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.

项目摘要/摘要 创新驱动的生物技术公司Abilita Bio（AB）要求SBIR I期资金 靶向G蛋白偶联受体（GPCR）的新型治疗抗体的发现 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）的免疫疗法可以实现持续的免疫 临床结果改善的反应。但是，这种方法在管理时特别有效 与靶向疗法的联合疗法结合疗法，旨在针对肿瘤扩散的基本组成部分 并特别在转移性癌症中生长。转移是癌症患者中限制生存的主要因素。 因此，干扰控制肿瘤细胞转移行为的分子机制是 成功治疗癌症至关重要。 实验和临床证据支持这样一种观念，即最重要的机制之一 在转移中运行涉及稳态趋化因子及其接收器。乳腺癌是 以涉及区域淋巴结，骨髓，肺和 肝。肿瘤细胞从原发性肿瘤部位迁移到转移性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，有几个 研究表明，乳腺癌细胞分泌前列腺素E2（PGE2）可以更新 CCR7表达通过前列腺素结合GPCR，EP2和EP4的激活。该提议重点 靶向关键GPCR参与肿瘤细胞转移性迁移和器官特异性侵袭的目标GPCR使用 治疗性抗体。 将理论抗体用于靶向GPCR具有巨大的癌症潜力 由于其高亲和力，相对于小分子的独家特异性和低毒性。然而， GPCR热抗体发现由于其继承低稳定性而极为挑战 在自然膜环境之外的表达和构象异质性。实际上，只有1 迄今为止，仅在日本就已经批准了靶向GPCR的抗体。为了克服附近 提到的障碍，Abilita Bio通过启用膜提供了创新的解决方案 蛋白质（EMPS™），以结构和功能增强的天然受体的增强版本 显着改善的表达和热稳定性。高度表达，稳定和 功能折叠的EMP将可以使用体外抗体分离新型治疗抗体 显示方法。 要求（a）生成拟议目标的突变基因库，然后选择增强的基因库 使用我们的定向演化系统的变体； （b）表征用于表达，功能和的变体 哺乳动物细胞的稳定性增强； （c）使用进化的EMP作为抗原开发生物植物测定工具 和屏幕噬菌体显示抗体库，用于隔离初步抗体 并在第二阶段开发。