Development of a Surgical Drug Delivery System for Enhancement of CAR T Cell Activity

开发增强 CAR T 细胞活性的外科药物输送系统

• 批准号: 10688135
• 负责人: Eric Bressler
• 金额: $ 5.27万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-09 至 2025-09-08
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10688135
• 关键词: Adaptor Signaling Protein; Address; Adverse effects; Adverse event; Animal Model; Antibodies; Antigen Targeting; Antigens; Beds; Biocompatible Materials; Biodistribution; Biological Availability; Bone Marrow; Breast Cancer Model; CAR T cell therapy; CD3 Antigens; CRISPR/Cas technology; Cell Line; Cell Therapy; Cells; Chitosan; Clinical Medicine; Clinical Skills; Communication; Coupling; Data Analyses; Deposition; Development; Development Plans; Drug Delivery Systems; ERBB2 gene; Encapsulated; Excision; Exhibits; Goals; Growth; Hematologic Neoplasms; Immune response; Immunology; Immunotherapy; Implant; In Vitro; Individual; Interdisciplinary Study; Interleukin-15; Knock-out; Knowledge; Laboratories; Leucine Zippers; Logic; Malignant Neoplasms; Malignant neoplasm of ovary; Mentors; Methods; Modality; Modeling; Molecular Biology; Monitor; Monoclonal Antibodies; Mus; Operative Surgical Procedures; Organ; Physicians; Population; Prevention; Proliferating; Proteins; ROR1 gene; Reporting; Research; Research Personnel; Research Project Grants; Risk Reduction; SKBR3; Safety; Scientist; Shapes; Signal Transduction; Site; Solid Neoplasm; Specificity; Spleen; Surface; Surgical Mesh; Surgical sutures; Surgically-Created Resection Cavity; System; T cell anergy; T-Cell Activation; T-Cell Proliferation; T-Lymphocyte; Techniques; Testing; Therapeutic; Tissues; Toxic effect; Training; Tumor Antigens; Tumor Debulking; bioluminescence imaging; chimeric antigen receptor; chimeric antigen receptor T cells; cytokine; cytokine release syndrome; cytotoxic; density; design; draining lymph node; efficacy evaluation; exhaustion; genetically modified cells; implantation; improved; in vivo; insight; malignant breast neoplasm; mouse model; multidisciplinary; nanofiber; nanopolymer; novel; novel therapeutics; prevent; programmed cell death protein 1; scaffold; skills; spatiotemporal; stem cells; success; therapy development; tumor; tumor microenvironment; tumor-immune system interactions

Project Summary and Abstract Chimeric antigen receptor (CAR) T cells are genetically engineered T lymphocytes designed to sense antigens and mount an immune response. Though CAR T cells have received FDA approval for the treatment of several hematologic malignancies, success in solid tumors is limited by a lack of specific antigens, the immunosuppressive tumor microenvironment, and treatment-limiting adverse effects such as on-target, off-tumor toxicity and cytokine release syndrome. Though investigators report strategies for mitigating these limitations such as biomaterials for reshaping the tumor microenvironment, and logic-gated CAR T cells to prevent non- specific toxicity, no proposed strategy has overcome each of these barriers. To surmount these limitations, I propose the use of a novel surgical mesh for implantation into the tumor resection cavity. This mesh will be used in conjunction with a split CAR T cell called a zipCAR, which uses a detached adaptor protein (a “zipFv”) to sense antigens. The mesh is composed of polymeric nanofibers with a matrix of chitosan deposited within the pores. The mesh supplies the zipFv adaptor protein, cytokines (IL-15), and T cell stimulatory antibodies (α- CD3/28). I hypothesize that the use of this surgical mesh will overcome the barriers to CAR T cell therapy in solid tumors by: (1) opposing T cell anergy and promoting proliferation in the resection cavity, (2) preventing antigen escape via encapsulation of zipFvs targeting multiple antigens, and (3) imparting spatiotemporal control over CAR T cell activity. Aim 1 of this proposal demonstrates the proliferation advantage of the mesh by monitoring CAR T cell proliferation in a murine model of HER2+ breast cancer. Aim 2 of this proposal demonstrates the efficacy and safety advantages of the meshes in a model of operative debulking of ovarian cancer. To demonstrate prevention of antigen escape, ROR1- and HER2-deficient OVCAR3 cell lines will be created using CRISPR-Cas9 knockouts. In a murine model of antigen escape, these cells will be used to demonstrate superior efficacy in mice treated with zipCAR T cells and meshes loaded with zipFvs against both antigens. To demonstrate superior safety, meshes will be utilized in the same model of ovarian cancer with mice that are irradiated to upregulate ROR1 expression in non-hematopoietic stem cells in the bone marrow and spleen, allowing observation of on-target, off-tumor toxicity. This proposal builds around four key components of critical research and clinical skills to support my development into an independent physician scientist: (1) an interdisciplinary research project focusing on novel surgical biomaterials for enhancement of CAR T cell activity; (2) multi-disciplinary mentoring from Drs. Grinstaff (biomaterials), Wong (immunotherapy); and, Colson (clinical medicine, animal models, and immunology), (3) academic physician scientist training in research conduct and communication skills, (4) commitment to an individual development plan (IDP) to guide my training goals.

项目概要和摘要 嵌合抗原受体 (CAR) T 细胞是基因工程 T 淋巴细胞，旨在感知 抗原并引发免疫反应。尽管 CAR T 细胞已获得 FDA 批准用于治疗 在几种血液系统恶性肿瘤中，实体瘤的成功受到缺乏特异性抗原的限制， 免疫抑制性肿瘤微环境，以及治疗限制性副作用，例如靶向、脱瘤 毒性和细胞因子释放综合征。尽管研究人员报告了减轻这些限制的策略 例如用于重塑肿瘤微环境的生物材料，以及用于预防非肿瘤发生的逻辑门控CAR T细胞。 由于特定的毒性，目前还没有提出的策略能够克服这些障碍。为了克服这些限制，我 提出使用新型手术网植入肿瘤切除腔。将使用该网格 与称为 zipCAR 的分裂 CAR T 细胞结合使用，该细胞使用分离的接头蛋白（“zipFv”）来 感觉抗原。该网由聚合物纳米纤维组成，其内沉积有壳聚糖基质 毛孔。该网格提供 zipFv 接头蛋白、细胞因子 (IL-15) 和 T 细胞刺激抗体 (α- CD3/28）。我假设使用这种手术网片将克服 CAR T 细胞治疗的障碍 在实体瘤中，通过：(1) 对抗 T 细胞无反应性并促进切除腔内的增殖，(2) 通过封装靶向多种抗原的 zipFv 来防止抗原逃逸，以及 (3) 对 CAR T 细胞活性的时空控制。该提案的目标 1 表明了扩散 通过监测 HER2+ 乳腺癌小鼠模型中的 CAR T 细胞增殖，利用网格的优势。目标2 该提案证明了网格在手术模型中的有效性和安全性优势 卵巢癌的减灭。为了证明防止抗原逃逸，ROR1 和 HER2 缺陷的 OVCAR3 将使用 CRISPR-Cas9 敲除技术创建细胞系。在抗原逃逸的小鼠模型中，这些细胞将 用于证明使用 zipCAR T 细胞和装载 zipFv 的网格治疗的小鼠具有卓越功效 针对两种抗原。为了证明卓越的安全性，网片将用于相同的卵巢模型 小鼠接受辐射后骨骼中非造血干细胞中 ROR1 表达上调，从而导致癌症 骨髓和脾脏，可以观察靶点、肿瘤外毒性。 该提案围绕关键研究和临床技能的四个关键组成部分来支持我 发展成为一名独立的医师科学家：（1）一个跨学科的研究项目，重点关注 用于增强 CAR T 细胞活性的新型外科生物材料； (2) 博士的多学科指导。 Grinstaff（生物材料）、Wong（免疫疗法）；以及 Colson（临床医学、动物模型和 免疫学），（3）学术医师科学家研究行为和沟通技巧培训，（4） 致力于个人发展计划 (IDP) 来指导我的培训目标。