Methods for identifying, detecting, and targeting cancer neoepitopes

Abstract

The invention provides methods to identify, detect, and target aberrantly expressed cancer neoepitopes associated with tumors and neoplasias, for example, breast cancer, colorectal cancer, cervical cancer, liver cancer, prostate cancer, lung cancer, and gastric cancer. Included herewith are methods of identifying and detecting zinc transporter proteins aberrantly expressed at the cell surface, for example SLC39A7 in breast cancer. Further, this disclosure relates to methods of treating a subject having detected cell surface expression of SLC39A7 indicative of cancer, such as breast cancer.

Description

METHODS FOR IDENTIFYING, DETECTING, AND TARGETING CANCER NEOEPITOPESINCORPORATION BY REFERENCE

[0001] This application claims priority to US provisional application Serial No. 63 / 560,196, filed March 1, 2024, which is incorporated by reference herein in its entirety.

[0002] All documents cited or referenced herein (“herein cited documents”), and all documents cited or referenced in herein cited documents, together with any manufacturer’s instructions, descriptions, product specifications, and product sheets for any products mentioned herein or in any document incorporated by reference herein, are hereby incorporated herein by reference, and may be employed in the practice of the invention. More specifically, all referenced documents are incorporated by reference to the same extent as if each individual document was specifically and individually indicated to be incorporated by reference.SEQUENCE LISTING

[0003] This application as originally filed includes / contains a Sequence Listing filed in electronic form in extensible Markup Language (XML) format entitled JI 385-99001. xml, created on February 25, 2025 and having a size of 4,332 bytes. The contents of the Sequence Listing are incorporated herein in its entirety.FIELD OF THE INVENTION

[0004] This disclosure relates to the identification, detection, and targeting of aberrantly expressed cancer neoepitopes associated with tumors and neoplasias, for example for example breast cancer, colorectal cancer, cervical cancer, liver cancer, prostate cancer, lung cancer, and gastric cancer. In particular, this invention relates to identifying, detecting, and targeting zinc transporter proteins which include two major protein families: the solute carrier 39A / Zrt- and Irt- like protein (SLC39 / ZIP) family encompassed by ZIPs 1-14, and the solute carrier 30 / zinc transporter (SLC30 / ZnT) family encompassed by ZnTs 1-10. Further, this disclosure relates to methods of treating a subject having a detected aberrantly expressed ZIP or ZnT protein indicative of cancer, in particular breast, colorectal, cervical, hepatocellular, prostate, or gastric cancer.BACKGROUND OF THE INVENTION

[0005] The following includes information that may be useful in understanding the present invention. It is not an admission that any of the information, publications or documents specificallyor implicitly referenced herein is prior art, or essential, to the presently described or claimed inventions. All publications, patents, related applications, and other written or electronic materials mentioned or identified herein are hereby incorporated herein by reference in their entirety. The information incorporated is as much a part of the application as filed as if all of the text and other content was repeated in the application, and should be treated as part of the text and content of the application as filed. Citation or identification of any document in this application is not an admission that such document is available as prior art to the present invention.

[0006] Zinc transporter proteins are membrane transport proteins controlling the membrane transport of zinc and regulating its intracellular and cytoplasmic concentrations. There are two major groups of zinc transporter proteins which comprise the two major protein families: the solute carrier 39A / Zrt- and Irt-like protein (SLC39A / ZIP) family and the solute carrier 30 / Zinc transporter (SLC30 / ZnT) family.

[0007] SLC39A7 / ZIP7 (SLC39A7 or ZIP7), a member of the SLC39A / ZIP family, is a specialized zinc transporter primarily located in the endoplasmic reticulum and Golgi apparatus (Taylor, K. The LIV-1 Subfamily of Zinc Transporters: From Origins to Present Day Discoveries. Int J Mol Sci. 24(2): 1255 (2023); Huang, L. et al. The ZIP7 gene (Slc39a7) encodes a zinc transporter involved in zinc homeostasis of the Golgi apparatus. J Biol Chem. 280(15): 15456-63 (2005); Ohashi W, et al. Zinc transporter SLC39A7 / ZIP7 promotes intestinal epithelial selfrenewal by resolving ER stress. PLoS Genet. 12:el006349 (2016); Bin B-H, et al. Requirement of zinc transporter SLC39A7 / ZIP7 for dermal development to fine-tune endoplasmic reticulum function by regulating protein disulfide isomerase. , / . Invest. Dermatol. 137: 1682-1691 (2017)). Its distinctive role is to facilitate the transport of zinc (Zn) from intracellular compartments to the cytoplasm, contributing significantly to cellular zinc homeostasis and influencing critical processes such as gene transcription, cell growth, and metabolism (Taylor KM, et al. Structurefunction analysis of ZIP7, a member of the new LIV-1 subfamily of zinc transporters. Biochem J. 377: 131-139 (2004)). ZIP7 is one of two zinc transporters in the SLC37A / ZIP family that does not reside on the plasma membrane, residing instead on the endoplasmic reticulum as the transporter responsible for releasing Zn from that store (Taylor, K. The LIV-1 Subfamily of Zinc Transporters: From Origins to Present Day Discoveries. Int J Mol Sci. 24(2): 1255 (2023); Nimmanon, T. et al. Phosphorylation of zinc channel ZIP7 drives MAPK, PI3K and mTOR growth and proliferation signaling. Metallomics, 9(5): 741-481 (2017)).

[0008] ZIP7 activity has been shown to be dependent on casein kinase II (CK2) phosphorylation to release Zn ions from intracellular reserves (Taylor, KM et al. Protein Kinase CK2 Triggers Cytosolic Zinc Signaling Pathways by Phosphorylation of Zinc Channel ZIP7. Sci Signal. 5(210):ral 1 (2012)). Phosphorylation on residues S275 and S276 lead to the activation and ZIP7-mediated Zn release which in turn activates multiple signaling pathways related to cell proliferation and growth such as MAPK, AKT, PI3K, and mTOR, and the inhibition of GSK3-0 (Taylor, K. The LIV-1 Subfamily of Zinc Transporters: From Origins to Present Day Discoveries. IntJMolSci. 24(2): 1255 (2023)).

[0009] Many cell signaling pathways are implicated in cancer, including the MAPK, PI3K- AKT, Notch, and mTOR pathways. Nimmanon et al. demonstrated that in response to ZIP7- mediated secretion of zinc ions, MAPK, PI3K-AKT, and mTOR signaling were all activated as primary downstream targets of ZIP7 (Nimmanon, T. et al. Phosphorylation of zinc channel ZIP7 drives MAPK, PI3K and mTOR growth and proliferation signaling. Metallomics, 9(5):741-481 (2017)). Nolin et al. identified a direct ZIP7, NVS-ZP7-4, that demonstrably disrupts Notch trafficking and signaling which induces apoptosis through ER stress and altered Zn levels in the endoplasmic reticulum (Nolin, E. et al. Discovery of aZIP7 inhibitor from a Notch pathway screen. Nat Chem Biol. 15 (2) : 179- 188 (2019)).

[0010] Breast cancers are classified by molecular subtypes according to the expression of certain hormone receptors or lack thereof: estrogen (ER), progesterone (PR), and human epidermal growth factor (HER2). Cancers that lack useful targets such as, for example, triple-negative breast cancer which lacks ER, PR, and HER2, or cancers that lose expression of targets such as treatmentresistant cancers remain difficult to treat or are untreatable. New targets, methods, and compositions are needed for treatment of cancers and neoplasias including first-line treatments of cancers as well treatments for recurrent cancers or cancers that have become untreatable.SUMMARY OF THE INVENTION

[0011] The expression of SLC39A7 / ZIP7 (SLC39A7) anywhere outside of the endoplasmic reticulum or Golgi apparatus has not been previously documented in any cancer cell type. Previous studies on SLC39A7 expression are limited to evaluating mRNA and using intracellular or wholeprotein techniques. Accordingly, SLC39A7 has not been considered as a cell surface antigen suitable for targeting through therapeutic antibody strategies such as Antibody Drug Conjugates (ADCs), bispecific antibodies, Chimeric Antigen Receptor (CAR) therapies, or cancer vaccines.Applicants’ research marks a significant milestone as the first to reveal the cell surface expression of SLC39A7 in various types of breast cancer cells, distinctly absent in healthy breast epithelial cells and peripheral blood mononuclear cells (PBMC) from individuals without the disease. Importantly, Applicants’ findings implicitly demonstrate the therapeutic targeting SLC39A7 on various breast cancer types using ADCs (Antibody Drug Conjugates), offering a precise and selective approach that avoids unintended interactions with adjacent healthy breast epithelial cells and other non-cancerous cell types. Furthermore, the identification of SLC39A7's cell surface expression across diverse breast cancer cell types establishes it as a promising biomarker for breast cancer diagnosis. This pivotal discovery not only advances comprehension of the disease but also paves the way for the development of targeted antibody therapies specifically designed to combat cancer cells expressing SLC39A7. Interestingly, elevated zinc levels have been observed in human breast tumors, and animal models show significant zinc accumulation within mammary tumors, emphasizing the importance of zinc in cancer biology (Qu Z, Liu Q, Kong X, Wang X, Wang Z, Wang J, Fang Y. A Systematic Study on Zinc-Related Metabolism in Breast Cancer. Nutrients. 15(7) (2023).

[0012] In one aspect, this disclosure relates to methods for detecting a neoplasm comprising contacting the neoplasm with an agent that detects cell surface expressed solute carrier family 39 member 7 (SLC39A7 aka ZIP7) in the subject or cell or tissue sample. In some embodiments, the agent binds to SLC39A7 and the method comprises detecting the bound agent. In some aspects, this disclosure provides for methods of detecting a neoplasm expressing SLC39A7 at the cell surface with an antibody that specifically binds to cell-surface-expressed SLC39A7 and detecting the bound antibody. In some embodiments, the agent comprises a monoclonal antibody that binds to SLC39A7. In some embodiments, the agent comprises a polyclonal antibody that specifically binds to SLC39A7.

[0013] In some aspects, this disclosure provides for methods of detecting neoplasms that include tumor types and neoplasias such as breast cancer, colorectal cancer, cervical cancer, liver cancer, prostate cancer, lung cancer, and gastric cancer. In some aspects, this disclosure provides for methods of detecting neoplasm including a cancer cell, such as a breast cancer cell. In some aspects, this disclosure provides for methods of detecting neoplasms in a subject having or suspected of having a neoplasm such as a breast cancer cell.

[0014] In one aspect, this disclosure provides for methods of treating a neoplasm in a subject. In some aspects, this disclosure provides for methods of treating neoplasms expressing SLC39A7 or a portion thereof on its surface. In some aspects, this disclosure provides for a method of treating a neoplasm expressing SLC39A7 or a portion thereof on its surface, the method comprising administering to the subject a binding protein that specifically binds to the surface-expressed SLC37A7 or a portion thereof. In some aspects, the method comprises administering a binding protein that is an antibody or antigen binding fragment thereof. In some aspects, the method comprises administering an antineoplastic agent. In some aspects, the method comprises administering a binding protein comprises a drug conjugate. In some aspects, the method further comprises administering to the subject a secondary agent that binds to the binding protein targeting SLC39A7 or portion thereof and delivers an antineoplastic agent to the neoplasm. In some aspects, the method comprises administering a binding protein with an antineoplastic agent. In some aspects, the antineoplastic agent comprises radiation, chemotherapy, a hormone therapy, an immunotherapy, an antibody-drug conjugate, or a combination thereof. In some aspects, the hormone therapy or immunotherapy comprises an enzymatic inhibitor, small molecule inhibitor, an antibody, or targeted vaccines.

[0015] As used herein, hormone therapy is an enzymatic inhibitor or small molecule directly or indirectly targeting ESRI, PR, HER2, EGFR, LH, CDK4, CDK6, PARP, PI3K, AKT, or a combination thereof. In some aspects, the hormone therapy inhibitor is selected from: tamoxifen, toremifene, fulvestrant, elacestrant, exemestane, letrozole, anastrozole, lapatinib, neratinib, palbociclib, ribociclib, abemaciclib, trilaciclib, tucatinib / irbinitinib, olaparib, talazoparib, alpelisib, capivasertib, or a combination thereof. In some aspects, the hormone therapy is an antibody that binds ESRI, PR, HER2, or EGFR, selected from: pertuzumab, traztuzumab, margetuximab, or a combination thereof. In some aspects, the immunotherapy comprises an antibody selected from: pembrolizumab, atezolizumab, avelumab, or a combination thereof. In some aspects, the antineoplastic agent comprises an antibody-drug conjugate selected from: sacituzumab govitecan, ado-trastuzumab emtansine, fam-trastuzumab deruxtecan, or aOIgG-NC- MMAF.

[0016] In one aspect, this disclosure provides for a method of targeting a cell that expresses SLC39A7 or a portion thereof at the cell surface, the method comprising contacting the cell with a binding protein that binds to the extracellular region of cell-surface expressed SLC39A7. In someaspects, the extracellular region of cell-surface expressed SLC39A7 comprises the amino acid sequence PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof, orKFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3) or a portion thereof. In some aspects, the binding protein in the method comprises a drug conjugate. In some aspects, the binding protein in the method comprises an antibody or antigen binding portion thereof.

[0017] In one aspect, this disclosure provides for a method of inducing an immune response against a target cell that expresses SLC39A7 or a portion thereof on its surface in a subject in need thereof, which comprises administering to the subject an immunogenic composition which comprises an SLC39A7 polypeptide. In some aspects, the target cell is a cancer cell. In some aspects, the target cell is a breast cancer cell. In some aspects, the method comprises administering to the subject an immunogenic composition comprising a B cell epitope of SLC39A7. In some aspects, the immunogenic composition comprises one or more polypeptides, wherein each of the one or more polypeptides comprises 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids of SLC39A7. In some aspects, the immunogenic composition comprises 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids of PHALEPHSHHTLEQPGHGHSHSGQ (aal90-213) (SEQ ID NO:2) orKFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (aa235-380) (SEQ ID NO:3). In some aspects, the immunogenic composition comprises a polypeptide comprising 8 or more contiguous acids of a polypeptide of Table 3. In some aspects, the immunogenic composition comprises a T cell epitope linked to the B cell epitope.

[0018] In one aspect, this disclosure provides for a method of treating a subject having or suspected of having a neoplasm with cell surface-expressed SLC39A7 or a portion thereof, which comprises obtaining or having obtained a sample from the subject, detecting or having detected surface-expressed SLC39A7, and administering to the subject a binding protein that specifically binds to the surface-expressed SLC39A7 or portion thereof. In some aspects, the method comprises administering a binding protein that is an antibody or antigen binding fragment thereof.In some aspects, the method comprises administering an antineoplastic agent. In some aspects, the method comprises administering a binding protein comprises a drug conjugate. In some aspects, the method further comprises administering to the subject a secondary agent that binds to the binding protein targeting SLC39A7 or portion thereof and delivers an antineoplastic agent to the neoplasm. In some aspects, the method comprises administering a binding protein with an antineoplastic agent. In some aspects, the antineoplastic agent comprises radiation, chemotherapy, a hormone therapy, an immunotherapy, an antibody-drug conjugate, or a combination thereof. In some aspects, the hormone therapy or immunotherapy comprises an enzymatic inhibitor, small molecule inhibitor, an antibody, or targeted vaccines.

[0019] In one aspect, this disclosure provides a SLC39A7 binding agent for use in detecting cell surface expression of solute carrier family 39 member 7 (SLC39A7) in a subject or cell or tissue sample, or for use in treating a cell or neoplasm or cancer that expresses cell surface SLC39A7. In some aspects, the cell or neoplasm comprises a breast cancer cell. In some aspects, the agent comprises a binding protein or a monoclonal antibody or polyclonal antibody or an antigen binding portion thereof that binds to SLC39A7 on the surface of a cell. In some aspects, the agent binds to an SLC39A7 polypeptide which comprises PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3) or a portion thereof. In some aspects, the agent comprises a drug conjugate. In some aspects, the agent comprises an antineoplastic agent.

[0020] Accordingly, it is an object of the invention not to encompass within the invention any previously known product, process of making the product, or method of using the product such that Applicants reserve the right and hereby disclose a disclaimer of any previously known product, process, or method. It is further noted that the invention does not intend to encompass within the scope of the invention any product, process, or making of the product or method of using the product, which does not meet the written description and enablement requirements of the USPTO (35 U.S.C. §112, first paragraph) or the EPO (Article 83 of the EPC), such that Applicants reserve the right and hereby disclose a disclaimer of any previously described product, process of making the product, or method of using the product. It may be advantageous in the practice of the inventionto be in compliance with Art. 53(c) EPC and Rule 28(b) and (c) EPC. All rights to explicitly disclaim any embodiments that are the subject of any granted patent(s) of applicant in the lineage of this application or in any other lineage or in any prior filed application of any third party is explicitly reserved. Nothing herein is to be construed as a promise.

[0021] It is noted that in this disclosure and particularly in the claims and / or paragraphs, terms such as "comprises", "comprised", "comprising" and the like can have the meaning attributed to it in U.S. Patent law; e.g., they can mean "includes", "included", "including", and the like; and that terms such as "consisting essentially of' and "consists essentially of' have the meaning ascribed to them in U. S. Patent law, e.g., they allow for elements not explicitly recited, but exclude elements that are found in the prior art or that affect a basic or novel characteristic of the invention.

[0022] These and other embodiments are disclosed or are obvious from and encompassed by, the following Detailed Description.BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0024] The following detailed description, given by way of example, but not intended to limit the invention solely to the specific embodiments described, may best be understood in conjunction with the accompanying drawings.

[0025] FIG. 1. Targeting SLC39A7 / ZIP7 Kills Cancer Cells While Sparing Normal Cells. Cytotoxic capacity of Total Anti-Breast Cancer Serum (before filtration), Oncotope Platform Filtered Anti -Breast Cancer Serum and Polyclonal antibodies 1 and 2 against Antigen ONCA-001 (SLC39A7 / ZIP7). Sera or Antibodies were added to cultures of both Cancer Cells (Human Breast Cancer Cells: Hs578T from ATCC) and corresponding Normal Cells from the same tissue type from the same individual (Breast Epithelial Cells: Hs578Bst from ATCC) in the presence of a Secondary Antibody Drug Conjugate (ADC). The ADC used was aOIgG-NC-MMAF (from Moradec), an anti-rabbit IgG (H+L) specific antibody conjugated to monomethyl auristatin F (MMAF) with a non-cleavable linker. Cells were cultured for 48hrs, and the viability assayed using CellTiter Gio. Percent Cell Death was calculated using GraphPad Prism, ns, not significant. ** p < 0.01.

[0026] FIG. 2A-2F. Gating Strategy to Test Binding Capacity of 2-Antigen Targeting Antibodies on Hs578T. Fig. 2A shows the initial capture of live cells with no gate (forward and side scatter only). 84.2% of cells were then gated for doublets. Fig. 2B shows the percentage of doublets captured. Fig. 2C shows the counts of doublets having isotype control or total anti-breast cancer serum (pTAB) labeled with AF488 using a 1 :400 dilution of antibody. Fig. 2D shows the counts of doublets having isotype control or pTAB labeled with AF488 using a 1 :800 dilution of antibody. Fig. 2E shows the quantitation of cells by percent of gated cells and geometric mean of the 1 :400 dilution data. Fig. 2F shows the quantitation of cells by percent of gated cells and geometric mean of the 1:800 dilution data.

[0027] FIG. 3A-3D. Gating Strategy to Test Binding Capacity of 2-Antigen Targeting Antibodies on Hs578T and Hs578T with varying dilutions. Fig. 3 A shows the counts of normal breast epithelial cell line Hs578Bst doublets having isotype control or pTAB labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 : 1600. Fig. 3B shows the counts of TNBC cell line Hs578T doublets having isotype control or pTAB labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 : 1600. Fig. 3C shows the quantitation of cells by percent of gated cells for each antibody dilution between the Hs578Bst and Hs578T cell lines. Fig. 3D shows the quantitation of cells by geometric mean for each antibody dilution between the Hs578Bst and Hs578T cell lines.

[0028] FIG. 4A-4D. Gating Strategy to Test Binding Capacity of 2-Antigen Targeting Antibodies on Hs578T and Hs578T with varying dilutions. Fig. 4A shows the counts of normal breast epithelial cell line Hs578Bst doublets having isotype control or pF AB labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 :1600. Fig. 4B shows the counts of TNBC cell line Hs578T doublets having isotype control or pF AB labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 :1600. Fig. 4C shows the quantitation of cells by percent of gated cells for each antibody dilution between the Hs578Bst and Hs578T cell lines. Fig. 4D shows the quantitation of cells by geometric mean for each antibody dilution between the Hs578Bst and Hs578T cell lines.

[0029] FIG. 5. Antibodies to SLC39A7 / ZIP7 Bind Cancer Cells But Not Normal Cells. Flow cytometric analysis of binding capacity of Total Anti-Breast Cancer Serum (before fdtration), Oncotope Platform Filtered Anti-Breast Cancer Serum and Polyclonal antibody 1 against Antigen ONCA-001 (SLC39A7 / ZIP7). Sera or Antibodies were added to cultures of bothCancer Cells (Human Breast Cancer Cells: Hs578T from ATCC) and corresponding Normal Cells from the same tissue type from the same individual (Breast Epithelial Cells: Hs578Bst from ATCC) and incubated for 1 hour followed by addition of Alexa-488 conjugated Secondary Donkey anti-Rabbit antibody. After 30 minutes Incubation with the secondary antibody, cells were analyzed on an Attune Flow cytometer. Binding capacity of the sera or antibodies was calculated using FCS express flow cytometry analysis software.

[0030] FIG. 6. Binding Capacity (Flow Cytometry) of Anti-ONCA 001 (SLC39A7 / ZIP7) on Multiple Breast Cell Lines.

[0031] FIG. 7. Cytotoxic Capacity of Anti-ONCA 001 (SLC39A7 / ZIP7) on Multiple Breast Cell Lines.

[0032] FIG. 8A-8B. Binding Capacity Comparison of Syngeneic Normal and TNBC Breast Cell Lines. Fig. 8A shows comparison of binding capacity of total anti-breast cancer serum, Oncotope platform fdtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for HTB125 cells. Fig. 8B shows comparison of binding capacity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for HTB126 cells.

[0033] FIG. 9A-9B. Binding Capacity Comparison of ESR1+ Breast Cell Lines. Fig. 9A shows comparison of binding capacity of total anti -breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for MDA-MB-415 cells. Fig. 9B shows comparison of binding capacity of total antibreast cancer serum, Oncotope platform filtered anti -breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for CAMA1 cells.

[0034] FIG. 10A-10D. Binding Capacity Comparison of HER2+ Breast Cell Lines. Fig. 10A shows comparison of binding capacity of total anti -breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400,1 :800, and 1 :1600 for HCC1419 cells. Fig. 10B shows comparison of binding capacity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for HCC202 cells. Fig. 10C shows comparison of binding capacity of total anti -breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for UACC893 cells. Fig. 10D shows comparison of binding capacity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 :1600 for HCC1954 cells.

[0035] FIG. 11A-11C. Binding Capacity Comparison of TNBC Breast Cell Lines. Fig. 11A shows comparison of binding capacity of total anti -breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 :1600 for BT549 cells. Fig. 11B shows comparison of binding capacity of total antibreast cancer serum, Oncotope platform filtered anti -breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for MDA-MB-436 cells. Fig. 11C shows comparison of binding capacity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for MDA-MB-436 cells.

[0036] FIG. 12A-12B. Binding Capacity Comparison of Healthy Breast Cell Lines. Fig. 12A shows comparison of binding capacity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 :1600 for MCF-10A cells. Fig. 12B shows comparison of binding capacity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody in dilutions of 1 :400, 1 :800, and 1 : 1600 for healthy donor PBMCs.

[0037] FIG. 13A-13B. Cytotoxic Capacity Comparison of Syngeneic Breast Cell Lines. Fig. 13A shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti -breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti- SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in HTB125 cells. Fig. 13B shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in HTB126 cells.

[0038] FIG. 14A-14B. Cytotoxic Capacity Comparison of ESR1+ Breast Cell Lines. Fig. 14A shows comparison of percent cytotoxicity of total anti -breast cancer serum, Oncotope platform filtered anti -breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti- SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in MDA-MB-415 cells. Fig. 14B shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in CAMA1 cells.

[0039] FIG. 15A-15D. Cytotoxic Capacity Comparison of HER2+ Breast Cell Lines. Fig. 15A shows comparison of percent cytotoxicity of total anti -breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti- SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in HCC1419 cells. Fig. 15B shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in HCC202 cells. Fig. 15C shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in UACC893 cells. Fig. 15D shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti -breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in HCC1954 cells.

[0040] FIG. 16A-16C. Cytotoxic Capacity Comparison of TNBC Breast Cell Lines. Fig. 16A shows comparison of percent cytotoxicity of total anti -breast cancer serum, Oncotope platform filtered anti -breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti- SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in BT549 cells. Fig. 16B shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in MDA-MB-436 cells. Fig. 16C shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in MDA-MB-468 cells.

[0041] FIG. 17A-17B. Cytotoxic Capacity Comparison of Healthy Breast Cell Lines. Fig. 17A shows comparison of percent cytotoxicity of total anti -breast cancer serum, Oncotope platform filtered anti -breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti- SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in MCF-10A cells. Fig. 17B shows comparison of percent cytotoxicity of total anti-breast cancer serum, Oncotope platform filtered anti-breast cancer serum, anti-SLC39A7 Proteintech primary antibody, anti-SLC39A7 BIOSS primary antibody, and anti-SLC39A7 Invitrogen primary antibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in healthy donor PBMCs.

[0042] FIG. 18. SLC39A7 sequence and primary antibody target regions. Fig. 18 shows the canonical amino acid sequence of SLC39A7 (SEQ ID NO: 1) is shown. The epitope on SLC39A7 for each primary antibody (Proteintech, BIOSS, Invitrogen, and Atlas Antibodies) is shown with highlight or different colored text.

[0043] FIG. 19A-19D. Test Binding Capacity of 2-Antigen Targeting Antibodies on Hs578T and Hs578T with varying dilutions (Proteintech Primary Antibody). Fig. 19A shows the counts of normal breast epithelial cell line Hs578Bst doublets having isotype control or Proteintech Anti-SLC39A7 antibody recognized by secondary antibody labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 : 1600. Fig. 19B shows the counts of TNBC cell line Hs578T doublets having isotype control or pF AB labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 : 1600. Fig. 19C shows the quantitation of cells by percent of gated cells for each antibody dilution between the Hs578Bst and Hs578T cell lines. Fig. 19D shows the quantitation of cells by geometric mean for each antibody dilution between the Hs578Bst and Hs578T cell lines.

[0044] FIG. 20A-20D. Test Binding Capacity of 2-Antigen Targeting Antibodies on Hs578T and Hs578T with varying dilutions (Atlas Antibodies Primary Antibody). Fig. 20A shows the counts of normal breast epithelial cell line Hs578Bst doublets having isotype control or Atlas Antibodies Anti-SLC39A7 antibody (Cat# HPA053999) recognized by secondary antibody labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 : 1600. Fig. 20B shows the counts of TNBC cell line Hs578T doublets having isotype control or pF AB labeled with AF488 using dilutions of antibody in: 1 :400, 1 :800, and 1 : 1600. Fig. 20C shows the quantitation of cells by percent of gated cells for each antibody dilution between the Hs578Bst and Hs578T cell lines. Fig. 20D shows the quantitation of cells by geometric mean for each antibody dilution between the Hs578Bst and Hs578T cell lines.

[0045] FIG. 21. Summary of Comparison between Binding Capacity of 2-Antigen Targeting Antibodies from Proteintech and Atlas Antibodies on Hs578T and Hs578T with varying dilutions. Fig. 21 summarizes the comparison of binding capacity between the Proteintech and Atlas Antibodies primary anti-SLC39A7 antibodies with varying primary antibody dilutions.

[0046] FIG. 22A-22D. Cytotoxic Capacity Comparison of Syngeneic Breast Cell Lines with Proteintech and Atlas Antibodies Primary Antibodies. Fig. 22A shows comparison of percent cytotoxicity of total anti-breast cancer serum; Fig. 22B shows comparison of percent cytotoxicity of the Oncotope platform filtered anti-breast cancer serum; Fig. 22C shows the comparison of percent cytotoxicity of the anti-SLC39A7 Proteintech primary antibody; and Fig. 22D shows the comparison of percent cytotoxicity of the anti-SLC39A7 Atlas Antibodies primaryantibody at varying antibody concentrations: 4 ug / ml, 2 ug / ml, and 1 ug / ml in HTB125 cells (Hs578Bst; blue bars) and in HTB126 cells (Hs578T; red bars).

[0047] FIG. 23. Test Efficacy of NVS-ZP7-4 on Breast Cancer Cell Lines. Fig 23 shows the percent viability of breast cancer cell lines with varying drug concentrations of NVS-ZP7-4: O.OlmM; 0.03uM; O.luM; 0.3uM; luM; 3uM and lOuM. Data is shown in log(Concentration (uM)), and the control was 0.2% DMSO without NVS-ZP7-4.

[0048] FIG. 24. TABLE of Characteristics of Breast Cancer Cell Lines. Table listing each breast cancer cell line, SLC39A7 / ZIP7 mRNA expression status, subtype, and receptor expression are listed. Luminal A, ER+ PR+ / - HER2- (LA); Luminal B, ER+ PR- HER2+ (LB); HERZ Positive (H); Triple Negative A (TNA); and Triple Negative B (TNB).

[0049] FIG. 25. TABLE of Characteristics of Breast Cell Lines. Table listing each breast cancer cell line for binding capacity comparison experiments and cytotoxicity comparion experiments is listed with SLC39A7 / ZIP7 mRNA expression status, Notch mutation status, and receptor expression are listed. Estrogen receptor (ESRI); human epidermal growth factor receptor 2 (HERZ); no receptor express! on / triple-negative breast cancer (TNBC, negative for ESRI, HER2, and progesterone receptor (PR)); normal (Healthy).

[0050] FIG. 26. TABLE of Protein Sequences of SLC39A7 and epitopes.DETAILED DESCRIPTION OF THE INVENTION

[0051] Before the present compounds, compositions, articles, devices, and / or methods are disclosed and described, it is to be understood that they are not limited to specific synthetic methods or specific recombinant biotechnology methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0052] A. Certain Exemplary Definitions

[0053] In this specification and in the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings:

[0054] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a pharmaceutical carrier” includes mixtures of two or more such carriers, and the like.

[0055] “Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes embodiments where said event or circumstance occurs and embodiments where it does not.

[0056] Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value” and possible ranges between values are also disclosed. For example, if the value “10” is disclosed the “less than or equal to 10” as well as “greater than or equal to 10” is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point 15 are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as values between 10 and 15. For example, it is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed. In this application, if a data point range is disclosed, it is understood that each unit from the lowest data point to the highest stated datapoint, including the first (lowest) and last (highest) data point is disclosed. For example, if a data point range 1-20 is disclosed, it is understood that data points 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20, and each unit between any two particular units in the range are also disclosed. It is also understood that whenever a series of values are disclosed, that any range falling between any two of the recited values is also understood to be included.

[0057] As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, whenappropriate, fractions thereof (including one-tenth and one-hundredth of an integer), unless otherwise indicated.

[0058] As used herein, the term “substantially the same” means an amount of expression, level, and / or activity of a gene or gene product within 90% of baseline expression, level, and / or activity as in a subject unaffected by a disease, for example in a subject unaffected by a neoplasm. “Substantially the same” can also mean an amount of expression, level, and / or activity of a gene or gene product within 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, to 100% of the baseline expression, level, and / or activity as in a subject unaffected by unaffected by a disease, for example in a subject unaffected by neoplasm.

[0059] As used herein, the term “neoplasm” includes, but is not limited to, cancer, solid tumor, blood tumor, lymph tumor, a cancer cell, or a tumor cell. In some embodiments, a neoplasm is a cancer cell. In some embodiments, a neoplasm is a breast cancer cell. In some embodiments, a neoplasm is breast cancer.

[0060] Breast cancers are classified by molecular subtypes, according to the expression of certain hormone receptors or lack thereof. Luminal A breast cancer is ESRl-positive, PR-positive, HER2-negative, and has low levels of Ki-67, a protein controlling the proliferation of cancer cells. Luminal B breast cancer is ESRl-positive and HER2 -negative, and has either high levels of Ki-67 oris PR-negative. Luminal B-like breast cancer is ESRl-positive and HER2 -positive, has any level of Ki-67, and may be PR-positive or PR-negative. HER2 -enriched breast cancer is E SRI -negative, PR-negative, and HER2-positive. Triple-negative (or “basal -like”) breast cancer is negative for the receptors ESRI, PR, and HER2, Triple-negative breast cancer can further be characterized as triple-negative A or triple-negative B based on whether the cancer cells display epithelial characteristics and have association with BRCA1 gene signatures (triple-negative A) or display mesenchymal and stem / progenitor-like characteristics (triple-negative B).

[0061] As used herein, the term “genetic factors” includes, but is not limited to, gene alterations and variants such as single nucleotide polymorphisms / single nucleotide variants (SNPs / SNVs), deletions, insertions, copy number variations, translocations, inversions, and structural variations. SNPs / SNVs include synonymous and nonsynonymous mutations. Nonsynonymous mutations include missense mutations, frame-shift mutations, nonsense mutations, and readthrough mutations. As used herein, the terms “risk allele”, “risk variant”, and “risk gene variant” include, but are not limited to a gene variant or SNP / SNV that is associatedwith risk to develop a disease, e.g. a cancer. As used herein, the terms “protective allele”, “protective variant”, and “protective gene variant” include, but are not limited to a gene variant or SNP / SNV that is associated with protection against developing a disease, e.g. a cancer. In some embodiments, a subject having a risk allele or risk variant is a subject that has a mutation in BRCA1, BRCA2, ATM, PALB2, TP53, CHEK2, PTEN, CDH1, STK11.

[0062] As used herein, the term “cell surface” refers to the plasma membrane of a cell. A protein expressed at the cell surface is expressed at the plasma membrane of the cell and includes any portion of said protein, including any inside, outside, or transmembrane portions of the protein.

[0063] As used herein, the terms “neoantigen,” “tumor neoantigen,” and “cancer neoantigen” refers to a protein self-antigen generated by tumor cells as a result of genomic mutation, somatic mutation, dysregulated RNA splicing, disordered post-translational modification, or virally encoded open reading frames. Neoantigens have the advantage of being uniquely tumor-specific and absent in normal tissues. Some neoantigens are herein referred to as “tumor specific neoantigens.”

[0064] The term “subject” refers to any individual who is the target of administration or treatment. The subject can be a vertebrate, for example, a mammal. In one aspect, the subject can be human, non-human primate, bovine, equine, porcine, canine, or feline. The subject can also be a guinea pig, rat, hamster, rabbit, mouse, or mole. Thus, the subject can be a human or veterinary patient. The term subject, in some embodiments, refers to a “patient” under the treatment of a clinician, e.g., physician.

[0065] As used herein, the terms “treat,” “treating” and “treatment” include administering a compound prior to the onset of clinical symptoms of a disease state / condition so as to prevent any symptom, as well as administering a compound after the onset of clinical symptoms of a disease state / condition so as to reduce or eliminate any symptom, aspect or characteristic of the disease state / condition. “Treating” is also used herein to refer to ameliorating at least one symptom of, curing and / or preventing the development of a given disease or condition. Such treating need not be absolute to be useful. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared toexpected survival if not receiving treatment. Those in need of treatment include those already with the condition or disorder and those prone to have the condition or disorder or those in which the condition or disorder is to be prevented. “Treatment” referring to prophylactic or preventive measures, wherein the object is to prevent or decrease an undesired physiological change or disorder by using the invention is also contemplated.

[0066] As used herein, the term "therapeutically effective amount" means an amount of a compound of the present invention that (i) treats the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of or reduces the intensity of one or more symptoms of the particular disease, condition, or disorder described herein. In reference to treating a disease state / condition, the term “therapeutically effective amount” refers to an amount of a compound either alone or as contained in a pharmaceutical composition that is capable of having any detectable, positive effect on any symptom, aspect, or characteristics of a disease state / condition when administered as a single dose or in multiple doses. Such effect need not be absolute to be beneficial.

[0067] As used herein, the term “therapeutic agent” refers to any agent or material that has a beneficial effect on a subject, including a mammalian recipient. Thus, “therapeutic agent” embraces both therapeutic and prophylactic molecules having nucleic acid or protein components.

[0068] As used herein, the term “preventing” means preventing in whole or in part, or ameliorating or controlling.

[0069] In certain embodiments, the present therapeutic agent may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.

[0070] The tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, com starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices.

[0071] The active compound may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts may be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0072] The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient that are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will bepreferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0073] Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.

[0074] Useful dosages of the compounds of the present invention can be determined by comparing their in vitro activity, and in vivo activity in animal models. In certain embodiments, a useful dose is from about 0.1 mg / kg to about 5 mg / kg or from about 0.5 mg / kg to about 2 mg / kg. Methods for the extrapolation of effective dosages in humans and animals of different sizes are known to the art; for example, see U.S. Pat. No. 4,938,949.

[0075] The amount of the compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician.

[0076] In general, however, a suitable dose will be in the range of from about 0.5 to about 100 mg / kg, e.g., from about 10 to about 75 mg / kg of body weight per day, such as 3 to about 50 mg per kilogram body weight of the recipient per day, preferably in the range of 6 to 90 mg / kg / day, most preferably in the range of 15 to 60 mg / kg / day.

[0077] The compound is conveniently administered in unit dosage form; for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of active ingredient per unit dosage form.

[0078] Ideally, the active ingredient should be administered to achieve peak plasma concentrations of the active compound of from about 0.5 to about 75 pM, preferably, about 1 to 50 pM, most preferably, about 2 to about 30 pM. This may be achieved, for example, by the intravenous injection of a 0.05 to 5% solution of the active ingredient, optionally in saline, or orally administered as a bolus containing about 1-100 mg of the active ingredient. Desirable blood levelsmay be maintained by continuous infusion to provide about 0.01-5.0 mg / kg / hr or by intermittent infusions containing about 0.4-15 mg / kg of the active ingredient(s).

[0079] The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.

[0080] B. Certain Exemplary Methods

[0081] In one aspect, the invention provides an anti-SLC39A7 binding protein suitable for use in a mammal, for example, without limitation, a human. In some aspects, the binding protein is an antibody, antigen binding fragment thereof, or an antibody-drug conjugate.

[0082] The term “antibody,” as used herein, includes, without limitation, a glycoprotein immunoglobulin which binds specifically to an antigen and antigen-binding fragments of full antibody molecules. The terms “antigen-binding portion” of an antibody, “antigen-binding fragment” of an antibody, and the like, as used herein, include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex.

[0083] As used herein, the term “specifically binds” or “binds specifically” means that an SLC39A7 binding protein of the invention reacts or associates more frequently, more rapidly, with greater duration and / or with greater affinity with SLC39A7 than it does with alternative antigens.

[0084] In general, an antibody can comprise at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, or an antigen-binding portion thereof. Each H chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three constant domains, CHI, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region comprises one constant domain, CL. The VH and VL regions are further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL comprises three CDRs and four FRs, arranged from amino-terminus to carboxy -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the Abs may mediate the binding of theimmunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. An immunoglobulin may derive from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG and IgM. IgG subclasses also include but are not limited to human IgGl, IgG2, IgG3 and IgG4. “Isotype” refers to the antibody class or subclass (e.g., IgM or IgGl) that is encoded by the heavy chain constant region genes. The term “antibody” can include or exclude both naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or nonhuman antibodies; wholly synthetic antibodies; monovalent and divalent antibody fragments or portions; and single chain antibodies. A nonhuman antibody may be humanized by recombinant methods to reduce its immunogenicity in man.

[0085] Antigen-binding fragments of an antibody may be derived, e.g., from full antibody molecules using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable and optionally constant domains. Such DNA is known and / or is readily available from, e.g., commercial sources, DNA libraries (including, e.g., phage-antibody libraries), or can be synthesized. The DNA may be sequenced and manipulated chemically or by using molecular biology techniques, for example, to arrange one or more variable and / or constant domains into a suitable configuration, or to introduce codons, create cysteine residues, modify, add or delete amino acids, etc.

[0086] Polyclonal antibodies may be affinity purified or filtered to enrich for target antibodies and deplete unwanted binders. In some embodiments, an anti-SLC39A7 polyclonal antibody may be enriched for antibodies that bind to portions or regions or epitopes of SLC39A7 that are differentially cell surface-exposed on neoplastic cells or breast cancer cells or tissues as compared to normal cells or tissues. In some embodiments, an anti-SLC39A7 polyclonal antibody may be enriched for antibodies that bind to SLC39A7 derived polypeptides known or expected to be cellsurface expressed on neoplastic cells or breast cancer cells or tissues as compared to normal cells or tissues. In some embodiments, an anti-SLC39A7 polyclonal antibody may be depleted for antibodies that bind to SLC39A7 derived polypeptides known or expected not to be cell-surface expressed on neoplastic cells or breast cancer cells or tissues as compared to normal cells or tissues.

[0087] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a constrained FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain- deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g. monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the expression "antigen-binding fragment," as used herein.

[0088] In certain embodiments, an antigen-binding fragment of an antibody comprises at least one variable domain. The variable domain may be of any size or amino acid composition and will generally comprise at least one CDR which is adjacent to or in frame with one or more framework sequences. In antigen-binding fragments having a VH domain associated with a VL domain, the VH and VL domains may be situated relative to one another in any suitable arrangement. For example, the variable region may be dimeric and contain VH-VH, VH-VL or VL-VL dimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric VH or VL domain.

[0089] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting, exemplary configurations of variable and constant domains that may be found within an antigenbinding fragment of an antibody of the present invention include: (i) VH-CH1; (ii) VH-CH2; (iii) VH-CH3; (iv) VH-CH1-CH2; (V) VH-CH1-CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL-CH1; (ix) VL-CH2, (x) VL-CH3; (xi) VL-CH1-CH2; (xii) VL-CH1-CH2-CH3; (xiii) VL- CH2-CH3; and (xiv) VL-CL. In any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be either directly linked to one another or may be linked by a full or partial hinge or linker region. A hinge region may consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids which result in a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. Moreover, an antigen-binding fragment of an antibody of the present invention may comprise a homo-dimer or hetero-dimer (or other multimer) of any of the variableand constant domain configurations listed above in non-covalent association with one another and / or with one or more monomeric VH or VL domain (e g., by disulfide bond(s)).

[0090] The anti-SLC39A7 proteins described herein, including binding proteins, antibodies or fragments thereof, are useful for ameliorating, or reducing the symptoms of, or treating, or preventing, diseases or conditions associated, characterized by, or caused by aberrantly expressed SLC39A7. Such diseases or conditions include, without limitation, cancer. The anti-SLC39A7 proteins or fragments, as well as combinations with other agent, are to be administered in a therapeutically effective amount to subjects in need of such treatment in the form of a pharmaceutical composition as described herein.

[0091] In certain embodiments the method comprises ameliorating, or reducing the symptoms of, or treating, or preventing disease in a subject. In certain embodiments, treatment comprises administering the anti-SLC39A7 binding proteins, antibodies, fragments thereof, or ADCs alone. In certain embodiments, treatment comprises administering the anti-SLC39A7 proteins, antibodies, fragments thereof, or ADCs alone, in conjunction with a second agent used to treat, ameliorate, reduce symptoms of, or prevent the same disease or disorder or to treat cancer.

[0092] Nonlimiting examples of cancers the binding proteins, antibodies, antibody fragments, ADCs, and methods are used for ameliorating, or reducing the symptoms of, or treating, or preventing include cancers of the breast, bladder, blood, bone, bone marrow, brain, colon, esophagus, gastrointestinal tract, gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, testis, tongue, or uterus, or malignant neoplasm, carcinoma, undifferentiated; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; nonencapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceousadenocarcinoma; ceruminous; adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infdtrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; Paget's disease, mammary; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma w / squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; thecoma, malignant; granulosa cell tumor, malignant; and roblastoma, malignant; Sertoli cell carcinoma; Leydig cell tumor, malignant; lipid cell tumor, malignant; paraganglioma, malignant; extramammary paraganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignant melanoma; amelanotic melanoma; superficial spreading melanoma; malignant melanoma in giant pigmented nevus; epithelioid cell melanoma; blue nevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma, malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mixed tumor, malignant; mullerian mixed tumor; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant; hemangiosarcoma; hemangioendothelioma, malignant; kaposi's sarcoma; hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal chondrosarcoma; giant cell tumor of bone; Ewing's sarcoma; odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma, malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma; glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor; meningioma, malignant; neurofibrosarcoma; neurilemmoma, malignant; granular cell tumor, malignant; malignant lymphoma; Hodgkin's disease; Hodgkin's lymphoma; paragranuloma; malignant lymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-Hodgkin's lymphomas; malignant histiocytosis; multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphoid leukemia; plasma cell leukemia; erythroleukemia; lymphosarcoma cell leukemia;myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia; myeloid sarcoma; and hairy cell leukemia.

[0093] In certain embodiments, the treatment is used to treat breast cancers, including but not limited to, luminal A, luminal B, HER2 Positive, triple negative A, and triple negative B subtypes of breast cancer.

[0094] In certain embodiments, the anti-SLC39A7 binding proteins, antibodies, antibody fragments, or ADCs are optionally administered in combination with one or more active agents including antineoplastic agents such as radiation, chemotherapy, hormone therapy, or immunotherapy. In some embodiments, the hormone therapy or immune therapy is an enzymatic inhibitor, small molecule inhibitor, or an antibody. Non-limiting examples of hormone therapies include enzymatic inhibitors or small molecule inhibitors targeting ESRI, PR, HER2, EGFR, LH, CDK4, CDK6, PARP, PI3K, AKT such as tamoxifen, toremifene, fulvestrant, elacestrant, exemestane, letrozole, anastrozole, lapatinib, neratinib, palbociclib, ribociclib, abemaciclib, trilaciclib, tucatinib / irbinitinib, olaparib, talazoparib, alpelisib, capivasertib, or combinations thereof. Non-limiting examples of hormone therapies include antibodies binding to estrogen receptor, progesterone receptor, HER2, or EGFR, such as pertuzumab, traztuzumab, margetuximab, or combinations thereof. Non-limiting examples of immunotherapies include pembrolizumab, atezolizumab, avelumab, or a combination thereof. Non-limiting examples of ADCs include sacituzumab govitecan, ado-trastuzumab emtansine, fam -trastuzumab deruxtecan, or aOIgG-NC-MMAF.

[0095] In certain embodiments, the anti-SLC39A7 binding proteins, antibodies, antibody fragments, or ADCs are administered in combination with one or more active agents including other analgesic agents. Such active agents include analgesic, anti-histamine, antipyretic, antiinflammatory, antibiotic, antiviral, and anti-cytokine agents. Active agents include agonists, antagonists, and modulators of TNF-a, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-18, IFN-a, IFN- Y, BAFF, CXCL13, IP-10, VEGF, EPO, EGF, HRG, Hepatocyte Growth Factor (HGF), Hepcidin, including antibodies reactive against any of the foregoing, and antibodies reactive against any of their receptors. Active agents also include, without limitation, 2-arylpropionic acids, aceclofenac, acemetacin, acetylsalicylic acid (Aspirin), alclofenac, alminoprofen, amoxiprin, ampyrone, arylalkanoic acids, azapropazone, benorylate / benorilate, benoxaprofen, bromfenac, carprofen, celecoxib, choline magnesium salicylate, clofezone, COX-2 inhibitors, dexibuprofen,dexketoprofen, diclofenac, diflunisal, droxicam, ethenzamide, etodolac, etoricoxib, faislamine, fenamic acids, fenbufen, fenoprofen, flufenamic acid, flunoxaprofen, flurbiprofen, ibuprofen, ibuproxam, indometacin, indoprofen, kebuzone, ketoprofen, ketorolac, lomoxicam, loxoprofen, lumiracoxib, magnesium salicylate, meclofenamic acid, mefenamic acid, meloxicam, metamizole, methyl salicylate, mofebutazone, nabumetone, naproxen, n-arylanthranilic acids, nerve growth factor (NGF), oxametacin, oxaprozin, oxicams, oxyphenbutazone, parecoxib, phenazone, phenylbutazone, phenylbutazone, piroxicam, pirprofen, profens, proglumetacin, pyrazolidine derivatives, rofecoxib, salicyl salicylate, salicylamide, salicylates, sulfinpyrazone, sulindac, suprofen, tenoxicam, tiaprofenic acid, tolfenamic acid, tolmetin, and val decoxib.

[0096] Active agents also include aldosterone, beclometasone, betamethasone, corticosteroids, cortisol, cortisone acetate, deoxycorticosterone acetate, dexamethasone, fludrocortisone acetate, glucocorticoids, hydrocortisone, methylprednisolone, prednisolone, prednisone, steroids, and triamcinolone. Any suitable combination of these active agents is also contemplated.

[0097] Formulations and Methods of Administration

[0098] For in vivo use, a therapeutic agent as described herein is generally incorporated into a pharmaceutical composition prior to administration. Within such compositions, one or more therapeutic compounds as described herein are present as active ingredient(s) (i.e., are present at levels sufficient to provide a statistically significant effect on the symptoms of cystic fibrosis, as measured using a representative assay). A pharmaceutical composition comprises one or more such compounds in combination with any pharmaceutically acceptable carrier(s) known to those skilled in the art to be suitable for the particular mode of administration. In addition, other pharmaceutically active ingredients (including other therapeutic agents) may, but need not, be present within the composition.

[0099] The antibodies of the present invention can be formulated according to standard methods (see, for example, Remington's Pharmaceutical Science, latest edition, Mark Publishing Company, Easton, U.S.A), and may comprise pharmaceutically acceptable carriers and / or additives. The present invention relates to compositions (including reagents and pharmaceuticals) comprising the antibodies of the invention, and pharmaceutically acceptable carriers and / or additives. Exemplary carriers include surfactants (for example, PEG and Tween), excipients, antioxidants (for example, ascorbic acid), coloring agents, flavoring agents, preservatives, stabilizers, buffering agents (for example, phosphoric acid, citric acid, and other organic acids),chelating agents (for example, EDTA), suspending agents, isotonizing agents, binders, disintegrators, lubricants, fluidity promoters, and corrigents. However, the carriers that may be employed in the present invention are not limited to this list. In fact, other commonly used carriers can be appropriately employed: light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, carmelose calcium, carmelose sodium, hydroxypropylcellulose, hydroxypropylmethyl cellulose, polyvinylacetaldiethylaminoacetate, polyvinylpyrrolidone, gelatin, medium chain fatty acid triglyceride, polyoxyethylene hydrogenated castor oil 60, sucrose, carboxymethylcellulose, corn starch, inorganic salt, and so on. The composition may also comprise other low-molecular-weight polypeptides, proteins such as serum albumin, gelatin, and immunoglobulin, and amino acids such as glycine, glutamine, asparagine, arginine, and lysine. When the composition is prepared as an aqueous solution for injection, it can comprise an isotonic solution comprising, for example, physiological saline, dextrose, and other adjuvants, including, for example, D-sorbitol, D-mannose, D-mannitol, and sodium chloride, which can also contain an appropriate solubilizing agent, for example, alcohol (for example, ethanol), polyalcohol (for example, propylene glycol and PEG), and non-ionic detergent (polysorbate 80 and HCO-50).

[0100] If necessary, antibodies of the present invention may be encapsulated in microcapsules (microcapsules made of hydroxycellulose, gelatin, polymethylmethacrylate, and the like), and made into components of colloidal drug delivery systems (liposomes, albumin microspheres, microemulsions, nano-particles, and nano-capsules) (for example, see "Remington's Pharmaceutical Science 16th edition", Oslo Ed. (1980)). Moreover, methods for making sustained- release drugs are known, and these can be applied for the antibodies of the present invention (Langer et al., J. Biomed. Mater. Res. 15: 167-277 (1981); Langer, Chem. Tech. 12: 98-105 (1982); U.S. Pat. No. 3,773,919; EP Patent Application No. 58,481; Sidman et al., Biopolymers 22: 547- 556 (1983); EP: 133,988).

[0101] Embodiments of the Invention

[0102] In some embodiments, this disclosure provides for methods of detecting aberrantly expressed proteins in a neoplasm not previously detected in subjects with a neoplasm which can include or exclude breast cancer. In some embodiments, this disclosure provides for methods of treating a neoplasm, or a subject having or suspected of having a neoplasm. In some embodiments, this disclosure provides for methods of targeting a cell expressing SLC39A7 at the cell surface. Insome embodiments, this disclosure provides for methods of inducing an immune response against a cell expressing SLC39A7 at the cell surface.

[0103] Detecting Aberrantly Expressed Proteins

[0104] The present disclosure relates to a method of detecting aberrantly expressed SLC39A7. In some embodiments, the method comprises contacting an agent that detects SLC39A7 with a cell or tissue of a subject or a test cell or a test tissue to determine whether there is aberrant SLC39A7 expression. In some embodiments, aberrant SLC39A7 expression comprises cell surface expression of SLC39A7 or a portion thereof. In some embodiments, the agent that detects SLC39A7 comprises a monoclonal or polyclonal bound antibody that specifically binds to SLC39A7 or a portion thereof. In some embodiments the agent detects cell-surface expression of SLC39A7. In some embodiments, the agent specifically binds to a portion of SLC39A7 that is expressed on the surface of a neoplastic cell or tissue. In some embodiments the neoplastic cell or tissue comprises a breast cancer cell or tissue. In some embodiments, the agent specifically binds to a polypeptide of SLC39A7, comprising PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof, or KFVRHVKGGHGHSHGHGHAHSHTRG SHGHGRQERSTKEKQSSEEEEKETRGVQKRRGGSTVPKDGPVRPQNAEEEKRGLDLRV SGYLNLAADLAHNFTDGLAIGASFRGGRGLGILTTMTVLLHEVPHEVGDFAILVQSGCS KKQA (235-380) (SEQ ID NO:3) or a portion thereof.

[0105] Useful portions of SLC39A7 (190-213) to detect include, without limitation, 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids of PHALEPHSHHTLEQPGHGHSHSGQ (SEQ ID NO:2), which may comprise, e g., SLC39A7 (190-197), SLC39A7 (194-201), SLC39A7 (198- 205), SLC39A7 (202-209), or SLC39A7 (206-213). Useful portions of SLC39A7 (190-213) to detect include, without limitation, 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids of KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (SEQ ID NO:3), which may comprise SLC39A7 (235-242), SLC39A7 (239-246), SLC39A7 (243-250), SLC39A7 (247-254), SLC39A7 (251-258), SLC39A7 (255-262), SLC39A7 (259-266), SLC39A7 (263-270), SLC39A7 (267-274), SLC39A7 (271-278), SLC39A7 (275-282), SLC39A7 (279-286), SLC39A7 (283-290), SLC39A7 (287-294), SLC39A7 (291-298), SLC39A7 (295-302), SLC39A7 (299-306), SLC39A7 (303-310), SLC39A7 (307-314), SLC39A7 (311-318), SLC39A7 (315-322), SLC39A7 (319-326), SLC39A7(323-330), SLC39A7 (327-334), SLC39A7 (331-338), SLC39A7 (335-342), SLC39A7 (339-346), SLC39A7 (343-350), SLC39A7 (347-354), SLC39A7 (351-358), SLC39A7 (355-362), SLC39A7 (359-366), SLC39A7 (361-370), SLC39A7 (365-374), or SLC39A7 (369-378).

[0106] Methods of Treatment

[0107] The present disclosure also relates to methods of treating a subject having or suspected of having a neoplasm. In some embodiments, this disclosure relates to methods of treating a subject, e.g., patient having a neoplasm. In some embodiments, the methods of treating a neoplasm expressing SLC39A7 or a portion thereof on its surface, comprising administering to the subject a binding protein that specifically binds to the surface-expressed SLC39A7 or portion thereof. In some embodiments, the neoplasm is a breast cancer cell. In some embodiments, the binding protein binds to a polypeptide of SLC39A7, comprising PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or portion thereof, or KFVRHVKGGHGHSHGHGHAHSHTRG SHGHGRQERSTKEKQSSEEEEKETRGVQKRRGGSTVPKDGPVRPQNAEEEKRGLDLRV SGYLNLAADLAHNFTDGLAIGASFRGGRGLGILTTMTVLLHEVPHEVGDFAILVQSGCS KKQA (235-380) (SEQ ID NO:3) or portion thereof. In some embodiments, the binding protein comprises an antibody, antigen binding fragment thereof, an antineoplastic agent, an antineoplastic agent, or a drug conjugate. In some embodiments, the binding protein comprises administering to the subject a secondary agent that binds to SLC39A7 or portion thereof and delivers an antineoplastic agent to the neoplasm.

[0108] In some embodiments the binding protein is administered with an antineoplastic agent. In some embodiments, the method comprises administering a binding protein with an antineoplastic agent that comprises radiation, chemotherapy, a hormone therapy, an immunotherapy, an antibody-drug conjugate, or a combination thereof. In some embodiments, the hormone therapy or immunotherapy comprises an enzymatic inhibitor, small molecule inhibitor, an antibody, or targeted vaccines. In some embodiments, the hormone therapy is an enzymatic inhibitor or small molecule inhibitor targeting estrogen receptor (ESRI), progesterone receptor (PR), receptor tyrosine-protein kinase erbB2 (HER2), epidermal growth factor receptor (EGFR), lutenizing hormone (LH), cyclin-dependent kinase 4 (CDK4), cyclin-dependent kinase 6 (CDK6), poly (ADP-ribose) polymerase (PARP), phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), or a combination thereof. In some embodiments, the hormone therapy inhibitor is selected from: tamoxifen, toremifene, fulvestrant, elacestrant, exemestane, letrozole, anastrozole, lapatinib,neratinib, palbociclib, ribociclib, abemaciclib, trilaciclib, tucatinib / irbinitinib, olaparib, talazoparib, alpeli sib, capivasertib, or a combination thereof. In some aspects, the hormone therapy is an antibody that binds ESRI, PR, HER2, or EGFR, selected from: pertuzumab, traztuzumab, margetuximab, or a combination thereof. In some embodiments, the immunotherapy comprises an antibody selected from: pembrolizumab, atezolizumab, avelumab, or a combination thereof. In some embodiments, the antineoplastic agent comprises an antibody-drug conjugate selected from: sacituzumab govitecan, ado-trastuzumab emtansine, fam-trastuzumab deruxtecan, or aOIgG-NC- MMAF.

[0109] Methods of Targeting a Cell Expressing Extracellular SLC39A 7

[0110] The present disclosure also relates to methods of targeting a cell that expresses SLC39A7 or a portion thereof at the cell surface, the method comprising contacting the cell with a binding protein that binds to the extracellular portion of cell-surface expressed SLC39A7. In some embodiments, the extracellular portion of SLC39A7 comprises the amino acid sequence PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof, or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3) or a portion thereof. In some embodiments, the binding protein in the method comprises a drug conjugate. In some embodiments, the binding protein in the method comprises an antibody or antigen binding portion thereof.

[0111] Antibodies

[0112] Monoclonal antibodies suitable for use according to the invention can be produced by a variety of techniques. In a non-limiting example, an anti-SLC29A7 antibody or antigen binding fragment is isolated by screening and selecting from an artificial antibody library. In certain embodiments, the anti-SLC29A7 antibody isolated from the library is humanized. In certain embodiments, the antibody library comprises human heavy and light chain variable domains. In another non-limiting example, an anti-SLC29A7 antibody is generated by immunizing an animal such as a mouse, more preferably a transgenic animal that produces human antibodies. Suitable transgenic mice include XenoMouse®, HuMAb® Mouse, TC™ mouse, and KM™ mouse. (See, e.g., Davis et al., Production of Human Antibodies from Transgenic Mice. Methods in Molecular Biology, 2004, Vol.248: 191-200.

[0113] Methods of Inducing an Immune Response Against a Cell Expressing Extracellular SLC39A7

[0114] The present disclosure also relates to methods of inducing an immune response against a target cell that expresses SLC39A7 or a portion thereof on its surface in a subject in need thereof, which comprises administering to the subject an immunogenic composition which comprises an SLC39A7 polypeptide. In some embodiments, the target cell is a cancer cell. In some embodiments, the target cell is a breast cancer cell. In some embodiments, the method comprises administering to the subject an immunogenic composition comprising a B cell epitope of SLC39A7. In some embodiments, the immunogenic composition comprises one or more polypeptides, wherein each of the one or more polypeptides comprises 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids of SLC39A7. In some embodiments, the immunogenic composition comprises 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids ofPHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO: 2) orKFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3). In some embodiments, the immunogenic composition comprises a polypeptide comprising 8 or more contiguous acids of a polypeptide of Table 3. In some embodiments, the immunogenic composition comprises a T cell epitope linked to the B cell epitope.

[0115] The invention provides methods and compositions for inducing an immune response against an extracellular portion of cell-surface expressed SLC29A7 in a subject. In certain embodiments, a humoral response is induced that targets SLC29A7. In certain embodiments, the humoral response differentially targets disease cells that display surface-expressed SLC29A7 or portion thereof vs. normal cells that do not display SLC29A7 or portion thereof. In certain embodiments, disease cell comprise neoplastic cells and cancer cells, including but not limited to breast cancer cells, that express a portion of SLC29A7 on the cell surface.

[0116] In certain embodiments, an immunogenic composition is provided which comprises a B cell epitope (or “determinant”) of SLC29A7 or a B cell epitope mimic. Methods of predicting B cell epitopes have been described. A non-limiting method for predicting linear B-cell epitopes is described by Larsen et al., Improved method for predicting linear B-cell epitopes, Immunome Research, 2006 April 24, Vol. 2, article no. 2. Larsen combined a hidden Markov model with apropensity scale method to obtain a method designated the BepiPred method. BepiPred is available online, for example, at tools.immuneepitope.org / bcell / . Jespersen describes a B-cell epitope prediction method using conformational epitopes. (Jespersen et al., BepiPred-2.0: improving sequence-based B-cell epitope prediction using conformational epitopes. Nucleic Acids Res. 2017 July 3, Vol. 45, web server issue, W24-W29). BepiPred-2.0 can be accessed online, for example at tools.immuneepitope.org / bcell / . BepiPred-3.0 employs a protein language model (LM) to improve prediction accuracy. (See Clifford et al., BepiPred-3.0: Improved B-cell epitope prediction using protein language models. Protein Science, 2022 Dec, 3 l(12):e4497. BepiPred-2.0 is accessible online, for example at services. healthtech. dtu.dk / services / BepiPred-3.0 / . Ras-Carmona describes a BLAST-based method based on a dataset of known linear B cell epitopes (Ras-Carmona et al., Prediction of B cell epitopes in proteins using a novel sequence similaritybased method. Sci Rep. 2022 Aug 12, 12(1): 13739). The method is implemented at imath.med.ucm.es / bepiblast.

[0117] B cell epitopes from any region of SLC29A7 can be used. The Applicants have discovered that regions SLC29A7 that do not encompass putative transmembrane regions of SLC29A7 comprise preferred target epitopes. In certain embodiments, B cell epitopes comprise peptides from PHALEPHSHHTLEQPGHGHSHSGQ (amino acids 190-213) or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (amino acids 235-380). In some embodiments, the target antigen, or fragment thereof, including but not limited to a B cell epitope is from 8 to 35 amino acids in length. In some embodiments, the target antigen or fragment thereof is from 10 to 25 amino acids in length. In some embodiments, the target antigen or fragment thereof, such as a B cell epitope of SLC29A7 is composed of a peptide 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 amino acids in length. In certain non-limiting embodiments, the target antigen or antigen fragment or B cell epitope thereof comprises a sequence 8 or more contiguous amino acids of SLC29A7 comprising e.g, aa 196-203, 199-206, 200-207, 201-208, 258-265, 259-266, 260-267, 264-271, 269-276, 278-285, 287-294, 288-295, 291-298, 295-302, 297-304, 301-308, 304-311, 309-316, 328-335, 340-347, 357-364, or 371-378. In certain embodiments, mimotopes of the aforementioned B cell epitopes are employed.

[0118] For induction of a humoral response, an immunogen preferably comprises both a B cell determinant and a T helper (Th) cell determinant. In certain embodiments, an immunogen of the invention comprises a B cell determinant coupled to a carrier protein. In certain embodiments, an immunogen of the invention comprises a B cell determinant coupled to an artificial Th epitope. In certain embodiments, an immunogen of the invention comprises a B cell determinant expressed with or conjugated to a viral particle. In certain embodiments, an immunogen of the invention comprises a host cell engineered to expresses the B cell determinant at the cell surface.

[0119] C. Certain Exemplary Sequences

[0120] SLC37A9 / ZIP7 Full Sequence:

[0121] MARGLGAPHWVAVGLLTWATLGLLVAGLGGHDDLHDDLQEDFHGHSHRH SHEDFHHGHSHAHGHGHTHESIWHGHTHDHDHGHSHEDLHHGHSHGYSHESLYHRGH GHDHEHSHGGYGESGAPGIKQDLDAVTLWAYALGATVLISAAPFFVLFLIPVESNSPRH RSLLQILLSFASGGLLGDAFLHLIPHALEPHSHHTLEQPGHGHSHSGQGPILSVGLWVLSG IVAFLVVEKFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETR GVQKRRGGSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFR GGRGLGILTTMTVLLHEVPHEVGDFAILVQSGCSKKQAMRLQLLTAVGALAGTACALL TEGGAVGSEIAGGAGPGWVLPFTAGGFIYVATVSVLPELLREASPLQSLLEVLGLLGGVI MM VLIAHLE (SEQ ID NO: 1)

[0122] SLC37A9 / ZIP7 AA 190-213:

[0123] PHALEPHSHHTLEQPGHGHSHSGQ (SEQ ID NO:2)

[0124] SLC37A9 / ZIP7 AA 235-380:

[0125] KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRG VQKRRGGSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRG GRGLGILTTMTVLLHEVPHEVGDFAILVQSGCSKKQA (SEQ ID NO:3)

[0126] Although the present invention and its advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined in the appended claims.

[0127] The present invention will be further illustrated in the following Examples which are given for illustration purposes only and are not intended to limit the invention in any way.Examples

[0128] The specific methods and compositions described herein are representative of preferred embodiments and are exemplary and not intended as limitations on the scope of the invention. Other objects, aspects, and embodiments will occur to those skilled in the art upon consideration of this specification, and are encompassed within the spirit of the invention as defined by the scope of the claims. Thus, for example, in each instance herein, and in embodiments or examples of the present invention, any of the terms “comprising”, “consisting essentially of’, and “consisting of’ may be replaced with either of the other two terms in the specification. The methods and processes illustratively described herein suitably may be practiced in differing orders of steps, and that they are not necessarily restricted to the orders of steps indicated herein or in the claims. It is also that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. Under no circumstances may the patent be interpreted to be limited to the specific examples or embodiments or methods specifically disclosed herein. Under no circumstances may the patent be interpreted to be limited by any statement made by any Examiner or any other official or employee of the Patent and Trademark Office unless such statement is specifically and without qualification or reservation agreed to and expressly adopted in a responsive writing by Applicants.Example 1: SLC39A7 identified as a novel neoantigen for TNBC

[0129] Initially, Applicants initiated the process by injecting Triple Negative Breast Cancer (TNBC) cells (Hs578T) into rabbits to generate polyclonal xeno-antibodies. Subsequently, leveraging Applicants’ proprietary Oncotope platform, Applicants isolated antibodies that exhibited a specific affinity for TNBC cells while excluding those with any binding to normal breast epithelial cells derived from the same patient. This culminated in the acquisition of a serum enriched with polyclonal antibodies highly specific to TNBC. Next, Total Anti-Breast Cancer Serum (before filtration) and Oncotope Platform Filtered Anti-Breast Cancer Serum was then reacted with approximately 30,000 human proteins on a Protein Microarray, in collaboration with Yamaha and Tuning Fork Bio. Results of differential binding of the Total Anti-Breast Cancer Serum (before filtration) and the Oncotope Platform Filtered Anti-Breast Cancer Serum to the Protein Microarray revealed 8 unique antigens with significantly higher affinity binding toOncotope Platform Filtered Anti-Breast Cancer Serum. Among these antigens, SLC39A7 emerged as one of particular interest.Example 2: SLC39A 7 is aberrantly expressed on the cell surface of breast cancer cells

[0130] The presence of SLC39A7 / ZIP7 on the cell surface was determined across a diverse spectrum of breast cancer cell lines, encompassing the five distinct subtypes of breast cancers (Luminal A, ER+ PR+ / - HERZ- (LA); Luminal B, ER+ PR- HER2+ (LB); HER2 Positive (H); Triple Negative A (TNA); and Triple Negative B (TNB), as outlined in TABLE 1) (Dai X, Cheng H, Bai Z, Li J. Breast Cancer Cell Line Classification and Its Relevance with Breast Tumor Subtyping. J Cancer. 8( 16):3131-3141 (2017). Among the ten breast cancer cell lines scrutinized, two exhibited no mRNA expression of SLC39A7 (as indicated in TABLE 1). Additionally, Applicants assessed two variations of normal breast epithelial cells and included healthy Peripheral Blood Mononuclear Cells (PBMCs) in Applicants’ analysis to comprehensively examine the presence of SLC39A7 on the cell surface.Example 3: SLC39A7 can be selectively targeted with antibody drug conjugates

[0131] Applicants validated the presence of SLC39A7 exclusively on the surface of breast cancer cells, distinguishing it from normal cells. This verification was accomplished by using commercially available rabbit polyclonal anti-SLC39A7 antibodies added to cultures of both Cancer Cells (Human Breast Cancer Cells: Hs578T from ATCC) and corresponding Normal Cells from the same tissue type from the same individual (Breast Epithelial Cells: Hs578Bst from ATCC) in the presence of a Secondary Antibody Drug Conjugate (ADC). The ADC used was aOIgG-NC-MMAF (from Moradec), an anti-rabbit IgG (H+L) specific antibody conjugated to monomethyl auristatin F (MMAF) with a non-cleavable linker. Following a 48-hour incubation period, Applicants assessed cell viability using the CellTiter Gio assay. The results revealed statistically significant cell death exclusively in the breast cancer cell lines, while the corresponding healthy breast epithelial cells remained unaffected (FIG. 1).

[0132] Additionally, Applicants conducted flow cytometric analysis to evaluate the binding capacity of the polyclonal antibody against SLC39A7. The gating strategy to test the binding capacity of 2-antigen targeting antibodies on Hs578T is shown by FIGs. 2A-2F. The gating strategy to test the binding capacity of 2-antigen targeting antibodies on Hs578Bst and Hs578T with varying dilutions is shown by FIGs. 3A-3D using Total anti-Breast Cancer Serum and FIGs.4A-4D using Oncotope Filtered anti-Breast Cancer Serum. These analyses demonstrated robust binding of the antibody to the breast cancer cells, whereas it exhibited no significant binding to the healthy breast epithelial cells (FIG. 5). These findings further underscore the selective targeting capability of the anti-SLC39A7 antibodies, validating their potential as a precise therapeutic tool for breast cancer.Example 4: Flow cytometric analysis confirms SLC39A 7 expression on the cell surface of breast cancer cells and targeting SLC39A 7 results in cell death

[0133] Applicants provided additional confirmation of the robust cell surface expression of SLC39A7 in the eight breast cancer cell lines characterized by SLC39A7 mRNA expression (FIG. 6). This outcome underscores the selective targeting potential of SLC39A7 and its suitability for therapeutic applications with minimal off-target effects. The results of the cell cytotoxicity assays were complementarily validated with by flow cytometric analysis.

[0134] Cell cytotoxicity assays were used to validate the cell surface expression of SLC39A7 across the eight breast cancer cell lines that exhibited SLC39A7 mRNA expression. Notably, minimal cell death was observed in the case of the two breast cancer cell lines that lacked SLC39A7 mRNA expression, as well as in the two normal breast epithelial cell lines and PBMCs (FIG. 7).Example 5: Flow cytometric analysis to evaluate binding capacity of 2-antigen targeting on breast cancer cells

[0135] Applicants used flow cytometry to test the binding capacity of 2-antigen targeting on breast cancer cells using Alexa-488 conjugated Secondary Donkey anti-Rabbit antibody to detect antibody binding of sera or primary antibodies. Applicants compared binding capacity in syngeneic normal and TNBC breast cell lines (FIG. 8), ESR1+ breast cell lines (FIG. 9), HER2+ breast cell lines (FIG. 10), TNBC breast cell lines (FIG. 11), and healthy cells (FIG. 12).Example 6: Cytotoxic capacity of ADC targeting on breast cancer cells

[0136] Applicants used CellTiter Gio assays to test the cytotoxic capacity of ADC targeting on breast cancer cells using aOIgG-NC-MMAF. Applicants compared cytotoxic capacity in syngeneic normal and TNBC breast cell lines (FIG. 13), ESR1+ breast cell lines (FIG. 14), HER2+ breast cell lines (FIG. 15), TNBC breast cell lines (FIG. 16), and healthy cells (FIG. 17).

[0137] The epitopes of each primary antibody evaluated for Examples 4 and 5 are shown in FIG. 18.Example 7: Comparison of Proteintech and Atlas Antibodies primary anti-SLC 39A7 antibodies

[0138] Using syngeneic TNBC cell lines Hs578Bst and Hs578T, Applicants conducted flow cytometric analysis to compare the binding capacity of the Proteintech and Atlas Antibodies polyclonal primary antibody against SLC39A7. The same gating strategy used in Example 2 was used. The analysis of both antibodies revealed that the Proteintech antibody, as shown in FIGs. 19A-D, had more robust binding to the breast cancer cells than the Atlas Antibodies antibody, as shown in FIGs. 20A-D. Comparing the pTAB, pF AB, Proteintech antibody, and Atlas Antibodies antibody percent binding as shown in FIG. 21 reveals that the specificity of the Proteintech antibody for cell surface-expressed SLC39A7 is more pronounced and underscores the importance of selecting or designing antibodies targeting specific SLC39A7 epitopes to be efficacious.Example 8: Comparison of ADC efficacy between Proteintech and Atlas Antibodies primary anti- SLC39A 7 antibodies

[0139] Applicants compared the ADC efficacy between Proteintech and Atlas Antibodies primary anti-SLC39A7 antibodies in syngeneic normal and TNBC breast cell lines. As in Example 1, the ADC used was aOIgG-NC-MMAF (from Moradec), an anti-rabbit IgG (H+L) specific antibody conjugated to monomethyl auristatin F (MMAF) with a non-cleavable linker, and following a 48-hour incubation period, Applicants assessed cell viability using the CellTiter Gio assay. While both primary antibodies were effective, the Proteintech primary antibody appears more specific for the surface-expressed SLC39A7 epitope than the Atlas Antibodies primary antibody, again underscoring the the importance of selecting or designing antibodies targeting specific SLC39A7 epitopes to be efficacious.Example 9: SLC39A7 -specific inhibitor NVS-ZP7-4 is not effective for cell surface-expressing SLC39A7 cells

[0140] To determine whether the SLC39A7-specific inhibitor NVS-ZP7-4 is cytotoxic against breast cancer cells expressing SLC39A7 at the cell surface, Applicants performed an CellTiterGlo assay after 60 hours of drug incubation at varying concentrations. Applicants found that in all breast cancer cells tested, the compound showed no significant effect on cell viability. Some reduction in the viability of HCC1954 cells, but even at a high concentration of 10 uM the viabilitydid not drop below 50%. Interestingly, NVS-ZP7-4 appeared to reduce the viability of breast epithelial cell line MCF-10A, likely due to the SLC39A7 being expressed not at the cell surface but rather at the endoplasmic reticulum or Golgi apparatus. These results rule out direct drug targeting for cell surface-expressed SLC39A7 and highlights the importance of developing antibody and immune-cell therapeutic alternatives for certain tumor-specific antigens.* * *

[0141] Having thus described in detail preferred embodiments of the present invention, it is to be understood that the invention defined by the above paragraphs is not to be limited to particular details set forth in the above description as many apparent variations thereof are possible without departing from the spirit or scope of the present invention.

Claims

WHAT IS CLAIMED IS:

1. A method of detecting a neoplasm in a subj ect or a cell or tissue sample from a subject, which comprises detecting surface expression of solute carrier family 39 member 7 (SLC39A7) in the subject or cell or tissue sample from the subject.

2. The method of claim 1, which comprises contacting the neoplasm with an antibody that specifically binds to solute carrier family 39 member 7 (SLC39A7) and detecting the bound antibody.

3. The method of claim 1 or 2, wherein the neoplasm comprises a breast cancer cell.

4. A method of treating a neoplasm in a subject, wherein the neoplasm expresses SLC39A7 or a portion thereof on its surface, which comprises administering to the subject a binding protein that specifically binds to the surface-expressed SLC39A7 or portion thereof.

5. The method of claim 4, wherein the neoplasm comprises a breast cancer cell.

6. The method of claim 4 or 5, wherein the binding protein binds to an SLC39A7 polypeptide which comprises PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3) or a portion thereof.

7. The method of any one of claims 4 - 6, wherein the binding protein comprises an antibody or an antigen binding fragment thereof.

8. The method of any one of claims 4 - 7, wherein the binding protein is linked to an antineoplastic agent.

9. The method of any one of claims 4 - 8, wherein the binding protein comprises a drug conjugate.

10. The method of any one of claims 4 - 9, which further comprises administering to the subject a secondary agent that binds to the SLC39A7-specific binding protein or the SLC39A7 or portion thereof and delivers an antineoplastic agent to the neoplasm.

11. The method of any one of claims 4 - 10, which comprises administering the binding protein with an antineoplastic agent.

12. The method of claim 11, wherein the antineoplastic agent comprises radiation, chemotherapy, a hormone therapy, an immunotherapy, an antibody-drug conjugate or a combination thereof.

13. The method of claim 12, wherein the hormone therapy or immunotherapy comprises an enzymatic inhibitor, small molecule inhibitor, an antibody, or targeted vaccine.

14. The method of claim 13, wherein the hormone therapy is an enzymatic inhibitor or small molecule inhibitor targeting estrogen receptor, progesterone receptor, HER2, EGFR, luteinizing hormone, cyclin-dependent kinase 4, cyclin-dependent kinase 6, PARP, PI3K, AKT, or a combination thereof.

15. The method of claim 14, wherein the inhibitor comprises tamoxifen, toremifene, fulvestrant, elacestrant, exemestane, letrozole, anastrozole, lapatinib, neratinib, palbociclib, ribociclib, abemaciclib, trilaciclib, tucatinib / irbinitinib, olaparib, talazoparib, alpelisib, capivasertib, or a combination thereof.

16. The method of claim 13, wherein the hormone therapy comprises an antibody that binds estrogen receptor, progesterone receptor, HER2, EGFR selected from: pertuzumab, traztuzumab, margetuximab, or a combination thereof.

17. The method of claim 13, wherein the immunotherapy comprises an antibody selected from: pembrolizumab, atezolizumab, avelumab, or a combination thereof.

18. The method of claim 11, wherein the antineoplastic agent comprises an antibodydrug conjugate selected from: sacituzumab govitecan, ado-trastuzumab emtansine, famtrastuzumab deruxtecan, or aOIgG-NC-MMAF.

19. A method of targeting a cell that expresses SLC39A7 or a portion thereof at the cell surface, which comprises contacting the cell with a binding protein that binds to the extracellular region of cell-surface expressed SLC39A7.

20. The method of claim 19, wherein the extracellular region of cell-surface expressed SLC39A7 comprises PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3) or a portion thereof.

21. The method of claim 19 or 20, wherein the binding protein comprises a drug conjugate.

22. The method of any one of claims 19 - 21, wherein the binding protein comprises an antibody or antigen binding portion thereof.

23. A method of inducing an immune response against a target cell that expresses SLC39A7 or a portion thereof on its surface in a subject in need thereof, which comprises administering to the subject an immunogenic composition which comprises an SLC39A7 polypeptide.

24. The method of claim 23, wherein the target cell is a cancer cell.

25. The method of claim 23, wherein the target cell is a breast cancer cell.

26. The method of any one of claims 23 - 25, which comprises administering to the subject an immunogenic composition which comprises a B cell epitope of SLC39A7.

27. The method of claim 26, wherein immunogenic composition comprises one or more polypeptides, wherein each of the one or more polypeptides comprises 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids of SLC39A7.

28. The method of claim 26, wherein the immunogenic composition comprises 8, 10, 12, 14, 16, 18, 20, or 22 contiguous amino acids of PHALEPHSHHTLEQPGHGHSHSGQ (190- 213) (SEQ ID NO:2) or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3).

29. The method of claim 26, wherein the immunogenic composition comprises a polypeptide comprising 8 or more contiguous amino acids of a polypeptide of Table 3.

30. The method of any one of claims 26 - 29, wherein the immunogenic composition comprises a T cell epitope linked to the B cell epitope.

31. A method of treating a subject having or suspected of having a neoplasm expressing SLC39A7 or a portion thereof on its surface, the method comprising: a. obtaining or having obtained a sample from the subject; b. detecting or having detected surface-expressed SLC39A7; and c. administering to the subject a binding protein that specifically binds to the surface-expressed SLC39A7 or portion thereof.

32. The method of claim 31, wherein the neoplasm comprises a breast cancer cell.

33. The method of claim 31, wherein the binding protein binds to an SLC39A7 polypeptide comprising amino acids PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3) or a portion thereof.

34. The method of claim 31, wherein the binding protein comprises an antibody or an antigen binding fragment thereof.

35. The method of claim 31, which further comprises administering to the subject a secondary agent that binds to the SLC39A7 or portion thereof and delivers an antineoplastic agent to the neoplasm.

36. The method of claim 31, which comprises administering the binding protein with an antineoplastic agent.

37. The method of claim 36, wherein the antineoplastic agent comprises radiation, chemotherapy, a hormone therapy, an immunotherapy, an antibody-drug conjugate or a combination thereof.

38. The method of claim 37, wherein the hormone therapy or immunotherapy comprises an enzymatic inhibitor, small molecule inhibitor, an antibody, or targeted vaccine.

39. The method of claim 38, wherein the hormone therapy is an enzymatic inhibitor or small molecule inhibitor targeting estrogen receptor, progesterone receptor, HER2, EGFR, luteinizing hormone, cyclin-dependent kinase 4, cyclin-dependent kinase 6, PARP, PI3K, AKT, or a combination thereof.

40. The method of claim 39, wherein the inhibitor comprises tamoxifen, toremifene, fulvestrant, elacestrant, exemestane, letrozole, anastrozole, lapatinib, neratinib, palbociclib, ribociclib, abemaciclib, trilaciclib, tucatinib / irbinitinib, olaparib, talazoparib, alpelisib, capivasertib, or a combination thereof.

41. The method of claim 38, wherein the hormone therapy comprises an antibody that binds estrogen receptor, progesterone receptor, HER2, EGFR selected from: pertuzumab, traztuzumab, margetuximab, or a combination thereof.

42. The method of claim 38, wherein the immunotherapy comprises an antibody selected from: pembrolizumab, atezolizumab, avelumab, or a combination thereof.

43. The method of claim 36, wherein the antineoplastic agent comprises an antibodydrug conjugate selected from: sacituzumab govitecan, ado-trastuzumab emtansine, famtrastuzumab deruxtecan, or aOIgG-NC-MMAF.

44. A solute carrier family 39 member 7 (SLC39A7) binding agent for use in detecting cell surface expression of solute carrier family 39 member 7 (SLC39A7) in a subject or cell or tissue sample, or for use in treating a cell or neoplasm or cancer that expresses cell surface SLC39A7.

45. The agent of claim 44, wherein the cell or neoplasm comprises a breast cancer cell.

46. The agent of any one of claims 44 - 45, wherein the agent comprises a binding protein or a monoclonal antibody or polyclonal antibody or an antigen binding portion thereof that binds to SLC39A7 on the surface of a cell.

47. The agent of any one of claims 44 - 46 wherein the agent binds to an SLC39A7 polypeptide which comprises PHALEPHSHHTLEQPGHGHSHSGQ (190-213) (SEQ ID NO:2) or a portion thereof or KFVRHVKGGHGHSHGHGHAHSHTRGSHGHGRQERSTKEKQSSEEEEKETRGVQKRRG GSTVPKDGPVRPQNAEEEKRGLDLRVSGYLNLAADLAHNFTDGLAIGASFRGGRGLGIL TTMTVLLHEVPHEVGDFAILVQSGCSKKQA (235-380) (SEQ ID NO:3) or a portion thereof.

48. The agent of any one of claims 44 - 47 wherein the agent comprises a drug conjugate.

49. The agent of any one of claims 44 - 48 wherein the agent comprises an antineoplastic agent.

Images