Anti-MYCT1 antibodies and uses thereof

Abstract

In various embodiments of the present disclosure, compositions comprising anti-MYCT1 antibodies and methods for modulating MYCT1 activity are provided. One embodiment of the present disclosure provides a method for controlling tumor angiogenesis and / or anti-tumor immunity (which inhibit tumor growth) in a subject.

Description

[Technical Field]

[0001] cross reference This application claims the benefit of U.S. Provisional Patent Application No. 63 / 357,888, filed July 1, 2023, entitled "ANTI-MYCT1 ANTIBODIES AND USES THEREOF," the contents of which are incorporated herein by reference in their entirety.

[0002] Technical Field The present disclosure relates generally to compositions comprising anti-MYCT1 antibodies and methods for modulating MYCT1 activity. More specifically, the present disclosure provides compositions and methods for preventing or reducing tumor angiogenesis, reconstituting tumor immunity, and improving the outcome of immunotherapy.

[0003] Incorporating a sequence listing This application contains a Sequence Listing which has been submitted via the Patent Center in .XML format and is incorporated herein by reference in its entirety. The WIPO Sequence Listing was created on July 3, 2023; the XML copy is named 760394.xml and is 137 kilobytes in size. [Background technology]

[0004] Angiogenesis and immune tolerance are both normal physiological mechanisms that are hijacked by tumors. Angiogenesis involves the formation of new blood vessels from pre-existing vessels during development and wound healing. Regulation of angiogenesis is highly regulated by pro- and anti-angiogenic factors, but in cancer, this process is disrupted and dysregulated. Tumor-induced hypoxia increases the expression of pro-angiogenic factors, leading to the formation of new blood vessels essential for tumor survival and growth. The VEGF family [consisting of six growth factors (VEGFA-VEGF)] plays a key role in angiogenesis by binding to its receptors VEGFR1-VEGFFR3 and neuropin. Angiogenesis can also be mediated by the angiopoietin (Ang1-2) / Tie-2 pathway, independently of the VEGF pathway. Therefore, over the past decade, drug development has largely focused on antiangiogenesis as a strategy to deprive tumors of nutrients and inhibit tumor growth. However, despite the drug's modest activity as a single agent or in combination with chemotherapy, tumors can overcome its effects and become resistant.

[0005] Cancer immunotherapy has emerged as a potential treatment for various cancers due to the discovery of immune checkpoints. Numerous studies on immune checkpoint inhibitors (ICIs) have demonstrated long-term clinical activity against many malignancies. ICIs block "immune exhaustion," another mechanism hijacked by tumors, and release the control of effector immune cells against cancer. Primary resistance to ICIs has been reported in tumors lacking tumor-infiltrating lymphocytes. In addition, tumors that initially respond to ICIs may develop secondary resistance due to defects in antigen presentation mechanisms and overexpression of co-inhibitory molecules, among other factors.

[0006] Thus, there is a need in the art for compositions and methods that address angiogenesis and immune exhaustion in tumor cells. Summary of the Invention

[0007] In some aspects, provided herein is an isolated anti-MYCT1 antibody or fragment thereof that specifically binds to an epitope within the first 100 amino acids of SEQ ID NO: 145. In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region comprising: L1 having the amino acid sequence of SEQ ID NO:28, SEQ ID NO:64, SEQ ID NO:100, or SEQ ID NO:136; L2 having the amino acid sequence of SEQ ID NO:30, SEQ ID NO:66, SEQ ID NO:102, or SEQ ID NO:138; L3 having the amino acid sequence of SEQ ID NO:32, SEQ ID NO:68, SEQ ID NO:104, or SEQ ID NO:140, or any combination thereof; and / or a heavy chain variable region comprising: H1 having the amino acid sequence of SEQ ID NO:10, SEQ ID NO:46, SEQ ID NO:82, or SEQ ID NO:118; H2 having the amino acid sequence of SEQ ID NO:12, SEQ ID NO:48, SEQ ID NO:84, or SEQ ID NO:120; H3 having the amino acid sequence of SEQ ID NO:14, SEQ ID NO:50, SEQ ID NO:86, or SEQ ID NO:122, or any combination thereof.

[0008] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:36, SEQ ID NO:72, SEQ ID NO:108, or SEQ ID NO:144; and / or a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:18, SEQ ID NO:54, SEQ ID NO:90, or SEQ ID NO:126, or any combination thereof.

[0009] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 28, L2 having the amino acid sequence of SEQ ID NO: 30, L3 having the amino acid sequence of SEQ ID NO: 32, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 10, H2 having the amino acid sequence of SEQ ID NO: 12, H3 having the amino acid sequence of SEQ ID NO: 14, or any combination thereof.

[0010] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 64, L2 having the amino acid sequence of SEQ ID NO: 66, L3 having the amino acid sequence of SEQ ID NO: 68, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 46, H2 having the amino acid sequence of SEQ ID NO: 48, H3 having the amino acid sequence of SEQ ID NO: 50, or any combination thereof.

[0011] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 100, L2 having the amino acid sequence of SEQ ID NO: 102, L3 having the amino acid sequence of SEQ ID NO: 104, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 82, H2 having the amino acid sequence of SEQ ID NO: 84, H3 having the amino acid sequence of SEQ ID NO: 86, or any combination thereof.

[0012] In some aspects, the isolated anti-MYCT1 antibody or fragment thereof comprises a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 136, L2 having the amino acid sequence of SEQ ID NO: 138, L3 having the amino acid sequence of SEQ ID NO: 140, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 118, H2 having the amino acid sequence of SEQ ID NO: 120, H3 having the amino acid sequence of SEQ ID NO: 122, or any combination thereof.

[0013] In some embodiments, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO: 36 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO: 18.

[0014] In some embodiments, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO: 72 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO: 54.

[0015] In some embodiments, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO: 108 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO: 90.

[0016] In some embodiments, the isolated anti-MYCT1 antibody or fragment thereof comprises the amino acid sequence of the light chain variable region of SEQ ID NO: 144 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO: 126.

[0017] In some aspects, any isolated antibody or fragment thereof disclosed herein is a monoclonal antibody. In some aspects, the isolated antibody or fragment thereof is a humanized antibody, a single-chain variable fragment (scFv) antibody, an antibody fragment, or a chimeric antibody.

[0018] In some aspects, there is further provided a pharmaceutical composition comprising the disclosed isolated antibody or fragment thereof and a pharmaceutically acceptable carrier or excipient. In some aspects, the isolated antibody is used as a medicament.

[0019] The present disclosure further encompasses methods of treating cancer in a subject in need thereof, comprising administering an effective amount of a composition comprising the isolated antibody or fragment thereof disclosed herein. In some aspects, the method further comprises administering an immune checkpoint inhibitor, and optionally, an anti-VEGF targeted therapy.

[0020] In a further aspect, the disclosure includes a method of sensitizing a cancer in a subject to an immune checkpoint inhibitor, the method comprising: selecting a subject having cancer, wherein the cancer is resistant to an immune checkpoint inhibitor; and administering to the subject an effective amount of a composition comprising the disclosed isolated antibody or fragment thereof. In some aspects of this method, administration of the anti-MYCT1 antibody reduces angiogenesis in cancer cells in the subject. In some aspects of this method, administration of the anti-MYCT1 antibody increases anti-tumor immunity in the subject. In some aspects of this method, administration of the anti-MYCT1 antibody reduces tumor volume in the subject.

[0021] Further provided are methods of controlling tumor angiogenesis and / or increasing anti-tumor immunity in a subject with cancer, comprising administering to the subject a therapeutically effective amount of a composition comprising an isolated antibody or fragment thereof disclosed herein. In some aspects, the method comprises administering an immune checkpoint inhibitor, and optionally, an anti-VEGF inhibitor.

[0022] Those skilled in the art will understand that the drawings described below are for illustrative purposes only and are not intended to limit the scope of the present teachings in any way. [Brief explanation of the drawings]

[0023] [Figure 1]Figures 1A-1D show the generation of anti-MYCT1 antibodies. Figure 1A shows a sequence alignment between human MYCT1 isoform 1 (SEQ ID NO: 145) and the mouse MYCT1 amino acid sequence (SEQ ID NO: 150). Figure 1B shows the immunization scheme for generating MYCT1 monoclonal antibodies. LS, lumazine synthase (LC) from the hyperthermophilic bacterium Aquifex aeolicus (AaLS), keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), HUVEC, human umbilical vein endothelial cells (HUVEC), mouse cardiac endothelial cells (MCEC), enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting (FACS), knockdown (KD), overexpression (OE). Figure 1C shows a graph of a representative MYCT1 terminal serum titer for fusion no. 4941 (animal no. 8). MYCT1 antigen was added at 1 μg / ml. Figure ID shows a graph of a representative MYCT1 terminal serum titer for fusion number 4952 (animal number 2). MYCT1 antigen was added at 1 μg / ml. [Figure 2] 2A-2E show the plasmid maps of the plasmids used in the studies disclosed herein. [Figure 3] Figures 3A-3D show the initial screening results of the generated monoclonal antibodies. Figure 3A shows the results of an ELISA assay. Figure 3B shows the results of fluorescence-activated cell sorting (FACS) of mouse cardiac endothelial cells (MCECs) overexpressing IgG (OE). Figures 3C and 3D show representative results and images of a scratch assay at 3 hours. [Figure 4] Figures 4A to 4C show the results of screening using the plasmid pcDNA3.1-Myct1Flag shown in Figure 2A. Figure 4A shows the results of FACS. Figures 4B and 4C show representative images of the scratch assay. [Figure 5]Figures 5A-5B show the identification of anti-MYCT1 antibody clones. Figure 5A shows the identification of clones 2B4 (IgG2b isotype) and 11B6 (IgG1 isotype) based on scratch assay, FACS staining, and ELISA. Figure 5B shows the identification of clones 3E3, 8D12, 9E2, and 9F8 based on scratch assay, FACS staining, and ELISA. [Figure 6] Figures 6A-6C show the characterization of antibody clone 2B4F9D9. Figure 6A shows FACS analysis of hybridoma supernatant containing candidate antibody 2B4F9D9 in HA-mouse Myct1-overexpressing (HA-mMOE) mouse cardiac endothelial cells (MCECs). Figures 6B and 6C show representative images and results of a scratch assay using antibody clone 2B4F9D9. [Figure 7] Figures 7A-7C show the results of a 15-hour sprouting assay in induced mouse pulmonary endothelial cells (iMLEC) treated with purified antibody clones. Figure 7A shows representative images of sprouting assays in iMLEC and iMLEC Mytc1 KO cells. Figure 7B shows the number of sprouts per bead in iMLEC and iMLEC Mytc1 KO cells. Figure 7C shows the sprout length in iMLEC and iMLEC Mytc1 KO cells. **p<0.01, ***p<0.001, ****p<0.0001 compared to IgG-treated cells, one-way ANOVA. [Figure 8] Figures 8A-8C show the results of a 30-hour sprouting assay in MCECs treated with purified antibody clones. Figure 8A shows a representative image of the sprouting assay in MCEC cells. Figure 8B shows the number of sprouts per bead in MCEC cells. Figure 8C shows the sprout length in MCEC cells. **p<0.01 compared to IgG-treated cells, one-way ANOVA. [Figure 9]Figures 9A-9G show the results of a sprouting assay in MCEC cells or induced human umbilical vein endothelial cells (iHUVECs) treated with purified antibody clones 3EC12, 8D2C12, 9E2H5, and 2B4F9D3. Figure 9A shows representative images of the sprouting assay in MCEC cells. Figure 9B shows the number of sprouts per bead in MCEC cells. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 compared to the HM-treated group, one-way ANOVA. Figure 9C shows the sprout length in MCEC cells. ***p<0.001, ****p<0.0001 compared to the HM-treated group, one-way ANOVA. Figure 9D shows representative images of a sprouting assay in iHUVEC cells treated with increasing concentrations of antibody clone 11B6 or 2B4F9D3 for 24 hours. Figure 9E shows representative images of a sprouting assay in iHUVEC cells treated with increasing concentrations of antibody clone 11B6 or 2B4F9D3 for 48 hours. Figure 9F shows a graph of the number and length of sprouts in iHUVEC cells treated with increasing concentrations of antibody clone 11B6 or 2B4F9D3 for 24 hours. *p<0.05 compared to IgG-treated cells, two-way ANOVA. Figure 9G shows a graph of the number and length of sprouts in iHUVEC cells treated with increasing concentrations of antibody clone 11B6 or 2B4F9D3 for 48 hours. *p<0.05, ***p<0.001, ****p<0.0001 compared to IgG-treated cells, two-way ANOVA. [Figure 10]Figures 10A-10E show the results of a tube formation assay using purified antibody clones. Figure 10A shows representative images of tube formation in iMLEC cells treated with 4 μg / well of 11B6 and 2B4F9D3 for 4 hours. Figure 10B shows representative images of tube formation in MCEC cells treated with 2B4F9D3, 3EC12, 8D2C12, and 9E2H5. Figure 10C shows the number of loops in MCEC cells treated with each treatment. **p<0.01, ***p<0.001 compared to HM-treated cells, one-way ANOVA. Figure 10D shows tube formation in iHUVEC cells treated with increasing concentrations of 11B6 or 2B4F9D3 for 16 hours. Scale bar = 100 μm. Figure 10E shows the number of loops and branches in treated iHUVEC cells. *p<0.05, **p<0.01 compared with IgG-treated cells, two-way ANOVA. [Figure 11]Figures 11A-11F show the results of scratch assays from cells treated with hybridoma supernatants. Figure 11A shows representative images of wound closure in MCEC cells treated with hybridoma supernatants of antibody clones 11B6, 2B4F9D3, or IgG added at 32 μg / well at 0, 8, and 10 hours. Figure 11B shows a graph showing relative tube closure in MCEC cells treated with hybridoma supernatants of 11B6, 2B4F9D3, or IgG. *p<0.05, **p<0.01 compared to IgG-treated cells, two-way ANOVA. Figure 11C shows representative images of wound closure in iMLEC cells and iMLEC Myct1 knockout (MKO) cells treated with hybridoma supernatants of 11B6, 2B4F9D3, or IgG added at 32 μg / well at 0, 9, and 11 hours. Figure 11D shows a graph depicting the relative number of closed tubes in iMLEC cells treated with 11B6, 2B4F9D3, or IgG hybridoma supernatants, and iMLEC MKO cells. *p<0.05, **p<0.01 compared to IgG-treated cells, two-way ANOVA. Figure 11E shows representative images of wound closure in MCEC cells treated with 3E3C12, 8D2C12, 9E2H5, 2B4F9D3, or IgG hybridoma supernatants added at 50 μg / well at 0 and 9 hours. Figure 11F shows a graph depicting the relative number of closed tubes in MCEC cells treated with 3E3C12, 8D2C12, 9E2H5, 2B4F9D3, or HM hybridoma supernatants. **p<0.01, ****p<0.0001 compared to HM-treated cells, one-way ANOVA. [Figure 12]Figures 12A-12F show in vivo characterization of 2B4F9D3 and 11B6 using tumor models. Figures 12A and 12C show the scheme of 11B6 or 2B4F9D3 injection with α-PD1 antibody treatment using the 1956 sarcoma line. Figures 12B and 12D show tumor growth in mice treated with PBS injection (control), α-PD1 alone, 2B4F9D3 + α-PD1, or 11B6 + α-PD1. Figure 12E shows the scheme of 11B6 or 2B4F9D3 injection with α-PD1 antibody treatment using the PyMT-BO1 orthotopic breast tumor model. Figure 12F shows tumor growth in mice treated with PBS injection (control), α-PD1 alone, 2B4F9D3 + α-PD1, or 11B6 + α-PD1. Figure 12G shows a composite line graph of tumor growth. α-PD1, 250 μg / injection / mouse, 2B4 and 11B6, 330 μg / injection / mouse. All antibodies were injected ip. DETAILED DESCRIPTION OF THE INVENTION

[0024] Both immune checkpoint inhibitors (ICIs) and antiangiogenic agents are changing the current landscape of cancer treatment. While antiangiogenic agents have demonstrated activity in highly angiogenic tumors, such as renal cell carcinoma and colorectal cancer, the effects of ICIs have primarily been seen in immunologically recognized tumors with a high concentration of immune-infiltrating lymphocytes. A major challenge in ICI drug development is shifting their activity to non-inflammatory tumors and overcoming resistance, which is partly caused by an immunosuppressive microenvironment. Angiogenic factors promote immunosuppression by directly suppressing antigen-presenting cells and immune effector cells or by enhancing the effects of regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), and tumor-associated macrophages (TAMs). These suppressive immune cells may also promote angiogenesis, creating a vicious cycle that impairs immune activation. Applicants have discovered that compositions and certain methods comprising MYCT1 reduce or prevent tumor growth, reduce or prevent tumor angiogenesis (i.e., the formation and development of the vascular system required for tumors to grow and progress), and / or increase tumor immune responses.

[0025] The MYCT1 protein appears to be dispensable for constitutive angiogenic signaling but is required for the maintenance of an immunosuppressive tumor vasculature and tumor microenvironment (Kabir et al., Sci. Transl. Med. 13, eabb6731 (2021), the contents of which are incorporated herein by reference). The present disclosure is based, in part, on the identification of an anti-MYCT1 monoclonal antibody that specifically binds to an epitope within the extracellular region of the MYCT1 protein. The MYCT1 protein exists as two isoforms in humans. One isoform has a sequence of 235 amino acids (referred to herein as isoform 1), and the other, shorter, isoform has a sequence of 187 amino acids (referred to herein as isoform 2). Amino acids 1-19 of mouse MYCT1 are identical to amino acids 48-67 of human MYCT1 isoform 1 (Figure 1A) and amino acids 1-19 of human MYCT1 isoform 2. This conserved region of MYCT1 was targeted for the generation of antibodies herein. The antibodies of the present disclosure generated using this extracellular region of MYCT1 have broad applicability, including neutralizing activity against both human and mouse MYCT1 proteins. Therefore, the present disclosure encompasses the use of compositions and methods comprising anti-MYCT1 antibodies for treating tumors or cancer in a subject. Specifically, the present disclosure demonstrates that MYCT1 expression and / or activity is required for the control of tumor angiogenesis and the reprogramming of tumor immunity. Further provided herein are compositions and methods for downregulating MYCT1 activity and / or expression in cells (e.g., endothelial cells). Downregulation of MYCT1 is associated with reduced angiogenesis, enhanced endothelial venule formation, and a promotion of an anti-tumor immune environment, limiting tumor progression. Accordingly, the present disclosure provides compositions and methods for reducing or preventing angiogenesis and increasing tumor immune responses.

[0026] Other aspects and iterations of the invention are described in more detail below.

[0027] I. Terminology In order that the present invention may be more readily understood, certain terms are defined. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the present invention pertain. Many methods and materials similar to, modified from, or equivalent to those described herein can be used in the practice of embodiments of the present invention without undue experimentation, and the preferred materials and methods are described herein. In describing and claiming embodiments of the present invention, the following terms will be used in accordance with the definitions set out below.

[0028] The terms "a" or "an" entity refer to one or more of that entity; for example, a "polypeptide subunit" is understood to refer to one or more polypeptide subunits. Thus, the terms "a" (or "an"), "one or more," and "at least one" may be used interchangeably herein.

[0029] The term "about," as used herein, refers to variations in quantity that may occur, for example, through typical measurement techniques and equipment, with respect to any quantifiable variable, including, but not limited to, mass, volume, time, distance, and amount. Furthermore, given real-world solid and liquid handling procedures, some degree of inadvertent error is expected, and variations may occur due to differences in the manufacture, source, or purity of ingredients used, for example, to make a composition or perform a method. The term "about" also encompasses these variations, which may be up to ±5%, but may also be ±4%, 3%, 2%, 1%, etc. Whether modified by the term "about," the claims include equivalents to these quantities.

[0030] The term "and / or," as used in the specification and claims, should be understood to mean "either or both" of the elements so coordinated (elements that are sometimes present conjunctively and in other cases present disjunctively). Multiple elements listed with "and / or" should be construed similarly, i.e., "one or more" of the elements so coordinated. Other elements, whether related or unrelated to the term specifically characterized, may optionally be present other than the elements specifically identified by the "and / or" clause. Thus, as a non-limiting example, a reference to "A and / or B," when used in conjunction with open-ended language such as "comprising," may, for example, in one embodiment refer to A only (including elements other than B, as appropriate); in another embodiment, refer to B only (including elements other than A, as appropriate); or in yet another embodiment, refer to both A and B (including other elements, as appropriate).

[0031] As used in the specification and claims, "or" should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" shall be construed as inclusive, i.e., including at least one but also more than one of a number or list of elements, and including additional unlisted items, as appropriate. Only in language clearly indicating the contrary, such as "only one of" or "exactly one of," or when used in the claims, "consisting of" shall refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or," when used herein, shall only be construed to indicate exclusive alternatives (i.e., "one or the other, but not both") when preceded by terms of exclusivity, such as "either," "one of," "only one of," or "exactly one of." "Consisting essentially of," when referred to in the claims, shall have its ordinary meaning as used in the field of patent law.

[0032] As used in this specification and claims, the phrase "at least one" in reference to a list of one or more elements should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, nor excluding any combination of elements in the list of elements. This definition also allows for elements other than the specifically identified elements in the list of elements to which the phrase "at least one" refers, whether related to the specifically identified elements or not, as appropriate. Thus, as a non-limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B," or, equivalently, "at least one of A and / or B" may, in one aspect, refer to, for example, at least one (including, where appropriate, multiple) A's (and, where appropriate, including elements other than B) in the absence of B; in another aspect, it may refer to at least one (including, where appropriate, multiple) B's (and, where appropriate, including elements other than A) in the absence of A; in yet another embodiment, it may refer to at least one (including, where appropriate, multiple) A's and at least one (including, where appropriate, multiple) B's (and, where appropriate, including other elements).

[0033] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Any reference to "or" herein is intended to encompass "and / or" unless expressly stated otherwise. Whenever the term "at least," "more than," or "more than or equal to" precedes the first number in a series of two or more numbers, the term "at least," "more than," or "more than or equal to" applies to each number in the series. For example, 1, 2, or 3 or more is equivalent to 1 or more, 2 or more, or 3 or more.

[0034] Whenever the first number in a series of two or more numbers is preceded by the terms "no more than," "less than," "less than or equal to," or "up to," the terms "no more than," "less than," "less than or equal to," or "up to" apply to each number in the series. For example, 3, 2, or 1 or less is equivalent to 3 or less, 2 or less, or 1 or less.

[0035] Where values are described as ranges, such disclosure is understood to include disclosure of all possible subranges within such ranges, and specific numerical values included in such ranges, whether or not a specific numerical value or specific subrange is explicitly stated.

[0036] Components used to prepare the compositions of the present disclosure, and the compositions themselves used in the methods disclosed herein. These and other materials are disclosed herein, and combinations, subsets, interactions, groups, etc. of these materials are disclosed; however, while specific reference to each of the various individual and collective combinations and permutations of these compounds may not be explicitly disclosed, it is understood that each is specifically contemplated and described herein. For example, if a particular compound is disclosed and discussed, and multiple modifications that can be made to multiple molecules of this compound are discussed, all combinations and permutations of this compound and possible modifications are specifically contemplated unless expressly stated to the contrary. Thus, if a class of molecules A, B, and C is disclosed, and a class of molecules D, E, and F, as well as an example of molecular combination A-D, each is individually and collectively contemplated, even if not individually listed, meaning that combinations AE, AF, BD, BE, BF, CD, CE, and CF are considered disclosed. Similarly, any subset or combination of these is also disclosed. Thus, for example, the subgroups AE, BF, and CE would be considered disclosed. This concept applies to all aspects of this application, including, but not limited to, steps in methods of making and using the disclosed compositions. Thus, where there are various additional steps that may be performed, it is understood that each of these additional steps may be performed with any specific aspect or combination of aspects of the disclosed methods.

[0037] As used herein, the term "non-naturally occurring" substance, composition, object, and / or any combination of substances, compositions, or objects, or any grammatical variations thereof, is a qualified term that expressly excludes, but only excludes, those forms of substance, composition, object, and / or any combination of substances, compositions, or objects that are well understood by those of ordinary skill in the art as "naturally occurring" or that have been or are at any time likely to be determined or interpreted as "naturally occurring" by a judge, or administrative body, or judicial body.

[0038] "Specifically binds" means that a binding molecule (e.g., an antibody or antigen-binding fragment thereof) binds to an epitope via its antigen-binding domain, and that this binding necessarily involves some recognition between the antigen-binding domain and the epitope. According to this definition, a binding molecule is said to "specifically bind" to an epitope if it binds to the epitope via an antigen-binding domain that it binds to more readily than it would bind to a random, unrelated epitope.

[0039] The portion of a target polypeptide that specifically interacts with the antigen-binding domain of an antibody is an "epitope." It should be noted that an "epitope" on MYCT1 may be a linear epitope or a conformational epitope, and in either case may contain non-polypeptide elements; for example, the epitope may contain carbohydrate or lipid side chains. The term "affinity" refers to a measure of the strength of binding between an individual epitope and the antigen-binding site of an antibody.

[0040] The term "antibody," as used herein, is used in the broadest sense and encompasses a variety of antibody and antibody-like structures, including, but not limited to, full-length monoclonal antibodies, polyclonal antibodies, and multispecific (e.g., bispecific, trispecific, etc.) antibodies, as well as heavy chain antibodies and antibody fragments, provided they exhibit the desired antigen-binding activity. The domain of an antibody involved in binding to an antigen is referred to as the "variable region" or "variable domain," and is described in further detail below. A single variable domain may be sufficient to confer antigen-binding specificity. Desirably, although not necessarily, antibodies useful for discovery are recombinantly produced. Antibodies may be glycosylated or non-glycosylated, although glycosylated antibodies may be preferred. An "isolated" antibody is an antibody that has been separated from components of its natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity, as determined by methods known in the art.

[0041] In addition to the antibodies described herein, it may be possible to design antibody mimetics or aptamers with substantially the same function as the antibodies of the present invention using methods known in the art. An "antibody mimetic" refers to a polypeptide or protein that can specifically bind to an antigen but is not structurally related to antibodies. Antibody mimetics have a mass of approximately 3 kDa to approximately 20 kDa. Non-limiting examples of antibody mimetics include affibody molecules, affilins, affimers, alphabodies, anticalins, avimers, DARPins, and monobodies. Aptamers are a type of small nucleic acid ligand composed of RNA or single-stranded DNA oligonucleotides and have high specificity and affinity for targets. Aptamers interact and bind to targets through structural recognition, a process similar to that of antigen-antibody reactions. Aptamers have a lower molecular weight than antibodies, typically approximately 8 to 25 kDa.

[0042] The terms "full-length antibody" and "intact antibody" may be used interchangeably and refer to antibodies having a structure substantially similar to that of a native antibody or to antibodies having heavy chains including an Fc region as defined herein. The basic structural unit of a native antibody comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" chain (approximately 25 kDa) and one "heavy" chain (approximately 50-70 kDa). Light chains are classified as gamma, mu, alpha, and lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which define the antibody's isotype as IgG, IgM, IgA, IgD, and IgE, respectively. The amino-terminal portion of each light and heavy chain contains a variable region (VL and VH, respectively) of approximately 100-110 amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function. Within light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with heavy chains also including a "D" region of about 10 more amino acids. Intact antibodies are suitably cross-linked by disulfide bonds, as is known in the art.

[0043] The variable domains of antibody heavy and light chains generally have similar structures, with each domain containing four conserved framework regions (FRs) and three hypervariable regions (HVRs). [See, for example, Kindt et al., Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007)]. A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a specific antigen can be isolated by screening a library of complementary VL or VH domains, respectively, using a VH or VL domain derived from an antibody that binds to that antigen. See, for example, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0044] "Framework region" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences generally appear in the following order: FR1-HVR1-FR2-HVR2-FR3-HVR3-FR4. The FR domains of the heavy and light chains can differ, as is known in the art.

[0045] The term "hypervariable region" or "HVR", as used herein, refers to each region of a variable domain that is hypervariable in sequence (also commonly referred to as "complementarity-determining region" or "CDR") and / or forms structurally defined loops (hypervariable loops) and / or contains antigen-contacting residues ("antigen contacts"). Generally, antibodies contain six HVRs: three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). As used herein, an "HVR from a variable region" refers to an HVR that has no more than two amino acid substitutions compared to the corresponding HVR from the original variable region. Exemplary HVRs herein include: (a) hypervariable loops occurring at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) [Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)]; (b) CDRs occurring at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) [Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)]; (c) antigenic contacts occurring at amino acid residues 27c–36 (L1), 46–55 (L2), 89–96 (L3), 30–35b (H1), 47–58 (H2), and 93–101 (H3) [MacCallum et al. J. Mol. Biol.262: 732-745 (1996)]; (d) the CDR1-IMGT region (positions 27-38), the CDR2-IMGT region (positions 56-65), and the CDR3-IMGT region (positions 105-116 or 105-117), which are numbered according to the IMGT unique numbering [Lefranc, “The IMGT unique numbering for immunoglobulins, T cell receptors, and Ig-like domains,” The Immunologist, 1999, 7: 132-136; Lefranc et al., Nucleic Acids Research, 2009, 37(Database issue): D1006-D1012; Ehrenmann et al., “Chapter 2: Standardized Sequence and Structure Analysis of Antibody Using IMGT,” in Antibody Engineering Volume 2, Eds. Roland E. Kontermann and Stefan Dubel, 2010, Springer-Verlag Berlin Heidelberg, doi: 10.1007 / 978-3-642-01147-4; imgt.org / IMGTScientificChart / Nomenclature / IMGT-FRCDRdefinition], and (e) combinations of (a), (b), (c), and / or (d), as defined below for various antibodies of the disclosure. Unless otherwise specified, HVR residues and other residues (e.g., FR residues) within variable domains that are assigned sequence identification numbers are numbered according to the IMGT unique numbering system (see above).

[0046] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain comprising at least a portion of the constant region. This term includes native sequence Fc regions and variant Fc regions. In one aspect, a human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, numbering of amino acid residues within the Fc region or constant region follows the EU numbering system (also referred to as the EU index) as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

[0047] A "variant Fc region" comprises an amino acid sequence that may differ from that of a native Fc region by one or more amino acid substitutions and / or an altered glycosylation pattern, compared to a native Fc region or the Fc region of a parent polypeptide. In one example, a variant Fc region may have from about one to about ten amino acid substitutions, or from about one to about five amino acid substitutions, in a native-sequence Fc region or the Fc region of a parent polypeptide. A variant Fc region herein may have at least about 80% homology, at least about 90% homology, or at least about 95% homology to a native-sequence Fc region and / or the Fc region of a parent polypeptide.

[0048] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Non-limiting examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; single-chain forms of antibodies and higher variants thereof; single-domain antibodies; and multispecific antibodies formed from antibody fragments.

[0049] Single-chain forms of antibodies and their higher-order forms may include, but are not limited to, single-domain antibodies, single-chain variant fragments (scFvs), bivalent scFvs (di-scFvs), trivalent scFvs (tri-scFvs), tetravalent scFvs (tetra-scFvs), diabodies, triabodies, and tetrabodies. ScFvs consist of a heavy chain variable region and a light chain variable region connected by a linker. In most (but not all) cases, the linker can be a peptide. The linker peptide is preferably 5 to 30 amino acids long, or 10 to 25 amino acids long. Typically, the linker allows for stabilization of the variable domains without interfering with the proper folding and formation of the active binding site. In a preferred embodiment, the linker peptide is rich in glycine and also rich in serine or threonine. ScFvs can be used to facilitate phage display, or they can be used in flow cytometry, immunohistochemistry, or as targeting domains. Methods for producing and using scFvs are known in the art. ScFvs can also be conjugated to human constant domains (e.g., heavy constant domains derived from IgG domains (e.g., IgG1, IgG2, IgG3, or IgG4), or heavy chain constant domains derived from IgA, IgM, or IgE). Diabodies, triabodies, and tetrabodies, as well as higher-order variants, are typically created by varying the length of the linker peptide from zero to several amino acids. Alternatively, it is known in the art that multivalent binding antibody variants can be generated using self-assembling units linked to variable domains.

[0050] A "single domain antibody" refers to an antibody fragment consisting of a single monomeric variable antibody domain.

[0051] Multispecific antibodies include bispecific, trispecific, or four or more specific antibodies. Multispecific antibodies can be made by combining the heavy and light chains of one antibody with the heavy and light chains of one or more other antibodies. The chains can be covalently linked.

[0052] The antibody of the present disclosure may be a Dual-Affinity Re-targeting Antibody (DART). The DART format is based on the diabody format, which separates the heavy and light chain cognate variable domains of two antigen-binding specificities on two separate polypeptide chains. While the diabody format involves non-covalent association of the two polypeptide chains, the DART format provides additional stabilization through a C-terminal disulfide bridge. DARTs can be manufactured in large quantities and with high quality, and they exhibit excellent stability in both formulation buffers and human serum.

[0053] As used herein, a "monoclonal antibody" refers to an antibody derived from a single copy or clone, such as, for example, any eukaryotic, prokaryotic, or phage clone. A "monoclonal antibody" is not limited to antibodies produced by hybridoma technology. Monoclonal antibodies may be produced using hybridoma technology known in the art, as well as recombinant, phage display, and synthetic technologies, or a combination of such technologies with other technologies readily known in the art. Furthermore, monoclonal antibodies may be labeled with a detectable label, immobilized on a solid phase, and / or conjugated to a heterologous compound (e.g., an enzyme or toxin) according to methods known in the art.

[0054] As used herein, "heavy chain antibody" refers to an antibody consisting of two heavy chains. The heavy chain antibody may be an IgG-like antibody derived from camel, llama, alpaca, shark, etc., or may be a traditional IgNAR from cartilaginous fish.

[0055] As used herein, a "humanized antibody" refers to a non-human antibody that has been modified to reduce the risk of eliciting an immune response in humans after administration, while retaining the same binding specificity and affinity as the original non-human antibody. A humanized antibody binds to the same or similar epitope as a non-human antibody. The term "humanized antibody" includes antibodies composed partially or completely of amino acid sequences derived from human antibody germlines by altering the sequence of an antibody with non-human hypervariable regions ("HVRs"). The simplest such alteration may consist solely of replacing the constant regions of a human antibody with mouse constant regions, resulting in a human / mouse chimera that may have sufficiently low immunogenicity to be acceptable for pharmaceutical use. Preferably, the variable regions of the antibody are also humanized using techniques currently known in the art. For example, the framework regions of the variable regions may be replaced with corresponding human framework regions while retaining one, some, or all six non-human HVRs. Some framework residues may be replaced with corresponding residues from a non-human VL or VH domain (e.g., the non-human antibody from which the HVR residues are derived) to, for example, restore or improve the specificity or affinity of the humanized antibody. A substantially human framework region has at least about 75% homology (i.e., at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% sequence identity) with a known human framework sequence. HVRs may also be randomly mutated such that binding activity and affinity for the antigen are maintained or enhanced relative to fully human germline framework regions or substantially human framework regions. As noted above, it is sufficient to utilize antibody fragments for use in the discovery methods. Furthermore, as used herein, the term "humanized antibody" refers to an antibody comprising substantially human framework regions, at least one HVR derived from a non-human antibody, and any constant regions present are substantially human. A substantially human constant region is at least about 90% identical to a known human constant sequence (ie, about 90%, about 95%, or about 99% sequence identity).Hence, all parts of a humanized antibody (except possibly the HVRs) are substantially identical to corresponding pairs of one or more germline human immunoglobulin sequences.

[0056] If desired, humanized immunoglobulin design can be performed as described below or using similar methods known to those skilled in the art (see, for example, Almagro, et al. Front. Biosci. 2008, 13(5):1619-33). The variable regions of the mouse antibody are aligned with the most similar human germline sequence (e.g., using BLAST or a similar algorithm). CDR residues from the mouse antibody sequence are grafted onto the similar human "acceptor" germline sequence. Subsequently, to achieve a humanized antibody with binding affinity similar to that of the original mouse antibody, one or more positions near the CDRs or within the framework (e.g., Vernier positions) can be reverted to the original mouse amino acid. Typically, several versions of the humanized antibody with various backmutations are generated and experimentally tested for activity. The humanized antibody variant with properties most similar to the parent mouse antibody and with the least reversion of the mouse framework is selected as the final humanized antibody candidate.

[0057] The term "specifically binds," as used herein with respect to an epitope binding agent, means that the epitope binding agent does not cross-react to any significant extent with other epitopes on the protein of interest (e.g., MYCT1) or on other proteins in general.

[0058] The terms "Myc target 1," "MYC target protein 1," "MTLC," "Myc target in myeloid cells protein 1," or "MYCT1" encompass all MYCT isoforms and orthologs, whether full-length, truncated, or post-translationally modified. In many animals (e.g., but not limited to, humans, non-human primates, rodents, fish, cows, frogs, goats, and chickens), MYCT1 is encoded by the Myct1 gene (also known as FLJ21269 and MTLC). In humans, the gene encoding MYCT1 is located on chromosome 6 (band q25.2; chromosomal position (bp) 152697895-152724567). MYCT1 was first identified as a novel target of the c-Myc oncogene in myeloid cells. Previous reports suggest that overexpression of MYCT1 in cancer cells may promote apoptosis, morphological changes, enhanced anchorage-independent cell growth, tumorigenic transformation, genomic instability, and inhibition of hematopoietic differentiation. MYCT1 was originally thought to be a transcription factor that binds to the promoters of several c-Myc-regulated genes; however, it has been suggested that the phenotypes observed in MYCT1-overexpressing cells result from deregulation of these genes. In humans, the MYCT1 protein exists in two isoforms: isoform 1, which has a sequence of 235 amino acids, and isoform 2, which is shorter and has a sequence of 187 amino acids. Amino acids 1–19 of mouse MYCT1 are identical to amino acids 48–67 of human MYCT1 isoform 1 (Figure 1A) and amino acids 1–19 of human MYCT1 isoform 2.

[0059] In an exemplary embodiment, the full-length MYCT1 polypeptide human isoform 1 comprises the amino acid sequence of SEQ ID NO: 145 below. MRTQVYEGLCKNYFSLAVLQRDRIKLLFFDILVFLSVFLLFLFLVDIMANNTTSLGSPWPENFWEDLIMSFTVSMAIGLVLGGFIWAVFICLSRRRRASAPISQWSSSRRSRSSYT HGLNRTGFYRHSGCERRSNLSLASLTFQRQASLEQANSFPRKSSFRASTFHPFLQCPPLPVETESQLVTLPSSNISPTISTSHSLSRPDYWSSNSLRVGLSTPPPPAYESIIKAFPDS

[0060] In some embodiments, the MYCT1 polypeptide human isoform 2 comprises the amino acid sequence of SEQ ID NO: 148 below. MANNTTSLGSPWPENFWEDLIMSFTVSMAIGLVLGGFIWAVFICLSRRRRASAPISQWSSSRRSRSSYTHGLNRTGFYRHSGCERRSNLSLASLTFQRQASLEQANSFPRKSSFRASTFHPFLQCPPLPVETESQLVTLPSSNISPTISTSHSLSRPDYWSSNSLRVGLSTPPPPAYESIIKAFPDS

[0061] In some embodiments, the human MYCT1 polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 149 below. ATGCGAACACAAGTATATGAGGGGTTGTGTAAAAAATTATTTTTCTCTTGCTGTACTACAAAGAGATAGAATCAAACTGCTTTTTTTCGACATACTGGTTTTCTTTCTGTTTTTCTTCTCTTTCTTCTATTTCTTGGATATTATGGCTAATAACACAACAAGTTTAGGGAGTCCA TGGCCAGAAAACTTTTGGGAGGACCTTATCATGTCCTTCACTGTATCCATGGCAATCGGGCTGGTACTTGGAGGATTTATTTGGGCTGTGTTCATTTGTCTGTCTCGAAGAAGAAGAGCCAGTGCTCCCATCTCACAGTGGAGTTCAAGCAGGAGATCTAGGTCTTCTTACACCCAC GGCCTCAACAGAACTGGATTTTACCGCCACAGTGGCTGTAACGTCGAAGCAACCTCAGCCTGGCCAGTCTCACCTTCCAGCGACAAGCTTCCCTGGAACAAGCAAATTCCTTTCCAAGAAAATCAAGTTTCAGAGCTTCTACTTTCCATCCCTTTCTGCAATGTCCACCACTTCCT GTGGAAACTGAGAGTCAGCTGGTGACTCTCCCTTCTTCCAATATCTCTCCCACCATCAGCACTTCCCACAGTCTGAGCCGTCCTGACTACTGGTCCAGTACAGTCTTCGAGTGGGCCTTTCAACACCGCCCCACCTGCCTATGAGTCCATCATCAAGGCATCCCCAGATTCCTGA

[0062] In some embodiments, the mouse MYCT1 polypeptide contains the following sequence number 150. MANNTTSLGSPWPENFWEDLIMSFTVSVAIGLAIGGFLWALFVFLSRRRRASAPISQWSPTRRPRSSYNHGLNRTGFYRHSGYERRSNLSLASLTFQRQASMELVNSFPRKSSFRASTFHPFLQCPPLVETESQLMTLSASTTPSTLSTAHSPSPRPDFRWSSNSLRMGLSTPPPPAYESIIKAFPDS

[0063] In exemplary embodiments, the full-length MYCT1 mRNA transcript (3030 base pairs in length) includes the NCBI reference sequence: NM_025107.3. Additional reference MYCT1 mRNAs include, but are not limited to, NM_001371624.1, NM_001371625.1, NM_001371626.1, and NM_025107.3. In exemplary embodiments, regulatory elements regulating the expression of MYCT1 include those described herein and those disclosed in Gene Card ID: GC06P152697.

[0064] The "percent identity" of two amino acid sequences can be determined using the algorithm of Karlin and Altschul Proc. Natl. Acad. Sci. USA 87:2264-68, 1990, as modified by Karlin and Altschul Proc. Natl. Acad. Sci. USA 90:5873-77, 1993. Such an algorithm has been incorporated into the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. J. Mol. Biol. 215:403-10, 1990. BLAST protein searches can be performed with the XBLAST program (score=50, wordlength=3) to obtain amino acid sequences homologous to a protein molecule of interest. When gaps exist between the two sequences, Gapped BLAST can be utilized as described in Altschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0065] As used herein, "nucleic acid," "nucleic acid molecule," and "polynucleotide" are used interchangeably herein. The terms "nucleic acid encoding" or "nucleic acid molecule encoding" should be understood to refer to a sequence of nucleotides that encodes a polypeptide.

[0066] A polynucleotide described herein can include one or more nucleic acids, each encoding a polypeptide, all of which are operably linked (i.e., in a functional relationship) to one or more regulatory sequences, such as a promoter. Such a polynucleotide can alternatively be referred to herein as a "nucleic acid construct" or "construct."

[0067] As used herein, "polypeptide" and "protein" are used interchangeably to refer to a polymer of amino acid residues.

[0068] Each amino acid sequence described herein is expressed as a percentage of identity or similarity to a given amino acid sequence, respectively, and in more preferred embodiments, is at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75% with the given nucleotide or amino acid sequence, respectively. , at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity or similarity. The terms "homology," "sequence identity," and the like are used interchangeably herein. Sequence identity is described herein as the relationship between two or more amino acid (polypeptide or protein) sequences or two or more nucleic acid (polynucleotide) sequences, as determined by comparing the sequences. In preferred embodiments, sequence identity is calculated based on the full length (amino acid or nucleotide) of two given SEQ ID NOs, or based on a portion thereof. A portion of a full-length sequence may be referred to as a fragment, and preferably refers to at least 50%, 60%, 70%, 80%, 90%, or 100% of the length (amino acids or nucleotides) of the reference sequence. "Identity" also refers to the degree of sequence relatedness between two amino acid sequences or two nucleic acid sequences, as the case may be, as determined by the match between strings of such sequences. The degree of sequence identity between two sequences can be determined, for example, by comparing the two sequences using a computer program commonly employed for this purpose, such as a global alignment algorithm or a local alignment algorithm.Non-limiting examples include BLASTp, BLASTn, Clustal W, MAFFT, Clustal Omega, AlignMe, Praline, GAP, BESTFIT, or another suitable method or algorithm. The Needleman and Wunsch global alignment algorithm can be used to align two sequences over their entire length or over a portion (where a portion can mean at least 50%, 60%, 70%, 80%, 90% of the length of the sequences) to maximize the number of matches and minimize the number of gaps. Default settings may be used, with preferred programs being Needle for pairwise alignments (in one embodiment, EMBOSS Needle 6.6.0.0, gap open penalty 10, gap extent penalty: 0.5, end gap penalty: false, end gap open penalty: 10, end gap extent penalty: 0.5 are used) and MAFFT for multiple sequence alignments (in one embodiment, MAFFT v7 default values are BLOSUM62 [bl62], gap open: 1.53, gap extension: 0.123, order: aligned, number of tree rebuilds: 2, guide tree output: ON [true], maximum iterations: 2, FFTS enforcement: not used).

[0069] "Similarity" between two amino acid sequences is determined, for example, by comparing the amino acid sequence and its conserved amino acid substitutes of one polypeptide with the sequence of another polypeptide. Similar algorithms used to determine sequence identity can be used to determine sequence similarity. Where appropriate, in determining the degree of amino acid similarity, those skilled in the art may also take into account so-called conservative amino acid substitutions. As used herein, "conservative" amino acid substitutions refer to the interchangeability of residues with similar side chains. Examples of classes of amino acid residues for conservative substitutions are shown in the table below:

[0070] TIFF2025525447000002.tif40155

[0071] Alternative conservative amino acid residue substitution classes: TIFF2025525447000003.tif39155

[0072] Alternative physical and functional classifications of amino acid residues: TIFF2025525447000004.tif68158

[0073] For example, the group of amino acids having aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; the group of amino acids having aliphatic hydroxyl side chains is serine and threonine; the group of amino acids having amide-containing side chains is asparagine and glutamine; the group of amino acids having aromatic side chains is phenylalanine, tyrosine, and tryptophan; the group of amino acids having basic side chains is lysine, arginine, and histidine; and the group of amino acids having sulfur-containing side chains is cysteine and methionine. Preferred conservative amino acid substitution groups are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, and asparagine-glutamine. Substitution variants of the amino acid sequences disclosed herein are those in which at least one residue in the disclosed sequence has been removed and a different residue inserted in its place. Preferably, the amino acid changes are conservative. Preferred conservative substitutions for each of the naturally occurring amino acids are as follows: Ala to Ser; Arg to Lys; Asn to Gln or His; Asp to Glu; Cys to Ser or Ala; Gln to Asn; Glu to Asp; Gly to Pro; His to Asn or Gln; Ile to Leu or Val; Leu to Ile or Val; Lys to Arg; Gln or Glu; Met to Leu or Ile; Phe to Met, Leu, or Tyr; Ser to Thr; Thr to Ser; Trp to Tyr; Tyr to Trp or Phe; and Val to Ile or Leu.

[0074] As used herein, "fragment" refers to a portion of a polypeptide that preferably comprises at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more of the full length of the reference polypeptide. In certain embodiments, the fragment has the same or a similar biological activity as the reference polypeptide.

[0075] The terms "treat," "treating," or "treatment," as used herein, refer to both therapeutic treatment and prophylactic or preventative measures, where the objective is to prevent or delay (alleviate) undesirable physiological changes or diseases / disorders. Beneficial or desired clinical results include, but are not limited to, the alleviation of symptoms, reduction in extent of disease, stable (i.e., not worsening) state of disease, delay or slowing of disease progression, improvement or palliation of disease state, and remission (partial or complete), whether detectable or undetectable. "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the disease, condition, or disorder, as well as those prone to have the disease, condition, or disorder, or those in whom the disease, condition, or disorder is to be prevented.

[0076] As used herein, "cancer," "tumor," or "malignant tumor" may refer to one or more neoplasms or cancers. Neoplasms can be malignant or benign, cancers can be primary or metastatic, and neoplasms or cancers can be early stage or late stage. Non-limiting examples of neoplasms or cancers can include acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal cancer, appendix cancer, astrocytoma (pediatric cerebellar or cerebral), basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumors (cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma), breast cancer, bronchial adenoma / carcinoid, Burkitt's lymphoma, calcitonin, leukemia ... Tinoid tumor (childhood, gastrointestinal), cancer of unknown primary, central nervous system lymphoma (primary), cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disorder, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, Ewing sarcoma in Ewing family tumors, extracranial germ cell tumor (childhood), extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer (intraocular melanoma, retinoblastoma), gallbladder cancer, stomach Cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, germ cell tumor (pediatric extracranial, extragonadal, ovarian), gestational trophoblastic tumor, glioma (adult, pediatric brainstem, pediatric cerebral astrocytoma, pediatric visual pathway and hypothalamus), gastric carcinoid, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, Hodgkin's lymphoma, hypopharyngeal cancer, hypothalamic and visual pathway glioma (pediatric), intraocular melanoma, pancreatic islet cell carcinoma, Kaposi's sarcoma, kidney cancer (renal cell carcinoma), laryngeal cancer, leukemia (acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myeloid) , hairy cell), lip and oral cancer, liver cancer (primary), lung cancer (non-small cell, small cell), lymphoma (AIDS-related, Burkitt's, cutaneous T-cell, Hodgkin's, non-Hodgkin's, primary central nervous system), macroglobulinemia (Waldenstrom), malignant fibrous histiocytoma / osteosarcoma of bone, medulloblastoma (childhood), melanoma, intraocular melanoma, Merkel cell carcinoma, mesothelioma (adult malignant, childhood), metastatic squamous cell carcinoma of the neck of unknown primary, oral cancer, multiple endocrine neoplasia syndrome (childhood), multiple melanoma / plasmacytoma,Mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative disorders, myeloid leukemia (chronic), myeloid leukemia (acute adult, acute childhood), multiple myeloma, myeloproliferative disorders (chronic), nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer (superficial epithelial-stromal tumor), ovarian germ cell tumor , ovarian low malignant potential tumor, pancreatic cancer, pancreatic cancer (islet cell), paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germinoma, pineoblastoma and supratentorial primitive neuroectodermal tumor (childhood), pituitary adenoma, plasma cell tumor, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma (kidney cancer), renal pelvis and ureteral transitional cell carcinoma, retinoblastoma, Rhabdomyosarcoma (childhood), salivary gland cancer, sarcoma (Ewing family tumors, Kaposi's, soft tissue, uterine), Sézary syndrome, skin cancer (non-melanoma, melanoma), skin tumor (Merkel cell), small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell carcinoma of the cervix of unknown primary (metastatic), gastric cancer, supratentorial primitive neuroectodermal tumor (childhood), T-cell lymphoma (skin), testicular cancer, pharyngeal cancer , thymoma (childhood), thymoma and thymic carcinoma, thyroid cancer, thyroid cancer (childhood), transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor (gestational), unknown primary site (adult, child), transitional cell carcinoma of the ureter and renal pelvis, urethral cancer, uterine cancer (endometrium), uterine sarcoma, vaginal cancer, visual pathway and hypothalamic glioma (childhood), vulvar cancer, Waldenstrom's macroglobulinemia, and Wilms' tumor (childhood).

[0077] As used herein, treating cancer can include increasing inhibition of cancer progression and / or metastasis, inhibiting tumor volume increase, decreasing tumor volume and / or growth, slowing tumor growth rate, eradicating or killing tumor and / or cancer cells, or any combination thereof. In some embodiments, the treatment can also extend the subject's survival, improve the prognosis, and / or improve the subject's quality of life.

[0078] As used herein, "resistant" or "insensitive" cancers, tumor cells, and tumors refer to cancers that have become resistant to cancer treatments (e.g., immune checkpoint inhibitors) and may lead to increased tumor volume, cancer metastasis, cancer recurrence, or reduced life expectancy.

[0079] The subject may be a rodent, a human, a livestock animal, a companion animal, or a zoo animal. In one aspect, the subject may be a rodent, such as a mouse, rat, guinea pig, etc. In another aspect, the subject may be a livestock animal. Non-limiting examples of suitable livestock animals may include pigs, cows, horses, goats, sheep, llamas, and alpacas. In yet another aspect, the subject may be a companion animal. Non-limiting examples of companion animals may include pets such as dogs, cats, rabbits, and birds. In yet another aspect, the subject may be a zoo animal. As used herein, "zoo animal" refers to an animal that may be found in a zoo. Such animals may include non-human primates, big cats, wolves, and bears. In a preferred aspect, the subject is a human.

[0080] As used herein, a "control sample" or "control cells" may be obtained from a healthy subject and / or from a subject with cancer obtained before the start of treatment (baseline). A control subject may be a healthy subject or a subject not receiving treatment. In some embodiments, a parameter measured during treatment may be the average of several control subjects or may be the population average. In some embodiments, a control sample may include non-cancerous cells. In some embodiments, the non-cancerous cells may be derived from the same tissue type as the cancer cells. For example, if the cancer cells are from breast cancer, the non-cancerous cells may be derived from healthy breast tissue. In some embodiments, a control may consist of the average level of an analyte in a sample from a subject before the onset of cancer. In some embodiments, a control sample may be a sample from a subject prior to diagnosis or treatment. In certain embodiments, an analyte may be measured in a person other than the subject with cancer. In some embodiments, a control may be a person with characteristics similar to the subject with cancer. In some embodiments, the control can be an average of a combination of analyte levels of the present disclosure from various healthy sources (e.g., multiple healthy control subjects, and / or multiple subjects prior to the start of treatment (baseline)). In some embodiments, the control sample can be a pooled sample.

[0081] As used herein, a biological sample can be any biological tissue, body fluid, or cell from a subject. The sample can be solid or liquid. The sample can be a heterogeneous cell population. Non-limiting examples of suitable biological samples include sputum, serum, blood, blood cells (e.g., white blood cells), biopsy, urine, ascites, pleural effusion, or cells derived therefrom. The biopsy can be fine needle aspiration biopsy, core needle biopsy, vacuum-assisted biopsy, surgical biopsy, shave biopsy, punch biopsy, incision biopsy, scraping biopsy, or deep shave biopsy. Biological samples can also include tissue sections, such as frozen sections or formalin-fixed sections taken for histological purposes. The sample can be tumor tissue, peritumoral tissue, or non-tumor tissue. Methods for obtaining biological samples from subjects are known in the art.

[0082] As used herein, the term "vector" is understood as a nucleic acid means comprising a nucleotide sequence that can be introduced into a host cell and recombined and inserted into the genome of the host cell, or can replicate autonomously as an episome. Vectors may include linear nucleic acids, plasmids, phagemids, cosmids, RNA vectors, viral vectors, and analogs thereof. Examples of viral vectors may include, but are not limited to, retroviruses, adenoviruses, and adeno-associated viruses.

[0083] As used herein, "immune checkpoint inhibitors" or "ICIs" refer to drugs that block immune checkpoints. These checkpoints are a normal part of the immune system and prevent overly aggressive immune responses. By blocking them, these drugs allow immune cells to respond more aggressively to cancer and other conditions. Immune checkpoint inhibitors work by preventing cancer cells from turning off T cells (white blood cells that detect infection and abnormalities). Non-limiting examples of immune checkpoint inhibitors include inhibitors of PD-1, PD-L1, TIM-3, LAG-3, CTLA-4, and CSF-1R, as well as any combination thereof. Immune checkpoint receptors can be present on tumor cells or on immune cells such as T cells, monocytes, microglia, and macrophages, but are not limited to these. Agents that perform immune checkpoint blockade can be small chemicals or polymers, antibodies, antibody fragments, single-chain antibodies, or other antibody constructs (e.g., but not limited to, bispecific antibodies and diabodies). Immune checkpoint inhibitors that can be used according to the present disclosure include those that disrupt the inhibitory interaction between cytotoxic T cells and tumor cells. These include, but are not limited to, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA4 antibodies, anti-LAG-3 antibodies, and anti-TIM-3 antibodies. The inhibitor need not be an antibody, but can be a small molecule or other polymer. If the inhibitor is an antibody, it can be polyclonal, monoclonal, fragment, single-chain, or other antibody variant construct.The inhibitor may target any immune checkpoint known in the art, including, but not limited to, CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, CD28, CD86, CD69, CD48, CD113, CEACAM-1, Galectin-1, TIGIT, GPR56, CD48, GARP, PD1H, LAIR1, TIM1, TIM4, and the B-7 family of ligands. Combinations of inhibitors directed against a single target immune checkpoint or combinations of different inhibitors directed against different immune checkpoints may be used. Specific examples of immune checkpoint inhibitors include CTLA-4 blocking antibodies [ipilimumab (Yervoy), tremelimumab (Imjuno)], PD-1 inhibitors [pembrolizumab (Keytruda), nivolumab (Opdivo), cemiplimab (Libtayo), CT-011 (pidilizumab), AMP224], PD-L1 inhibitors [atezolizumab (tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), BMS-936559], Lag3 inhibitors (relatulimab), and combinations of Lag3 and PD1 inhibitors [PD-1 inhibitor nivolumab (Opdualag), OX40 inhibitors (MEDI6469), CD160 inhibitors (BY55)]. Non-limiting examples of inhibitors of CSF-1R include PLX3397, PLX486, RG7155, AMG820, ARRY-382, FPA008, IMC-CS4, JNJ-40346527, and MCS 110. The terms "ICI treatment," "ICI therapy," "ICI compound," and the like refer to one or more ICIs (or uses thereof) disclosed herein or known to those of skill in the art.

[0084] As used herein, "angiogenesis" refers to the growth of blood vessels from pre-existing vasculature. Angiogenesis, as used herein, may be further defined as involving: (i) endothelial cell activation; (ii) increased vascular permeability; (iii) the formation of a provisional fibrin gel extracellular matrix followed by basement membrane dissolution and extravasation of plasma components; (iv) endothelial cell proliferation and recruitment; (v) the remodeling of recruited endothelial cells to form functional capillaries; (vi) the formation of capillary loops; (vi) the deposition of basement membranes and recruitment of perivascular cells to newly formed blood vessels, or any combination thereof. Normal angiogenesis is active during tissue growth from embryonic development to maturity, followed by a relatively quiescent period in adulthood. Normal angiogenesis is also activated during wound healing or at certain stages of the female reproductive cycle. Inappropriate or pathological angiogenesis has been associated with several disease states, such as various retinopathies, ischemic diseases, atherosclerosis, chronic inflammatory disorders, and cancer. Pathological angiogenesis can be the formation or neovascularization of blood vessels that are not physiologically normal. Such vessels may be immature and leaky, resulting in vessels that cannot be perfused with blood. Similarly, pathological angiogenesis can also be angiogenesis or neovascularization in a manner that differs from physiologically normal. For example, the term can refer to the formation of new blood vessels in response to stimuli such as inflammation.

[0085] As used herein, "angiogenesis-related diseases or disorders" refers to any disease or disorder that results in the aforementioned "pathological angiogenesis or neovascularization" or causes dysfunction in an individual due to "pathological angiogenesis or neovascularization." Non-limiting examples of such diseases or disorders include age-related macular degeneration, ischemic retinopathy, intraocular neovascularization, corneal neovascularization, retinal neovascularization, choroidal neovascularization, diabetic macular edema, diabetic retinal ischemia, diabetic retinal edema, diabetic retinopathy, cancer, rheumatoid arthritis, endometriosis, and alopecia. In some embodiments, the disease or disorder is cancer.

[0086] As used herein, a "VEGF inhibitor" describes an inhibitor of VEGF synthesis or activity, as the case may be, at the protein level, to prevent or downregulate the production of the protein or at least one biological activity of the produced protein. In some embodiments, the VEGF inhibitor may be selected from the group consisting of an anti-VEGF immunoglobulin, an anti-VEGF antibody, an anti-VEGF monoclonal antibody, and a humanized anti-VEGF monoclonal antibody.

[0087] As used herein, a "formulation" refers to a preparation of a drug in a form suitable for administration to a subject, such as a human. As such, a "formulation" can include a pharmaceutically acceptable excipient, such as a diluent or carrier.

[0088] As used herein, "pharmaceutically acceptable" describes a substance or component that does not cause an unacceptable loss of pharmacological activity or unacceptable adverse side effects. Examples of pharmaceutically acceptable components may be those listed in the United States Pharmacopeia (USP 29) and National Formulary (NF 24), United States Pharmacopeial Convention, Inc., Rockville, Maryland, 2005 ("USP / NF"), or those bearing the monogram in more recent editions, as well as those listed in the FDA's continually updated Inactive Ingredient Search online database. Other useful components not described in USP / NF, etc., may also be used.

[0089] As used herein, "pharmaceutically acceptable excipients" may include any solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, or absorption delaying agents. The use of such media and agents for pharmaceutical active substances is well known in the art [see generally, Remington's Pharmaceutical Sciences (AR Gennaro, Ed.), 21st edition, ISBN: 0781746736 (2005)]. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. The compositions may also incorporate supplementary active ingredients.

[0090] As used herein, a "stable" formulation or composition can refer to a composition that has sufficient stability to permit storage at a convenient temperature, e.g., from about 0°C to about 60°C, for a commercially reasonable period of time, e.g., at least about 1 day, at least about 1 week, at least about 1 month, at least about 3 months, at least about 6 months, at least about 1 year, or at least about 2 years.

[0091] As used herein, "expression," or "expression level," or "level of expression" refers to the amount of a particular analyte (e.g., MYCT1) present in a sample. The amount can be a concentration, number, ratio, proportion, or percentage of the analyte compared to a control sample or determined using a standard curve. The amount can be an absolute amount or a relative amount.

[0092] As used herein, "combination therapy" refers to the administration of two or more therapeutic agents to treat a therapeutic condition or disorder described in this disclosure. Such administration encompasses the co-administration of these therapeutic agents substantially simultaneously, such as in a single capsule having a fixed ratio of active ingredients, or in multiple separate capsules, one for each active ingredient. In addition, such administration also encompasses the sequential use of various therapeutic agents. In either case, the treatment regimen provides the beneficial effects of the drug combination in treating the condition or disorder described herein.

[0093] As used herein, "therapeutically effective" or "effective" is intended to qualify the amount of active ingredient used in the treatment of a disease or disorder or the effect of a clinical endpoint. An effective amount can be an amount that achieves the intended purpose. For example, an effective amount of an antibody can be an amount that reduces tumor volume (e.g., an amount that shrinks or eliminates a tumor, or an amount that reduces the number of circulating cancer cells). Determining an effective amount is well within the capabilities of one of ordinary skill in the art, especially in light of the present disclosure provided herein.

[0094] II. Anti-MYCT1 antibody The present disclosure provides antibodies that specifically bind to MYCT1 protein. In some embodiments, the antibodies of the present disclosure bind to an epitope of the MYCT1 protein. In some embodiments, the antibodies described herein can be isolated antibodies that bind to an epitope of the MYCT1 protein. In some embodiments, the isolated antibodies herein can bind to an epitope of the MYCT1 protein, where the MYCT1 protein is human MYCT1 protein. In some embodiments, the isolated antibodies herein can bind to an epitope of the MYCT1 protein, where the MYCT1 protein is mouse MYCT1 protein.

[0095] In certain embodiments, provided herein are antibodies that bind to the extracellular domain of MYCT1. In some embodiments, provided herein are antibodies that specifically bind to an epitope within the first 150 amino acids of the MYCT1 protein. In some embodiments, the antibody binds to an epitope within the first 100 amino acids of MYCT1. In some embodiments, the antibody binds to an epitope within the first 70 amino acids of MYCT1. In some embodiments, the antibody binds to an epitope within the first 67 amino acids of MYCT1 comprising the amino acid sequence of SEQ ID NO: 145. In some embodiments, the antibody binds to an epitope within the first 30 amino acids of MYCT1 comprising the amino acid sequence of SEQ ID NO: 148 or SEQ ID NO: 150. In some embodiments, the antibody binds to an epitope within the first 20 amino acids of MYCT1 comprising the amino acid sequence of SEQ ID NO: 148 or SEQ ID NO: 150. In some embodiments, the epitope comprises the amino acid sequence of SEQ ID NO: 146.

[0096] In certain aspects, the antibodies described herein may have suitable binding affinity for a target antigen (e.g., a MYCT1 protein or epitope thereof). As used herein, "binding affinity" refers to the apparent association constant or K A K A is the dissociation constant (K D In some embodiments, the antibodies described herein have a binding affinity (K) for a MYCT1 protein or epitope thereof of at least about 100 nM, at least about 10 nM, at least about 1 nM, at least about 0.1 nM, or lower. D In some embodiments, the antibodies described herein may have a binding affinity (K) of about 0.1 pM to about 10 μM, preferably about 0.1 pM to about 1 μM, and more preferably about 0.1 pM to about 100 nM. DIn some embodiments, the antibodies described herein may have a binding affinity (K) of about 100 nM to about 0.1 nM (e.g., about 100 nM, about 75 nM, about 50 nM, about 25 nM, about 10 nM, about 5 nM, about 1 nM, about 0.75 nM, about 0.5 nM, about 0.25 nM, about 0.1 nM) to a MYCT1 protein or epitope thereof. D In some embodiments, the antibodies described herein may have a binding affinity (K) of about 50 nM to about 40 nM (e.g., about 50 nM, about 49 nM, about 48 nM, about 47 nM, about 46 nM, about 45 nM, about 44 nM, about 43 nM, about 42 nM, about 41 nM, about 40 nM) to a MYCT1 protein or epitope thereof. D In some embodiments, the antibodies described herein may have a binding affinity (K) of about 50 nM to about 40 nM (e.g., about 50 nM, about 49 nM, about 48 nM, about 47 nM, about 46 nM, about 45 nM, about 44 nM, about 43 nM, about 42 nM, about 41 nM, about 40 nM) to a MYCT1 protein or epitope thereof. D In some embodiments, binding affinity (or binding specificity) may be determined by various methods, such as equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance, and / or spectroscopy (e.g., using a fluorescence assay).

[0097] In certain embodiments, the antibodies described herein may have neutralizing activity against MYCT1 protein. In some embodiments, the antibodies described herein may have neutralizing activity against MYCT1 protein with an IC50 of less than about 150 ng / mL. In some embodiments, the antibodies described herein may have neutralizing activity against MYCT1 protein with an IC50 in the range of about 0.1 ng / mL to about 150 ng / mL (e.g., about 0.1 ng / mL, about 0.25 ng / mL, about 0.5 ng / mL, about 1 ng / mL, about 2.5 ng / mL, about 5 ng / mL, about 7.5 ng / mL, about 10 ng / mL, about 25 ng / mL, about 50 ng / mL, about 75 ng / mL, about 100 ng / mL, about 125 ng / mL, about 150 ng / mL).

[0098] In some embodiments, the antibodies described herein can reduce the expression and / or activity of MYCT1 compared to the expression and / or activity of MYCT1 in the absence of the antibodies described herein. In certain embodiments, the antibodies described herein can reduce the expression and / or activity of MYCT1 by about 5% to about 99% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%) compared to the expression and / or activity of MYCT1 in the absence of the antibodies described herein.

[0099] In some embodiments, MYCT1 expression can be measured using methods known in the art. As non-limiting examples, MYCT1 expression levels can be measured using RNA-seq, nanopore sequencing, nanostring, multiplex RT-PCR, singleplex RT-PCR, NASBA, fluorometry, or spectrophotometry. MYCT1 interacts with the tight junction protein Zona Occludens 1 and regulates Rho GTPase-dependent actin cytoskeleton dynamics, thereby promoting endothelial cell motility. Therefore, in certain embodiments, actin cytoskeleton dynamics can be measured as an indicator of MYCT1 activity. Actin cytoskeleton dynamics and MYCT1 activity can be measured using standard methods in the art, as described in the Examples below. In yet another embodiment, tubular structure formation in Matrigel assays, Boyden chamber tumor chemotaxis assays, wound closure assays, and cell morphology assays can be measured as indicators of MYCT1 activity.

[0100] In certain aspects, the antibodies described herein may have the same heavy chain and / or light chain CDRs as any of the exemplary antibodies described herein (e.g., antibody clones 2B4, 9F8, 8D2). Two antibodies with identical heavy chain and / or light chain CDRs mean that these CDRs are identical as determined by the same approach (e.g., the Kabat approach, the Chothia approach, the AbM approach, the Contact approach, or the IMGT approach). In certain aspects, the antibodies described herein may have the same heavy chain and / or light chain as any of the exemplary antibodies described herein. In certain embodiments, the antibodies disclosed herein may share one or more amino acid sequences set forth in Table 7.

[0101] In some aspects, an antibody herein may comprise a heavy chain that, individually or collectively, has at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity when compared to the heavy chain of an exemplary antibody described herein. In some aspects, an antibody herein may comprise a light chain that, individually or collectively, has at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity when compared to the light chain of an exemplary antibody described herein. In some aspects, an antibody herein may comprise heavy chain CDRs that, individually or collectively, have at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity when compared to the heavy chain CDRs of an exemplary antibody described herein. In some aspects, the antibodies herein may comprise light chain CDRs that, individually or collectively, have at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) sequence identity when compared to the light chain CDRs of the exemplary antibodies described herein.

[0102] In some embodiments, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region (VH) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 18, 54, 90, or 126. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 54. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 90. In some embodiments, the anti-MYCT1 antibody comprises a VH that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:126.

[0103] In some embodiments, the isolated anti-MYCT1 antibody may comprise a VH comprising the amino acid sequence of any one of SEQ ID NOs: 18, 54, 90, or 126. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 18. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 54. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 90. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 126.

[0104] In some embodiments, an isolated anti-MYCT1 antibody may comprise a light chain variable region (VL) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 36, 72, 108, or 144. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 36. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 72. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 108. In some embodiments, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 144.

[0105] In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the amino acid sequence of any one of SEQ ID NOs: 36, 72, 108, or 144. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the amino acid sequence of SEQ ID NO: 36. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the amino acid sequence of SEQ ID NO: 72. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the amino acid sequence of SEQ ID NO: 108. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the amino acid sequence of SEQ ID NO: 144.

[0106] In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 18, 54, 90, or 126; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 36, 72, 108, or 144, or any combination thereof. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 18; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 36. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 54; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 72. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 90; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 108. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 126; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 144.

[0107] In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising the amino acid sequence of any one of SEQ ID NOs: 18, 54, 90, or 126; and a VL comprising the amino acid sequence of any one of SEQ ID NOs: 36, 72, 108, or 144, or any combination thereof. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising the amino acid sequence of SEQ ID NO: 18; and a VL comprising the sequence of SEQ ID NO: 36. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising the amino acid sequence of SEQ ID NO: 54; and a VL comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising the sequence of SEQ ID NO: 90; and a VL comprising the amino acid sequence of SEQ ID NO: 108. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising the amino acid sequence of SEQ ID NO: 126; and a VL comprising the amino acid sequence of SEQ ID NO: 144.

[0108] In some embodiments, an isolated anti-MYCT1 antibody may comprise a heavy chain variable region 1 (H1) that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 10. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 46. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82. In some embodiments, the isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO:118.

[0109] In some aspects, an isolated anti-MYCT1 antibody may comprise a heavy chain variable region 1 (H1) comprising the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the amino acid sequence of SEQ ID NO: 10. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the amino acid sequence of SEQ ID NO: 46. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the amino acid sequence of SEQ ID NO: 82. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the amino acid sequence of SEQ ID NO: 118.

[0110] In some embodiments, an isolated anti-MYCT1 antibody may comprise a light chain variable region 1 (L1) that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 64. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100. In some embodiments, the isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136.

[0111] In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the amino acid sequence of SEQ ID NO: 28. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the amino acid sequence of SEQ ID NO: 64. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the amino acid sequence of SEQ ID NO: 100. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the amino acid sequence of SEQ ID NO: 136.

[0112] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 10; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 46; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 64. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 118; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136.

[0113] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118; and an L1 comprising the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the amino acid sequence of SEQ ID NO: 10; and an L1 comprising the amino acid sequence of SEQ ID NO: 28. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the amino acid sequence of SEQ ID NO: 46; and an L1 comprising the amino acid sequence of SEQ ID NO: 64. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the amino acid sequence of SEQ ID NO: 82; and an L1 comprising the amino acid sequence of SEQ ID NO: 100. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the sequence of SEQ ID NO: 118; and an L1 comprising the amino acid sequence of SEQ ID NO: 136.

[0114] In some embodiments, an isolated anti-MYCT1 antibody may comprise a heavy chain variable region 2 (H2) that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 48. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84. In some embodiments, the isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120.

[0115] In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the amino acid sequence of SEQ ID NO: 12. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the amino acid sequence of SEQ ID NO: 48. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the amino acid sequence of SEQ ID NO: 84. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the amino acid sequence of SEQ ID NO: 120.

[0116] In some embodiments, an isolated anti-MYCT1 antibody may comprise a light chain variable region 2 (L2) that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 30. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 66. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 102. In some embodiments, the isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 138.

[0117] In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the amino acid sequence of SEQ ID NO: 30. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the amino acid sequence of SEQ ID NO: 66. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the amino acid sequence of SEQ ID NO: 102. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the amino acid sequence of SEQ ID NO: 138.

[0118] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 30. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 48; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 66. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 102. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 138.

[0119] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120; and an L2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the amino acid sequence of SEQ ID NO: 12; and an L2 comprising the amino acid sequence of SEQ ID NO: 30. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the sequence of SEQ ID NO: 48; and an L2 comprising the amino acid sequence of SEQ ID NO: 66. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the amino acid sequence of SEQ ID NO: 84; and an L2 comprising the sequence of SEQ ID NO: 102. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the amino acid sequence of SEQ ID NO: 120; and an L2 comprising the amino acid sequence of SEQ ID NO: 138.

[0120] In some embodiments, an isolated anti-MYCT1 antibody may comprise a heavy chain variable region 3 (H3) that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 14. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 50. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 86. In some embodiments, the isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 122.

[0121] In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 14. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 50. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 86. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 122.

[0122] In some embodiments, an isolated anti-MYCT1 antibody may comprise a light chain variable region 3 (L3) that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 32. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 68. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 104. In some embodiments, the isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 140.

[0123] In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the amino acid sequence of SEQ ID NO: 32. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the amino acid sequence of SEQ ID NO: 68. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the amino acid sequence of SEQ ID NO: 104. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the amino acid sequence of SEQ ID NO: 140.

[0124] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 14; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 32. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 50; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 68. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 86; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 104. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 122; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 140.

[0125] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122; and an L3 comprising the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 14; and an L3 comprising the sequence of SEQ ID NO: 32. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 50; and an L3 comprising the amino acid sequence of SEQ ID NO: 68. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 86; and an L3 comprising the amino acid sequence of SEQ ID NO: 104. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the amino acid sequence of SEQ ID NO: 122; and an L3 comprising the amino acid sequence of SEQ ID NO: 140.

[0126] In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118; an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120; and an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 10; an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12; and an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 14, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 46; an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 48; and an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 50, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82; an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84; and an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 86, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 118; an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120; and an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 122, or any combination thereof.

[0127] In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118; an H2 comprising the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120; and an H3 comprising the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the amino acid sequence of SEQ ID NO: 10; an H2 comprising the amino acid sequence of SEQ ID NO: 12; and an H3 comprising the amino acid sequence of SEQ ID NO: 14, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the amino acid sequence of SEQ ID NO: 46; an H2 comprising the amino acid sequence of SEQ ID NO: 48; and an H3 comprising the amino acid sequence of SEQ ID NO: 50, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the amino acid sequence of SEQ ID NO: 82, an H2 comprising the amino acid sequence of SEQ ID NO: 84, and an H3 comprising the amino acid sequence of SEQ ID NO: 86, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the amino acid sequence of SEQ ID NO: 118, an H2 comprising the amino acid sequence of SEQ ID NO: 120, and an H3 comprising the amino acid sequence of SEQ ID NO: 122, or any combination thereof.

[0128] In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 30; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 32, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 64; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 66; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 68, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 102; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 104, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 138; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 140, or any combination thereof.

[0129] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136; an L2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138; and an L3 comprising the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the amino acid sequence of SEQ ID NO: 28; an L2 comprising the amino acid sequence of SEQ ID NO: 30; and an L3 comprising the amino acid sequence of SEQ ID NO: 32, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the amino acid sequence of SEQ ID NO: 64; an L2 comprising the sequence of SEQ ID NO: 66; and an L3 comprising the amino acid sequence of SEQ ID NO: 68, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise L1 comprising the amino acid sequence of SEQ ID NO: 100, L2 comprising the sequence of SEQ ID NO: 102, and L3 comprising the amino acid sequence of SEQ ID NO: 104, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise L1 comprising the amino acid sequence of SEQ ID NO: 136, L2 comprising the sequence of SEQ ID NO: 138, and L3 comprising the amino acid sequence of SEQ ID NO: 140, or any combination thereof.

[0130] In some embodiments, the isolated anti-MYCT1 antibodies herein include any of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 10, 46, 82, or 118; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 12, 48, 84, or 120; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 14, 50, 86, or 122. 3; L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 28, 64, 100, or 136; L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 30, 66, 102, or 138; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of any one of SEQ ID NOs: 32, 68, 104, or 140, or any combination thereof. In some embodiments, the isolated anti-MYCT1 antibodies herein include: H1, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 10; H2, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 12; and H3, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the amino acid sequences of SEQ ID NO: 14. L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 28; L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 30; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 32, or any combination thereof.In some embodiments, the isolated anti-MYCT1 antibodies herein include: H1, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 46; H2, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 48; or H3, which comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the amino acid sequences of SEQ ID NO: 50. L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 64; L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 66; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 68, or any combination thereof. In some embodiments, the isolated anti-MYCT1 antibodies herein include any of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 82; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 84; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the amino acid sequences of SEQ ID NO: 86. L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 100; L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 102; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 104, or any combination thereof.In some embodiments, the isolated anti-MYCT1 antibodies herein include any of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 118; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 120; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the amino acid sequences of SEQ ID NO: 122. The amino acid sequence may include H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 136; L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 138; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the amino acid sequence of SEQ ID NO: 140, or any combination thereof.

[0131] In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include H1 comprising any one of the amino acid sequences set forth in SEQ ID NOs: 10, 46, 82, or 118; H2 comprising any one of the amino acid sequences set forth in SEQ ID NOs: 12, 48, 84, or 120; H3 comprising any one of the amino acid sequences set forth in SEQ ID NOs: 14, 50, 86, or 122; L1 comprising any one of the amino acid sequences set forth in SEQ ID NOs: 28, 64, 100, or 136; L2 comprising any one of the amino acid sequences set forth in SEQ ID NOs: 30, 66, 102, or 138; and L3 comprising any one of the amino acid sequences set forth in SEQ ID NOs: 32, 68, 104, or 140, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the amino acid sequence of SEQ ID NO: 10; H2 comprising the amino acid sequence of SEQ ID NO: 12; H3 comprising any one of the amino acid sequences of SEQ ID NO: 14; L1 comprising the amino acid sequence of SEQ ID NO: 28; L2 comprising the sequence of SEQ ID NO: 30; and L3 comprising the sequence of SEQ ID NO: 32, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the amino acid sequence of SEQ ID NO: 46; H2 comprising the amino acid sequence of SEQ ID NO: 48; H3 comprising any one of the sequences of SEQ ID NO: 50; L1 comprising the amino acid sequence of SEQ ID NO: 64; L2 comprising the sequence of SEQ ID NO: 66; and L3 comprising the amino acid sequence of SEQ ID NO: 68, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the amino acid sequence of SEQ ID NO: 82; H2 comprising the sequence of SEQ ID NO: 84; H3 comprising the amino acid sequence of any one of SEQ ID NO: 86; L1 comprising the sequence of SEQ ID NO: 100; L2 comprising the amino acid sequence of SEQ ID NO: 102; and L3 comprising the amino acid sequence of SEQ ID NO: 104, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the amino acid sequence of SEQ ID NO: 118; H2 comprising the amino acid sequence of SEQ ID NO: 120; H3 comprising the amino acid sequence of any one of SEQ ID NO: 122; L1 comprising the amino acid sequence of SEQ ID NO: 136; L2 comprising the amino acid sequence of SEQ ID NO: 138; and L3 comprising the amino acid sequence of SEQ ID NO: 140, or any combination thereof.

[0132] In another exemplary embodiment, the anti-MYCT1 antibody is one or more of the antibodies listed in Table 1.

[0133] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11]

[0134] In certain aspects, the heavy chain of any of the antibodies disclosed herein may further comprise a heavy chain constant region (CH) or a portion thereof (e.g., CH1, CH2, CH3, or a combination thereof). In some aspects, the heavy chain constant regions used herein may be of any suitable origin (e.g., human, mouse, rat, or rabbit). In some aspects, alternatively or additionally, the light chain of any of the antibodies disclosed herein may further comprise a light chain constant region (CL), which may be any CL known in the art. In some aspects, the CL may be a kappa light chain. In some embodiments, the CL may be a lambda light chain. Antibody heavy and light chain constant regions are known in the art, such as those provided in the IMGT database (www.imgt.org) or www.vbase2.org / vbstat.php, both of which are incorporated herein by reference.

[0135] In some embodiments, the isolated anti-MYCT1 antibodies herein can be full-length antibodies or antigen-binding fragments thereof. In some embodiments, the isolated antibodies herein can be full-length antibodies, and the full-length antibodies are IgG molecules. In some embodiments, the isolated antibodies herein can be Fab, (Fab')2, and / or single-chain antibodies. In some embodiments, the antibodies disclosed herein can be single-chain antibodies (scFv). In some embodiments, the scFv antibodies herein can comprise a VH fragment and a VL fragment, which can be linked by a linker. In accordance with these embodiments, the linker incorporated between the two variable regions herein can be a flexible linker, a rigid linker, a cleavable linker, or any combination thereof. In some embodiments, the linker incorporated between the two variable regions herein can be a flexible peptide linker, a rigid peptide linker, a cleavable peptide linker, or any combination thereof. In accordance with these embodiments, the peptide linker incorporated between the two variable regions herein can be at least one amino acid. In some embodiments, the peptide linker incorporated between two variable regions herein can be from about 1 amino acid to about 50 amino acids (e.g., about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 32, about 34, about 36, about 38, about 40, about 42, about 44, about 46, about 48, or about 50). In some embodiments, the scFv antibodies herein may comprise a VH fragment and a VL fragment, which may be linked by a flexible peptide linker.

[0136] In some embodiments, the scFv antibodies herein may be in a VH-VL orientation (N- to C-terminus). In some embodiments, the scFv antibodies herein may be in a VL-VH orientation (N- to C-terminus).

[0137] In some embodiments, the isolated antibodies herein can be human or humanized. Humanized antibodies refer to forms of non-human (e.g., murine) antibodies that are specific chimeric immunoglobulins, immunoglobulin chains, or antigen-binding fragments thereof that contain minimal sequence derived from non-human immunoglobulin. In some embodiments, each of the exemplary antibodies described above can also include a variant Fc region.

[0138] As exemplified herein, recombinant techniques can be used to prepare any one of the antibodies exemplified herein (e.g., an anti-MYCT1 antibody). In certain aspects, nucleic acids (i.e., polynucleotides) encoding the heavy and light chains of an antibody described herein can be cloned into a single expression vector, with each nucleotide sequence operably linked to a suitable promoter. In some aspects, each polynucleotide sequence encoding the heavy and light chains can be operably linked to a separate promoter. In some aspects, the polynucleotide sequences encoding the heavy and light chains can be operably linked to a single promoter, such that both the heavy and light chains are expressed from the same promoter. In some embodiments, an internal ribosome entry site (IRES) can be inserted between the heavy and light chain coding sequences, if necessary.

[0139] In certain aspects, the polynucleotide sequences encoding the heavy and / or light chains of the antibodies described herein may share one or more polynucleotide sequences provided in Table 7.

[0140] In some embodiments, the isolated anti-MYCT1 antibody may comprise a heavy chain variable region (VH) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89, or 125. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 17. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 53. In some embodiments, the anti-MYCT1 antibody comprises a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 89. In some embodiments, the anti-MYCT1 antibody comprises a VH that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:125.

[0141] In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising the polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89, or 125. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the polynucleotide sequence of SEQ ID NO: 17. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the polynucleotide sequence of SEQ ID NO: 53. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the polynucleotide sequence of SEQ ID NO: 89. In some aspects, the anti-MYCT1 antibody comprises a VH comprising the polynucleotide sequence of SEQ ID NO: 125.

[0142] In some embodiments, an isolated anti-MYCT1 antibody may comprise a light chain variable region (VL) comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107, or 143. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 35. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 71. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 107. In some embodiments, the isolated anti-MYCT1 antibody may comprise a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 143.

[0143] In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107, or 143. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the polynucleotide sequence of SEQ ID NO: 35. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the polynucleotide sequence of SEQ ID NO: 71. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the polynucleotide sequence of SEQ ID NO: 107. In some aspects, an isolated anti-MYCT1 antibody may comprise a VL comprising the polynucleotide sequence of SEQ ID NO: 143.

[0144] In some aspects, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89, or 125; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107, or 143, or any combination thereof. In some aspects, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 17; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 35. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the sequence of SEQ ID NO: 53; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 71. In some embodiments, an isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 89; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 107. In some aspects, the isolated anti-MYCT1 antibody may comprise a VH comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 125; and a VL comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 143.

[0145] In some aspects, an isolated anti-MYCT1 antibody may comprise a VH comprising the polynucleotide sequence of any one of SEQ ID NOs: 17, 53, 89, or 125; and a VL comprising the polynucleotide sequence of any one of SEQ ID NOs: 35, 71, 107, or 143, or any combination thereof. In some aspects, an isolated anti-MYCT1 antibody may comprise a VH comprising the polynucleotide sequence of SEQ ID NO: 17; and a VL comprising the polynucleotide sequence of SEQ ID NO: 35. In some aspects, an isolated anti-MYCT1 antibody may comprise a VH comprising the polynucleotide sequence of SEQ ID NO: 53; and a VL comprising the polynucleotide sequence of SEQ ID NO: 71. In some aspects, an isolated anti-MYCT1 antibody may comprise a VH comprising the polynucleotide sequence of SEQ ID NO: 89; and a VL comprising the polynucleotide sequence of SEQ ID NO: 107. In some aspects, an isolated anti-MYCT1 antibody may comprise a VH comprising the polynucleotide sequence of SEQ ID NO: 125; and a VL comprising the polynucleotide sequence of SEQ ID NO: 143.

[0146] In some embodiments, an isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 9. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 45. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81. In some embodiments, the isolated anti-MYCT1 antibody may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:117.

[0147] In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 9. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 45. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 81. In some aspects, an isolated anti-MYCT1 antibody may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 117.

[0148] In some embodiments, an isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 27. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 63. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 99. In some embodiments, the isolated anti-MYCT1 antibody may comprise an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135.

[0149] In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 27. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 63. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 99. In some aspects, an isolated anti-MYCT1 antibody may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 135.

[0150] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 9; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 27. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 45; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 63. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 99. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 117; and an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135.

[0151] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117; and an L1 comprising the polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 9; and an L1 comprising the polynucleotide sequence of SEQ ID NO: 27. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 45; and an L1 comprising the polynucleotide sequence of SEQ ID NO: 63. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 81; and an L1 comprising the polynucleotide sequence of SEQ ID NO: 99. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H1 comprising the polynucleotide sequence of SEQ ID NO: 117; and an L1 comprising the polynucleotide sequence of SEQ ID NO: 135.

[0152] In some embodiments, an isolated anti-MYCT1 antibody may comprise a heavy chain variable region 2 (H2) that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 11. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 47. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83. In some embodiments, the isolated anti-MYCT1 antibody may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:119.

[0153] In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the polynucleotide sequence of any one of SEQ ID NO: 11, 47, 83, or 119. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 11. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 47. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 83. In some aspects, an isolated anti-MYCT1 antibody may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 119.

[0154] In some embodiments, an isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 29. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 65. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 101. In some embodiments, the isolated anti-MYCT1 antibody may comprise an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137.

[0155] In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the polynucleotide sequence of SEQ ID NO: 29. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the polynucleotide sequence of SEQ ID NO: 65. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the polynucleotide sequence of SEQ ID NO: 101. In some aspects, an isolated anti-MYCT1 antibody may comprise an L2 comprising the polynucleotide sequence of SEQ ID NO: 137.

[0156] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 11; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 29. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 47; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 65. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 101. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 119; and an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137.

[0157] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119; and an L2 comprising the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 11; and an L2 comprising the polynucleotide sequence of SEQ ID NO: 29. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 47; and an L2 comprising the polynucleotide sequence of SEQ ID NO: 65. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 83; and an L2 comprising the polynucleotide sequence of SEQ ID NO: 101. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H2 comprising the polynucleotide sequence of SEQ ID NO: 119; and an L2 comprising the polynucleotide sequence of SEQ ID NO: 137.

[0158] In some embodiments, an isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 13. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 49. In some embodiments, an isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 85. In some embodiments, the isolated anti-MYCT1 antibody may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:121.

[0159] In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the polynucleotide sequence of any one of SEQ ID NO: 13, 49, 85, or 121. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 13. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 49. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 85. In some aspects, an isolated anti-MYCT1 antibody may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 121.

[0160] In some embodiments, an isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 31. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 67. In some embodiments, an isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 103. In some embodiments, the isolated anti-MYCT1 antibody may comprise an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 139.

[0161] In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the polynucleotide sequence of SEQ ID NO: 31. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the polynucleotide sequence of SEQ ID NO: 67. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the polynucleotide sequence of SEQ ID NO: 103. In some aspects, an isolated anti-MYCT1 antibody may comprise an L3 comprising the polynucleotide sequence of SEQ ID NO: 139.

[0162] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 13; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 31. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 49; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 67. In some aspects, an isolated anti-MYCT1 antibody herein may comprise an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 85; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 103. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 121; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 139.

[0163] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121; and an L3 comprising the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 13; and an L3 comprising the polynucleotide sequence of SEQ ID NO: 31. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 49; and an L3 comprising the polynucleotide sequence of SEQ ID NO: 67. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 85; and an L3 comprising the polynucleotide sequence of SEQ ID NO: 103. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an H3 comprising the polynucleotide sequence of SEQ ID NO: 121; and an L3 comprising the polynucleotide sequence of SEQ ID NO: 139.

[0164] In some aspects, the isolated anti-MYCT1 antibodies of the present specification may include an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117; an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119; and an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 9; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 11; and H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 13, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 45; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 47; and H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 49, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83; and H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 85, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 117; an H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 119; and an H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 121, or any combination thereof.

[0165] In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117; an H2 comprising the polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119; and an H3 comprising the polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the polynucleotide sequence of SEQ ID NO: 9; an H2 comprising the polynucleotide sequence of SEQ ID NO: 11; and an H3 comprising the polynucleotide sequence of SEQ ID NO: 13, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an H1 comprising the polynucleotide sequence of SEQ ID NO: 45; an H2 comprising the sequence of SEQ ID NO: 47; and an H3 comprising the polynucleotide sequence of SEQ ID NO: 49, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the polynucleotide sequence of SEQ ID NO: 81; H2 comprising the polynucleotide sequence of SEQ ID NO: 83; and H3 comprising the polynucleotide sequence of SEQ ID NO: 85, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the polynucleotide sequence of SEQ ID NO: 117; H2 comprising the polynucleotide sequence of SEQ ID NO: 119; and H3 comprising the polynucleotide sequence of SEQ ID NO: 121, or any combination thereof.

[0166] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135; an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137; and an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 27; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 29; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 31, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 63; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 65; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 67, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibodies herein may include an L1 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 99; an L2 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 101; and an L3 that comprises at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 103, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135; an L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137; and an L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0167] In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135; an L2 comprising the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137; and an L3 comprising the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 27; an L2 comprising the polynucleotide sequence of SEQ ID NO: 29; and an L3 comprising the polynucleotide sequence of SEQ ID NO: 31, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 63; an L2 comprising the polynucleotide sequence of SEQ ID NO: 65; and an L3 comprising the polynucleotide sequence of SEQ ID NO: 67, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 99, an L2 comprising the polynucleotide sequence of SEQ ID NO: 101, and an L3 comprising the polynucleotide sequence of SEQ ID NO: 103, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may comprise an L1 comprising the polynucleotide sequence of SEQ ID NO: 135, an L2 comprising the polynucleotide sequence of SEQ ID NO: 137, and an L3 comprising the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0168] In some embodiments, the isolated anti-MYCT1 antibodies herein include any of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 9, 45, 81, or 117; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 11, 47, 83, or 119; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 13, 49, 85, or 121. 3; L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 27, 63, 99, or 135; L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 29, 65, 101, or 137; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of any one of SEQ ID NOs: 31, 67, 103, or 139, or any combination thereof.In some embodiments, the isolated anti-MYCT1 antibodies herein include any of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:9; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:11; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the polynucleotide sequences of SEQ ID NO:13. The polynucleotide sequences may include H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:27; L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:29; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:31, or any combination thereof. In some embodiments, the isolated anti-MYCT1 antibodies herein include any of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 45; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 47; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the polynucleotide sequences of SEQ ID NO: 49. The polynucleotide sequences may include H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 63; L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 65; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 67, or any combination thereof.In some embodiments, the isolated anti-MYCT1 antibodies herein comprise one of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 81; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 83; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the polynucleotide sequences of SEQ ID NO: 85; ) identity to the sequence of SEQ ID NO:99; L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:101; L2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:103; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO:103, or any combination thereof. In some embodiments, the isolated anti-MYCT1 antibodies herein include any of the following: H1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 117; H2 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 119; H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to any one of the polynucleotide sequences of SEQ ID NO: 121. The polynucleotide sequences may include H3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 135; L1 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 137; and L3 comprising at least about 80% (e.g., about 85%, about 90%, about 95%, about 98%) identity to the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0169] In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising any one of the polynucleotide sequences set forth in SEQ ID NOs: 9, 45, 81, or 117; H2 comprising any one of the polynucleotide sequences set forth in SEQ ID NOs: 11, 47, 83, or 119; H3 comprising any one of the polynucleotide sequences set forth in SEQ ID NOs: 13, 49, 85, or 121; L1 comprising any one of the polynucleotide sequences set forth in SEQ ID NOs: 27, 63, 99, or 135; L2 comprising any one of the polynucleotide sequences set forth in SEQ ID NOs: 29, 65, 101, or 137; and L3 comprising any one of the polynucleotide sequences set forth in SEQ ID NOs: 31, 67, 103, or 139, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the polynucleotide sequence of SEQ ID NO:9; H2 comprising the sequence of SEQ ID NO:11; H3 comprising the polynucleotide sequence of any one of SEQ ID NOs:13; L1 comprising the polynucleotide sequence of SEQ ID NO:27; L2 comprising the sequence of SEQ ID NO:29; and L3 comprising the polynucleotide sequence of SEQ ID NO:31, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the polynucleotide sequence of SEQ ID NO:45; H2 comprising the polynucleotide sequence of SEQ ID NO:47; H3 comprising the sequence of any one of SEQ ID NOs:49; L1 comprising the polynucleotide sequence of SEQ ID NO:63; L2 comprising the sequence of SEQ ID NO:65; and L3 comprising the polynucleotide sequence of SEQ ID NO:67, or any combination thereof. In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the polynucleotide sequence of SEQ ID NO: 81; H2 comprising the polynucleotide sequence of SEQ ID NO: 83; H3 comprising any one of the polynucleotide sequences of SEQ ID NO: 85; L1 comprising the polynucleotide sequence of SEQ ID NO: 99; L2 comprising the polynucleotide sequence of SEQ ID NO: 101; and L3 comprising the polynucleotide sequence of SEQ ID NO: 103, or any combination thereof.In some aspects, the isolated anti-MYCT1 antibodies herein may include H1 comprising the polynucleotide sequence of SEQ ID NO: 117; H2 comprising the polynucleotide sequence of SEQ ID NO: 119; H3 comprising any one of the polynucleotide sequences of SEQ ID NO: 121; L1 comprising the polynucleotide sequence of SEQ ID NO: 135; L2 comprising the polynucleotide sequence of SEQ ID NO: 137; and L3 comprising the polynucleotide sequence of SEQ ID NO: 139, or any combination thereof.

[0170] In certain aspects, the expression and / or production of an antibody in a host cell can be facilitated by expressing a leader peptide preceding the variable region of the heavy or light chain of the antibody. In some embodiments, the leader peptide can be optimized for expression in a particular host cell, such as E. coli.

[0171] In some embodiments, genetically engineered antibodies (e.g., single-chain antibodies) can be produced, for example, by conventional recombinant techniques or any method known in the art. In some embodiments, DNA encoding monoclonal antibodies specific for a target antigen can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of binding specifically to genes encoding the heavy and light chains of the monoclonal antibody). Once isolated, this DNA can be inserted into one or more expression vectors and then introduced into host cells such as E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not normally produce immunoglobulin proteins, to synthesize the monoclonal antibody in the recombinant host cells. This DNA can then be modified, for example, by substituting coding sequences for human heavy and light chain constant domains for the homologous murine sequences, or by covalently linking the immunoglobulin coding sequences to all or part of the coding sequence for a non-immunoglobulin polypeptide. In some embodiments, engineered antibodies (eg, chimeric or hybrid antibodies) can be prepared that have the binding specificity of a target antigen.

[0172] In some aspects, the single-chain antibodies herein can be prepared recombinantly by linking a nucleotide sequence encoding a heavy chain variable region with a nucleotide sequence encoding a light chain variable region. In some aspects, a linker can be incorporated between these two variable regions. In some embodiments, the techniques described for producing single-chain antibodies can be adapted to generate phage or yeast scFv libraries, and scFv clones specific for MYCT1 can be identified from the libraries according to routine procedures. In some embodiments, positive clones can be subjected to further screening to identify clones that bind to specific epitopes on MYCT1.

[0173] In some aspects, one or more vectors (e.g., expression vectors) carrying a nucleic acid encoding any of the antibodies herein may be introduced into a host cell suitable for producing the antibody. In some aspects, the host cells may be cultured under conditions suitable for expression of the antibody or any polypeptide chains thereof. In some aspects, the antibody or polypeptide chains thereof may be recovered from the cultured cells (e.g., the cells or culture supernatant) by conventional methods such as, for example, affinity purification. In some aspects, the polypeptide chains of an antibody herein may be incubated under suitable conditions for a suitable period of time to allow for production of the antibody.

[0174] In some embodiments, methods for preparing antibodies described herein can include a recombinant expression vector encoding both the heavy and light chains of an antibody that binds to MYCT1, as also described herein. In some embodiments, the recombinant expression vector can be introduced into suitable host cells (e.g., dhfr-CHO cells) by conventional methods, such as calcium phosphate-mediated transfection. In some embodiments, positively transformant host cells can be selected and cultured under appropriate conditions to allow expression of the two polypeptide chains that form the antibody, and the antibody can be recovered from the cells or culture medium. In some embodiments, the two chains recovered from the host cells can be incubated under appropriate conditions to allow antibody formation.

[0175] In certain aspects, two recombinant expression vectors may be provided, one encoding the heavy chain of a disclosed antibody and the other encoding the light chain of a disclosed antibody. In some aspects, both of these recombinant expression vectors may be introduced into suitable host cells (e.g., dhfr-CHO cells) by conventional methods such as calcium phosphate-mediated transfection. In some aspects, each expression vector may be introduced into a suitable host cell. In some aspects, positive transformants may be selected and cultured under appropriate conditions that allow for expression of the antibody polypeptide chains. In some aspects, if these two expression vectors are introduced into the same host cell, the antibody produced therein may be recovered from the host cell or from the culture medium. In some aspects, the polypeptide chain may be recovered from the host cell or from the culture medium and then incubated under appropriate conditions to allow for antibody formation. In some aspects, if these two expression vectors are introduced into different host cells, each of them may be recovered from the corresponding host cell or from the corresponding culture medium. In some embodiments, the two polypeptide chains may then be incubated under conditions that allow for antibody formation.

[0176] In certain aspects, standard molecular biology techniques can be used to prepare the recombinant expression vector, transfect the host cells, select for transformants, culture the host cells, and recover the antibody from the culture medium. In some embodiments, some antibodies can be isolated by affinity chromatography on a Protein A or Protein G-coupled matrix.

[0177] In an additional aspect, the antibodies herein may be characterized by identifying the epitope or epitopes to which the antigen binds (i.e., by "epitope mapping"). Many methods are known in the art for mapping and characterizing the location of epitopes on proteins, including, but not limited to, solving the crystal structure of an antibody-antigen complex, competition assays, gene fragment expression assays, and synthetic peptide-based assays. In some embodiments, epitope mapping may be used to determine the sequence to which the antibody binds.

[0178] Chimeric Antigen Receptor In some aspects, the present disclosure encompasses chimeric antigen receptor (CAR) immune effector cells (CAR immune effector cells) comprising an antigen binding domain comprising an antibody or fragment thereof that specifically binds to MYCT1 as disclosed herein. In one aspect, the CAR immune effector cell comprising an antigen binding domain comprising an antibody or fragment thereof that specifically binds to MYCT1 comprises an amino acid sequence or polynucleotide sequence listed in Table 7.

[0179] The hinge domain is a structure between the antigen-recognition domain (e.g., an antibody that binds to MYCT1) and the cell plasma membrane. These sequences are generally derived from IgG subclasses (e.g., IgG1 and IgG4), IgD, and CD8 domains, of which IgG1 is the most widely used for CAR construction. Currently, research on hinge domains has focused mainly on the following four aspects: 1) reducing the binding affinity to Fcγ receptors, thereby eliminating off-target activation; 2) enhancing the flexibility of single-chain variable fragments (scFvs), thereby alleviating the spatial constraint between tumor antigens and CARs and subsequently facilitating synapse formation between CAR immune effector cells and target cells; 3) shortening the distance between scFvs and the target epitope; and 4) facilitating the detection of CAR expression using anti-Fc reagents.

[0180] In one aspect, the hinge domain includes a hinge domain of a human protein selected from CD28, 4-1BB (CD137), OX-40 (CD134), CD3ζ, T-cell receptor alpha or beta chain, CD45, CD4, CD5, CD8, CD8α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, ICOS, CD154, functional derivatives and / or combinations thereof.

[0181] Typically, the extracellular domain, i.e., the antigen recognition domain (e.g., an antibody that binds to MYCT1), or targeting element, is linked to the intracellular domain (i.e., the costimulatory and signaling domains of a chimeric antigen receptor) by a transmembrane domain. The transmembrane domain spans the cell membrane, anchoring the CAR to the immune effector cell surface and connecting the extracellular domain to the intracellular domain, affecting the expression of the CAR on the immune effector cell surface.

[0182] Transmembrane domains include hydrophobic polypeptides that span the cell membrane, specifically from one side of the cell membrane (extracellular) to the other side of the cell membrane (intracellular or cytoplasmic).

[0183] The transmembrane domain may be in the form of an alpha helix or a beta barrel, or a combination thereof. The transmembrane domain may include polytopic proteins having many transmembrane segments (each alpha helical, beta sheet, or a combination thereof).

[0184] In one embodiment, a transmembrane domain that naturally associates with one of the domains in the CAR is used. In another embodiment, the transmembrane domain is selected or modified by amino acid substitution to avoid binding of such domain to transmembrane domains of the same or different surface membrane proteins to minimize interactions with other members of the receptor complex.

[0185] For example, the transmembrane domain is selected from the transmembrane domains of the T cell receptor alpha or beta chain, the CD3 zeta chain, CD28, CD3s, CD45, CD4, CD5, CD7, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD68, OX-40 (CD134), 4-1BB (CD137), ICOS, CD41, CD154, functional derivatives and / or combinations thereof.

[0186] The chimeric antigen receptors of the present disclosure may comprise one or more costimulatory domains and / or one or more spacers derived from costimulatory proteins that enhance cytokine production, proliferation, cytotoxicity, and / or persistence in vivo.

[0187] In one aspect, the costimulatory domain is selected from OX-40 (CD134), CD27, CD28, CD30, CD40, PD-1, CD2, CD7, CD258, natural killer group 2 member C (NKG2C), natural killer group 2 member D (NKG2D), B7-H3, a ligand that binds to at least one of the following: CD83, ICAM-1, LFA-1 (CD11a / CD18), ICOS, and 4-1BB (CD137), CDS, ICAM-1, LFA-1 (CD1a / CD18), CD40, CD27, active fragments thereof, functional derivatives thereof, and combinations thereof.

[0188] The chimeric antigen receptor of the present disclosure also contains a signaling domain that confers an intracellular signal to an immune effector cell upon binding of an antigen to the antigen recognition domain. The signaling domain of the chimeric antigen receptor of the present disclosure is involved in activating at least one of the effector functions of the immune effector cell in which the chimeric receptor is expressed. The term "effector function" refers to the specialized function of a differentiated cell. The effector function of an immune cell can be, for example, cytolytic activity or helper activity, including cytokine secretion. The effector function of naive, memory, or memory-type immune cells can also include antigen-dependent proliferation. Thus, the term "intracellular domain" refers to the intracellular portion of a CAR that transmits an effector function signal upon binding of an antigen to the extracellular domain, inducing the immune cell to perform a specialized function. Non-limiting examples of suitable signaling domains include the zeta chain or its homologs (e.g., eta, delta, gamma, or epsilon) of immune cell receptors, MB-1 chain, 829, FcRIII, FcRI, and combinations of signaling molecules such as CD3ζ, and CD28, CD27, 4-1BB (CD137), DNAX activation protein 10 (DAP10), OX-40 (CD134), and combinations thereof, as well as other similar molecules and fragments. Signaling domains of other activation proteins, such as FcγRIII and FcεRI, may also be used. While the entire signaling domain is typically utilized, in many cases it is not necessary to use the entire intracellular polypeptide. To this extent, truncated portions of the signaling domain may be used as long as they still transduce an effector function signal. The term intracellular domain is also meant to include any truncated portion of the intracellular domain sufficient to transduce an effector function signal. Alternatively, or in addition, the CAR immune effector cells encompassed by the present disclosure may further comprise one or more suicide genes. As used herein, a "suicide gene" refers to a nucleic acid sequence that is introduced into a CAR immune effector cell by standard methods known in the art and that, when activated, results in the death of the CAR immune effector cell.The suicide gene can facilitate effective tracking and elimination of CAR immune effector cells in vivo, if desired. Activation of the suicide gene can facilitate killing by methods known in the art. Suitable suicide gene therapy systems known in the art include, but are not limited to, various herpes simplex virus thymidine kinase (HSVtk) / ganciclovir (GCV) suicide gene therapy systems, or inducible caspase-9 protein. In an exemplary embodiment, the suicide gene is a CD34 / thymidine kinase chimeric suicide gene.

[0189] Methods for designing CARs, delivering them to and expressing them in immune cells, and producing clinical-grade CAR immune effector cell populations are known in the art. See, for example, Lee et al., Clin. Cancer Res., 2012, 18(10): 2780-90 (incorporated herein by reference in its entirety). For example, engineered CARs can be introduced into immune effector cells using retroviruses, which efficiently and stably integrate the nucleic acid sequence encoding the chimeric antigen receptor into the target cell genome. Exemplary methods for viral vector production are described in the Examples section. Other methods known in the art include, but are not limited to, lentiviral transduction, transposon-based systems, direct RNA transfection, and CRISPR / Cas systems (e.g., Cas3, Cas4, Cas5, Cas5e (or CasD), Cas6, Cas6e, Cas6f, Cas7, Cas8a1, Cas8a2, Cas8b, Cas8c, Cas9, Cas10, Cas10d, CasF, CasG, CasH, Csy1, Csy2, Csy3, Csel (or CasA), Type I, Type II, or Type III systems using an appropriate Cas protein such as Cse2 (or CasB), Cse3 (or CasE), Cse4 (or CasC), Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csz1, Csx15, Csf1, Csf2, Csf3, Csf4, and Cul966.

[0190] In some embodiments, the immune effector cells are selected from the group consisting of T cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTLs), and regulatory T cells.

[0191] In some aspects, immune effector cells include any white blood cell involved in protecting the body from infectious diseases and foreign substances. For example, immune effector cells can include lymphocytes, monocytes, macrophages, dendritic cells, mast cells, neutrophils, basophils, eosinophils, or any combination thereof. In some aspects, immune effector cells include T lymphocytes.

[0192] As disclosed herein, T cells can be any subset of T cells, including, but not limited to, T helper cells (TH cells), cytotoxic T cells (TC cells or CTL), memory T cells, regulatory T cells (Treg cells), or natural killer T (NKT) cells.

[0193] T helper cells (TH cells) assist other white blood cells in immune processes, such as the maturation of B cells into plasma cells and memory B cells, and the activation of cytotoxic T cells and macrophages. These cells are also known as CD4+ T cells because they express the CD4 glycoprotein on their surface. Helper T cells are activated when peptide antigens are presented by MHC class II molecules expressed on the surface of antigen-presenting cells (APCs). Once activated, they divide rapidly and secrete small proteins called cytokines that control or support active immune responses. These cells can differentiate into one of several subtypes, such as TH1, TH2, TH3, TH17, TH9, or TFH, and secrete various cytokines to promote different types of immune responses.

[0194] Cytotoxic T cells (TC cells or CTLs) destroy virus-infected and tumor cells and are involved in transplant rejection. These cells are also known as CD8+ T cells because they express the CD8 glycoprotein on their surface. These cells recognize their targets by binding to antigens associated with MHC class I molecules, which are present on the surface of all nucleated cells. CD8+ cells can be inactivated into an anergic state via IL-10, adenosine, and other molecules secreted by regulatory T cells, thereby preventing autoimmune disease.

[0195] Memory T cells are a subset of antigen-specific T cells that persist long after an infection has subsided. Upon re-exposure to their cognate antigen, these memory T cells rapidly expand into large numbers of effector T cells, thus providing the immune system with a "memory" of past infection. Memory cells can be either CD4+ or CD8+. Memory T cells typically express the cell surface protein CD45RO.

[0196] Regulatory T cells (Treg cells), formerly known as suppressor T cells, are essential for maintaining immune tolerance. Their primary role is to shut down T cell-mediated immunity at the end of an immune response and to suppress autoreactive T cells that have escaped the negative selection process in the thymus. Two major classes of CD4+ Treg cells have been reported: naturally occurring Treg cells and adaptive Treg cells.

[0197] Natural killer T (NKT) cells (not to be confused with natural killer (NK) cells) bridge the gap between the adaptive and innate immune systems. Unlike conventional T cells, which recognize peptide antigens presented by major histocompatibility complex (MHC) molecules, NKT cells recognize glycolipid antigens presented by MHC molecules, such as CD1d. In some embodiments, NKT cells are invariant natural killer T (iNKT) cells. iNKT cells recognize CD1d and are restricted by their T cell receptor (TCR), which in humans is Vα24Jα18 and typically pairs with Vβ11. iNKT cells also ameliorate graft-versus-host disease (GVHD).

[0198] In some aspects, the T cells comprise a mixture of CD4+ cells. In other aspects, the T cells are enriched for one or more subsets based on cell surface expression. For example, in some cases, the T cells comprise cytotoxic CD8+ T lymphocytes. In some embodiments, the T cells comprise γδ T cells, which have distinct T cell receptors (TCRs) with one γ chain and one δ chain instead of α and β chains.

[0199] Natural killer (NK) cells are CD56+CD3- large granular lymphocytes that can kill virus-infected and transformed cells and constitute an important cell subset of the innate immune system. Unlike cytotoxic CD8+ T lymphocytes, NK cells exert cytotoxic effects against tumor cells without the need for prior sensitization and can eradicate MHC-I-negative cells.

[0200] In some embodiments, immune effector cells can be obtained from the subject to be treated (i.e., autologous) and administered as an adoptive transfer after being genetically modified to express a CAR having an anti-MYCT1 antigen recognition domain of the present disclosure (e.g., an anti-MYCT1 antibody or fragment thereof disclosed herein). In such embodiments, adoptive transfer involves procuring immune effector cells from the subject, subsequently genetically modifying the cells to express a CAR having an anti-MYCT1 antigen recognition domain of the present disclosure, and infusing the modified cells back into the subject. In some embodiments, immune effector cell lines or donor effector cells (allogeneic) are used. Immune effector cells for allogeneic therapy can be harvested from a single subject or multiple subjects. Immune effector cells can be obtained from many sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumor. Immune effector cells can be obtained from blood drawn from a subject using any number of techniques known to those skilled in the art, such as Ficoll™ separation. For example, cells from an individual's circulating blood can be obtained by apheresis. In some embodiments, immune effector cells can be isolated from peripheral blood lymphocytes by lysing red blood cells and depleting monocytes, for example, by centrifugation through a PERCOLL™ gradient or by counterflow centrifugal elutriation. Specific subpopulations of immune effector cells can be further separated by positive or negative selection techniques. For example, immune effector cells can be isolated using a combination of antibodies directed against surface markers characteristic of positively selected cells, e.g., by incubating with antibody-conjugated beads for a period sufficient to positively select the desired immune effector cells. Alternatively, enrichment of immune effector cell populations can be achieved by negative selection using a combination of antibodies directed against surface markers characteristic of negatively selected cells.

[0201] In some embodiments, expression of a nucleic acid encoding a CAR can be achieved by operably linking a nucleic acid encoding a CAR polypeptide comprising the anti-MYCT1 antigen recognition domain of the present disclosure to a promoter and incorporating this construct into an expression vector. Typical cloning vectors contain transcription and translation terminators, initiation sequences, and promoters useful for controlling expression of the desired nucleic acid sequence. In some embodiments, the immune effector cells further comprise a modification of the endogenous T cell receptor alpha chain (TRAC), such that signaling through the endogenous T cell receptor is blocked in the CAR-T cells.

[0202] In some embodiments, the CAR comprising the anti-MYCT1 antigen recognition domain of the present disclosure can be cloned into many types of vectors.For example, the CAR comprising the anti-MYCT1 antigen recognition domain of the present disclosure can be cloned into vectors, including but not limited to, plasmids, phagemids, phage derivatives, animal viruses, and cosmids.In some embodiments, non-limiting examples of vectors include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.

[0203] Many virus-based systems have been developed for gene transfer into mammalian cells.For example, the CAR comprising the anti-MYCT1 antigen recognition domain of the present disclosure can be inserted into vector and packaged into retroviral particles using techniques known in the art.Then, this recombinant virus can be isolated and delivered to target cells either in vivo or ex vivo.

[0204] Bispecific, multispecific, or hybrid antibodies In some embodiments, the present disclosure further encompasses bispecific, multispecific, or hybrid antibodies having at least two different binding domains with varying specificities, wherein at least one domain comprises an H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed in Table 7. In some embodiments, the bispecific, multispecific, or hybrid antibody comprises an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein and at least one immune cell engager (e.g., a T cell engager, an NK cell engager, an iNKT cell engager, a B cell engager, a dendritic cell engager, or a macrophage cell engager). Such antibody constructs are recombinant protein constructs made from two flexibly linked antibody-derived binding domains: one binding domain specific for a selected tumor antigen on target tumor cells, and another binding domain that binds to and / or activates T cells, NK cells, iNKT cells, B cells, dendritic cells, or macrophage cells. In some embodiments, the antibodies of the present disclosure can transiently target immune cells to target tumor cells and / or activate the intrinsic cytolytic ability of immune cells against target tumor cells. In some embodiments, bispecific, multispecific, or hybrid antibodies comprise at least one domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL as disclosed herein, and another domain comprising a binding domain specific for an antigen expressed by B cell and / or plasma cell neoplasm cells. Bispecific, multispecific, or hybrid antibody constructs can be produced by a variety of methods, including fusion of hybridomas, linking of Fab' fragments, or other methods known in the art.

[0205] In some embodiments, the bispecific, multispecific, or hybrid antibody comprises a T cell engager (e.g., a bispecific T cell engager (BiTE)) comprising a first antigen-recognition domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein, and a second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment that binds to or activates a T cell. In such embodiments, the second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of CD2, CD3, CD4, CD5, CD6, CD8, CD25, CD27, CD28, CD30, CD40, CD40L, CD44, CD45, CD69, CD90, CRa, TCRp, TCRy, TCRC, ICOS, HVEM, LIGHT, 4-1BB, OX40, DR3, GITR, TIM1, SLAM, or CD226.

[0206] In some embodiments, a bispecific, multispecific, or hybrid antibody has a binding domain that binds to NK cells. In such embodiments, the bispecific antibody comprises a first antigen-recognition domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein, and a second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment that binds to or activates NK cells. In such embodiments, the second antigen recognition domain comprising the antibody or antigen-binding antibody fragment binds to one or more of CD16 (e.g., CD16a, CD16b, or both), NKp30, NKp40, NKp44, NKp46, NKG2D, DNAM1, DAP10, CRTAM, CD27, PSGL1, CD96, CD100 (SEMA4D), NKp80, CD244 (also known as SLAMF4 or 2B4), SLAMF6, SLAMF7, KIR2DS2, KIR2DS4, KIR3DS1, KIR2DS3, KIR2DS5, KIR2DS1, CD94, NKG2C, NKG2E, or CD160.

[0207] In some embodiments, a bispecific, multispecific, or hybrid antibody has a binding domain that binds to iNKT cells. In such embodiments, the bispecific antibody comprises a first antigen-recognition domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein, and a second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment that binds to or activates iNKT cells. In such embodiments, the second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment binds to CD3.

[0208] In some embodiments, a bispecific antibody, multispecific antibody, or hybrid antibody has a binding domain that binds to B cells. In such embodiments, the bispecific antibody comprises a first antigen-recognition domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein, and a second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment that binds to or activates B cells. In such embodiments, the second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of OX40, CD40, or CD70.

[0209] In some embodiments, a bispecific antibody, a multispecific antibody, or a hybrid antibody has a binding domain that binds to dendritic cells. In such embodiments, the bispecific antibody comprises a first antigen-recognition domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein, and a second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment that binds to or activates dendritic cells. In such embodiments, the second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of CD2, OX40, 4-IBB, TLR4, CD47, or STING.

[0210] In some embodiments, a bispecific antibody, multispecific antibody, or hybrid antibody has a binding domain that binds to macrophage cells. In such embodiments, the bispecific antibody comprises a first antigen-recognition domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL disclosed herein, and a second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment that binds to or activates macrophage cells. In such embodiments, the second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of OX40, CD40, CD70, TLR4, TLR9, CD47, or a STIN agonist.

[0211] In some embodiments, the bispecific, multispecific, or hybrid antibody comprises a first antigen-recognition domain comprising an anti-MYCT1 H1, H2, H3, L1, L2, L3, VH, and / or VL as disclosed herein, and a second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment that binds to an antigen expressed by a tumor cell. In some embodiments, the antigen is an antigen expressed by a sarcoma and / or breast cancer. In such embodiments, the second antigen-recognition domain comprising an antibody or antigen-binding antibody fragment binds to one or more of MAGE-A4, NY-ESO-1, PRAME, TRAG-3 / CSAGE, SSX, CTA, HLTF, ITGA10, PLCG1, TTC3, HER2, MUC-1, CEA, hTERT, STn, WT1, or p53.

[0212] III. Composition In certain embodiments, the present disclosure further encompasses compositions comprising anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies. In some embodiments, the compositions of the present disclosure include compositions that modulate MYCT1. Specifically, a composition that modulates MYCT1 can be a composition that downregulates the expression and / or activity of MYCT1. In certain embodiments, the compositions of the present disclosure include one or more of the antibodies, fragments thereof, or polynucleotides encoding the antibodies disclosed herein.

[0213] The composition of the present invention may further comprise a pharmaceutically acceptable excipient, carrier, or diluent, such as a preservative, solubilizer, stabilizer, wetting agent, emulsifier, sweetener, colorant, odorant, salt (the substance of the present invention itself may be provided in the form of a pharmaceutically acceptable salt), buffer, coating agent, or antioxidant.

[0214] In certain aspects, the compositions provided herein are pharmaceutical compositions comprising one or more of the antibodies, fragments thereof, or polynucleotides encoding the antibodies disclosed herein. In some aspects, the pharmaceutical compositions herein can comprise one or more of the antibodies, fragments thereof, or polynucleotides encoding the antibodies disclosed herein and at least one pharmaceutically acceptable carrier and / or excipient. Pharmaceutically acceptable carriers or excipients suitable for the compositions described herein are known to those of skill in the art for use in storing and delivering antibodies or antibody fragments to any mammalian subject (e.g., humans and other mammals).

[0215] Compositions comprising the anti-MYCT1 antibodies or fragments thereof disclosed herein can be formulated for administration in any convenient manner, such as compositions for injection, infusion, or implantation. The compositions described herein can be formulated for subcutaneous, intradermal, intratumoral, intranodal, intramedullary, intramuscular, intravenous (iv), or intraperitoneal injection or administration. In some embodiments, the compositions of the present disclosure are formulated for administration by intradermal or subcutaneous injection. In some embodiments, the compositions of the present disclosure are formulated for administration by iv injection. The compositions can also be formulated for direct injection into a tumor, lymph node, or site of infection. In some embodiments, the compositions disclosed herein are formulated for intravenous administration. In some embodiments, the compositions disclosed herein are formulated for intraperitoneal injection.

[0216] In some embodiments, the compositions described herein may comprise liposomes containing the antibody, a fragment thereof, or a polynucleotide encoding the antibody. In some embodiments, the liposomes used herein may be generated by reverse phase evaporation using a lipid composition comprising phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE). In some embodiments, the liposomes used herein may be extruded through filters of defined pore size to generate liposomes of the desired diameter.

[0217] In some aspects, the antibodies herein, fragments thereof, or encoding polynucleotides may be encapsulated in microcapsules prepared, for example, by coacervation techniques or interfacial polymerization, e.g., hydroxymethylcellulose or gelatin microcapsules, and poly(methyl methacrylate) microcapsules, or macroemulsions in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), respectively.

[0218] In some aspects, the compositions described herein can be formulated in sustained-release form. In some aspects, the compositions herein used for in vivo administration can be sterile. In some aspects, this can be readily accomplished, for example, by filtration through sterile filtration membranes. In some aspects, the therapeutic antibody compositions herein can be placed, for example, in a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0219] In certain aspects, the compositions described herein may be present in unit dosage forms such as tablets, pills, capsules, powders, granules, solutions or suspensions, or suppositories for oral, parenteral, or rectal administration, or for administration by inhalation or insufflation.

[0220] In some aspects, the emulsion compositions herein may be prepared by mixing an antibody with Intralipid™ or its components (soybean oil, egg phospholipids, glycerol, and water).

[0221] Suitable emulsions can be prepared using commercially available fat emulsions such as Intralipid™, Liposyn™, Infonutrol™, Lipofundin™, and Lipiphysan™. The active ingredient can be dissolved in a premixed emulsion composition or in oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil, or almond oil) and mixed with phospholipids (e.g., egg phospholipids, soybean phospholipids, or soybean lecithin) and water to form an emulsion. It will be appreciated that other ingredients (e.g., glycerol or glucose) can be added to adjust the osmotic pressure of the emulsion. Suitable emulsions typically contain up to about 20% oil, for example, from about 5% to about 20%. The fat emulsion may contain fat droplets of about 0.1 μm to about 1.0 μm, particularly about 0.1 μm to 0.5 μm, and may have a pH in the range of about 5.5 to about 8.0.

[0222] In some embodiments, pharmaceutical compositions herein for inhaled or nasal administration may include solutions, suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. In some embodiments, the liquid or solid compositions herein may contain suitable pharmaceutically acceptable excipients as described above. In other embodiments, the compositions may be administered orally or via the nasal respiratory route to achieve a local or systemic effect.

[0223] In some aspects, compositions that may be in a sterile, pharmaceutically acceptable solvent may be nebulized by the use of a gas. In some aspects, nebulized solutions herein may be inhaled directly from the nebulizing device, or the nebulizing device may be attached to a face mask, tent, or intermittent positive pressure breathing machine. In some embodiments, solution, suspension, or powder compositions herein may be administered, preferably orally or nasally, from a device that delivers the formulation in an appropriate manner.

[0224] Components of the composition In certain embodiments, the compositions disclosed herein may be pharmaceutical compositions, which comprise, as an active ingredient, an anti-MYCT1 antibody as described above and at least one pharmaceutically acceptable excipient.

[0225] The pharmaceutically acceptable excipient may be a diluent, binder, filler, buffer, pH adjuster, disintegrant, dispersant, preservative, lubricant, taste masking agent, flavoring agent, or coloring agent. The amount and type of excipient utilized to form a pharmaceutical composition may be selected according to known principles of pharmacy.

[0226] (i) Diluent In one embodiment, the excipient can be a diluent. The diluent can be compressible (i.e., plastically deformable) or abrasively brittle. Non-limiting examples of suitable compressible diluents include microcrystalline cellulose (MCC), cellulose derivatives, cellulose powder, cellulose esters (i.e., mixed esters of acetic and butyric acids), ethyl cellulose, methyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, sodium carboxymethyl cellulose, corn starch, phosphorylated corn starch, pregelatinized corn starch, rice starch, potato starch, tapioca starch, starch-lactose, starch-calcium carbonate, sodium starch glycolate, glucose, fructose, lactose, lactose monohydrate, sucrose, xylose, lactitol, mannitol, malitol, sorbitol, xylitol, maltodextrin, and trehalose. Non-limiting examples of suitable abrasive brittle diluents include dicalcium phosphate (anhydrous or dihydrate), tricalcium phosphate, calcium carbonate, and magnesium carbonate.

[0227] (ii) Binder In another embodiment, the excipient can be a binder. Suitable binders include, but are not limited to, starch, pregelatinized starch, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamide, polyvinyloxoazolidone, polyvinyl alcohol, C 12 ~C 18 Fatty acid alcohols, polyethylene glycols, polyols, sugars, oligosaccharides, polypeptides, oligopeptides, and combinations thereof.

[0228] (iii) Filler In another embodiment, the excipient can be a filler.Suitable fillers include, but are not limited to, carbohydrates, inorganic compounds, and polyvinylpyrrolidone.By way of non-limiting example, the filler can be calcium sulfate (both dibasic and tribasic), starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, dibasic calcium phosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc, modified starch, lactose, sucrose, mannitol, or sorbitol.

[0229] (iv) buffer In yet another aspect, the excipient can be a buffer. Representative examples of suitable buffers include, but are not limited to, phosphate, carbonate, citrate, Tris buffer, and buffered saline salts (e.g., Tris-buffered saline or phosphate-buffered saline).

[0230] (v) pH adjuster In various aspects, the excipient can be a pH adjusting agent. By way of non-limiting example, the pH adjusting agent can be sodium carbonate, sodium bicarbonate, sodium citrate, citric acid, or phosphoric acid.

[0231] (vi) disintegrant In a further embodiment, the excipient may be a disintegrant. The disintegrant may be non-effervescent or effervescent. Suitable examples of non-effervescent disintegrants include, but are not limited to, starches such as corn starch, potato starch, pregelatinized and modified starches thereof, sweeteners, clays such as bentonite, microcrystalline cellulose, alginates, sodium starch glycolate, gums such as agar, guar, carob, karaya, pesitin, and tragacanth. Non-limiting examples of suitable effervescent disintegrants include sodium bicarbonate in combination with citric acid and sodium bicarbonate in combination with tartaric acid.

[0232] (vii) dispersants In yet another aspect, the excipient may be a dispersant or dispersion enhancer. Suitable dispersants may include, but are not limited to, starch, alginic acid, polyvinylpyrrolidone, guar gum, kaolin, bentonite, purified wood cellulose, sodium starch glycolate, isoamorphous silicate, and microcrystalline cellulose.

[0233] (viii) excipients In another alternative embodiment, the excipient can be a preservative. Non-limiting examples of suitable preservatives include antioxidants such as BHA, BHT, vitamin A, vitamin C, vitamin E, or retinyl palmitate, citric acid, sodium citrate; chelating agents such as EDTA or EGTA; and antimicrobial agents such as parabens, chlorobutanol, or phenol.

[0234] (ix) Lubricants In a further aspect, the excipient can be a lubricant. Non-limiting examples of suitable lubricants include minerals, such as talc or silica; and fats, such as vegetable stearin, magnesium stearate, or stearic acid.

[0235] (x) taste masking agent In yet another embodiment, the excipient can be a taste-masking agent, such as cellulose ethers, polyethylene glycols, polyvinyl alcohols, copolymers of polyvinyl alcohol and polyethylene glycol, monoglycerides or triglycerides, acrylic polymers, mixtures of acrylic polymers and cellulose ethers, cellulose acetate phthalates, and combinations thereof.

[0236] (xi) Flavoring agents In an alternative embodiment, the excipient may be a flavoring agent, which may be selected from synthetic flavor oils and flavoring aromatics and / or extracts from natural oils, plants, leaves, flowers, fruits, and combinations thereof.

[0237] (xii) Coloring agent In a further aspect, the excipient can be a colorant. Suitable color additives include, but are not limited to, Food, Drug, and Cosmetic Colors (FD&C), Drug and Cosmetic Colors (D&C), or External Drug and Cosmetic Colors (Ext. D&C).

[0238] The weight fraction of the excipient or combination of excipients in the composition can be about 99% or less, about 97% or less, about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, about 40% or less, about 35% or less, about 30% or less, about 25% or less, about 20% or less, about 15% or less, about 10% or less, about 5% or less, about 2%, or about 1% or less of the total weight of the composition.

[0239] The present disclosure encompasses pharmaceutical compositions comprising the anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies described herein to facilitate administration and promote stability of the active agent. For example, the anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies of the present disclosure can be mixed with at least one pharmaceutically acceptable carrier or excipient to obtain a pharmaceutical composition that is appropriately and effectively administered (given) to a living subject, such as a suitable subject (i.e., a "subject in need of treatment" or "subject in need"). Methods for preparing and administering the anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies disclosed herein to a subject in need thereof are known to, or can be readily determined by, those skilled in the art. The route of administration of the anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies can be, for example, peripheral, oral, parenteral, inhaled, or topical.

[0240] For effective administration, pharmaceutical compositions are purposefully designed to be suitable for a selected mode of administration and will use pharmaceutically acceptable excipients, such as compatible carriers, dispersing agents, buffers, surfactants, preservatives, solubilizers, isotonicity agents, stabilizers, and the like, as appropriate.

[0241] Non-limiting examples of pharmaceutically acceptable carriers include: saline, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, saturated vegetable fatty acids, partial glyceride mixtures of water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, wool fat, or combinations thereof.

[0242] Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, the composition will also include isotonic agents, for example, sugars, polyalcohols, such as mannitol, sorbitol, or sodium chloride.

[0243] Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0244] The compositions disclosed herein can be frozen or lyophilized for storage and reconstituted in a suitable sterile carrier prior to use.

[0245] In some embodiments, anti-MYCT1 antibodies may be formulated for parenteral administration. Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions, or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (e.g., those based on Ringer's dextrose), and the like. Preservatives and other additives (e.g., antimicrobials, antioxidants, chelating agents, and inert gases, and the like) may also be present. Parenteral formulations may be a single bolus dose, an infusion, or a loading bolus dose, followed by a maintenance dose. The composition may be administered for a specific fixed or variable period of time (eg, once daily), or may be administered on an "as needed" basis.

[0246] Certain pharmaceutical compositions disclosed herein may be orally administered in an acceptable dosage form, such as, for example, capsules, tablets, aqueous suspensions, or solutions. Certain pharmaceutical compositions may also be administered by nasal aerosol or inhalation. Such compositions may be prepared as a solution in saline, utilizing benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, and / or other conventional solubilizing or dispersing agents.

[0247] The amount of anti-MYCT1 antibody combined with the carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. The composition may be administered as a single dose, multiple doses, or as an infusion over a period of time. The dosage regimen may also be adjusted to obtain the optimum desired response (e.g., a therapeutic or prophylactic response).

[0248] In some aspects, the concentration of the antibodies disclosed herein can be a predetermined or standard concentration. In some aspects, the antibodies disclosed herein can be at a concentration of about 1 microgram / milliliter (mg / ml) to about 500 mg / ml, about 1 mg / ml to about 250 mg / ml, about 1 mg / ml to about 200 mg / ml, about 1 mg / ml to about 150 mg / ml, about 1 mg / ml to about 100 mg / ml, about 1 mg / ml to about 75 mg / ml, about 1 mg / ml to about 50 mg / ml, about 0.1 mg / ml to about 100 mg / ml, or other suitable concentration. In some embodiments, the antibodies are formulated to a concentration of about 30 mg / ml. In some embodiments, the antibodies are lyophilized. In some aspects, the antibodies are diluted with an appropriate solution to an appropriate concentration prior to administration (e.g., for therapeutic applications described below).

[0249] In some aspects, the compositions described herein may further comprise an active agent in addition to the antibodies, fragments thereof, or polynucleotides provided herein, including, but not limited to, antibiotics, antipyretics, antibacterial agents, antifungals, NSAIDs, chemotherapeutic agents, and anticancer agents.

[0250] In further embodiments, a composition comprising an anti-MYCT1 antibody of the present disclosure, a fragment thereof, or a polynucleotide encoding the antibody may be administered to a subject in need thereof. In some embodiments, a composition of the present disclosure may be administered to a subject in need thereof, and the subject is a human subject. In some embodiments, a composition of the present disclosure may be administered to a subject in need thereof, and the subject has, is suspected of having, or is at risk for an angiogenesis-related disease. In some embodiments, the angiogenesis-related disease is age-related macular degeneration, ischemic retinopathy, intraocular neovascularization, corneal neovascularization, retinal neovascularization, choroidal neovascularization, diabetic macular edema, diabetic retinal ischemia, diabetic retinal edema, diabetic retinopathy, cancer, rheumatoid arthritis, endometriosis, or alopecia.

[0251] In some embodiments, the compositions of the present disclosure can be administered to a subject in need thereof, the subject having, suspected of having, or at risk for cancer. In some embodiments, the cancer is a cancer comprising cancer cells with increased expression or activity of endogenous MYCT1. Such cancer cells express MYCT1 at a higher level than normal cells (e.g., control breast cells) or cancer cells when compared to cancer cells in an entire patient sample. Such cells express MYCT1 at a higher level than the average level expressed by a population of cancer cells (e.g., the average level of MYCT1 expression in breast cancer cells). In some embodiments, the cancer cells express MYCT1 at a level at least 10% higher than normal cells (e.g., control breast cells) or cancer cells when compared to cancer cells in an entire patient sample, e.g., express MYCT1 at a 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or higher level.

[0252] In some aspects, the cancer is a sarcoma. In some aspects, the cancer is Ewing's sarcoma, soft tissue sarcoma, liposarcoma, epithelioid sarcoma, malignant peripheral nerve sheath tumor, synovial sarcoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, dermatofibrosarcoma, progressive fibromatosis, rhabdomyosarcoma, leiomyosarcoma, undifferentiated pleomorphic sarcoma, chondrosarcoma, gastrointestinal stromal tumor, or uterine sarcoma. In some aspects, the cancer is breast cancer. In some aspects, the cancer is ductal carcinoma in situ, invasive ductal carcinoma, inflammatory breast cancer, or metastatic breast cancer.

[0253] In some embodiments, compositions comprising an anti-MYCT1 antibody of the present disclosure, a fragment thereof, or a polynucleotide encoding the antibody may reduce and / or inhibit the activity, function, expression, and / or accumulation of MYCT1. In some embodiments, administration of a composition comprising an anti-MYCT1 antibody of the present disclosure, a fragment thereof, or a polynucleotide encoding the antibody reduces the activity, function, expression, and / or accumulation of MYCT1 in a subject. In some embodiments, administration of a composition comprising an antibody, a fragment thereof, or a polynucleotide encoding an anti-MYCT1 antibody described herein reduces the activity, function, expression, and / or accumulation of MYCT1 by at least about 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to the activity, function, expression, and / or accumulation of MYCT1 prior to administration of the antibody, fragment thereof, or polynucleotide encoding the antibody (baseline), or compared to a control subject not receiving the treatment.

[0254] In some aspects, a composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces tumor volume in a subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to the tumor volume before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline) or compared to a control subject not receiving the treatment.

[0255] In some embodiments, a composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces tumor volume in a subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to the tumor volume before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline), or compared to a control subject not receiving the treatment. Tumor volume in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-ray, CT scan, bone marrow biopsy, serum protein electrophoresis (SPEP), or MRI.

[0256] In some embodiments, a composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces tumor angiogenesis in a subject by at least 10%, e.g., by 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to tumor angiogenesis before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline), or compared to a control subject not receiving the treatment. Tumor angiogenesis in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as counting the number of blood vessels in a given area using a stereomicroscope, or by alpha- or alpha-receptor immunohistochemistry. v These include measuring the expression of biomarkers associated with angiogenesis, such as β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR)2.

[0257] In some embodiments, a composition comprising an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody may increase the immune response or anti-tumor immunity against cancer cells in a subject by at least 10%, e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, when compared to the immune response or anti-tumor immunity before administration of the antibody, fragment thereof, or polynucleotide encoding the antibody (baseline), or compared to a control subject not receiving the treatment. The immune response against cancer cells may be measured using biomarkers that reflect the tumor immune microenvironment and intrinsic characteristics of tumor cells, such as PD-L1 expression and tumor-infiltrating lymphocyte (TIL) density.

[0258] In further aspects, compositions of the present disclosure may optionally include one or more immune checkpoint inhibitor compounds that may target any immune checkpoint known in the art, including, but not limited to, CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, CD28, CD86, CD69, CD48, CD113, CEACAM-1, Galectin-1, TIGIT, GPR56, CD48, GARP, PD1H, LAIR1, TIM1, TIM4, and the B-7 family of ligands. In some embodiments, the immune checkpoint inhibitor is a CTLA-4 blocking antibody [ipilimumab (Yervoy), tremelimumab (Imjuno)], a PD-1 inhibitor [pembrolizumab (Keytruda), nivolumab (Opdivo), cemiplimab (Libtayo), CT-011 (pidilizumab), AMP224], a PD-L1 inhibitor [atezolizumab (tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), BMS-936559], a Lag3 inhibitor (relatulimab), a combination of a Lag3 and PD1 inhibitor [the PD-1 inhibitor nivolumab (Opdualag), an OX40 inhibitor (MEDI6469), a CD160 inhibitor (BY55)]. Non-limiting examples of inhibitors of CSF-1R include PLX3397, PLX486, RG7155, AMG820, ARRY-382, FPA008, IMC-CS4, JNJ-40346527, MCS 110, or any combination thereof.

[0259] In certain embodiments, compositions of the present disclosure may include one or more PD-1 inhibitor and / or PD-L1 inhibitor compounds. Non-limiting examples of immune checkpoint compounds include monoclonal antibodies that target either PD-1 or PD-L1. In some embodiments, monoclonal antibodies that target PD-1 or PD-L1 include pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, durvalumab, and / or ipilimumab.

[0260] In some embodiments, a combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody, and an immune checkpoint inhibitor compound may be administered to a subject having a cancer that is initially sensitive to immune checkpoint inhibitor treatment but later becomes insensitive to immune checkpoint inhibitor treatment. In some embodiments, a combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody, and an immune checkpoint inhibitor compound may be administered to a subject having a cancer that does not respond to immune checkpoint inhibitor treatment. In some embodiments, the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody of the present disclosure sensitizes the cancer to an immune checkpoint inhibitor.

[0261] In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody, and an immune checkpoint inhibitor compound may reduce tumor volume in a subject by at least 10%, e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, when compared to the tumor volume before administration of the combination of the antibody, a fragment thereof, or a polynucleotide encoding the antibody, and an immune checkpoint inhibitor compound (baseline), a control subject not receiving the treatment, or a subject administered only the antibody, a fragment thereof, or a polynucleotide encoding the antibody, or only the immune checkpoint inhibitor compound. In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody, and an immune checkpoint inhibitor compound reduces tumor volume in a subject by at least 25%. Tumor volume in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-ray, CT scan, bone marrow biopsy, serum protein electrophoresis (SPEP), or MRI.

[0262] In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody with an immune checkpoint inhibitor compound may reduce tumor angiogenesis in a subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to tumor angiogenesis before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline), a control subject not receiving the treatment, or a subject administered only the antibody, a fragment thereof, or a polynucleotide encoding the antibody, or only the immune checkpoint inhibitor compound. Tumor angiogenesis in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as counting the number of blood vessels in a given area using a stereomicroscope, or alpha-antibody assays.v These include measuring the expression of biomarkers associated with angiogenesis, such as β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR)2.

[0263] In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody with an immune checkpoint inhibitor compound may increase the immune response or anti-tumor immunity against cancer cells in a subject by at least 10%, e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, when compared to the immune response or anti-tumor immunity (baseline) before administration of the combination of the antibody, fragment thereof, or polynucleotide encoding the antibody and the immune checkpoint inhibitor compound, a control subject not receiving the treatment, or a subject administered only the antibody, fragment thereof, or polynucleotide encoding the antibody or only the immune checkpoint inhibitor compound. The immune response against cancer cells may be measured using biomarkers reflecting the tumor immune microenvironment and intrinsic characteristics of tumor cells, such as PD-L1 expression and tumor-infiltrating lymphocyte (TIL) density.

[0264] Additionally, compositions of the present disclosure may optionally include one or more VEGF inhibitors. In a non-limiting example, compositions of the present disclosure may optionally include one or more of axitinib, bevacizumab, cabozantinib, lapatinib, lenvatinib, pazopanib, ponatinib, ramucirumab, ranibizumab, regorafenib, sorafenib, sunitinib, and / or vandetanib.

[0265] In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody, and a VEGF inhibitor may reduce tumor volume in a subject by at least 10%, e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, when compared to the tumor volume before administration of the combination of the antibody, fragment thereof, or polynucleotide encoding the antibody and the VEGF inhibitor (baseline), a control subject not receiving the treatment, or a subject administered only the antibody, fragment thereof, or polynucleotide encoding the antibody, or only the VEGF inhibitor. In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody, and a VEGF inhibitor compound reduces tumor volume in a subject by at least 25%. Tumor volume in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-ray, CT scan, bone marrow biopsy, serum protein electrophoresis (SPEP), or MRI.

[0266] In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody with a VEGF inhibitor reduces tumor angiogenesis in a subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to tumor angiogenesis before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline), a control subject not receiving the treatment, or a subject administered only the antibody, a fragment thereof, or a polynucleotide encoding the antibody, or only a VEGF inhibitor. Tumor angiogenesis in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as counting the number of blood vessels in a given area using a stereomicroscope, or α- vThese include measuring the expression of biomarkers associated with angiogenesis, such as β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR)2.

[0267] In some embodiments, the combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody with a VEGF inhibitor compound may increase the immune response or anti-tumor immunity against cancer cells in a subject by at least 10%, e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, when compared to the immune response or anti-tumor immunity (baseline) before administration of the combination of the antibody, fragment thereof, or polynucleotide encoding the antibody and the VEGF inhibitor, a control subject not receiving the treatment, or a subject receiving only the antibody, fragment thereof, or polynucleotide encoding the antibody or only the VEGF inhibitor. The immune response against cancer cells may be measured using biomarkers reflecting the tumor immune microenvironment and intrinsic characteristics of the tumor cells, such as PD-L1 expression and tumor-infiltrating lymphocyte (TIL) density.

[0268] As mentioned above, the agents or compositions described herein may also be used in combination with other therapeutic agents, as further described below. Thus, in addition to the treatments described herein, the subject may also be provided with other therapies known to be effective in treating the disease, disorder, or condition. In various examples, the method further includes administering to the patient an additional cancer treatment. In some examples, the additional cancer treatment is selected from the group including surgery, radiation therapy, chemotherapy, toxin therapy, immunotherapy, cryotherapy, gene therapy, and combinations thereof. Examples of anti-angiogenic therapeutic targets useful according to the present disclosure include EGF, VEGF, FGF, and matrix remodeling proteins. In various examples, the chemotherapeutic agent is a drug or drug formulation. Non-limiting examples of drug formulations include polymeric micelle formulations, liposome formulations, dendrimer formulations, polymer-based nanoparticle formulations, silica-based nanoparticle formulations, nanoscale coordination polymer formulations, nanoscale metal-organic framework formulations, inorganic nanoparticle formulations, and the like.

[0269] A variety of chemotherapeutic agents (e.g., chemotherapy drugs) may be used. Any FDA-approved chemotherapeutic agent (e.g., chemotherapy drug) may be used. Combinations of chemotherapeutic agents may be used. Chemotherapeutic agents and anti-cancer drugs commonly used to treat cancer and that may be used in combination with any of the compositions disclosed herein include, but are not limited to, platinum compounds (e.g., cisplatin or carboplatin) and taxane compounds, such as paclitaxel (Taxol®) or docetaxel (Taxotere®). Other chemotherapeutic agents that are used to treat cancer and that may be used in combination with any of the compositions disclosed herein include, but are not limited to, albumin-bound paclitaxel (nab-paclitaxel, Abraxane®), pemetrexed (Alimta®), irinotecan (CPT-11, Camptosar®), cyclophosphamide (Cytoxan®), liposomal doxorubicin (Doxil®), gemcitabine (Gemzar®), altretamine (Hexalen®), ifosfamide (Ifex®), melphalan (Alkeran), vinorelbine (Navelbine®), topotecan (Hycamtin), etoposide (VP-16), and capecitabine (Xeloda®).

[0270] In some embodiments, the additional drug or therapeutically active agent can be a genotoxic agent (e.g., a DNA damaging agent or drug). As used herein, "genotoxic therapy" refers to a tumor or cancer treatment that utilizes the destructive properties of this treatment to induce DNA damage in tumor or cancer cells. This treatment is traditionally part of a standardized treatment plan. Any damage caused to a tumor cancer is inherited by progeny cancer cells as they continue to grow. If this damage is severe, apoptosis is induced in the cells. By way of non-limiting example, genotoxic therapies may include gamma irradiation, alkylating agents such as nitrogen mustards (chlorambucil, cyclophosphamide, ifosfamide, melphalan), nitrosoureas (streptozocin, carmustine, lomustine), alkylsulfonates (busulfan), triazines (dacarbazine, temozolomide), and ethylenimines (thiotepa, altretamine), platinum drugs such as cisplatin, carboplatin, oxalaplatin, antimetabolites such as 5-fluorouracil, 6-mercaptopurine, capecitabine, cladribine, clofarabine, cytarabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, Rhea, methotrexate, pemetrexed, pentostatin, thioguanine, anthracyclines such as daunorubicin, doxorubicin, epirubicin, idarubicin, antitumor antibiotics such as actinomycin-D, bleomycin, mitomycin-C, mitoxantrone, topoisomerase inhibitors such as topoisomerase I inhibitors (topotecan, irinotecan), and topoisomerase II inhibitors (etoposide, teniposide, mitoxantrone), mitotic inhibitors such as taxanes (paclitaxel, docetaxel), epothilones (ixabepilone), vinca alkaloids (vinblastine, vincristine, vinorelbine), and estramustine.

[0271] Treatment according to the methods described herein may be performed before, simultaneously with, or after conventional treatment modalities for the cancer or tumor.

[0272] The dosage of the additional drug or therapeutically active agent can vary over a wide range, depending on the disease or disorder being treated and the age and condition of the subject being treated. In embodiments in which a composition further comprising at least one additional drug or therapeutically active agent is contacted with the sample, the concentration of the at least one additional drug or therapeutically active agent can be from about 0.01 μM to about 10 μM. Alternatively, the concentration of the at least one additional drug or therapeutically active agent can be from about 0.01 μM to about 5 μM. For example, the concentration of the at least one additional drug or therapeutically active agent can be about 0.01, about 0.05, about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 μM. Additionally, the concentration of the at least one additional drug or therapeutically active agent can be greater than 10 μM. For example, the concentration of the at least one additional agent can be about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, or about 100 μM.

[0273] In embodiments in which a composition further comprising at least one additional drug or therapeutically active agent is administered to a subject, the dose of the additional drug or therapeutically active agent can be about 0.1 mg / kg to about 500 mg / kg. For example, the dose of the at least one additional drug or therapeutically active agent can be about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, or about 25 mg / kg. Alternatively, the dose of the at least one additional drug or therapeutically active agent can be about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 125 mg / kg, about 150 mg / kg, about 175 mg / kg, about 200 mg / kg, about 225 mg / kg, or about 250 mg / kg. Additionally, the dose of the at least one additional drug or therapeutically active agent can be about 300 mg / kg, about 325 mg / kg, about 350 mg / kg, about 375 mg / kg, about 400 mg / kg, about 425 mg / kg, about 450 mg / kg, about 475 mg / kg, or about 500 mg / kg.

[0274] Generally, a safe and effective amount of the composition is administered, e.g., an amount that provides a desired therapeutic effect in a subject while minimizing undesirable side effects. In various embodiments, an effective amount of the compositions described herein can substantially reduce the growth or spread of cancer in a subject. In some embodiments, an effective amount is an amount that can treat cancer or tumors. In some embodiments, an effective amount is an amount that can treat one or more symptoms associated with cancer or tumors.

[0275] The amount of the compositions described herein that are combined with a pharmaceutically acceptable carrier to produce a single dosage form will vary depending on the host treated and the particular mode of administration. Those skilled in the art will understand that the unit content of a drug contained in an individual dose of each dosage form need not itself constitute a therapeutically effective amount, as the required therapeutically effective amount may be achieved by administering multiple individual doses.

[0276] The toxicity and therapeutic efficacy of the compositions described herein were evaluated using LD 50 (lethal dose for 50% of the population) and ED 50 The dose that is therapeutically effective in 50% of the population can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. The dose ratio between toxic and therapeutic effects is the LD 50 / ED 50 The therapeutic index can be expressed as a ratio, and it is generally understood in the art that a larger therapeutic index is optimal.

[0277] The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, weight, general health, sex, and diet of the subject; the time of administration; the route of administration; the rate of excretion of the composition employed; the duration of treatment; drugs used in combination with or concomitantly with the specific compound employed; and factors known in the medical arts (e.g., Koda-Kimble et al. (2004) Applied Therapeutics: The Clinical Use of Drugs, Lippincott Williams & Wilkins, ISBN 0781748453; Winter (2003) Basic Clinical Pharmacokinetics, 4th ed., Lippincott Williams & Wilkins, ISBN 0781741475; Sharqel (2004) Applied Biopharmaceutics & Pharmacokinetics, McGraw-Hill / Appleton & Lange, ISBN (See, e.g., 0071375503). For example, it is well within the skill of one in the art to start a dose of the composition at a lower level than necessary to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. If necessary, the effective daily dose may be divided into multiple doses for purposes of administration. Consequently, single dose compositions may contain such amounts or submultiples thereof to make up the daily dose. However, it will be understood that the total daily usage of the compounds and compositions of the present disclosure will be determined by the attending physician within the scope of sound medical judgment.

[0278] The administration of the compositions disclosed herein can be performed as a single event or can be performed during the course of treatment.For example, the compositions can be administered daily, weekly, biweekly, or monthly.For the treatment of acute conditions, the treatment period usually lasts for at least several days.In certain conditions, treatment can be extended from several days to several weeks.For example, treatment can last for 1 week, 2 weeks, or 3 weeks.For chronic conditions, treatment can last for several weeks to several months, or even more than a year.

[0279] When multiple administrations are performed in the present methods, the administrations can be spaced apart by 1 minute, 2 minutes, 3, 4, 5, 6, 7, 8, 9, 10, or more, and can be spaced apart by about 1 hour, 2 hours, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, etc. In the context of time, the term "about" means any time interval plus or minus up to 30 minutes. Administrations can also be spaced apart by 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, and any combination thereof. The present disclosure is not limited to administration intervals that are evenly spaced in time, and encompasses non-equally spaced doses, such as, by way of non-limiting example, a priming schedule consisting of administration on days 1, 4, 7, and 25.

[0280] For example, dosing schedules of once per week, twice per week, three times per week, four times per week, five times per week, six times per week, seven times per week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, and the like, can be used in the present disclosure. Dosing schedules encompass administration over an entire period, including, for example, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, and 12 months.

[0281] A cycle of the above-mentioned administration schedule is provided. This cycle can be repeated, for example, about every 7 days; every 14 days; every 21 days; every 28 days; every 35 days; every 42 days; every 49 days; every 56 days; every 63 days; every 70 days, and the like. Between cycles, non-administration intervals can occur, and these intervals can be, for example, about 7 days; 14 days; 21 days; 28 days; 35 days; 42 days; 49 days; 56 days; 63 days; 70 days, and the like. In this context, the term "about" means plus or minus 1 day, plus or minus 2 days, plus or minus 3 days, plus or minus 4 days, plus or minus 5 days, plus or minus 6 days, or plus or minus 7 days.

[0282] IV. Method The present disclosure further provides methods of treating a subject with cancer. In some embodiments, the methods described herein can include administering to a subject in need thereof an effective amount of one or more of the anti-MYCT1 antibodies, or fragments thereof, disclosed herein, or one or more of the pharmaceutical compositions disclosed herein. In some embodiments, the methods herein can include administering to a subject in need thereof, where the subject is a human subject. In some embodiments, the methods herein can include administering to a subject in need thereof, where the subject has, is suspected of having, or is at risk for an angiogenesis-related disease. In some embodiments, the angiogenesis-related disease is age-related macular degeneration, ischemic retinopathy, intraocular neovascularization, corneal neovascularization, retinal neovascularization, choroidal neovascularization, diabetic macular edema, diabetic retinal ischemia, diabetic retinal edema, diabetic retinopathy, cancer, rheumatoid arthritis, endometriosis, or alopecia.

[0283] In some embodiments, the methods herein can include administering to a subject having, suspected of having, or at risk for cancer. In some embodiments, the cancer is a cancer comprising cancer cells with increased expression of endogenous MYCT1. Such cancer cells express MYCT1 at a higher level than normal cells (e.g., control breast cells) or cancer cells when compared to cancer cells in an entire patient sample. Such cells express MYCT1 at a higher level than the average level expressed by a population of cancer cells (e.g., the average level of MYCT1 expression in breast cancer cells). In some embodiments, the cancer cells express MYCT1 at a level at least 10% higher than normal cells (e.g., control breast cells) or cancer cells when compared to cancer cells in an entire patient sample, e.g., express MYCT1 at a 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or higher level. In such an aspect, the method comprises administering to the subject a therapeutically effective amount of one or more of the anti-MYCT1 antibodies, or fragments thereof, disclosed herein, or one or more of the pharmaceutical compositions disclosed herein.

[0284] In some aspects of this method, the cancer is a sarcoma. In some aspects, the cancer is Ewing's sarcoma, soft tissue sarcoma, liposarcoma, epithelioid sarcoma, malignant peripheral nerve sheath tumor, synovial sarcoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, dermatofibrosarcoma, progressive fibromatosis, rhabdomyosarcoma, leiomyosarcoma, undifferentiated pleomorphic sarcoma, chondrosarcoma, gastrointestinal stromal tumor, or uterine sarcoma. In some aspects, the cancer is breast cancer. In some aspects, the cancer is ductal carcinoma in situ, invasive ductal carcinoma, inflammatory breast cancer, or metastatic breast cancer.

[0285] In some embodiments, administration of an anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody of the present disclosure may reduce and / or inhibit MYCT1 activity, function, expression, and / or accumulation. In some embodiments, administration of a composition comprising an anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody of the present disclosure reduces MYCT1 activity, function, expression, and / or accumulation in a subject. In some embodiments, administration of an antibody, fragment thereof, or polynucleotide encoding an anti-MYCT1 antibody described herein reduces MYCT1 activity, function, expression, and / or accumulation by at least about 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to MYCT1 activity, function, expression, and / or accumulation before administration of the antibody, fragment thereof, or polynucleotide encoding the antibody (baseline), or compared to a control subject not receiving the treatment.

[0286] In some aspects, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody reduces tumor volume in a subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to the tumor volume before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline) or compared to a control subject not receiving the treatment.

[0287] In some embodiments, administration of an anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody reduces tumor volume in the subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to the tumor volume before administration of the anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody (baseline) or compared to a control subject not receiving the treatment. Tumor volume in a subject can be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-ray, CT scan, bone marrow biopsy, serum protein electrophoresis (SPEP), or MRI.

[0288] In some embodiments, administration of an anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody reduces tumor angiogenesis in the subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to tumor angiogenesis before administration of the anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody (baseline), or compared to a control subject not receiving the treatment. Tumor angiogenesis in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as counting the number of blood vessels in a given area using a stereomicroscope, or alpha-antibody. v These include measuring the expression of biomarkers associated with angiogenesis, such as β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR)2.

[0289] In some embodiments, administration of an anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody may result in at least a 10% increase in immune response or anti-tumor immunity against cancer cells in a subject compared to the immune response or anti-tumor immunity (baseline) before administration of the antibody, fragment thereof, or polynucleotide encoding the antibody, or compared to a control subject not receiving the treatment, and may result in a 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or greater increase. The immune response against cancer cells may be measured using biomarkers that reflect the tumor immune microenvironment and intrinsic characteristics of tumor cells, such as PD-L1 expression and tumor-infiltrating lymphocyte (TIL) density.

[0290] In further aspects, the methods of the present disclosure may optionally include the administration of one or more immune checkpoint inhibitor compounds, which may, in some aspects, be administered prior to, during, or after the administration of an antibody, fragment thereof, or polynucleotide encoding the antibody of the present disclosure. In some aspects, the immune checkpoint inhibitor can be an inhibitor of CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, CD28, CD86, CD69, CD48, CD113, CEACAM-1, Galectin-1, TIGIT, GPR56, CD48, GARP, PD1H, LAIR1, TIM1, TIM4, and the B-7 family of ligands, or any combination thereof. In some embodiments, immune checkpoint inhibitors for use in the methods herein are CTLA-4 blocking antibodies [ipilimumab (Yervoy), tremelimumab (Imjuno)], PD-1 inhibitors [pembrolizumab (Keytruda), nivolumab (Opdivo), cemiplimab (Libtayo), CT-011 (pidilizumab), AMP224], PD-L1 inhibitors [atezolizumab (tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), BMS-936559], Lag3 inhibitors (relatulimab), combinations of Lag3 and PD1 inhibitors [PD-1 inhibitor nivolumab (Opdualag), OX40 inhibitors (MEDI6469), CD160 inhibitors (BY55)]. Non-limiting examples of inhibitors of CSF-1R include PLX3397, PLX486, RG7155, AMG820, ARRY-382, FPA008, IMC-CS4, JNJ-40346527, MCS 110, or any combination thereof.

[0291] In certain embodiments, the methods may include administering one or more PD-1 and / or PD-L1 inhibitor compounds. Non-limiting examples of PD-1 and / or PD-L1 inhibitor compounds include monoclonal antibodies that target either PD-1 or PD-L1. In some embodiments, monoclonal antibodies that target PD-1 or PD-L1 include pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, durvalumab, and / or ipilimumab.

[0292] In a further aspect, a method of treating cancer in a subject unresponsive or resistant to immune checkpoint inhibitor monotherapy is provided. In such aspects, the subject has been diagnosed with, is suspected of having, or develops a cancer unresponsive or resistant to immune checkpoint inhibitors. In such aspects, the combination therapy disclosed herein results in greater than additive inhibition of tumor growth, which may be determined using any suitable measurement, non-limiting examples of which include determining tumor volume. The combination therapy may be combined with any other conventional cancer treatment, including, but not limited to, surgical and chemotherapy approaches.

[0293] In some embodiments of the method, a combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody and an immune checkpoint inhibitor compound is administered to a subject having a cancer that is initially sensitive to immune checkpoint inhibitor treatment and later becomes insensitive to immune checkpoint inhibitor treatment. In some embodiments, a combination of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody and an immune checkpoint inhibitor compound may be administered to a subject having a cancer that is insensitive to immune checkpoint inhibitor treatment. In some embodiments, the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody of the present disclosure sensitizes the cancer to an immune checkpoint inhibitor.

[0294] In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound may reduce tumor volume in a subject by at least 10%, e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, when compared to the tumor volume before administration of the antibody, fragment thereof, or polynucleotide encoding the antibody in combination with the immune checkpoint inhibitor compound (baseline), a control subject not receiving the treatment, or a subject administered only the antibody, fragment thereof, or polynucleotide encoding the antibody or only the immune checkpoint inhibitor compound. In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound reduces tumor volume in a subject by at least 25%. Tumor volume in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-ray, CT scan, bone marrow biopsy, serum protein electrophoresis (SPEP), or MRI.

[0295] In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound may reduce tumor angiogenesis in a subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to tumor angiogenesis before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline), a control subject not receiving the treatment, or a subject administered only the antibody, a fragment thereof, or a polynucleotide encoding the antibody, or only the immune checkpoint inhibitor compound. Tumor angiogenesis in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as counting the number of blood vessels in a given area using a stereomicroscope, or alpha-antibody assays. v These include measuring the expression of biomarkers associated with angiogenesis, such as β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR)2.

[0296] In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with an immune checkpoint inhibitor compound may result in at least a 10% increase in immune response or anti-tumor immunity against cancer cells in a subject, compared to the immune response or anti-tumor immunity (baseline) before administration of the combination of the antibody, fragment thereof, or polynucleotide encoding the antibody and the immune checkpoint inhibitor compound, a control subject not receiving the treatment, or a subject administered only the antibody, fragment thereof, or polynucleotide encoding the antibody or only the immune checkpoint inhibitor compound. The immune response against cancer cells may be measured using biomarkers reflecting the tumor immune microenvironment and intrinsic characteristics of tumor cells, such as PD-L1 expression and tumor-infiltrating lymphocyte (TIL) density.

[0297] Additionally, the methods of treatment described herein may optionally include administering one or more VEGF inhibitors. In some embodiments, the VEGF inhibitors may be administered before, during, or after administration of the disclosed antibody, fragment thereof, or polynucleotide encoding the antibody. In a non-limiting example, the disclosed compositions may optionally include one or more of axitinib, bevacizumab, cabozantinib, lapatinib, lenvatinib, pazopanib, ponatinib, ramucirumab, ranibizumab, regorafenib, sorafenib, sunitinib, and / or vandetanib.

[0298] In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor may reduce tumor volume in a subject by at least 10%, e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, when compared to the tumor volume before administration of the antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor (baseline), a control subject not receiving the treatment, or a subject administered only the antibody, a fragment thereof, or a polynucleotide encoding the antibody, or only a VEGF inhibitor. In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor compound reduces tumor volume in a subject by at least 25%. Tumor volume in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as X-ray, CT scan, bone marrow biopsy, serum protein electrophoresis (SPEP), or MRI.

[0299] In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor reduces tumor angiogenesis in the subject by at least 10%, e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more, when compared to tumor angiogenesis before administration of the anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody (baseline), a control subject not receiving the treatment, or a subject receiving only the antibody, a fragment thereof, or a polynucleotide encoding the antibody, or only a VEGF inhibitor. Tumor angiogenesis in a subject may be assessed using any conventional method known in the art, non-limiting examples of which include diagnostic techniques such as counting the number of blood vessels in a given area using a stereomicroscope, or α vThese include measuring the expression of biomarkers associated with angiogenesis, such as β3 integrin, endoglin, and vascular endothelial growth factor (VEGF) receptor (VEGFR)2.

[0300] In some embodiments, administration of an anti-MYCT1 antibody, a fragment thereof, or a polynucleotide encoding the antibody in combination with a VEGF inhibitor compound may result in at least a 10% increase in immune response or anti-tumor immunity against cancer cells in a subject, compared to the immune response or anti-tumor immunity (baseline) before administration of the antibody, fragment thereof, or polynucleotide encoding the antibody in combination with a VEGF inhibitor, a control subject not receiving the treatment, or a subject receiving only the antibody, fragment thereof, or polynucleotide encoding the antibody or only the VEGF inhibitor. The immune response against cancer cells may be measured using biomarkers reflecting the tumor immune microenvironment and intrinsic characteristics of tumor cells, such as PD-L1 expression and tumor-infiltrating lymphocyte (TIL) density.

[0301] Among various aspects of the present disclosure, methods for modulating MYCT1 activity are provided. One aspect of the present disclosure provides a method for regulating tumor angiogenesis (anti-angiogenesis, Myct1-targeted vascular regulation) and / or immune stimulation that inhibits tumor growth in a subject. In some aspects, the method comprises administering an anti-MYCT1 antibody to a subject with cancer.

[0302] In further aspects, the methods disclosed herein include sensitizing a cancer in a subject to an immune checkpoint inhibitor. In such aspects, the methods include selecting a subject with cancer, wherein the cancer is resistant to an immune checkpoint inhibitor. In some aspects, the subject may be diagnosed, e.g., by a physician, with, is suspected of having, or has developed a cancer that is resistant or insensitive to an immune checkpoint inhibitor. In further aspects, the methods include administering to the subject an effective amount of a composition comprising an anti-MYCT1 antibody or fragment thereof disclosed herein. In some aspects, administration of the anti-MYCT1 antibody or fragment thereof in such a subject reduces angiogenesis and increases anti-tumor immunity in the subject. In further aspects, administration of the anti-MYCT1 antibody in such a subject reduces tumor volume.

[0303] Further embodiments include methods of controlling tumor angiogenesis and / or increasing anti-tumor immunity in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising an anti-MYCT1 antibody or fragment thereof.

[0304] In some embodiments, an effective amount of an anti-MYCT1 antibody can be an amount effective to substantially block or inhibit MYCT1 expression and / or activity in cells. In some embodiments, an effective amount of an anti-MYCT1 antibody can be an amount effective to substantially block or inhibit MYCT1 expression and / or activity in endothelial cells (ECs). In some embodiments, blocking MYCT1 in endothelial cells promotes an immunostimulatory microenvironment by enhancing CTL infiltration and preventing CTL apoptosis; contributes to an immunostimulatory microenvironment; provides an anti-tumor microenvironment; and / or promotes endothelial control of tumor immunity. In some embodiments, an effective amount of an anti-MYCT1 antibody can be an amount effective to reduce angiogenesis. In some embodiments, an effective amount of an anti-MYCT1 antibody can be an amount effective to promote high endothelial venule (HEV) formation. In some embodiments, administration of a composition of the present disclosure increases EC HEV. In some embodiments, after administration of an anti-MYCT1 antibody of the present disclosure, a subject experiences improved delivery of an anti-tumor drug and enhanced anti-tumor immunity. In some aspects, the subject has formed high endothelial venules and an anti-tumor immune environment, limiting tumor progression.

[0305] In some further embodiments, an effective amount of an anti-MYCT1 antibody can be an amount effective to enhance robust CTL infiltration. In some embodiments, an effective amount of an anti-MYCT1 antibody can be an amount effective to promote inflammatory M1 macrophage polarization. In some embodiments, administration of a composition of the present disclosure reduces EC motility. In some embodiments, administration of a composition of the present disclosure results in loss of tumor vasculature. In some embodiments, administration of a composition of the present disclosure downregulates Fas ligand expression in EC.

[0306] In some embodiments, the subject has downregulated MYCT1 in EC prior to administration of the anti-MYCT1 antibody. Suitable compositions, dosage forms, administrations, and administration schedules for use in the methods of the present disclosure are disclosed herein, for example, as described in Section III, which is incorporated herein by reference.

[0307] In some embodiments, the amounts of anti-MYCT1 antibody and immune checkpoint inhibitor and / or VEGF inhibitor are effective to reduce or prevent the depletion of infiltrating cytotoxic T lymphocytes (CTLs). In some embodiments, the combination of vascular regulation and immune regulation results in synergistic anti-tumor activity.

[0308] In another aspect, the present disclosure provides compositions disclosed herein for in vitro, in vivo, or ex vivo use.

[0309] Thus, in some aspects, the methods of the present disclosure are useful for inhibiting tumor or cancer progression or prolonging the survival of subjects with cancer or tumors. The present disclosure provides a combination of vascular and immune regulation that results in synergistic anti-tumor activity. For example, targeting MYCT1 using the anti-MYCT1 antibodies of the present disclosure improves response to anti-PD1 therapy in treatment-responsive and treatment-refractory tumors, and combining MYCT1 and VEGF targeting with anti-PD1 treatment results in superior tumor control, suggesting a synergistic effect between the MYCT1 and VEGF pathways.

[0310] In one aspect, the method generally includes the steps of providing a composition described herein and administering the composition to a subject or a cell (e.g., an endothelial cell).

[0311] In some aspects, the present disclosure provides methods for treating tumors or cancer cells at increased risk of angiogenesis. In some aspects, the present disclosure provides methods for treating cancer or tumors by administering a composition of the present disclosure sequentially or simultaneously with another cancer treatment. When administered sequentially, the additional agent may be administered before or after the compound of the present disclosure. In a preferred aspect, the additional agent is an immune checkpoint blockade, and may be an anti-VEGF therapy.

[0312] In other aspects, the compositions of the present disclosure can be delivered to cancer cells in vitro. The cancer cells can be a cancer cell line cultured in vitro. In some alternatives to this aspect, the cancer cell line can be a primary cell line that has not yet been described. Methods for preparing primary cancer cell lines utilize standard techniques known to those of skill in the art. In other alternatives, the cancer cell line can be an established cancer cell line. The cancer cell line can be adherent or non-adherent, or the cell line can be grown under conditions that promote adherent, non-adherent, or organotypic growth using standard techniques known to those of skill in the art. The cancer cell line can be contact inhibited or non-contact inhibited.

[0313] In some embodiments, the cancer cell line may be an established human cell line derived from a tumor. Non-limiting examples of cancer cell lines derived from a tumor include the MM cell lines MM.1S, H929, and RPMI, osteosarcoma cell lines 143B, CAL-72, G-292, HOS, KHOS, MG-63, Saos-2, and U-2 OS; prostate cancer cell lines DU145, PC3, and Lncap; breast cancer cell lines MCF-7, MDA-MB-438, and T47D; myeloid leukemia cell line THP-1; glioblastoma cell line U87; neuroblastoma cell line SHSY5Y; bone cancer cell line Saos-2; and colon cancer cell lines WiDr, COLO 320DM, HT29, DLD-1, COLO 205, and COLO 201, HCT-15, SW620, LoVo, SW403, SW403, SW1116, SW1463, SW837, SW948, SW1417, GPC-16, HCT-8, HCT 116, NCI-H716, NCI-H747, NCI-H508, NCI-H498, COLO 320HSR, SNU-C2A, LS 180, LS 174T, MOLT-4, LS513, LS1034, LS411N, Hs 675.T, CO 88BV59-1, Co88BV59H21-2, Co88BV59H21-2V67-66, 1116-NS-19-9, TA 99, AS 33, T.S. 106, Caco-2, HT-29, SK-CO-1, SNU-C2B, or SW480; B16-F10, RAW264.7, F8 cell lines, or the pancreatic cancer cell line Panc1.

[0314] Generally, the methods described herein involve administering to a subject a therapeutically effective amount of a composition of the present disclosure. The methods described herein are generally performed on a subject in need thereof.

[0315] The level of MYCT1 can be used as a predictive marker of the subject's angiogenesis status, and is therefore useful for making treatment decisions for subjects who may benefit from a particular treatment (e.g., an anti-angiogenic agent). For example, the MYCT1 level of a candidate subject can be compared with a reference value.

[0316] Thus, the present disclosure also provides anti-MYCT1 antibodies conjugated to a detectable signal (i.e., a measurable substance or a substance that generates a measurable signal). Non-limiting examples include various enzymes, prosthetic groups, fluorescent substances, luminescent substances, bioluminescent substances, radioactive substances, positron-emitting metals using various positron emission tomography techniques, and non-radioactive paramagnetic metal ions. See, e.g., U.S. Pat. No. 4,741,900 for metal ions that can be conjugated to antibodies for use as diagnostic agents according to the present disclosure. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin; examples of suitable radioactive materials include 125 I, 131 I, 111 In, or 99 Tc The signal generated by the agent can be measured, for example, by single photon emission computed tomography (SPECT) or positron emission tomography (PET).

[0317] Anti-MYCT1 antibodies conjugated to a detectable signal can be used diagnostically to monitor the development or progression of tumors or cancer, for example, as part of a clinical trial procedure to determine the effectiveness of a given treatment and / or prophylactic regimen. Patient treatment can be adjusted based on the level of progression of the neurodegenerative disease.

[0318] In some embodiments, methods are provided for assessing disease progression in a subject undergoing treatment for cancer or a tumor, the methods comprising: (a) administering an anti-MYCT1 antibody disclosed herein labeled with an agent that generates a measurable signal as described herein (i.e., a "labeled anti-MYCT1 antibody"), wherein the signal is measured in the patient after administration; (b) administering a labeled anti-MYCT1 or antigen-binding fragment thereof at one or more time intervals after administration of (a), wherein the signal is measured in the patient after administration; and (c) assessing disease progression in the patient based on a change in the signal measured in the patient at one or more time intervals after administration of (a), wherein an increase in the signal indicates progression of a neurodegenerative disease in the patient. In certain embodiments, the subject is treated with the same anti-MYCT1 antibody, but in an unlabeled form. In certain embodiments, the subject is treated with an anti-MYCT1 antibody that competitively inhibits binding of the labeled anti-MYCT1 antibody to human ApoE. In certain embodiments, the subject has been treated with an anti-MYCT1 antibody that does not competitively inhibit the binding of a labeled anti-MYCT1 antibody to human MYCT1, hi certain embodiments, the subject has been treated with other drugs known in the art.

[0319] In another aspect, a method for assessing disease progression in a subject undergoing treatment for cancer or a tumor is provided, the method comprising: (a) administering an anti-MYCT1 antibody disclosed herein labeled with an agent that generates a measurable signal as described herein (i.e., a "labeled anti-MYCT1 antibody"), where the signal is measured in the patient after administration; (b) assessing the disease state in the subject by comparing the signal measured in the subject with the signal measured after administration of the labeled antibody or antigen-binding fragment thereof to one or more control subjects, where an increase in the signal generated in the patient relative to the control subjects correlates with an increase in cerebral amyloid plaques; and (c) treating the patient with a therapy appropriate for the patient's disease state. A "control subject" refers to any normal, healthy subject (or pool of subjects), subjects with various degrees of disease, or actual test subjects at an early stage of disease.

[0320] The reference value may represent the same level of MYCT1 in a control subject or may represent the MYCT1 level in a control population. In some examples, the same level of MYCT1 in a control subject or control population may be determined by the same method used to determine the MYCT1 level in a candidate subject. In some cases, the control subject or control population may be a subject with cancer or a tumor, or a population of subjects with cancer or a tumor that responds to treatment. In other cases, the control subject or control population may be a subject with cancer or a tumor, or a population of subjects with cancer or a tumor that does not respond to treatment. Alternatively, the control subject or control population may refer to a healthy subject or a population of healthy subjects. In preferred embodiments, the control subject or control population is of the same species as the candidate subject (e.g., a human subject or a population of human subjects). As used herein, assessing "responsiveness" or "non-responsiveness" to a treatment refers to determining the likelihood that a subject will respond or not respond to a therapeutic agent. For example, a responsive subject will experience a therapeutic benefit as a result of administering the agent. In some embodiments, the therapeutic benefit includes prevention or delay (alleviation) of an undesirable physiological change or disease / disorder. Beneficial or desirable clinical outcomes include, but are not limited to, alleviation of symptoms, reduction in extent of disease, stable (i.e., not worsening) disease, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (partial or complete), or can also mean prolonged survival as compared to expected survival for a subject or population that does not receive or respond to treatment.

[0321] It should be understood that the methods provided herein do not require that a reference value be measured each time a candidate subject is tested. Rather, in some embodiments, it is contemplated that a reference value may be obtained and recorded, and that any test level may be compared to such a reference level. This reference level may be a single cutoff value or a range of values.

[0322] By comparing the levels of MYCT1 in a candidate subject disclosed herein with the reference values also described herein, the subject can be identified as responding or likely to respond, or not responding or unlikely to respond, to the treatment based on the assessment.

[0323] For example, if the reference value represents a level of MYCT1 that is responsive to a treatment, derivation from such a reference value would be indicative of non-responsiveness to the treatment. In another embodiment, if the reference value represents a level of MYCT1 that is responsive to a genotoxic therapy, approximately the same level of MYCT1 phosphorylation in the candidate subject would be indicative of responsiveness to the therapy.

[0324] Alternatively, if the reference value represents a level of MYCT1 that is non-responsive to treatment, derivation from such reference value would be indicative of responsiveness to treatment. In another embodiment, if the reference value represents a level of MYCT1 that is non-responsive to treatment, a substantially identical level of MYCT1 in a candidate subject would be indicative of non-responsiveness to treatment.

[0325] In some cases, derivation means that the level of MYCT1 in the candidate subject (e.g., represented by a value) is elevated or decreased compared to a reference value, e.g., by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 300%, 400%, 500%, or even above or below this reference value.

[0326] Thus, the present disclosure provides methods for assessing responsiveness to a treatment or detecting the angiogenic status of a cancer or tumor in a subject having cancer or a tumor. In one aspect, the method for assessing responsiveness to a treatment or detecting the angiogenic status may include (a) providing a biological sample from the subject and measuring the level of MYCT1 in cancer or tumor cells or endothelial cells of the sample; and (b) determining the responsiveness to a treatment or the angiogenic status of the subject if the measured MYCT1 level deviates from a reference value.

[0327] MYCT1 levels can be used in various mathematical calculations to assess reactivity or angiogenesis status. For example, MYCT1 values can be combined with other known biomarkers and used in various statistical models (e.g., linear regression, LME curve, LOESS curve, etc.). The selection of measurements and mathematical calculations can be optimized to maximize the specificity of the method. For example, diagnostic accuracy can be evaluated by the area under the receiver operating characteristic curve (ROC) curve, and in some embodiments, an ROC AUC value of 0.7 or greater is set as the threshold (e.g., 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, etc.).

[0328] V. Kit The present disclosure provides kits containing one or more anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies for use in the methods of the present disclosure. The kits may include a composition containing an anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody, or a pharmaceutical composition thereof, and instructions for administering the antibody fragment, polynucleotide encoding the antibody, or pharmaceutical composition to a subject in need thereof. The kits may further include an immune cell inhibitor and / or a VEGF inhibitor, which may be administered in combination with the anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody. In some embodiments, the kits may further include a sterile, pharmaceutically acceptable carrier, buffer, or other diluent. The kits provided herein generally include instructions for carrying out the methods. The instructions included in the kits may be affixed to packaging or included as a package insert. The instructions are typically written or printed, but are not limited thereto. Any medium capable of storing and transmitting such instructions to an end user is contemplated by the present disclosure. Such media include, but are not limited to, electronic storage media (e.g., magnetic discs, tapes, cartridges, chips), optical media (e.g., CD ROM), and the like. As used herein, the term "instructions" may include the address of an internet site that provides the instructions.

[0329] The kits of the present disclosure may have a single container containing the anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody (with or without any additional components), or may have a separate container for each desired agent. When a combination of therapeutic agents is provided, a single solution may be premixed in molar equivalent combinations, or premixed so that one component is in excess of the other. Alternatively, the antibody, fragment thereof, or polynucleotide encoding the antibody and each of the other anti-cancer drug components of the kit may be kept separate in separate containers prior to administration to a patient.

[0330] When the components of the kit are provided in one or more liquid solutions, the liquid solution is preferably an aqueous solution, with a sterile aqueous solution being particularly preferred. However, the components of the kit can also be provided as a dry powder. When a reagent or component is provided as a dry powder, the powder can be reconstituted by adding a suitable solvent. It is also contemplated that the solvent can be provided in a separate container.

[0331] The containers of the kit generally include at least one vial, test tube, flask, bottle, syringe, or other container means into which the anti-MYCT1 antibody, fragment thereof, or polynucleotide encoding the antibody of the present disclosure and any other desired agents can be placed, and preferably suitably aliquoted. When separate components are included, the kit also generally includes a second vial or other container for containing them, thereby allowing for the administration of separate, designed doses. The kit may also include second or third container means for containing a sterile, pharmaceutically acceptable buffer or other diluent.

[0332] The kits may also include a means for administering the disclosed anti-MYCT1 antibodies, fragments thereof, or polynucleotides encoding the antibodies to an animal or patient, such as one or more needles or syringes, or even an eyedropper, pipette, or other similar device, from which the formulation may be injected into an animal or applied to an affected area of the body. The kits of the present invention also typically include a means for containing vials or the like and other components in a closed condition for commercial sale, such as injection- or blow-molded plastic containers into which the desired vials and other devices may be placed and retained.

[0333] Since various modifications can be made to the materials and methods described above without departing from the scope of the present invention, all matter contained in the above description and the following examples is to be interpreted as illustrative and not in a limiting sense. [Example]

[0334] The following examples are included to illustrate various aspects of the present disclosure. Those of skill in the art should understand that the techniques disclosed in the examples below represent techniques discovered by the inventors to function well in the practice of the invention, and as such, can be considered to constitute preferred modes for practicing the invention. However, those of skill in the art should, in light of the present disclosure, understand that many changes can be made in the specific embodiments disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

[0335] Materials and Methods [Table 2]

[0336] Cultivation of hybridoma lines The media shown in Table 3 were prepared using the following reagents. [Table 3]

[0337] Approximately 1 x 10 cells 5 cells / ml ~ 1 x 10 6 The cells were maintained in culture at 100 cells / ml.

[0338] EC culture MCEC and its derivatives: MCEC cells were generated from FVB / N mouse hearts derived from mice with the MHC haplotype "q." Culture medium was prepared using DMEM high glucose, 10% FBS, 10 mmol / L Pen-Strep, and 10 mmol / L HEPES. Gelatin pre-coating is recommended but not required for this cell line. The cell doubling time was approximately 30 hours. The cells were subcultured at a passage ratio of approximately 1:6. Cells were cryopreserved at 1.5 million cells / ml in 10% DMSO in FBS.

[0339] MLEC, MLEC Myct1 KO, Myct1 KO MLEC:iMLEC, and iMLEC Myct1 KO cells overexpressing human short or long MYCT1 and its derivatives were generated from the lungs of wild-type and Cdh5-cre Myct1 f / f C57 / B6 mice, respectively. These cells were immortalized by DOX-inducible SV40 T with puromycin and blasticidin selection (pLV[Tet]-TRE3G>SV40-T(ns):T2A:Puro and pLV[Exp]-Bsd-{mCd144}>Tet3 constructs). Culture medium was prepared using DMEM high glucose supplemented with 10% FBS, 10 mmol / L Pen-Strep, 10 mmol / L HEPES, and DOX (40 ng / ml). Optimal growth requires coating the culture vessel with 0.1% gelatin (in dH2O) (30 min in a 37°C incubator >> vacuum >> dry in a TC hood for 30 min >> ready to use). After passaging, it takes approximately 3-4 days for the cells to fully repopulate the culture vessel. Subculture was performed at a split ratio of approximately 1:3. Cryopreservation was performed using 10% DMSO in FBS at 1.5 million cells / ml.

[0340] Mouse Myct1-overexpressing HUVECs, MYCT1-knockdown HUVECs, and MYCT1-knockdown HUVECs were purchased from Fisher (catalog number 08-774-388). HUVEC cells were immortalized using human TERT lentivirus (vector: LB12_FLAG_TERT_pCDH_MCS_EF1_NEO) followed by G418 selection. Endothelial Cell Growth Medium (Ready-to-Use) (Promo Cell, catalog: C-22010) was used as the culture medium, containing approximately 2% FBS and 10% serum. For optimal growth, coating the culture vessel with 0.1% gelatin (in dH2O) (30 min in a 37°C incubator >> vacuum >> dry in a TC hood for 30 min >> ready-to-use) was required. After passaging, it takes approximately 4–5 days for the cells to fully repopulate the culture vessel. Passage was performed at a split ratio of 1:2. We observed that HUVEC cells prefer to reside close to each other, and if cells are sparsely seeded or passaged, they may not grow at all. Cryopreservation was performed in 10% DMSO in FBS at 1.5 million cells / ml.

[0341] A HUVEC MYCT1 KD cell line was developed by transducing a HUVEC cell line with MYCT1 shRNA lentiviral particles followed by puromycin selection. A HUVEC MYCT1 KD Myct1-overexpressing cell line was generated by further transducing an iHUVEC MYCT1 KD cell line with Myct1-overexpressing lentiviral particles followed by blasticidin selection.

[0342] Endothelial tube formation assay Endothelial cells were serum-starved overnight. The next day, μ-Slide Angiogenesis plates (catalog number 81506, ibidi) were coated with Matrigel (catalog number 96992, Corning) (10 μl / well) and incubated at 37°C for 30–45 min until the Matrigel polymerized. Endothelial cells serum-starved overnight were seeded onto the Matrigel-coated plates at 8,000 cells / well and topped with 50 μl of culture medium. The plates were incubated at 37°C for 6–8 h and then photographed under a microscope. The total tube length, number of loops, and number of branches were quantified using the Angiogenesis Analyzer module of ImageJ software (NIH).

[0343] Endothelial sprouting assay The following reagents are used in the endothelial sprouting assay: 1. EGM-2 Bullet Kit (including FGF2 and VEGF-A; Lonza, Catalog No. CC-3162) 2.M199 3. DMEM High Glucose 4. Fetal Bovine Serum 5.HBSS 6. Ca2+- and Mg2+-free Dulbecco's Phosphate Buffered Saline (DPBS; Life Technologies, Cat. No. 14190-094) 7. Cytodex 3 microcarrier beads (GE Healthcare, catalog number 17-0485-01) 8. Fibrinogen type I from bovine plasma (Sigma-Aldrich, Cat. No. F8630) 9. Thrombin from bovine plasma (Sigma-Aldrich, Cat. No. T3399) 10. Aprotinin from bovine lung (Sigma-Aldrich, Catalog No. A1153) 11. Trypsin / EDTA, UV inactivated (PAA Laboratories, Catalog No. L11-660) 12.5ml, 15ml, and 50ml polypropylene tubes 13.5ml polystyrene tube (FACS tube) 14. Low-binding Eppendorf Chews (1.5ml) 15.PKH Kit 16.24-well glass bottom plate and #1.5 cover glass 17. Cell Counting Kit

[0344] Endothelial growth medium (EGM-2) was prepared and supplemented according to the supplier's instructions before use.

[0345] Fibroblast growth medium was prepared with DMEM containing 10% FBS, which was supplemented with 10% FBS before use.

[0346] Endothelial cells (YSE / MCEC cells) were grown to approximately 80% confluency in EGM-2 before the start of the assay.

[0347] Fibroblasts (NIH3T3 cells) were grown in fibroblast growth medium to approximately 80% confluency.

[0348] Microcarrier beads: 1 g of Cytodex3 microcarriers (3 x 10^6 beads) were incubated with Ca for at least 3 hours at room temperature. 2+ and Mg 2+ The microcarriers were swelled and hydrated in 50 ml of Ca-free PBS. The supernatant was decanted and the microcarriers were rehydrated in fresh Ca-free PBS. 2+ and Mg 2+ The cells were washed with Ca-free PBS (30-50 ml) for several minutes. The PBS was discarded and fresh Ca 2+ and Mg 2+ The solution was replaced with free PBS (30-50 ml / g), and the microcarriers were sterilized by autoclaving (115°C, 15 min, 15 psi). The pH of all solutions was adjusted to 7.4. When hydrating Cytodex 3, initial surface tension may prevent wetting and settling of the microcarriers. In such cases, Tween 80 can be added to the PBS used for the initial hydration rinse (2-3 drops of Tween 80 per 100 ml PBS).

[0349] Thrombin solution: Thrombin was dissolved in DPBS at 50 U / ml. Aliquots of 0.5 ml each were made. Aliquots can be stored at -20°C. Repeated freezing and thawing should be avoided.

[0350] Aprotinin solution: Aprotinin was dissolved in ddH2O at 4 U / ml. The solution was sterile filtered and aliquots of 0.5 ml were made. Aliquots can be stored at -20°C.

[0351] Endothelial sprouting assays were performed using the following procedure.

[0352] Step 0: Cell preparation [4-5 days] 0.1 Various types of YSE / MCEC cells and NIH3T3 cells are cultured in 10% FBS M199 1% P / S medium and 10% FBS DMEM 1 mM l-glutamine 1% P / S medium, respectively. The day before beading of 0.2 YSE / MCEC cells, switch the medium to EGM-2 (day 0).

[0353] Step 1: Preparation of NIH3T3 fibroblasts (Day 1) [10 min] 1. Remove and discard the culture medium from a T75 flask of NIH3T3 cells at approximately 80% confluency and replace with pre-warmed EGM-2. Incubate overnight in a cell culture incubator at 37°C, 5% CO2.

[0354] Step 2: Prepare various endothelial cells (Day 1) [35 min] This process should be done separately for all types of YSE / MCEC cells: 2.1 Harvest 80% confluent YSE / MCEC cells from a T75 flask by removing the medium, rinsing with prewarmed trypsin-EDTA, adding 3 ml of trypsin-EDTA, and incubating at 37°C for 2-3 minute...

Claims

1. An isolated anti-MYCT1 antibody or fragment thereof that specifically binds to an epitope within the first 100 amino acids of SEQ ID NO:

145.

2. 2. The isolated anti-MYCT1 antibody or fragment thereof of claim 1, comprising a light chain variable region comprising: L1 having the amino acid sequence of SEQ ID NO:28, SEQ ID NO:64, SEQ ID NO:100, or SEQ ID NO:136; L2 having the amino acid sequence of SEQ ID NO:30, SEQ ID NO:66, SEQ ID NO:102, or SEQ ID NO:138; L3 having the amino acid sequence of SEQ ID NO:32, SEQ ID NO:68, SEQ ID NO:104, or SEQ ID NO:140; or any combination thereof; and / or a heavy chain variable region comprising: H1 having the amino acid sequence of SEQ ID NO:10, SEQ ID NO:46, SEQ ID NO:82, or SEQ ID NO:118; H2 having the amino acid sequence of SEQ ID NO:12, SEQ ID NO:48, SEQ ID NO:84, or SEQ ID NO:120; H3 having the amino acid sequence of SEQ ID NO:14, SEQ ID NO:50, SEQ ID NO:86, or SEQ ID NO:122; or any combination thereof.

3. 2. The isolated anti-MYCT1 antibody or fragment thereof of claim 1, comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 36, SEQ ID NO: 72, SEQ ID NO: 108, or SEQ ID NO: 144; and / or a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 18, SEQ ID NO: 54, SEQ ID NO: 90, or SEQ ID NO: 126, or any combination thereof.

4. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 28, L2 having the amino acid sequence of SEQ ID NO: 30, L3 having the amino acid sequence of SEQ ID NO: 32, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 10, H2 having the amino acid sequence of SEQ ID NO: 12, H3 having the amino acid sequence of SEQ ID NO: 14, or any combination thereof.

5. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 64, L2 having the amino acid sequence of SEQ ID NO: 66, L3 having the amino acid sequence of SEQ ID NO: 68, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 46, H2 having the amino acid sequence of SEQ ID NO: 48, H3 having the amino acid sequence of SEQ ID NO: 50, or any combination thereof.

6. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 100, L2 having the amino acid sequence of SEQ ID NO: 102, L3 having the amino acid sequence of SEQ ID NO: 104, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 82, H2 having the amino acid sequence of SEQ ID NO: 84, H3 having the amino acid sequence of SEQ ID NO: 86, or any combination thereof.

7. An isolated anti-MYCT1 antibody or a fragment thereof, comprising a light chain variable region comprising L1 having the amino acid sequence of SEQ ID NO: 136, L2 having the amino acid sequence of SEQ ID NO: 138, L3 having the amino acid sequence of SEQ ID NO: 140, or any combination thereof; and / or a heavy chain variable region comprising H1 having the amino acid sequence of SEQ ID NO: 118, H2 having the amino acid sequence of SEQ ID NO: 120, H3 having the amino acid sequence of SEQ ID NO: 122, or any combination thereof.

8. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 36 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

18.

9. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 72 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

54.

10. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 108 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

90.

11. An isolated anti-MYCT1 antibody or a fragment thereof, comprising the amino acid sequence of the light chain variable region of SEQ ID NO: 144 and / or the amino acid sequence of the heavy chain variable region of SEQ ID NO:

126.

12. 10. The isolated antibody or fragment thereof of any one of the preceding claims, wherein the antibody is a monoclonal antibody.

13. 10. The isolated antibody or fragment thereof of any one of the preceding claims, wherein the antibody is a humanized antibody, a single chain variable fragment (scFv) antibody, an antibody fragment, or a chimeric antibody.

14. A pharmaceutical composition comprising the isolated antibody or fragment thereof of any one of the preceding claims and a pharmaceutically acceptable carrier or excipient.

15. 10. An isolated antibody according to any one of the preceding claims for use as a medicament.

16. 1. A method of treating cancer in a subject in need thereof, comprising: Administering an effective amount of a composition comprising the isolated antibody or fragment thereof of any one of the preceding claims. A method comprising:

17. Administering immune checkpoint inhibitors and, if appropriate, anti-VEGF targeted therapy.

17. The method of claim 16, further comprising:

18. 1. A method of sensitizing cancer to an immune checkpoint inhibitor in a subject, comprising: Selecting a subject having cancer, wherein the cancer is resistant to an immune checkpoint inhibitor; and administering to said subject an effective amount of a composition comprising the isolated antibody or fragment thereof of any one of claims 1 to 15. A method comprising:

19. The method of any one of claims 16 to 18, wherein administration of the anti-MYCT1 antibody reduces angiogenesis in cancer cells of the subject.

20. The method of any one of claims 16 to 18, wherein administration of the anti-MYCT1 antibody increases anti-tumor immunity in the subject.

21. The method of any one of claims 16 to 18, wherein administration of the anti-MYCT1 antibody reduces tumor volume in the subject.

22. 1. A method of controlling tumor angiogenesis and / or increasing anti-tumor immunity in a subject with cancer, comprising: administering to said subject a therapeutically effective amount of a composition comprising the isolated antibody or fragment thereof of any one of claims 1 to 15. A method comprising:

23. Administering immune checkpoint inhibitors and, if appropriate, anti-VEGF inhibitors.

28. The method of claim 27, further comprising:

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