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（应用程序提供）：基于抑制性和兴奋性趋化输入的智能决策过程中的T细胞；并通过固体组织屏障向肿瘤微环境外观。研究人员的长期目标是在R33阶段追求的，是将T细胞开发到平台矢量技术中，以将其用于实体瘤的主动药物运输。在此R21应用中，它们的目标是设计一种可靠的机制，以将自体T细胞转化为生物疗法，以在肿瘤细胞刺激后自主合成细胞性秘密蛋白。具有空间临时分辨率的蛋白质蛋白质（POI）的自主合成将使肿瘤燃烧以细胞分辨率和分子特异性的范围。它们的中心假设是，可以将T细胞遗传编码用于肿瘤部位的治疗蛋白的鲁棒调节和生物合成。他们选择了表达叶酸受体α（FRA）作为疾病模型的卵巢癌，以及其细胞毒性治疗价值的凋亡蛋白。但是，将T细胞作为蛋白质生物效果的开发理由是，这是一种平台技术，可以将其特异性修改为 靶向任何癌症，并自主合成任何原位POI。为了帮助这项R21的工作，他们开发了表达生物发光和荧光记者的细胞系，仅在被肿瘤细胞刺激时表达。以以下特定目标，他们将向该项目推进其 目标：（1）建立对蛋白质蛋白质生物合成的强大控制（POI）； （2）发展t 细胞为抗肿瘤凋亡蛋白的自主分泌的生物效果。预计该R21相会导致T细胞不会在健康组织中激活（增强的安全性）和 在肿瘤微环境中特别激活（提高效率）。这项研究的影响还将通过减少移植物抗宿主 - 疾病（GVHD）的INTODEAD（GVHD）的持续T细胞治疗试验的影响。这种开创性方法是基于正在进行的T细胞免疫疗法试验的，该试验目前仅限于控制最小残留疾病（MRD），并在化学疗法后进行给药。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，以取代系统性化疗。然后，这将用于减少肿瘤燃烧，而不是检查MRD。其次，非人类起源的治疗蛋白被免疫系统有效降解和去除。这是他们体内应用的障碍。自体T细胞在目标部位使用自体T细胞的治疗蛋白生物合成将消除任何免疫原性问题。预计会出现异常高的影响，因为T细胞的特异性可以重定向到不同癌症上的表面标记，因此可用于靶向众多癌症。