Defining Pre-treatment Correlates of Patient GD2 CAR T Cell Exhaustion and Memory Using Multi-Dimensional Immune Profiling

使用多维免疫分析定义患者 GD2 CAR T 细胞耗竭和记忆的预处理相关性

• 批准号: 10606473
• 负责人: Sneha Ramakrishna
• 金额: $ 21.21万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10606473
• 关键词: ATAC-seq; Advisory Committees; Affect; Aftercare; Agammaglobulinaemia tyrosine kinase; Antigens; B-Cell Acute Lymphoblastic Leukemia; Biological Assay; Biology; Blood Component Removal; CAR T cell therapy; CD28 gene; Cell Line; Cells; Cellular immunotherapy; Child; Childhood Solid Neoplasm; Clinical; Clinical Research; Clinical Trials; Clonality; Data; Data Analyses; Dimensions; Epigenetic Process; Failure; Foundations; Funding; Future; Gene Expression Profile; Hematologic Neoplasms; Immune; Immunology procedure; Immunotherapy; In Vitro; Learning; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; Memory; Mentors; Mentorship; Modeling; Molecular; Molecular Profiling; Mus; OX40; Outcome; Pathway interactions; Patients; Pediatric Oncology; Pharmaceutical Preparations; Phenotype; Pre-Clinical Model; Prior Chemotherapy; Production; Relapse; Research; Research Proposals; Sampling; Signal Transduction; Solid Neoplasm; T cell receptor repertoire sequencing; T memory cell; T-Cell Activation; T-Lymphocyte; TCF Transcription Factor; Technology; Testing; Training; Transcription Factor AP-1; Tumor Antigens; Tyrosine Kinase Inhibitor; Up-Regulation; Work; Xenograft Model; career; chemotherapy; chimeric antigen receptor T cells; cytokine; cytotoxicity; design; diffuse midline glioma; exhaustion; experience; fitness; immunoregulation; improved; innovation; insight; manufacture; multidimensional data; novel; osteosarcoma; overexpression; patient variability; pre-clinical; profiles in patients; response; single-cell RNA sequencing; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; translational research program; tumor; tumor immunology; young adult

PROJECT SUMMARY While chimeric antigen receptor T cells (CAR-Ts) have provided impressive responses in hematologic malignancies, children and young adults with metastatic or relapsed solid tumors have not yet benefited from CAR-Ts and continue to suffer dismal outcomes. A major barrier to CAR-T efficacy in solid tumors is inadequate CAR-T expansion, driven in part by CAR-T exhaustion and poor memory potential. Preclinically, CAR-T exhaustion results from excessive CAR signaling, and can be rescued by eliminating the CD28 T cell costimulatory domain from CAR design. Clinically, in samples from general pediatric oncology patients, poor memory potential in T cells is associated with chemotherapy exposure. Preclinical models further suggest that CAR-T exhaustion can be reduced by overexpressing a transcription factor, cJun, and that memory potential can be enhanced by exposing T cells to a drug, Ibrutinib. Based on these observations, Dr. Ramakrishna will apply novel single-cell and multi-dimensional technologies to CAR-T patient samples to assess the impact of CAR costimulatory domain or pre-apheresis chemotherapy exposure on CAR-T exhaustion and memory potential and ultimately on patient CAR-T fitness, defined as CAR-T molecular signature paired with functionality. To accomplish her aims, Dr. Ramakrishna will innovatively compare CAR-T samples across three GD2 CAR-T clinical trials. In Aim 1, with training in multi-dimensional data analysis, Dr. Ramakrishna will integrate phenotypic (CyTOF), epigenetic (ATACseq), and transcriptomic (RNAseq) molecular signature with CAR-T functional assessments to determine whether GD2.Ox40.CD28.z CAR-Ts (NCT02107963) confer exhaustion, thereby affecting CAR-T fitness, as compared to GD2.41BB.z CAR-Ts (NCT04539366) in osteosarcoma patients. In Aim 2, with training on cancer immune biology, Dr. Ramakrishna will use CyTOF, ATACseq, and in vitro cytokine production, to test the hypothesis that apheresis and CAR-T products from chemotherapy-naïve patients (diffuse midline glioma; NCT04196413) have improved CAR-T fitness as compared to those from chemotherapy-treated patients (osteosarcoma; NCT04539366), followed by single-cell RNAseq to track persistent CAR-T transcriptional profiles in patients. In Aim 3, with training in immune regulation, Dr. Ramakrishna will evaluate whether CAR-T fitness in chemotherapy-treated patient aphereses can be enhanced through modulating molecular pathways by cJun or Ibrutinib. To build upon her substantial prior CAR-T research and clinical experience, Dr. Ramakrishna has developed a strong training plan and mentorship team, including her primary mentor, Dr. Crystal Mackall, a pioneer in translational immunotherapy research; co-mentor, Dr. Sean Bendall, an innovator in multi-dimensional immune assays; advisory committee, Dr. David Miklos, Dr. Holden Maecker, and Dr. Michelle Monje; and collaborators, Dr. Rosie Kaplan and Dr. Steven Feldman. In completing her proposed plan with this team, Dr. Ramakrishna will be prepared to compete for R01 funding and to launch a translational research program identifying and overcoming CAR-T limitations for pediatric solid tumors patients.

项目概要 虽然嵌合抗原受体 T 细胞 (CAR-T) 在血液学方面提供了令人印象深刻的反应 恶性肿瘤、患有转移性或复发性实体瘤的儿童和年轻人尚未受益 CAR-T 并继续遭受惨淡的结果，这是 CAR-T 在实体瘤中疗效不足的一个主要障碍。 CAR-T 扩张，部分原因是 CAR-T 耗尽和临床前记忆潜力不佳。 过度的 CAR 信号传导导致疲劳，可以通过消除 CD28 T 细胞来挽救 临床上，在一般儿科肿瘤患者的样本中，CAR 设计的共刺激域效果较差。 T 细胞的记忆潜力与化疗暴露相关。 CAR-T 耗竭可以通过过度表达转录因子 cJun 来减少，并且记忆潜力可以 Ramakrishna 博士将通过将 T 细胞暴露于药物依鲁替尼 (Ibrutinib) 来增强这一效果。 利用新颖的单细胞和多维技术对 CAR-T 患者样本进行评估，以评估 CAR 的影响 共刺激域或血浆分离术前化疗暴露对 CAR-T 衰竭和记忆潜力的影响 最终取决于患者的 CAR-T 适应性，定义为 CAR-T 分子特征与功能配对。 为了实现她的目标，Ramakrishna 博士将创新性地比较三个 GD2 CAR-T 的 CAR-T 样本 在目标 1 中，Ramakrishna 博士将通过多维数据分析培训来整合表型。 具有 CAR-T 功能的 (CyTOF)、表观遗传学 (ATACseq) 和转录组 (RNAseq) 分子特征 评估以确定 GD2.Ox40.CD28.z CAR-T (NCT02107963) 是否会导致衰竭，从而 与骨肉瘤患者中的 GD2.41BB.z CAR-T (NCT04539366) 相比，影响 CAR-T 的适应性。 2、经过癌症免疫生物学培训，Ramakrishna博士将使用CyTOF、ATACseq和体外细胞因子 生产，以检验来自未接受过化疗的患者的单采术和 CAR-T 产品（弥漫性 中线神经胶质瘤；NCT04196413）与化疗治疗的患者相比，CAR-T 适应性有所改善 患者（骨肉瘤；NCT04539366），然后通过单细胞 RNAseq 追踪持久性 CAR-T 在目标 3 中，Ramakrishna 博士将通过免疫调节方面的培训来评估患者的转录谱。 是否可以通过调节来增强化疗患者血浆分离术中的 CAR-T 适应性 cJun 或 Ibrutinib 的分子途径以她之前大量的 CAR-T 研究和临床为基础。 经验丰富，罗摩克里希纳博士制定了强大的培训计划和指导团队，包括她的主要 导师 Crystal Mackall 博士，转化免疫疗法研究的先驱，共同导师 Sean Bendall 博士， 多维免疫测定的创新者；顾问委员会 David Miklos 博士、Holden Maecker 博士、 和 Michelle Monje 博士；以及合作者 Rosie Kaplan 博士和 Steven Feldman 博士完成了她的研究。 Ramakrishna 博士将与该团队一起提出计划，准备竞争 R01 资金并启动 转化研究项目确定并克服儿童实体瘤患者的 CAR-T 局限性。