Optimization of CAR-Bacteria for Oral Cancer

口腔癌 CAR 细菌的优化

• 批准号: 10648292
• 负责人: Jack D Bui
• 金额: $ 22.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10648292
• 关键词: Affinity; Animal Model; Antigen Targeting; Bacteria; Bilateral; Binding; Cell Death; Cell Line; Cells; Cervical; Complex; Cytolysis; Data; Dose; Engineering; Engraftment; Evolution; Future; Goals; Growth; HLA-A2 Antigen; Head and Neck Cancer; Human; Human Cell Line; Human Papillomavirus; Innate Immune Response; Kinetics; Label; Libraries; Major Histocompatibility Complex; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Measures; Mediating; Methods; Molecular Evolution; Mouthwash; Mus; Oncoproteins; Pattern; Peptide/MHC Complex; Peptides; Pharmaceutical Preparations; Physiologic pulse; Population Density; Production; Proteins; Research; Specificity; T-Lymphocyte; Therapeutic; Toxic effect; Treatment Efficacy; Tumor Specific Peptide; Viral; Work; Xenograft Model; antigen challenge; cancer cell; chimeric antigen receptor; experimental study; human model; improved; in vivo; innovation; intraperitoneal; invention; knock-down; malignant mouth neoplasm; nanobodies; neoplastic cell; novel; oral bacteria; overexpression; prevent; response; systemic toxicity; technology platform; tumor; tumor growth; tumor immunology

ABSTRACT The goal of this project is to explore the feasibility of generating Chimeric Antigen Receptors (CARs) expressed by bacteria. Instead of using T cells, we propose to explore the idea and use of bacteria to mediate the targeted destruction of cancer cells. In addition, we explore the possibility that we could target CAR-bacteria to specific major histocompatibility complex (MHC)-neopeptide complexes (pMHC) on tumor cells. We call these engineered bacteria cells CAR-bacteria. Current CAR-expressing T cells provide durable responses but have three main limitations: specificity, toxicity, and feasibility. This study will address all three concerns. In this proposal, we target engineered bacterial lysis to head and neck or cervical cancer cells expressing HLA-A2-E7, a known pMHC on tumor cells infected by human papilloma virus (HPV). The targeting enables anti-tumor protein production only when a predefined population density of bacteria is reached. This method should dramatically reduce bacterial colony size and greatly lowers/prevents systemic toxicities. The approach has the potential to be a therapeutic in HPV-positive oral cancer as it could be administered and controlled as a bacterial mouthwash. This project combines two innovations leveraging a Synthetically-Evolved Nanobody (SEN) library proven capable of selectively binding MHC-peptide complexes and combining this specific cancer cell targeting with synchronized circuit lysis to create CAR-bacteria. Our proposed studies provide proof-of-concept that (1) CARs can be produced that recognize pMHC, (2) CAR-bacteria localization and colonization can be controlled by tumor expression of certain pMHC and (3) CAR-bacteria have therapeutic activity against tumor cells that express specific pMHC. We accomplish these proof-of-concept studies via the following two objectives: (1) Optimize binding and specificity of CAR-bacteria to HLA-A2-E7 and (2) Check efficacy of CAR-bacteria in an animal model of head and neck and cervical cancer.

抽象的 该项目的目标是探索生成嵌合抗原受体（CAR）的可行性 由细菌表达。我们建议不使用 T 细胞，而是探索细菌介导的想法和用途 有针对性地破坏癌细胞。此外，我们还探索了针对 CAR 细菌的可能性 肿瘤细胞上特定的主要组织相容性复合物（MHC）-新肽复合物（pMHC）。我们称这些为 工程细菌细胞CAR-细菌。目前表达 CAR 的 T 细胞可提供持久的反应，但 三个主要局限性：特异性、毒性和可行性。这项研究将解决所有三个问题。在这个 建议，我们针对表达 HLA-A2-E7 的头颈或宫颈癌细胞进行工程细菌裂解， 已知的 pMHC 与人乳头瘤病毒 (HPV) 感染的肿瘤细胞有关。靶向使抗肿瘤蛋白成为可能 仅当达到预定的细菌种群密度时才进行生产。该方法应该显着 减少细菌菌落大小并大大降低/防止全身毒性。该方法有潜力 可以作为 HPV 阳性口腔癌的治疗剂，因为它可以作为细菌漱口水进行施用和控制。 该项目结合了两项创新，利用经过验证的合成进化纳米抗体 (SEN) 库 能够选择性地结合 MHC-肽复合物，并将这种特定的癌细胞靶向与 同步电路裂解以产生 CAR 细菌。我们提出的研究提供了以下概念验证：(1) CAR 可以生产识别pMHC，（2）CAR-细菌定位和定植可以由肿瘤控制 (3) CAR-细菌对表达某些 pMHC 的肿瘤细胞具有治疗活性 特异性 pMHC。我们通过以下两个目标完成这些概念验证研究：(1) 优化 CAR-细菌与 HLA-A2-E7 的结合和特异性以及 (2) 检查 CAR-细菌在动物模型中的功效 头颈癌和宫颈癌。