Discovery of therapeutic antibodies targeting breast cancer metastasis using Enabled G-Protein-Coupled Receptors

使用启用的 G 蛋白偶联受体发现针对乳腺癌转移的治疗抗体

• 批准号: 9256138
• 负责人: Mauro Mileni
• 金额: $ 30万
• 依托单位: ABILITA BIO, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-19 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9256138
• 关键词: Address; Affinity; Agonist; Animals; Antibodies; Antigens; Automobile Driving; Bacteriophages; Behavior; Binding; Biological Assay; Biological Response Modifier Therapy; Biotechnology; Bone Marrow; Breast Cancer Cell; Breast cancer metastasis; Cancer Patient; Cell Line; Cell Proliferation; Clinical; Clinical Research; Combined Modality Therapy; Complement; Complex; Cytotoxic T-Lymphocyte-Associated Protein 4; Development; Dinoprostone; Directed Molecular Evolution; Disseminated Malignant Neoplasm; Diversity Library; Drug Targeting; Ectopic Expression; Environment; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Funding; G-Protein-Coupled Receptors; Gene Library; Generations; Growth; Heterogeneity; Homing; Homology Modeling; Immune; Immune response; Immunization; Immunotherapy; In Vitro; Japan; Lead; Leukocyte Trafficking; Libraries; Liver; Lung; Malignant Neoplasms; Mammalian Cell; Maps; Membrane; Membrane Proteins; Metastatic breast cancer; Metastatic to; Methods; Modality; Molecular; Molecular Conformation; Mutagenesis; Mutation; Neoplasm Metastasis; Organ; Outcome; PDCD1LG1 gene; Pattern; Phage Display; Pharmacology; Phase; Primary Neoplasm; Process; Property; Prostaglandin Receptor; Prostaglandins; Proteins; Signal Transduction; Site; Small Business Innovation Research Grant; Specificity; System; Technology; Testing; Therapeutic; Therapeutic antibodies; Toxic effect; Tumor Cell Migration; Variant; WFDC2 gene; abstracting; antibody libraries; base; cancer cell; cancer therapy; chemokine; chemokine receptor; genetic variant; improved; in vivo Model; inhibitor/antagonist; innovation; lymph nodes; malignant breast neoplasm; migration; neoplastic cell; new technology; novel therapeutics; personalized medicine; precision medicine; pressure; receptor; small molecule; targeted treatment; therapy design; thermostability; tool; tumor

Project Summary/Abstract Abilita Bio (AB), an innovation driven biotechnology company, requests SBIR Phase I funding for the discovery of novel therapeutic antibodies targeting G protein-coupled receptors (GPCRs) for the treatment of metastatic breast cancer. In cancer therapy each treatment modality has its own limitations. Clinical studies clearly indicate that immunotherapy with checkpoint inhibitors (PD-1/PD-L1, CTLA-4 etc.) can achieve a persistent immune response with improved clinical outcomes. However, this approach is especially effective when administered in combination therapy with targeted therapy aiming at essential components of tumor spreading, homing and growth especially in metastatic cancer. Metastasis is the major factor limiting survival in cancer patients. Therefore, interfering with the molecular mechanisms controlling the metastatic behavior of tumor cells is essential for successfully treating cancer. Experimental and clinical evidence supports the notion that one of the most important mechanisms operating in metastasis involves homeostatic chemokines and their receptors. Breast cancer is characterized by a distinct metastatic pattern involving the regional lymph nodes, bone marrow, lung and liver. Tumor cell migration from the primary tumor site to the metastatic homing sites shares many similarities with leukocyte trafficking, which is critically regulated by chemokine-binding GPCRs. Interestingly, several studies have established that prostaglandin E2 (PGE2) secretion by breast cancer cells can upregulate CCR7 expression via activation of prostaglandin-binding GPCRs, EP2 and EP4. This proposal focuses on targeting key GPCRs critically involved in tumor cells metastatic migration and organ-specific invasion using therapeutic antibodies. The use of therapeutic antibodies to target GPCRs has a tremendous potential for the treatment of cancer because of their high affinity, exquisite specificity and low toxicity relative to small molecules. However, GPCRs therapeutic antibody discovery is extremely challenging because of their inherent low stability, low expression and conformational heterogeneity outside their natural membrane environment. In fact, only 1 GPCR-targeting antibody has been approved to date, and only in Japan. In order to overcome the afore mentioned barriers, Abilita Bio offers an innovative solution through the generation of Enabled Membrane Proteins (EMPs™), representing structurally and functionally enhanced versions of natural receptors with significant improved expression and thermostability. The availability of highly expressing, stable and functionally folded EMPs will enable the isolation of novel therapeutic antibodies using in vitro antibody display methods. Funds are requested to (a) generate mutational gene libraries for proposed targets and select for enhanced variants using our directed evolution system; (b) characterize variants for expression, function and enhanced stability in mammalian cells; (c) develop biopanning assay tools using evolved EMPs as antigens and screen phage display antibody libraries for the isolation of preliminary antibody leads to be optimized and developed in Phase II.

项目概要/摘要 Abilita Bio (AB) 是一家创新驱动的生物技术公司，请求 SBIR 第一阶段资金用于 发现针对 G 蛋白偶联受体 (GPCR) 的新型治疗抗体用于治疗 转移性乳腺癌。 在癌症治疗中，每种治疗方式都有其自身的局限性。临床研究清楚地表明 检查点抑制剂（PD-1/PD-L1、CTLA-4等）免疫治疗可实现持续免疫 反应并改善临床结果。然而，这种方法在施用时特别有效 与靶向治疗联合治疗，针对肿瘤扩散、归巢的重要组成部分 和生长，尤其是在转移性癌症中。转移是限制癌症患者生存的主要因素。 因此，干扰控制肿瘤细胞转移行为的分子机制是 对于成功治疗癌症至关重要。 实验和临床证据支持这样的观点：最重要的机制之一 转移中的作用涉及稳态趋化因子及其受体。乳腺癌是 其特点是具有明显的转移模式，涉及区域淋巴结、骨髓、肺和 肝。肿瘤细胞从原发肿瘤部位迁移到转移归巢部位有许多相似之处 白细胞运输受到趋化因子结合 GPCR 的严格调节。有趣的是，几个 研究证实乳腺癌细胞的前列腺素 E2 (PGE2) 分泌可以上调 CCR7 通过激活前列腺素结合 GPCR、EP2 和 EP4 表达。该提案的重点是 靶向与肿瘤细胞转移迁移和器官特异性侵袭密切相关的关键 GPCR 治疗性抗体。 使用靶向 GPCR 的治疗性抗体在治疗癌症方面具有巨大潜力 因为它们相对于小分子具有高亲和力、精细的特异性和低毒性。然而， GPCR 治疗性抗体的发现极具挑战性，因为其固有的低稳定性、低 天然膜环境之外的表达和构象异质性。事实上，只有1 GPCR 靶向抗体迄今为止已获得批准，且仅在日本获得批准。为了克服上述问题 提到的障碍，Abilita Bio 通过生成启用膜提供了创新的解决方案 蛋白质 (EMPs™)，代表天然受体的结构和功能增强版本 显着改善表达和热稳定性。具有高表达、稳定和 功能性折叠的 EMP 将能够使用体外抗体分离新型治疗抗体 显示方法。 需要资金（a）为拟议的目标生成突变基因库并选择增强的 使用我们的定向进化系统的变体； (b) 表征表达、功能和的变体 增强哺乳动物细胞的稳定性； (c) 使用进化的电磁脉冲作为抗原开发生物淘选测定工具 并筛选噬菌体展示抗体库，以优化初步抗体引线的分离 并在二期开发。