GPR160 antibody development for cancer treatment

用于癌症治疗的 GPR160 抗体开发

• 批准号: 10711206
• 负责人: James D Brien
• 金额: $ 21.25万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-16 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711206
• 关键词: Affinity; Affinity Chromatography; Alloys; Amino Acid Sequence; Antibodies; Antibody Therapy; Avidity; Binding; Biological Assay; Biological Products; Breast; Breast Cancer Cell; Breast Cancer cell line; Cancer cell line; Cause of Death; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Chemosensitization; Cisplatin; Clinical; Colon; Colon Carcinoma; Combined Modality Therapy; Cytotoxin; Data; Development; Dose; Dose Limiting; Elements; Esophagus; Flow Cytometry; G-Protein-Coupled Receptors; Genes; Goals; Human; Human Cell Line; Hybridomas; Immunize; Immunodeficient Mouse; In Vitro; Keyhole Limpet Hemocyanin; Ligands; Light; Lung; MDA MB 231; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Malignant neoplasm of prostate; Measures; Mediating; Modeling; Monoclonal Antibodies; Monoclonal Antibody Therapy; Mus; Oryctolagus cuniculus; Paclitaxel; Patients; Prostate; Publications; Role; SW480; Stomach; Testing; Therapeutic; Therapeutic Monoclonal Antibodies; Toxic effect; Transgenic Mice; Translating; Trastuzumab; Treatment-Related Cancer; Treatment-related toxicity; Uncertainty; Work; Xenograft Model; Xenograft procedure; anti-cancer; anticancer treatment; cancer cell; cancer therapy; clinical effect; clinical efficacy; clinical predictors; colon cancer cell line; cytotoxic; experimental study; extracellular; human monoclonal antibodies; improved; in vitro Model; in vivo; malignant breast neoplasm; mouse model; neoplastic cell; overexpression; oxaliplatin; pharmacologic; polyclonal antibody; programs; receptor; response; scale up; side effect; small molecule; success; therapy development; triple-negative invasive breast carcinoma; tumor

We discovered that a rabbit polyclonal antibody raised against the second extracellular loop of human G-protein- coupled receptor 160 (hGPR160/ECL2) has two remarkable actions: (1) direct anti-cancer effects on human colon cancer and triple negative breast cancer (TNBC) cell lines that express the receptor without altering the viability of a normal colon cancer cells line, and (2) great potentiation of the cytotoxic effects of the small molecule chemotherapeutic, oxaliplatin, at doses that by themselves have no significant activity. This potentiation indicates that combination with the hGPR160/ECL2 antibody might yield a considerable improvement in the clinical effect with cytotoxin doses that are lower and produce fewer side-effects than those in current use. To translate these observations into a practical therapy, we propose to develop a human monoclonal hGPR160/ECL2 antibody for further characterization of its anticancer effects and as a step towards developing a therapeutically useful human monoclonal hGPR160/ECL2 antibody. The current proposal is in response to PAR-22-216 with aims to develop and test a new biologic agent that both treats cancer and mitigates cancer treatment-related toxicities. We are unaware of any commercially available small molecule ligands for GPR160. Our recent publication1 and preliminary data presented here indicate that an hGPR160/ECL2 monoclonal antibody is a viable chemotherapeutic. However, our preliminary studies used a rabbit polyclonal antibody, which has an irreducible element of uncertainty concerning the locus of binding. Accordingly, a human monoclonal hGPR160/ECL2 antibody will address this issue and permit unambiguous in vitro experiments on anticancer effects in human cancer cell lines and in vivo experiments on human cell line-derived xenografts (CDX) in immunodeficient mice. Success with the work proposed here in three Aims would lead to further work to develop a therapeutic monoclonal antibody. In Aim 1, we will develop human monoclonal antibodies to hGPR160/ECL2 by immunizing transgenic mice expressing human heavy and light genes with the hGPR160’s ECL2 amino acid sequence (or subsequence thereof) conjugated to KLH and developing hybridomas via PEG-fusion. We will screen clonal antibodies by flow cytometry and down-select using a blocking-of-binding assay to a panel of ~10 hGPR160/ECL2 mAbs clones for further in vitro and in vivo studies. We will then test the anti-cancer cell activity of our antibodies with in vitro studies. In Aim 2, the top 2-3 candidates (best binding affinity against the ligand and LC50 in the in vitro cell viability assay) will be advanced for (1) in vivo orthotopic cell line-derived xenograft (CDX) mouse models of colon cancer and TNBC. In Aim 3, the top candidate will be tested in in vitro oxaliplatin and paclitaxel potentiation studies. An hGPR160/ECL2 mAb may prove to be a significant improvement to current anti-cancer treatments that are inadequate for a large percentage of patients. However, small molecule cytotoxins will continue to have an important role and combination therapy with an hGPR160/ECL2 antibody may allow their cytotoxic effects to be maximized while using doses with far fewer side effects.

我们发现了一种兔多克隆抗体，可以抵抗人g蛋白-的细胞外第二环