

[0264] In some embodiments, the transmembrane domain or a portion thereof is derived from a sorting protein (SORT). In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORT, a continuous portion thereof, or a variant thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORT. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORT. An exemplary SORT transmembrane domain is shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-26, or a variant thereof having at least 80% (e.g., at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:1-26.

[0265] In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1, or a variant having at least 80% (e.g., at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORT transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:1 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO:2-4, or a variant having at least 80% (e.g., at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in any of SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:2). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:3). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:4).

[0266] In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of at least one amino acid (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23) in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:1). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 22 amino acids in length in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:5). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 21 amino acids in length in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:6. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:6). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 20 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:7. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:7). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 19 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:8. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:8). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 18 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:9. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:9). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 17 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:10.In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO: 10). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 16 amino acids in length as shown in SEQ ID NO: 1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 11. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO: 11). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 15 amino acids in length as shown in SEQ ID NO: 11. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 12. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO: 12). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 14 amino acids in length as shown in SEQ ID NO: 11. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 13. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:13. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 13 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:14. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:14. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 12 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:15. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:15. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 11 amino acids in length as shown in SEQ ID NO:11. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:16. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:16. In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 10 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:17.In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:17). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 9 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:18. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:18). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 8 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:19. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:19). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 7 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:20. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:20). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 6 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:21. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:21). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 5 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:22. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:22). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 4 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:23. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:23). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 3 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence (SVP) shown in SEQ ID NO:24. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., of) the amino acid sequence shown in SEQ ID NO:24.In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 2 amino acids in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence (SV) shown in SEQ ID NO:25. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:25). In some embodiments, the transmembrane region or a portion thereof comprises a continuous portion of 1 amino acid in length as shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence (S) shown in SEQ ID NO:26. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., the amino acid sequence shown in SEQ ID NO:26).

[0267] In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:2, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) with the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:3, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:3. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:4, a variant thereof, or a continuous portion thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:4.In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:4. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:5, a variant of, or a continuous portion thereof, having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:6, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:6. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:6. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:6.

[0268] In some implementations, the transmembrane domain or a portion thereof from SORT contains the sequence "LI".

[0269] In some embodiments, the transmembrane domain or a portion thereof is derived from SORL. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORL, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORL. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORL. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:27 or 28, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:27 or 28, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:27. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:27. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORL transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:27 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:28, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:28. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:28. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:28.

[0270] In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS1. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS1, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORCS1. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS1. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:29 or 30, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:29 or 30, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:29. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:29. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORCS1 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:29 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:30, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:30. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:30. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:30.

[0271] In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS2, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORCS2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:31 or 32, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:31 or 32, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:31. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:31. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORCS2 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:31 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:32, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:32. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:32. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:32.

[0272] In some embodiments, the transmembrane domain or a portion thereof is derived from SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS3, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SORCS3. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:33 or 34, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:33 or 34, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:33. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:33. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SORCS3 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:33 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:34, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:34. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:34. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:34.

[0273] In some embodiments, the transmembrane domain or a portion thereof is derived from SCARB2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SCARB2, its continuous portions, or variants thereof. In some embodiments, the transmembrane domain or a portion thereof comprises a continuous portion of a transmembrane domain from SCARB2. In some embodiments, the transmembrane domain or a portion thereof comprises a transmembrane domain from SCARB2. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:35 or 36, a variant having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:35 or 36, or a continuous portion thereof. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:35. In some embodiments, the transmembrane domain or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:35. In some embodiments, the transmembrane domain or a portion thereof contains a leucine (L) or isoleucine (I) mutant near the C-terminus of the SCARB2 transmembrane domain. In some embodiments, leucine (L) or isoleucine (I) is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof contains a length-maintaining variant in which leucine (L) or isoleucine (I) near the C-terminus of SEQ ID NO:35 is mutated to alanine (A). In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:36, a variant thereof having at least 80% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) of the amino acid sequence shown in SEQ ID NO:36. In some embodiments, the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:36. In some embodiments, the transmembrane domain or a portion thereof consists substantially of (e.g., constitutes) the amino acid sequence shown in SEQ ID NO:36.

[0274] The experimental results disclosed herein fully demonstrate that even when the cytoplasmic tail region of the shuttle receptor is mutated or completely deleted, chimeric shuttle receptors containing transmembrane domains derived from Golgi-associated shuttle receptor molecules, or parts thereof or variants thereof, still have the function of mediating target protein degradation, maintaining both high degradation efficiency and good degradation stability. The experimental results strongly demonstrate that this type of chimeric shuttle receptor possesses good environmental tolerance, further validating the reliability of chimeric shuttle receptors as a "protein expression regulation tool."

[0275] B. Cytoplasmic tail region

[0276] In some embodiments, the chimeric shuttle receptor does not include a cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor also includes a cytoplasmic tail region or a portion thereof. In some embodiments, the cytoplasmic tail region or a portion thereof is not derived from a Golgi-associated shuttle receptor. In some embodiments, the cytoplasmic tail region or a portion thereof is derived from a Golgi-associated shuttle receptor, such as any cytoplasmic tail region or a portion thereof described in Section II.A herein. In some embodiments, the cytoplasmic tail region or a portion thereof is derived from a Golgi-associated shuttle receptor that is the same as or different from a transmembrane domain or a portion thereof.

[0277] In some implementations, the chimeric shuttle receptor comprises the cytoplasmic tail region or a portion thereof as described in Section II.A of this document.

[0278] In some embodiments, the cytoplasmic tail region or a portion thereof originates from a Golgi-associated shuttle receptor. In some embodiments, the Golgi-associated shuttle receptor includes sorting protein (SORT), Sortilin-related receptor (SORL), Sortilin-related receptor central nervous system-expressed 1 (SORCS1), Sortilin-related receptor 2 (SORCS2), Sortilin-related receptor 3 (SORCS3), or lysosome membrane protein 2 (SCARB2). In some embodiments, the Golgi-associated shuttle receptor is a VPS10P domain receptor.

[0279] In some embodiments, the cytoplasmic tail region includes a cytoplasmic tail region or a portion thereof originating from SORT, SORL, SORCS1, SORCS2, SORCS3, SCARB2, or a portion thereof, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 4 2, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 consecutive amino acids, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a portion of the cytoplasmic tail region from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, having a length of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 4 2, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 consecutive amino acids, or variants thereof. In some embodiments, the cytoplasmic tail region or a portion thereof comprises a variant of the cytoplasmic tail region from SORT, SORL, SORCS1, SORCS2, SORCS3, or SCARB2, for example, an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of its cytoplasmic tail regions or a portion thereof.In some embodiments, the cytoplasmic tail region or a portion thereof comprises a cytoplasmic tail region from SORT, SORL, SORCS1, SORCS2, SORCS3, or ScarB2. In some embodiments, the cytoplasmic tail region or a portion thereof is substantially composed of (e.g., composed of) cytoplasmic tails from SORT, SORL, SORCS1, SORCS2, SORCS3, or ScarB2.

[0280] In some embodiments, the cytoplasmic tail region or a portion thereof is not derived from the Golgi-associated shuttle receptor; for example, the cytoplasmic tail region or a portion thereof is derived from low-density lipoprotein receptor-associated protein (LRP). In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:115, or an amino acid sequence having at least 75% (e.g., at least about 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity with it. In some embodiments, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:115. In some embodiments, the cytoplasmic tail region or a portion thereof is substantially composed of (e.g., composed of) the amino acid sequence shown in SEQ ID NO:115.

[0281] C. Heterogeneous combination part

[0282] In some embodiments, the chimeric shuttle receptor also includes a heterologous binding portion, such as any heterologous binding portion described in Section II.B herein.

[0283] D. Hinge domain

[0284] In some embodiments, the chimeric shuttle receptor described herein does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor further includes a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof may be derived from or not derived from a Golgi-associated shuttle receptor. In some embodiments, the hinge domain or a portion thereof may be derived from a Golgi-associated shuttle receptor that is the same as or different from the transmembrane domain or a portion thereof and / or the cytoplasmic tail region or a portion thereof.

[0285] In some implementations, a hinge domain or a portion thereof includes any hinge domain or a portion thereof described in Section II.D of this document.

[0286] E. signal peptide

[0287] In some embodiments, the chimeric shuttle receptor does not contain a signal peptide. In some embodiments, the chimeric shuttle receptor also contains a signal peptide, which can be subsequently cleaved upon expression. In some embodiments, the signal peptide may or may not be derived from a Golgi-associated shuttle receptor. In some embodiments, the signal peptide may be derived from a Golgi-associated shuttle receptor that is the same as or different from a transmembrane domain or a portion thereof and / or a cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor contains any signal peptide described in Section II.E herein.

[0288] F. Exemplary Implementation

[0289] In some respects, this document provides exemplary chimeric shuttle receptors. In some embodiments, a chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, such as any heterologous binding portion described in Section III.C; and (b) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor, such as any transmembrane domain or a portion thereof described in Section III.A.

[0290] In some embodiments, the chimeric shuttle receptor further comprises: (c) a cytoplasmic tail region or a portion thereof, such as any portion described in Section III.B, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, such as any heterologous binding portion described in Section III.C; (b) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor, such as any transmembrane domain or a portion thereof described in Section III.A; and (c) a cytoplasmic tail region or a portion thereof, such as any cytoplasmic tail region or a portion thereof described in Section III.B, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor.

[0291] In some embodiments, the chimeric shuttle receptor further comprises a hinge domain or a portion thereof, such as any hinge domain or a portion thereof described in Section III.D. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, (b) a hinge domain or a portion thereof, and (c) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor.

[0292] In some embodiments, the chimeric shuttle receptor further comprises a signal peptide, such as any signal peptide described in Section III.E. In some embodiments, the chimeric shuttle receptor comprises: (a) a signal peptide, (b) a heterologous binding portion capable of specifically binding to a target protein, and (c) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises: (a) a signal peptide, (b) a heterologous binding portion capable of specifically binding to a target protein, (c) a hinge domain or a portion thereof, and (d) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor.

[0293] In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, (a) a heterobinding moiety, and (b) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, (a) a signal peptide, (b) a heterobinding moiety, and (c) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, (a) a signal peptide, (b) a heterobinding moiety, (c) a hinge domain or a portion thereof, and (d) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor does not include a cytoplasmic tail region or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a cytoplasmic tail region or a portion thereof, such as any cytoplasmic tail region or a portion thereof described in Section III.B herein, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor.

[0294] Exemplary chimeric shuttle receptor constructs are shown in Tables E1, E3, E4, E5, and E9. In some embodiments, the chimeric shuttle receptor comprises the amino acid sequence shown in any one of SEQ ID NO: 116-121, 125-142, 143-160, 161-188, and 207-214, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it. In some embodiments, the chimeric shuttle receptor comprises the amino acid sequence shown in any one of SEQ ID NO: 116-121, 125-142, 143-160, 161-188, and 207-214. In some embodiments, the chimeric shuttle receptor is essentially composed of (e.g., composed of) the amino acid sequences shown in any of SEQ ID NO: 116-121, 125-142, 143-160, 161-188, 207-214.

[0295] In some aspects, this document provides a chimeric shuttle receptor comprising a transmembrane domain or a portion thereof derived from SORT, such as a transmembrane domain or a portion thereof comprising a transmembrane domain or a portion thereof derived from SORT. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof derived from SORT (e.g., any transmembrane domain or a portion thereof derived from SORT described in Section III.A herein). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof (comprising the sequence shown in any of SEQ ID NO: 1-26). In some embodiments, the chimeric shuttle receptor further comprises a cytoplasmic tail region or a portion thereof (e.g., a cytoplasmic tail region or a portion thereof described in Section III.B herein, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor). In some embodiments, the chimeric shuttle receptor does not include a cytoplasmic tail region or a portion thereof. In some embodiments, the heterologous binding portion includes scFv. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof.

[0296] In some aspects, this document provides a chimeric shuttle receptor comprising a transmembrane domain or a portion thereof derived from SORL, such as a transmembrane domain or a portion thereof comprising a transmembrane domain or a portion thereof derived from SORL. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof derived from SORL (e.g., any transmembrane domain or a portion thereof derived from SORL described in Section III.A herein). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof (comprising the sequence shown in SEQ ID NO: 27 or 28). In some embodiments, the chimeric shuttle receptor further comprises a cytoplasmic tail region or a portion thereof (e.g., a cytoplasmic tail region or a portion thereof described in Section III.B herein, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor). In some embodiments, the chimeric shuttle receptor does not include a cytoplasmic tail region or a portion thereof. In some embodiments, the heterologous binding portion includes scFv. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof.

[0297] In some aspects, this document provides a chimeric shuttle receptor comprising a transmembrane domain or a portion thereof derived from SORCS1, such as a transmembrane domain or a portion thereof comprising a transmembrane domain or a portion thereof derived from SORCS1. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof derived from SORCS1 (e.g., any transmembrane domain or a portion thereof derived from SORCS1 described in Section III.A herein). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof (comprising the sequence shown in SEQ ID NO: 29 or 30). In some embodiments, the chimeric shuttle receptor further comprises a cytoplasmic tail region or a portion thereof (e.g., a cytoplasmic tail region or a portion thereof described in Section III.B herein, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor). In some embodiments, the chimeric shuttle receptor does not include a cytoplasmic tail region or a portion thereof. In some embodiments, the heterologous binding portion includes scFv. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof.

[0298] In some aspects, this document provides a chimeric shuttle receptor comprising a transmembrane domain or a portion thereof derived from SORCS2, such as a transmembrane domain or a portion thereof comprising a transmembrane domain or a portion thereof derived from SORCS2. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof derived from SORCS2 (e.g., any transmembrane domain or a portion thereof derived from SORCS2 described in Section III.A herein). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof (comprising the sequence shown in SEQ ID NO: 31 or 32). In some embodiments, the chimeric shuttle receptor further comprises a cytoplasmic tail region or a portion thereof (e.g., a cytoplasmic tail region or a portion thereof described in Section III.B herein, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor). In some embodiments, the chimeric shuttle receptor does not include a cytoplasmic tail region or a portion thereof. In some embodiments, the heterologous binding portion includes scFv. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof.

[0299] In some aspects, this document provides a chimeric shuttle receptor comprising a transmembrane domain or a portion thereof derived from SORCS3, such as a transmembrane domain or a portion thereof comprising a transmembrane domain or a portion thereof derived from SORCS3. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof derived from SORCS3 (e.g., any transmembrane domain or a portion thereof derived from SORCS3 described in Section III.A herein). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof comprising SEQ ID NO. (Sequence shown in NO:33 or 34). In some embodiments, the chimeric shuttle receptor further comprises a cytoplasmic tail region or a portion thereof (e.g., the cytoplasmic tail region or a portion thereof described in Section III.B herein, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor). In some embodiments, the chimeric shuttle receptor does not comprise a cytoplasmic tail region or a portion thereof. In some embodiments, the heterologous binding portion comprises scFv. In some embodiments, the chimeric shuttle receptor does not comprise a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor comprises a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof.

[0300] In some aspects, this document provides a chimeric shuttle receptor comprising a transmembrane domain or a portion thereof derived from SCARB2, such as a transmembrane domain or a portion thereof comprising a transmembrane domain or a portion thereof derived from SCARB2. In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof derived from SCARB2 (e.g., any transmembrane domain or a portion thereof derived from SCARB2 described in Section III.A herein). In some embodiments, the chimeric shuttle receptor comprises, from N-terminus to C-terminus, a heterologous binding portion (e.g., any heterologous binding portion described in Section III.C herein) and a transmembrane domain or a portion thereof (comprising the sequence shown in SEQ ID NO: 35 or 36). In some embodiments, the chimeric shuttle receptor further comprises a cytoplasmic tail region or a portion thereof (e.g., a cytoplasmic tail region or a portion thereof described in Section III.B herein, or a cytoplasmic tail region or a portion thereof not derived from a Golgi-associated shuttle receptor). In some embodiments, the chimeric shuttle receptor does not include a cytoplasmic tail region or a portion thereof. In some embodiments, the heterologous binding portion includes scFv. In some embodiments, the chimeric shuttle receptor does not include a hinge domain or a portion thereof located between the heterologous binding portion and the transmembrane domain or a portion thereof. In some embodiments, the chimeric shuttle receptor includes a hinge domain or a portion thereof between the heterologous binding portion and the transmembrane domain.

[0301] IV. Target Protein

[0302] In some embodiments, the chimeric shuttle receptor includes a binding moiety capable of specifically binding to a target protein. In some embodiments, the heterologous binding moiety is capable of specifically binding to a target protein, such as any of the target proteins described below. In some embodiments, the chimeric shuttle receptor downregulates the target protein. In some embodiments, the chimeric shuttle receptor mediates the delivery of the target protein to an endosomal system, such as an endosome or lysosome. In some embodiments, the chimeric shuttle receptor causes the degradation of the target protein. In some embodiments, expression of the chimeric shuttle receptor results in the downregulation (e.g., knockdown or knockout) of a specific target protein. In some embodiments, the chimeric shuttle receptor reduces the level or expression of the target protein in cells compared to control cells without the chimeric shuttle receptor.

[0303] The chimeric shuttle receptors provided herein offer a variety of benefits and can be used to degrade specific target proteins described herein, depending on different needs. For example, in some embodiments, the target protein includes any disease-related protein, such as any protein whose overexpression leads to disease or symptom. In some embodiments, the target protein includes a target protein expressed in or on a cell. In some embodiments, the target protein is an extracellular protein, a membrane protein, or an intracellular protein.

[0304] In some implementations, the target protein is an inhibitory immune receptor, a ligand of an inhibitory immune receptor, or an immune checkpoint molecule.

[0305] In some implementations, the target protein is a suppressive immune receptor. In some implementations, the suppressive immune receptor is a T-cell receptor (TCR).

[0306] In some implementations, the target protein is a tumor antigen.

[0307] In some implementations, the target protein is an antigen recognition receptor (such as any antigen recognition receptor described in Section VI.A).

[0308] In some embodiments, the target protein can be any protein that is desired to be downregulated. In some embodiments, the target protein is one or more components of the major histocompatibility complex (MHC) or the CD3 / T cell receptor (TCR) complex. In some embodiments, the target protein comprises MHC class I molecules (HLA-A, HLA-B, HLA-C, HLA-G, or HLA-E), MHC class II molecules (HLA-DP, HLA-DQ, HLA-DR), or MHC class III molecules. In some embodiments, the target proteins include the T cell receptor (TCR) or its subunits, the CD3 / TCR complex or its subunits (CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, and CD3ζ), programmed cell death protein 1 (PD-1), programmed cell death ligand 1 (PD-L1), CTLA-4, CTLA-1, TIM3, LAG3, TIGIT, epidermal growth factor receptor (EGFR), glucocorticoid receptor (GR), CD38, CD7, CD25, CD69, CD70, CD56, CD22, FcγRII-B, 4-1BB, 4-BBL, and β2-microglobulin (B2M). ), CIITA, NKG2A, NKG2D, CD94, TGF-β receptor, killer immunoglobulin-like receptor (KIR)2DL1, KIR2DL2, KIR2DL3, HLA, BCMA, CFH, ARMS2, MYOC, OPTN, VEGF, HIF1A, RB1, MUC16, TGFBR1, TGFBR2, OPN1SW, OPN1MW, MTTP, FOXC2, MSX1, KERA, SLC4A11, PAX6, OTX2, RHO, P23H, USH2A, MYO7A, LXS, CRYAA, CRYAB, ABCA4, CEP290 or MHCI.In some embodiments, the target protein is selected from one of the following: MHC class I molecules (HLA-A, HLA-B, HLA-C, HLA-G, or HLA-E), MHC class II molecules (HLA-DP, HLA-DQ, HLA-DR), MHC class III molecules, T cell receptor (TCR) or its subunits, CD3 / TCR complex or its subunits (CD3ε, TCRα, TCRβ, TCRγ, TCRδ, CD3δ, CD3γ, and CD3ζ), programmed cell death protein 1 (PD-1), programmed cell death ligand 1 (PD-L1), CTLA-4, CTLA-1, TIM3, LAG3, TIGIT, epidermal growth factor receptor (EGFR), glucocorticoid receptor (GR), CD38, CD7, CD25, CD69, C D70, CD56, CD22, FcγRII-B, 4-1BB, 4-BBL, β2-microglobulin (B2M), CIITA, NKG2A, NKG2D, CD94, TGF-β receptor, cytotoxic immunoglobulin-like receptor (KIR)2DL1, KIR2DL2, KIR2DL3, HLA, BCMA, CFH, ARMS2, MYOC, OPTN, VEGF, HIF1A, RB1, MUC16, TGFBR1, TGFBR2, OPN1SW, OPN1MW, MTTP, FOXC2, MSX1, KERA, SLC4A11, PAX6, OTX2, RHO, P23H, USH2A, MYO7A, LXS, CRYAA, CRYAB, ABCA4, CEP290, and MHCI. In some implementations, the heterogeneous binding portion is bound to NR3C1, Perforin1, GUCY2C, CLEC12A, PRLR, ERBB2, IMPDH, ITK, TMEM30A, Regnase-1, IFNGR, CD58, HSP90B1, or PIM3.

[0309] In some embodiments, the target protein includes CD3ε. In some embodiments, the target protein includes CD38. In some embodiments, the target protein includes NKG2A. In some embodiments, the target protein includes BCMA. In some embodiments, the target protein includes CD7. In some embodiments, the target protein includes CD25. In some embodiments, the target protein includes HLA-A (e.g., HLA-A2). In some embodiments, the target protein includes B2M.

[0310] V. Nucleic Acids and Vectors

[0311] This document also provides nucleic acids (including a nucleic acid genome) encoding the chimeric shuttle receptor described herein (e.g., the chimeric shuttle receptor described in Section II or Section III). In some embodiments, the nucleic acid (or nucleic acid genome) encodes the chimeric shuttle receptor. In some embodiments, the nucleic acid encodes the chimeric shuttle receptor or a polypeptide portion thereof.

[0312] In some embodiments, the nucleic acid encodes a chimeric shuttle receptor, such as any chimeric shuttle receptor described in Section II or Section III. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, (b) a transmembrane domain or a portion thereof, and (c) a cytoplasmic tail region or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the chimeric shuttle receptor comprises: (a) a heterologous binding portion capable of specifically binding to a target protein, and (b) a transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor. In some embodiments, the Golgi-associated shuttle receptor is a sorting protein (SORT), a Sortilin-related receptor (SORL), a VPS10P domain-containing receptor 1 (SORCS1), a VPS10P domain-containing receptor 2 (SORCS2), a VPS10P domain-containing receptor 3 (SORCS3), or a lysosomal membrane protein 2 (SCARB2).

[0313] In some embodiments, the chimeric shuttle receptor further includes a hinge domain or a portion thereof. In some embodiments, the hinge domain or a portion thereof may or may not be derived from a Golgi-associated shuttle receptor. In some embodiments, the hinge domain or a portion thereof may be derived from a Golgi-associated shuttle receptor that is the same as or different from one from which the transmembrane domain or a portion thereof originates and / or one from which the cytoplasmic tail region or a portion thereof originates. In some embodiments, the chimeric shuttle receptor further includes a signal peptide. In some embodiments, the signal peptide may or may not be derived from a Golgi-associated shuttle receptor. In some embodiments, the signal peptide may be derived from a Golgi-associated shuttle receptor that is the same as or different from one from which the transmembrane domain or a portion thereof originates and / or one from which the cytoplasmic tail region or a portion thereof originates.

[0314] In some embodiments, the chimeric shuttle receptor comprises an amino acid sequence represented by any one of SEQ ID NO: 116-121, 125-194, 196, 198-214, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it. In some embodiments, the chimeric shuttle receptor comprises the amino acid sequence shown in any one of SEQ ID NO: 116-121, 125-142, 143-160, 161-188, 207-214, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it. In some embodiments, the chimeric shuttle receptor comprises an amino acid sequence represented by any one of SEQ ID NO: 116-121, 125-142, 143-156, 158-160, 189-194, 196, 198-206, 207-214, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with it.

[0315] In some embodiments, the nucleic acid (or nucleic acid set) also encodes an antigen recognition receptor (e.g., any antigen recognition receptor described in Section VI.A of this document). In some embodiments, the antigen recognition receptor is selected from engineered T-cell receptors (TCRs), chimeric antigen receptors (CARs), T-cell antigen conjugates (TACs), chimeric TCRs (cTCRs), T-cell antigen conjugate-like (TAC) chimeric receptors, chimeric switch receptors, signal transduction receptors, induced-regulated dimerization-activated receptors (DARICs), chimeric cytokine receptors, co-stimulatory receptors, dominant-negative receptors, or portions thereof. In some embodiments, the antigen recognition receptor includes an engineered TCR. In some embodiments, the antigen recognition receptor includes a CAR. In some embodiments, the antigen recognition receptor includes a TAC. In some embodiments, the antigen recognition receptor specifically recognizes BCMA, DLL3, GPC3, GD2, GU2CYC, and Claudin. 6. Claudin 18.2, CD3, CD5, CD7, CD19, CD20, CD22, CD30, CD33, CD38, CD44V6, CD47, CD52, CD70, CD138, CD123 / IL-3Rα, IL-13Rα, c-Met, CLL1, CS1, gp100, MUC1, MUC16, EpCAM, EGFR, ALK, HER2, IGF1R, FGFR4, mesothelin, PSMA, ER / PR, ROR1, CEA, NY-ESO-1, glycolipid F77, PD-1, PD-L1, PD-L2, TGF-β, VEGF, GRP78, PSCA, PSMA, NKG2D, NKG2DL, LILRB4, PRLR, ALPP, TROP2, tissue factor (Tissue) Factor (TF), Nectin-4, FGFR2B, FLT3, GPRC5D, EGFRvIII, B7H4 and / or B7H3. In some embodiments, the antigen recognition receptor comprises: (a) an extracellular antigen-binding domain comprising a single-domain antibody (sdAb) domain binding to GUCY2C; (b) a transmembrane domain; and (c) an intracellular signaling domain. In some embodiments, the antigen recognition receptor comprises the amino acid sequence shown in any of SEQ ID NO:95, 241-261, or an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the sequence shown in any of SEQ ID NO:95, 241-261, or is composed of said amino acid sequence.

[0316] In some embodiments, the nucleic acid (or nucleic acid group) encodes a chimeric shuttle receptor and an antigen recognition receptor on the same nucleic acid. In some embodiments, the nucleic acid (or nucleic acid group) encodes a chimeric shuttle receptor upstream of an antigen recognition receptor (e.g., CAR). In some embodiments, the nucleic acid (or nucleic acid group) encodes a chimeric shuttle receptor downstream of an antigen recognition receptor (e.g., CAR). In some embodiments, the chimeric shuttle receptor and the antigen recognition receptor are separated by a self-cleaving linker, such as any 2A peptide (e.g., P2A, T2A, E2A, or F2A peptide). In some embodiment...

Claims

A chimeric shuttle receptor, comprising: (a) It can specifically bind to the hetero-binding portion of the target protein, and (b) A polypeptide derived from the Golgi-associated shuttle receptor. According to claim 1, the chimeric shuttle receptor, wherein the polypeptide derived from the Golgi-associated shuttle receptor includes a transmembrane domain or a portion thereof. The chimeric shuttle receptor according to claim 2, wherein the chimeric shuttle receptor further includes a cytoplasmic tail region or a portion thereof. According to claim 3, the chimeric shuttle receptor, wherein the cytoplasmic tail region or a portion thereof is derived from or not derived from the Golgi-associated shuttle receptor. According to claim 3 or 4, the chimeric shuttle receptor, wherein the cytoplasmic tail region or a portion thereof originates from a Golgi-associated shuttle receptor that is the same as or different from the transmembrane domain or a portion thereof. According to claim 1, the chimeric shuttle receptor, wherein the polypeptide derived from the Golgi-associated shuttle receptor includes a cytoplasmic tail region or a portion thereof. According to claim 6, the chimeric shuttle receptor further includes a transmembrane domain or a portion thereof located between the heterologous binding portion and the cytoplasmic tail region or a portion thereof. According to claim 7, the chimeric shuttle receptor, wherein the transmembrane domain or a portion thereof is derived from or not derived from the Golgi-associated shuttle receptor. According to claim 7 or 8, the chimeric shuttle receptor, wherein the transmembrane domain or a portion thereof is derived from a Golgi-associated shuttle receptor that is the same as or different from the cytoplasmic tail region or a portion thereof. The chimeric shuttle receptor according to any one of claims 1-9, wherein the Golgi-associated shuttle receptor comprises sorting protein (SORT), Sortilin-associated receptor (SORL), centrally expressed Sortilin-associated receptor 1 (SORCS1), centrally expressed Sortilin-associated receptor 2 (SORCS2), centrally expressed Sortilin-associated receptor 3 (SORCS3), and lysosomal membrane protein 2 (SCARB2). The chimeric shuttle receptor according to any one of claims 1-9, wherein the Golgi-associated shuttle receptor comprises a VPS10P domain receptor; optionally, wherein the VPS10P domain receptor comprises SORT, SORL, SORCS1, SORCS2, and SORCS3. The chimeric shuttle receptor according to any one of claims 2-5 and 7-11, wherein the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:1-36, SEQ ID NO:99, SEQ ID NO:101 and SEQ ID NO:114, or an amino acid sequence having at least 75% sequence identity with any of the aforementioned amino acid sequences. According to claim 12, the chimeric shuttle receptor, wherein the transmembrane domain or a portion thereof is derived from a Golgi-associated shuttle receptor, the transmembrane domain or a portion thereof comprising the amino acid sequence shown in any of SEQ ID NO: 1-36, its continuous portions, or variants thereof, wherein the variants have at least 75% sequence identity with any of the aforementioned amino acid sequences; optionally, wherein the variants have at least 80% sequence identity with any of the aforementioned sequences. According to claim 12 or 13, the chimeric shuttle receptor, wherein the transmembrane domain or a portion thereof comprises a leucine (L) or isoleucine (I) mutant near the C-terminus of the transmembrane domain or a portion thereof derived from a Golgi-associated shuttle receptor; optionally, the leucine (L) or isoleucine (I) is mutated to alanine (A). The chimeric shuttle receptor according to any one of claims 12-14, wherein the transmembrane domain or a portion thereof comprises a continuous portion of at least 5 amino acids derived from a transmembrane domain or a portion thereof of a Golgi-associated shuttle receptor; optionally, a continuous portion of at least 10 amino acids; more optionally, a continuous portion of at least 20 amino acids. The chimeric shuttle receptor according to any one of claims 12-15, wherein the transmembrane domain or a portion thereof is derived from SORT, and the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:1-26, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the aforementioned sequences. According to claim 16, the chimeric shuttle receptor, wherein the transmembrane domain or a portion thereof comprises the amino acid sequence shown in any of SEQ ID NO: 1-6, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 12-15, wherein the transmembrane domain or a portion thereof is derived from SORL, and the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO: 27 or 28, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 12-15, wherein the transmembrane domain or a portion thereof is derived from SORCS1, and the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:29 or 30, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 12-15, wherein the transmembrane domain or a portion thereof is derived from SORCS2, and the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:31 or 32, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 12-15, wherein the transmembrane domain or a portion thereof is derived from SORCS3, and the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:33 or 34, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 12-15, wherein the transmembrane domain or a portion thereof is derived from SCARB2, and the transmembrane domain or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:35 or 36, its continuous portions, or variants thereof, wherein the variants have an amino acid sequence with at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 3-22, wherein the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:37-59 and SEQ ID NO:115, or an amino acid sequence having at least 75% identity with any of the aforementioned amino acid sequences. According to claim 23, the chimeric shuttle receptor, wherein the cytoplasmic tail region or a portion thereof is derived from a Golgi-associated shuttle receptor, the cytoplasmic tail region or a portion thereof comprising the amino acid sequence shown in any of SEQ ID NO:37-59, its continuous portions, or variants thereof, the variant having at least 75% sequence identity with any of the aforementioned amino acid sequences; optionally, wherein the variant has at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to claim 23 or 24, wherein the cytoplasmic tail region or a portion thereof contains an AP binding motif. The chimeric shuttle receptor according to any one of claims 23-25, wherein the cytoplasmic tail region or a portion thereof comprises a tyrosine-based motif. The chimeric shuttle receptor of any one of claims 23-26, wherein the cytoplasmic tail or portion thereof comprises wherein X is any amino acid, It is L, I, M, V or F; optionally, the cytoplasmic tail region or a portion thereof comprises YSVL (SEQ ID NO:60), YAQM (SEQ ID NO:64) or YAQV (SEQ ID NO:65). The chimeric shuttle receptor according to any one of claims 23-27, wherein the cytoplasmic tail region or a portion thereof comprises a CK-2 site; optionally, wherein the cytoplasmic tail region or a portion thereof comprises DDSDED (SEQ ID NO:61), DEDLL (SEQ ID NO:62), or DDLGEDDEDAPMI (SEQ ID NO:63). The chimeric shuttle receptor according to any one of claims 23-28, wherein the cytoplasmic tail region or a portion thereof comprises a bisleucine-like site; optionally, wherein the cytoplasmic tail region or a portion thereof comprises the amino acid sequence LL or DEDLL (SEQ ID NO:62). The chimeric shuttle receptor according to any one of claims 23-29, wherein the cytoplasmic tail region or a portion thereof comprises a GGA motif; optionally, wherein the cytoplasmic tail region or a portion thereof comprises an amino acid sequence LL or comprises (D / E)XXLL, wherein X is any amino acid and (D / E) is D or E. The chimeric shuttle receptor according to any one of claims 23-30, wherein the cytoplasmic tail region or a portion thereof comprises a continuous portion of at least 5 amino acids derived from the cytoplasmic tail region or a portion thereof of a Golgi-associated shuttle receptor; optionally, a continuous portion of at least 10, 20, 30, 40, 50, 60, 70 or 80 amino acids. The chimeric shuttle receptor according to any one of claims 23-31, wherein the cytoplasmic tail region or a portion thereof is derived from SORT, and the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:37-51, its continuous portion, or a variant thereof, the variant having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 23-32, wherein the cytoplasmic tail region or a portion thereof is derived from SORL, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:52, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the preceding sequences. Chimeric shuttle receptor according to any one of claims 23-32, wherein the cytoplasmic tail region or a portion thereof is derived from SORCS1, the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:53-56, its continuous portion, or a variant thereof, the variant having an amino acid sequence with at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 23-32, wherein the cytoplasmic tail region or a portion thereof is derived from SORCS2, and the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:57, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 23-32, wherein the cytoplasmic tail region or a portion thereof is derived from SORCS3, and the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:58, its continuous portion, or a variant thereof, the variant having at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 23-32, wherein the cytoplasmic tail region or a portion thereof is derived from SCARB2, and the cytoplasmic tail region or a portion thereof comprises the amino acid sequence shown in SEQ ID NO:59, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the preceding sequences. The chimeric shuttle receptor according to any one of claims 1-37, wherein the heterologous binding portion is a polypeptide, ligand, aptamer, designed ankyrin repeat protein (DARPin), artificially designed binding protein, or D-Domain. The chimeric shuttle receptor according to any one of claims 1-37, wherein the heterologous binding portion comprises an antibody or an antigen-binding fragment thereof; optionally, the heterologous binding portion is an antibody. According to claim 39, the chimeric shuttle receptor, wherein the antibody or its antigen-binding fragment is IgG, a single-chain variable fragment (scFv), a single-domain antibody (sdAb), VHH, Fab, F(ab')2, (scFv')2, or a nanobody. The chimeric shuttle receptor according to any one of claims 1-40, wherein the heterologous binding portion binds to extracellular protein molecules, membrane protein molecules, or intracellular molecules. The chimeric shuttle receptor according to any one of claims 1-41, wherein the heterologous binding portion binds to an inhibitory immune receptor, a ligand of an inhibitory immune receptor, or an immune checkpoint molecule. The chimeric shuttle receptor according to any one of claims 1-42, wherein the heterologous binding portion binds to major histocompatibility complex (MHC) class I molecules (HLA-A, HLA-B, HLA-C, HLA-G, or HLA-E), MHC class II molecules (HLA-DP, HLA-DQ, HLA-DR), MHC class III molecules, T cell receptors (TCRs) or their subunits, CD3 / TCR complexes or their subunits (CD3ε, TCRα, T... CRβ, TCRγ, TCRδ, CD3δ, CD3γ and CD3ζ), programmed cell death protein 1 (PD-1), programmed cell death ligand 1 (PD-L1), CTLA-4, CTLA-1, TIM3, LAG3, TIGIT, epidermal growth factor receptor (EGFR), glucocorticoid receptor (GR), CD38, CD7, CD25, CD69, CD70, CD56, CD22, FcγRII-B, 4-1BB, 4 -BBL, B2M, CIITA, NKG2A, NKG2D, CD94, TGF-β receptor, cytotoxic immunoglobulin-like receptor (KIR)2DL1, KIR2DL2, KIR2DL3, HLA, BCMA, CFH, ARMS2, MYOC, OPTN, VEGF, HIF1A, RB1, MUC16, TGFBR1, TGFBR2, OPN1SW, OPN1MW, MTTP, FOXC2, MSX1, K ERA, SLC4A11, PAX6, OTX2, RHO, P23H, USH2A, MYO7A, LXS, CRYAA, CRYAB, ABCA4, CEP290, MHCI, NR3C1, Per forin1, GUCY2C, CLEC12A, PRLR, ERBB2, IMPDH, ITK, TMEM30A, Regnase-1, IFNGR, CD58, HSP90B1, or PIM3. According to claim 43, the chimeric shuttle receptor, wherein the heterologous binding portion binds to molecules selected from the group consisting of CD3ε, CD38, CD7, NKG2A, BCMA, 41BB, and CD25. The chimeric shuttle receptor of claim 43, wherein the heterologous binding portion binds to B2M or HLA-A (e.g., HLA-A2). The chimeric shuttle receptor according to any one of claims 1-45, wherein the chimeric shuttle receptor further comprises a hinge domain or portion thereof located between the heterologous binding portion and the transmembrane domain or portion thereof. The chimeric shuttle receptor of claim 46, wherein the hinge domain or a portion thereof is derived from or not derived from a Golgi-associated shuttle receptor. According to claim 47, the chimeric shuttle receptor, wherein the hinge domain or a portion thereof is derived from a Golgi-associated shuttle receptor that is the same as or different from the transmembrane domain or a portion thereof and / or the cytoplasmic tail region or a portion thereof. The chimeric shuttle receptor according to any one of claims 46-48, wherein the hinge domain or a portion thereof comprises an artificial connector. According to claim 49, the chimeric shuttle receptor, wherein the hinge domain or a portion thereof comprises a GS adapter, an α-helical adapter, a glycine-alanine polymer adapter, an alanine-serine polymer adapter, and an IgG4-Fc adapter. According to claim 49 or 50, the chimeric shuttle receptor, wherein the hinge domain or a portion thereof comprises GG, GS, SG, SS, GSS, SSG, GSG, SGS, SGG, G4S, (G4S)2, (G4S)3 or (G4S)4. The chimeric shuttle receptor according to any one of claims 46-48, wherein the hinge domain or a portion thereof comprises an amino acid sequence shown in any one of SEQ ID NO:66-88, SEQ ID NO:100, 102 or 239, or an amino acid sequence having at least 75% identity with any of the aforementioned amino acid sequences. According to claim 52, the chimeric shuttle receptor, wherein the hinge domain or a portion thereof is derived from a Golgi-associated shuttle receptor, the hinge domain or a portion thereof comprising an amino acid sequence shown in any of SEQ ID NO: 66-88, its continuous portions, or variants thereof, the variant having at least 75% sequence identity with any of the aforementioned amino acid sequences; optionally, wherein the variant has at least 80% sequence identity with any of the aforementioned sequences. According to claim 52 or 53, the chimeric shuttle receptor, wherein the hinge domain or a portion thereof comprises a continuous portion of at least one amino acid derived from the cytoplasmic tail region or a portion thereof of a Golgi-associated shuttle receptor; optionally, a continuous portion of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 amino acids. The chimeric shuttle receptor according to any one of claims 52-54, wherein the hinge domain or a portion thereof is derived from SORT, and the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:66-78, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 52-54, wherein the hinge domain or a portion thereof is derived from SORL, and the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:79 or 80, its continuous portion, or a variant thereof, the variant having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 52-54, wherein the hinge domain or a portion thereof is derived from SORCS1, and the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO: 81 or 82, its continuous portion, or a variant thereof, the variant having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 52-54, wherein the hinge domain or a portion thereof is derived from SORCS2, and the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:83 or 84, its continuous portion, or a variant thereof, the variant having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 52-54, wherein the hinge domain or a portion thereof is derived from SORCS3, and the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:85 or 86, its continuous portion, or a variant thereof, the variant having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 52-54, wherein the hinge domain or a portion thereof is derived from SCARB2, and the hinge domain or a portion thereof comprises the amino acid sequence shown in any one of SEQ ID NO:87 or 88, its continuous portions, or variants thereof, the variants having at least 80% sequence identity with any of the aforementioned sequences. The chimeric shuttle receptor according to any one of claims 1-60, wherein the chimeric shuttle receptor further comprises a signal peptide. The chimeric shuttle receptor of claim 61, wherein the signal peptide is derived from or not derived from a Golgi-associated shuttle receptor. According to claim 62, the chimeric shuttle receptor, wherein the signal peptide is derived from a Golgi-associated shuttle receptor that is the same as or different from the transmembrane domain or a portion thereof and / or the cytoplasmic tail region or a portion thereof. The chimeric shuttle receptor according to any one of claims 61-63, wherein the signal peptide comprises the amino acid sequence shown in any one of SEQ ID NO: 89-94, 97, 113 and 240, or an amino acid sequence having at least 75% identity with any of the aforementioned amino acid sequences. According to claim 64, the chimeric shuttle receptor, wherein the signal peptide is derived from a Golgi-associated shuttle receptor, the signal peptide comprising an amino acid sequence shown in any of SEQ ID NO:89-94, its continuous portions, or variants thereof, the variant having at least 75% sequence identity with any of the aforementioned amino acid sequences; optionally, the variant having at least 80% sequence identity with any of the aforementioned sequences. A polypeptide or combination of polypeptides comprising the chimeric shuttle receptor according to any one of claims 1-65. The polypeptide or combination of polypeptides according to claim 66 further comprises an antigen recognition receptor. The polypeptide or combination of polypeptides according to claim 67, wherein the antigen recognition receptor is an engineered T-cell receptor (TCR), a chimeric antigen receptor (CAR), a T-cell antigen conjugate (TAC), a chimeric TCR (cTCR), a T-cell antigen conjugate (TAC) chimeric receptor, a chimeric switch receptor, a signal transduction receptor, an inducible regulation dimerization activation receptor (DARIC), a chimeric cytokine receptor, a co-stimulatory receptor, a dominant-negative receptor, or a portion thereof. The polypeptide or combination of polypeptides according to claim 68, wherein the antigen recognition receptor is a chimeric antigen receptor (CAR). The polypeptide or combination of polypeptides according to claim 69, wherein the CAR comprises: (a) an extracellular antigen-binding domain comprising a single-domain antibody (sdAb) domain that binds to GUCY2C; (b) a transmembrane domain; and (c) an intracellular signal transduction domain. The polypeptide or combination of polypeptides according to claim 70, wherein the sdAb domain comprises an amino acid sequence shown in any of SEQ ID NO:215-235, or an amino acid sequence having at least 75% identity with any of the aforementioned amino acid sequences. The polypeptide or combination of polypeptides according to claim 70 or 71, wherein the CAR comprises the amino acid sequence shown in any one of SEQ ID NO: 95, 241-261, or an amino acid sequence having at least 75% identity with any of the aforementioned amino acid sequences. The polypeptide or combination of polypeptides according to any one of claims 66-72 further comprises exogenous inosine monophosphate dehydrogenase (IMPDH). The polypeptide or combination of polypeptides according to claim 73, wherein the exogenous IMPDH is resistant to purine biosynthesis inhibitors. The polypeptide or combination of polypeptides according to claim 73 or 74, wherein the exogenous IMPDH is derived from prokaryotic IMPDH; optionally, the exogenous IMPDH is derived from bacterial IMPDH or a variant thereof. The polypeptide or combination of polypeptides according to any one of claims 73-75, wherein the exogenous IMPDH is derived from Gram-positive bacterial IMPDH or a variant thereof or Gram-negative bacterial IMPDH or a variant thereof; optionally, wherein the Gram-positive bacterial IMPDH is derived from Bacillus subtilis, Lactobacillus plantarum or Staphylococcus aureus; and / or the Gram-negative bacterial IMPDH is derived from Escherichia coli or Flora intercalation. The polypeptide or combination of polypeptides according to any one of claims 73-76, wherein the exogenous IMPDH comprises an amino acid sequence selected from any one of SEQ ID NO:262-268, or an amino acid sequence having at least 75% identity with any of the aforementioned amino acid sequences. The polypeptide or combination of polypeptides according to any one of claims 73-77, wherein the chimeric shuttle receptor is not connected to the antigen recognition receptor and / or the exogenous IMPDH. The polypeptide or combination of polypeptides according to any one of claims 73-77, wherein the chimeric shuttle receptor is linked to the antigen recognition receptor and / or the exogenous IMPDH; optionally, wherein the amino acid sequence of the chimeric shuttle receptor is located upstream or downstream of the amino acid sequence of the antigen recognition receptor and / or the exogenous IMPDH. According to claim 79, the polypeptide or combination of polypeptides, wherein the chimeric shuttle receptor is separated from the antigen recognition receptor and / or the exogenous IMPDH by a self-cleavable peptide, an internal ribosome entry site, or an enzyme cleavage site; optionally, wherein the self-cleavable peptide comprises a 2A peptide; more optionally, the 2A peptide comprises a P2A, T2A, E2A, or F2A peptide; optionally, wherein the enzyme cleavage site comprises a trypsin cleavage site or a thrombin cleavage site. The polypeptide or combination of polypeptides according to any one of claims 66-80 further comprises a detection marker. The polypeptide or combination of polypeptides according to claim 81, wherein the detection marker comprises an antibody or ligand, a resistance gene (including antibiotic resistance genes, such as puromycin resistance gene, G418 resistance gene (Neo), or blast fungicide resistance gene (BSD)) or a selection marker (including an affinity tag, such as a histidine tag), or a fluorescent protein. An isolated nucleic acid or nucleic acid group encoding a chimeric shuttle receptor as described in any one of claims 1-65, or encoding a polypeptide or combination of polypeptides as described in any one of claims 76-92. The nucleic acid or nucleic acid group according to claim 83 further comprises a second nucleic acid encoding an antigen recognition receptor. The nucleic acid or nucleic acid group according to claim 84, wherein the antigen recognition receptor is the antigen recognition receptor as defined in any one of claims 68-72. The nucleic acid or nucleic acid group according to any one of claims 83-85 further comprises a third nucleic acid encoding exogenous inosine monophosphate dehydrogenase (IMPDH). The nucleic acid or nucleic acid genome according to claim 86, wherein the exogenous IMPDH is the exogenous IMPDH as defined in any one of claims 73-77. The nucleic acid or nucleic acid group according to any one of claims 83-87, wherein the nucleic acid encoding the chimeric shuttle receptor, the nucleic acid encoding the antigen recognition receptor, and / or the nucleic acid encoding the exogenous IMPDH are separated by nucleic acids encoding a self-cleavable peptide, an internal ribosome entry site, or an enzyme cleavage site; optionally, wherein the self-cleavable peptide comprises a 2A peptide; more optionally, the 2A peptide comprises a P2A, T2A, E2A, or F2A peptide; optionally, wherein the enzyme cleavage site comprises a trypsin cleavage site or a thrombin cleavage site. The nucleic acid or nucleic acid group according to any one of claims 83-88, wherein the nucleic acid encoding the chimeric shuttle receptor is located upstream or downstream of the nucleic acid encoding the antigen recognition receptor and / or the nucleic acid encoding the exogenous IMPDH. A vector comprising the nucleic acid or nucleic acid genome as described in any one of claims 83-89. The vector according to claim 90, wherein the vector is a viral vector; optionally, wherein the viral vector is a retroviral vector, a lentiviral vector, an adenovirus vector, or an adeno-associated virus vector. An engineered cell comprising a chimeric shuttle receptor as described in any one of claims 1-65, a polypeptide or combination of polypeptides as described in any one of claims 66-82, a nucleic acid or nucleic acid group as described in any one of claims 83-89, or a vector as described in claims 90 or 91. The engineered cell of claim 92, wherein the cell is a mammalian cell; optionally, wherein the cell is a human cell. The engineered cell according to claim 92 or 93, wherein the cell is an immune cell. The engineered cells according to claim 92 or 93, wherein the cells are T cells, natural killer (NK) cells, γδT cells, αβT cells, cytotoxic T cells (CTL), regulatory T (Treg) cells, natural killer T (NKT) cells, peripheral blood mononuclear cells (PBMCs), hematopoietic stem cells, pluripotent stem cells, embryonic stem cells, memory T cells, T memory stem cells (TSCM), naive T cells, effector T cells, T helper cells, stem cells, induced pluripotent stem (iPSC) cells, ocular tissue cells, hepatocytes, kidney cells, cardiac cells, bone marrow cells, or tumor-infiltrating lymphocytes (TILs). The engineered cell according to any one of claims 92-95, wherein the cell is an allogeneic cell or an autologous cell. A method for producing engineered cells according to any one of claims 92-96, the method comprising introducing a nucleic acid or nucleic acid group according to any one of claims 83-89, or a vector according to claims 90 or 91, into the cells. A method for regulating target proteins in cells includes introducing a nucleic acid or nucleic acid group as described in any one of claims 83-89, or a vector as described in claims 90 or 91, into a cell, wherein the heterologous binding portion is capable of specifically binding to the target protein. A pharmaceutical composition comprising the chimeric shuttle receptor of any one of claims 1-65; the polypeptide or combination of polypeptides of any one of claims 66-82; the nucleic acid or nucleic acid group of any one of claims 83-89; the vector of any one of claims 90 or 91; or the engineered cell of any one of claims 92-96. The pharmaceutical composition according to claim 99 further comprises a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition according to claim 99 or 100 further comprises additional therapeutic agents or pharmaceutical agents. The pharmaceutical composition according to any one of claims 99-101 further comprises an agent that inhibits the function of antigen-presenting cells (APCs). The pharmaceutical composition of claim 102, wherein the pharmaceutical agent is abatacept, tofacitinib, secukinumab, ipilimumab, cyclosporine A, fingolimod, methotrexate, or equivalents thereof. The pharmaceutical composition according to any one of claims 99-103 further comprises a purine biosynthesis inhibitor. The pharmaceutical composition according to claim 104, wherein the purine biosynthesis inhibitor is mycophenolic acid (MPA), mycophenolic ester, mycophenolate mofetil (MMF), sodium mycophenolate (MPS), calcium mycophenolate, potassium mycophenolate, or derivatives or equivalents thereof, ribavirin, mizoribine, tiazofurin, AVN-944 (VX-944), FF-10501, AS2643361, or BMS-986126, or pharmaceutically acceptable esters, salts, or prodrugs thereof. A method of treating an individual disease or symptom, comprising administering to the individual an effective amount of any chimeric shuttle receptor as claimed in any one of claims 1-65; a polypeptide or combination of polypeptides as claimed in any one of claims 66-82; a nucleic acid or nucleic acid group as claimed in any one of claims 83-89; a carrier as claimed in claim 90 or 91; engineered cells as claimed in any one of claims 92-96; or a pharmaceutical composition as claimed in any one of claims 99-105. The method according to claim 106, wherein the method is allogeneic cell therapy or autologous cell therapy. A method for treating graft-versus-host disease (GVHD) or host-versus-graft reaction (HVGR) in a subject, comprising administering to an individual an effective amount of the chimeric shuttle receptor of any one of claims 1-65; a polypeptide or combination of polypeptides of any one of claims 66-82; a nucleic acid or nucleic acid set of any one of claims 83-89; a vector of any one of claims 90 or 91; engineered cells of any one of claims 92-96; or a pharmaceutical composition of any one of claims 99-105. The method of claim 108, wherein treating GVHD or HVGR is to prevent GVHD or HVGR, or to reduce the risk of developing GVHD or HVGR; optionally, wherein the host-versus-graft reaction (HVGR) includes acute HVGR and chronic HVGR; and / or, wherein the graft-versus-host disease (GVHD) includes acute GVHD and chronic GVHD; more optionally, the graft-versus-host disease (GVHD) is steroid-refractory acute GVHD.

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