Cellular biofactories for therapeutic protein synthesis in tumor microenvironment

肿瘤微环境中治疗性蛋白质合成的细胞生物工厂

• 批准号: 9114273
• 负责人: Parijat Bhatnagar
• 金额: $ 27.13万
• 依托单位: SRI INTERNATIONAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9114273
• 关键词: Address; Amidohydrolases; Anabolism; Antigen Targeting; Antigens; Apoptotic; Autologous; Cell Line; Cytoplasm; Decision Making; Development; Disease model; Drug Delivery Systems; Drug Transport; Engineering; Foundations; Future; Goals; IL6 gene; Immune system; Immunosuppressive Agents; Immunotherapy; In Situ; Incidence; Injection of therapeutic agent; Intercellular Fluid; Lead; Malignant Neoplasms; Malignant neoplasm of ovary; Mission; Molecular; Penicillin G; Permeability; Pharmaceutical Preparations; Phase; Process; Prodrugs; Protein Biosynthesis; Proteins; Public Health; Regulation; Reporter; Reporting; Research; Research Personnel; Residual Neoplasm; Resolution; Response Elements; Safety; Site; Solid; Solid Neoplasm; Specificity; Surface; System; T cell response; T cell therapy; T-Cell Development; T-Cell Immunologic Specificity; T-Lymphocyte; Technology; Therapeutic; Therapy trial; Tissues; Transgenes; Tumor Antigens; Tumor Burden; Work; amidase; base; cellular engineering; chemotherapy; chimeric antigen receptor; cytotoxic; graft vs host disease; human FOLR1 protein; immunogenicity; in vivo; interest; nanoparticle; neoplastic cell; nuclear factors of activated T-cells; pressure; prevent; promoter; public health relevance; secretory protein; spatiotemporal; success; therapeutic protein; tumor; tumor microenvironment; tumor specificity; vector

 DESCRIPTION (provided by applicant): T cells undergo an intelligent decision-making process based on the inhibitory and excitatory chemotactic inputs; and extravagate to the tumor microenvironment through solid tissue barriers. Investigators' long-term goal, to be pursued in R33 phase, is to develop T cells into a platform vector technology for active drug- transport to the solid tumors. In this R21 application, their objective is to engineer a robust mechanism to transform autologous T cells into biofactories for autonomous synthesis of cytocidal secretory proteins upon stimulation by the tumor cells. Autonomous synthesis of Protein-of-Interest (PoI) with spatiotemporal resolution will enable targeting of tumor burden with cellular resolution and molecular specificity. Their central hypothesis is that T cells can be genetically encoded for robust modulation and biosynthesis of therapeutic proteins at the tumor site. They have chosen ovarian cancer that expresses Folate Receptor alpha (FRa) as disease model, and an apoptotic protein for its cytotoxic therapeutic value. However, the rationale for developing T cells as protein biofactories is that this is a platform technology and their specificity can be modified to target any cancer and autonomously synthesize any PoI in situ. To aid this R21 effort, they have developed cell lines expressing bioluminescent and fluorescent reporters expressed only when stimulated by tumor cells. With following specific aims, they will advance this project towards its goals: (1) To develop robust control on biosynthesis of protein-of-interest (PoI); (2) To develop T cells as biofactories for autonomous secretion of antitumor apoptotic protein. It is expected that this R21 phase will lead to T cells that will not activate in healthy tissues (enhanced safety) and hyperactivate specifically in the tumor microenvironment (enhanced efficacy). The impact of this research will also be felt on ongoing T-cell therapy trials by reducing the incidences of graft-versus-host-disease (GVHD). This pioneering approach builds upon ongoing T-cell immunotherapy trials, which are currently limited to control minimal residual disease (MRD) and is administered after chemotherapy. Proposed research is transformative, firstly because it will expand the scope of T-cell engineering to replace systemic chemotherapies. This will then be used for reduction of tumor burden rather than keeping MRD in check. Secondly, therapeutic proteins of non-human origin are efficiently degraded and removed by the immune system. This is a barrier to their in vivo application. The use of autologous T cells for biosynthesis of therapeutic proteins at the target site will eliminate any immunogenicity issues. Unusually high impact is expected because the specificity of T cells can be redirected towards the surface markers on different cancers and therefore be used to target multitude of cancers.

 描述（由申请人提供）：T 细胞根据抑制性和兴奋性趋化输入进行智能决策过程；并通过实体组织屏障扩展到肿瘤微环境，这是研究人员在 R33 阶段追求的长期目标。旨在将 T 细胞开发成一种平台载体技术，用于将药物主动转运至实体瘤。在该 R21 应用中，他们的目标是设计一种强大的机制，将自体 T 细胞转化为生物工厂，以自主合成药物。具有时空分辨率的目标蛋白（PoI）的刺激后的杀细胞分泌蛋白将能够以细胞分辨率和分子特异性靶向肿瘤负荷，他们的中心假设是T细胞可以通过基因编码来实现稳健。他们选择表达叶酸受体α（FRa）的卵巢癌作为疾病模型，并选择凋亡蛋白作为其细胞毒性治疗价值。然而，将 T 细胞开发为蛋白质生物工厂的基本原理是，这是一种平台技术，其特异性可以修改为 靶向任何癌症并在原位自主合成任何 PoI 为了帮助 R21 的努力，他们开发了仅在受到肿瘤细胞刺激时表达的生物发光和荧光产生剂的细胞系，他们将推动该项目朝着其目标迈进。 目标：(1) 开发对目标蛋白 (PoI) 生物合成的稳健控制；(2) 开发 T 细胞作为自主分泌抗肿瘤凋亡蛋白的生物工厂，预计该 R21 阶段将导致 T 细胞不会在健康组织中激活（增强安全性）和 这项研究的影响也将通过减少移植物抗宿主病（GVHD）的发生率而体现在正在进行的 T 细胞治疗试验上。 -细胞免疫治疗试验，目前仅限于控制微小残留病（MRD）并在化疗后进行。拟议的研究具有变革性，首先是因为它将扩大 T 细胞工程的范围以取代全身化疗。其次，非人类来源的治疗蛋白会被免疫系统有效降解和去除，这是其在体内应用的障碍。在靶位点生物合成治疗性蛋白质将消除任何免疫原性问题，因为 T 细胞的特异性可以重定向到不同癌症的表面标记，因此可用于靶向多种癌症。