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: 10351423
• 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 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10351423
• 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; Cell physiology; Cells; Cellular immunotherapy; Child; Childhood Solid Neoplasm; Clinical; Clinical Research; Clinical Trials; Clonality; Crystallization; Data; Data Analyses; 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; Metastatic to; 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; Research Training; Sampling; Signal Transduction; Solid Neoplasm; T cell receptor repertoire sequencing; T memory cell; T-Lymphocyte; TCF Transcription Factor; Technology; Testing; Training; Transcription Factor AP-1; Tumor Antigens; Tyrosine Kinase Inhibitor; Up-Regulation; Work; Xenograft Model; base; career; chemotherapy; chimeric antigen receptor T cells; cytokine; cytotoxicity; design; diffuse midline glioma; exhaustion; experience; fitness; immunoregulation; improved; innovation; insight; multidimensional data; neoplastic cell; novel; osteosarcoma; overexpression; 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.

项目摘要 而嵌合抗原受体T细胞（CAR-TS）在血液学中提供了令人印象深刻的反应 恶性，儿童和年轻人患有转移性或接力实体瘤的儿童和年轻人尚未从中受益 汽车TS并继续遭受令人沮丧的结果。实体瘤中CAR-T效率的主要障碍不足 CAR-T扩展，部分是由Car-T耗尽和记忆力差的部分驱动。临床上，car-t 疲惫是由于过量的汽车信号传导而导致的，可以通过消除CD28 T细胞来救出 汽车设计的共刺激域。临床上，在一般小儿肿瘤患者的样本中， T细胞中的记忆潜力与化学疗法暴露有关。临床前模型进一步表明 通过过表达转录因子CJUN可以减少CAR-T疲惫，并且记忆力可以 通过将T细胞暴露于药物IBRUTINIB中可以增强。基于这些观察结果，Ramakrishna博士将适用 新型的单细胞和多维技术针对CAR-T患者样品评估汽车的影响 在汽车-T衰竭和记忆潜力上的共同刺激域或投票前化学疗法暴露，并且 最终，在患者CAR-T适应性上，定义为与功能配对的CAR-T分子特征。到 达到她的目标，Ramakrishna博士将在三个GD2 CAR-T中进行创新比较CAR-T样品 临床试验。在AIM 1中，通过多维数据分析的培训，Ramakrishna博士将整合表型 （胞质），表观遗传（ATACSEQ）和具有CAR-T功能的转录组（RNASEQ）分子特征 评估以确定GD2.OX40.CD28.Z CAR-TS（NCT02107963）会议疲惫 与骨肉瘤患者的GD2.41BB.Z CAR-TS（NCT04539366）相比，影响CAR-T的适应性。目标 2，在癌症免疫生物学的培训中，Ramakrishna博士将使用Cytof，Atacseq和体外细胞因子 生产，以检验以下假设，即从未化学疗法的患者的置换和CAR-T产物（弥漫性） 中线胶质瘤；与化学疗法治疗的nct04196413）相比之提高了CAR-T适合度 患者（骨肉瘤； NCT04539366），然后是单细胞RNASEQ，以跟踪持续的CAR-T 患者的转录曲线。在AIM 3中，通过对免疫调节的培训，Ramakrishna博士将评估 通过调节，是否可以在化学疗法治疗的患者疗程中进行CAR-T适应性。 CJUN或IBRUTINIB的分子途径。建立在她先前的CAR-T研究和临床上的基础上 经验，Ram​​akrishna博士制定了一个强大的培训计划和心态，包括她的主要 导师Crystal Mackall博士，转化免疫疗法研究的先驱；联合官员，肖恩·本德尔博士， 多维免疫测定中的创新者；咨询委员会，David Miklos博士，Holden Maecker博士， 和米歇尔·蒙耶（Michelle Monje）博士；以及合作者Rosie Kaplan博士和Steven Feldman博士。完成她 与该团队的拟议计划，Ramakrishna博士将准备竞争R01资金并发起 转化研究计划确定和克服儿科实体瘤患者的CAR-T限制。