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: 10513306
• 负责人: Eric Bressler
• 金额: $ 5.18万
• 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/10513306
• 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; Communication Research; Coupling; Data Analyses; Deposition; Development; Development Plans; Drug Delivery Systems; ERBB2 gene; 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; Polymers; Population; Prevention; Proteins; ROR1 gene; Reporting; Research; Research Personnel; Research Project Grants; Risk; SKBR3; Safety; Scientist; 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; novel; novel therapeutics; prevent; programmed cell death protein 1; programs; scaffold; skills; spatiotemporal; 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.

项目摘要和摘要 嵌合抗原受体（CAR）T细胞是基因设计的T淋巴细胞 抗原并安装免疫响应。尽管汽车T细胞已获得FDA批准用于治疗 在几种血液学恶性肿瘤中，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和限制治疗限制不良反应，例如靶向肿瘤 毒性和细胞因子释放综合征。尽管调查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，以及逻辑门控的CAR T细胞，以防止非 - 具体的毒性，没有提出的策略克服了这些障碍中的每一个。为了克服这些限制，我 提出使用新型手术网格将其植入肿瘤切除腔中。该网格将被使用 与称为Zipcar的分裂汽车T细胞结合使用，该单元使用分离的适配器蛋白（ZIPFV”） 感觉抗原。网格由聚合物纳米纤维组成，该纳米纤维具有沉积在 毛孔。网格提供ZipFV衔接蛋白，细胞因子（IL-15）和T细胞刺激抗体（α-- CD3/28）。我假设使用此手术网格将克服CAR T细胞疗法的障碍 在实体瘤中：（1）在切除腔中反对T细胞消极和促进增殖，（2） 通过封装靶向多种抗原的ZipFV，防止抗原逃脱，（3） 对汽车T细胞活性的时空控制。该提案的目标1证明了扩散 通过监测HER2+乳腺癌的鼠模型中的CAR T细胞增殖的优势。目标2 该提案证明了在操作模型中网格的效率和安全优势 缓解卵巢癌。为了证明预防抗原逃脱，ROR1-和HER2缺乏率OVCAR3 将使用CRISPR-CAS9敲除创建细胞系。在抗原逃生的鼠模型中，这些细胞将 用于证明用Zipcar T细胞和ZipFVS的网眼处理的小鼠的效率较高 针对两种抗原。为了证明优越的安全性，将在同一卵巢模型中使用网格 癌症的癌症被照射为上调骨骼中非山摩托质干细胞中的ROR1表达 骨髓和脾脏，允许观察到靶标，肿瘤的毒性。 该建议建立了批判性研究和临床技能的四个关键组成部分，以支持我 发展成独立的物理科学家：（1）跨学科研究项目的重点 新型手术生物材料，可增强CAR T细胞活性； （2）DRS的多学科指导。 Wong（免疫疗法）Grinstaff（生物材料）；还有科尔森（临床医学，动物模型和 免疫学），（3）研究行为和沟通技巧的学术物理科学家培训，（4） 对个人发展计划（IDP）的承诺，以指导我的培训目标。