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.

项目总结/摘要 Abilita Bio（AB）是一家创新驱动的生物技术公司，要求SBIR第一阶段资助， 发现了靶向G蛋白偶联受体（GPCR）的新型治疗性抗体， 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， 使用检查点抑制剂（PD-1/PD-L1，CTLA-4等）进行免疫治疗可以获得持久免疫 改善临床结果。然而，这种方法在给药时特别有效。 在与靶向治疗的联合治疗中，针对肿瘤扩散、归巢 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， 是成功治疗癌症的关键 实验和临床证据支持这样一种观点，即最重要的机制之一 在转移中起作用涉及稳态趋化因子及其受体。乳腺癌是 其特征在于涉及区域淋巴结、骨髓、肺和淋巴结的独特转移模式。 肝脏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 与白细胞运输，这是由趋化因子结合GPCR关键调控。有趣的是，几个 研究已经确定乳腺癌细胞分泌的前列腺素E2（PGE 2）可以上调 CCR 7表达通过激活结合胰高血糖素的GPCR，EP 2和EP 4。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 使用治疗性抗体靶向GPCR对于癌症的治疗具有巨大的潜力 因为它们相对于小分子具有高亲和性、精确的特异性和低毒性。然而，在这方面， GPCR治疗性抗体的发现是极具挑战性的，因为它们固有的低稳定性、低生物学活性和低生物学活性。 在其天然膜环境外的表达和构象异质性。事实上，只有1 迄今为止，GPCR靶向抗体仅在日本获得批准。为了克服前 提到的障碍，Abilita生物提供了一个创新的解决方案，通过生成启用膜 蛋白质（EMPs™），代表天然受体的结构和功能增强版本， 显著改善的表达和热稳定性。高表达、稳定、 功能性折叠的EMP将使得能够使用体外抗体分离新的治疗性抗体 显示方法 要求提供资金，以（a）为拟议的目标建立突变基因文库，并为增强的 使用我们的定向进化系统的变体;（B）表征变体的表达、功能和 增强哺乳动物细胞中的稳定性;（c）开发使用进化的EMP作为抗原的生物淘选分析工具 并筛选噬菌体展示抗体库，对分离的初步抗体进行了优化 并在第二阶段开发。