Next-generation Ligand-dependent Transcriptional Switch Receptors for the Control and Customization of Cell Therapeutics

用于细胞治疗的控制和定制的下一代配体依赖性转录开关受体

• 批准号: 10370305
• 负责人: Iowis Zhu
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370305
• 关键词: Address; Affinity; Antigens; Antitumor Response; Behavior; Binding; CAR T cell therapy; California; Cell Culture Techniques; Cells; Cellular immunotherapy; Clinical; Collaborations; Custom; Development; Doctor of Philosophy; ERBB2 gene; Educational Curriculum; Educational process of instructing; Engineering; Enzyme-Linked Immunosorbent Assay; Equilibrium; Faculty; Feedback; Flow Cytometry; Future; Genetic Transcription; Glioblastoma; Half-Life; Head; Human; IL2 gene; IL7 gene; Immune response; Immune system; Immunotherapeutic agent; Immunotherapy; Institutes; Institution; Integral Membrane Protein; Interleukin-12; Interleukin-15; Lesion; Ligands; Location; Malignant - descriptor; Malignant Neoplasms; Measurement; Measures; Mediating; Medical; Modification; Mutation; Normal tissue morphology; Nucleic Acid Regulatory Sequences; Output; Patients; Pharmaceutical Preparations; Physicians; Population; Preceptorship; Preparation; Production; Prognosis; Protein Engineering; Proteolysis; Regimen; Regulation; Research; Research Personnel; Resources; Risk; Running; Safety; San Francisco; Scientist; Signal Transduction; Site; Standardization; Stimulus; Surface; Synthetic immunology; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Therapeutic Uses; Time; Tissues; Toxic effect; Training; Treatment Efficacy; Tumor Antigens; Universities; base; cancer cell; cancer immunotherapy; career; chimeric antigen receptor; chimeric antigen receptor T cells; clinical efficacy; clinically relevant; cytokine; cytotoxic; design; doctoral student; dosage; efficacy testing; epidermal growth factor receptor VIII; experience; extracellular; gamma secretase; improved; malignant breast neoplasm; medical schools; mouse model; next generation; notch protein; novel; patient derived xenograft model; receptor; receptor binding; receptor expression; response; scaffold; screening; side effect; tool; tumor; tumor heterogeneity

ABSTRACT Cytokine immunotherapeutics and chimeric antigen receptor-T cells (CAR-Ts) carry great potential for inducing a patient’s own immune system to clear a malignant lesion. Unfortunately, many of these potential immunotherapies are hindered by poor efficacies and unacceptable toxicities, such as “on-target, off-tumor effects”, where healthy tissues become unintended CAR-T targets by expressing tumor antigens at low levels. In contrast to constitutive CAR expression, inducing a T cell to only express an immunotherapeutic in the presence of a second tumor antigen adds additional regulation over the cytotoxic response. A recent development, synthetic Notch (SynNotch) receptor circuits allow for immunotherapeutic expression to be restricted to tumor sites. However, these receptors do not provide control over the amount of expressed immunotherapeutic, nor do they allow for regulation of an output’s half-life. The ability to tune the level and duration of an induced cell-based immunotherapeutic would further improve on the safety of cell-based immunotherapies and provide a platform for robust tuning of an immunotherapeutic regimen. To date in my graduate training, several novel receptor designs that produce distinct levels and half-lives of transcriptional output in response to a surface-expressed antigen have been engineered. Through receptor domain modulation, these receptor scaffolds can prompt the delivery of a set amount of output to a set tissue type, for a set amount of time. In addition, certain novel receptors act as “OR” gates, being activatable by either ligand or T-cell activation. For this proposal, these receptors will be tested for their ability to induce the expression of various potential immunotherapeutics, including CARs and cytokines in human primary T-cells, as measured by ELISA and flow cytometry. In addition, receptors will be tested as part of receptor-CAR positive feedback circuits for their ability to prime CAR-T cells to better clear a heterogenous tumor by inducing a more sustained anti-tumor response, both in cell culture and in mouse model. The University of California, San Francisco (UCSF) is a leading institution in synthetic immunology, and its faculty are well experienced in physician-scientist training. As a MD/PhD student in the Roybal lab at UCSF, I have access to support and collaboration from world-renowned investigators, as well as resources from the Parker Institute for Cancer Immunotherapy and the Chan Zuckerberg BioHub. In addition to my PhD research, my training plan includes medical preceptorships with world-class physicians during my graduate training, additional teaching opportunities through the UCSF School of Medicine, and further clinical training through UCSF resources, including the UCSF Bridges medical curriculum. Overall, this training proposal seeks to further develop synthetic receptor circuits as a tool for improving the safety and efficacy of potential immunotherapies while providing me a diverse set of experiences in preparation for a future career as a physician-scientist.

摘要 细胞因子免疫治疗剂和嵌合抗原受体-T细胞（CAR-Ts）具有巨大的潜力， 诱导病人自身的免疫系统清除恶性病变不幸的是，许多潜在的 免疫疗法受到不良功效和不可接受的毒性的阻碍，例如“中靶、脱瘤”， 在这种情况下，健康组织通过以低水平表达肿瘤抗原而成为非预期的CAR-T靶标。 与组成型CAR表达相反，诱导T细胞仅表达免疫球蛋白， 第二肿瘤抗原的存在增加了对细胞毒性应答的额外调节。最近的一 发展，合成Notch（SynNotch）受体电路允许免疫表达， 仅限于肿瘤部位。然而，这些受体不提供对表达的量的控制。 免疫系统，也不允许调节输出的半衰期。能够调整电平， 诱导的基于细胞的免疫应答的持续时间将进一步改善基于细胞的免疫应答的安全性。 本发明提供了一种用于免疫治疗的方法，并提供了用于免疫治疗方案的稳健调整的平台。 到目前为止，在我的研究生培训中，几种新颖的受体设计，产生不同的水平和半衰期， 已经工程化了响应于表面表达的抗原的转录输出。通过受体 域调节，这些受体支架可以促使将设定量的输出递送到设定的组织 类型，一段时间。此外，某些新的受体充当“或”门，可被以下任一者激活： 配体或T细胞活化。对于这个提议，这些受体将被测试它们诱导表达的能力。 各种潜在的免疫治疗剂，包括人原代T细胞中的汽车和细胞因子，如测量的 通过ELISA和流式细胞术。此外，受体将作为受体-CAR正反馈的一部分进行检测 他们能够通过诱导更持续的CAR-T细胞活化来更好地清除异质性肿瘤。 在细胞培养物和小鼠模型中的抗肿瘤应答。 加州大学弗朗西斯科分校（UCSF）是合成免疫学的领先机构， 其教员在医生-科学家培训方面经验丰富。作为加州大学旧金山分校罗伊巴尔实验室的一名MD/PhD学生， 我可以获得世界知名调查人员的支持和合作，以及来自 帕克癌症免疫治疗研究所和陈·扎克伯格生物中心。除了我的博士研究， 我的培训计划包括在我的研究生培训期间与世界一流的医生进行医学指导， 通过UCSF医学院提供额外的教学机会，并通过 UCSF资源，包括UCSF桥梁医学课程。总的来说，这一培训建议旨在进一步 开发合成受体回路，作为提高潜在免疫疗法安全性和有效性的工具 同时也为我将来成为一名物理学家和科学家提供了丰富的经验。