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 虽然嵌合抗原受体T细胞（CAR-T）在血液学中提供了令人印象深刻的反应， 患有转移性或复发性实体瘤的恶性肿瘤、儿童和年轻人尚未受益于 CAR-T并继续遭受令人沮丧的结果。CAR-T在实体瘤中疗效的主要障碍是不足 CAR-T扩增，部分由CAR-T耗竭和记忆潜力差驱动。临床前，CAR-T 过度的CAR信号转导导致衰竭，并且可以通过消除CD 28 T细胞来挽救。 来自CAR设计的共刺激结构域。临床上，在普通儿科肿瘤患者的样本中， T细胞的记忆潜力与化疗暴露有关。临床前模型进一步表明， CAR-T耗竭可以通过过度表达转录因子cJun来减少，记忆潜力可以 通过将T细胞暴露于一种药物--依鲁替尼来增强。根据这些观察结果，罗摩克里希纳博士将申请 新的单细胞和多维技术用于CAR-T患者样本，以评估CAR的影响 共刺激结构域或单采前化疗暴露对CAR-T耗竭和记忆潜力的影响， 最终取决于患者的CAR-T适应性，其定义为与功能性配对的CAR-T分子特征。到 为了实现她的目标，Ramakrishna博士将创新性地比较三个GD 2 CAR-T的CAR-T样本。 临床试验在目标1中，通过多维数据分析的培训，Ramakrishna博士将整合表型 具有CAR-T功能性的CyTOF、表观遗传（ATACseq）和转录组（RNAseq）分子标记 评估以确定GD2.Ox40.CD28.z CAR-Ts（NCT 02107963）是否导致耗竭，从而 影响CAR-T适应性，与骨肉瘤患者中的GD2.41BB.z CAR-T（NCT 04539366）相比。在Aim中 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，以检验来自化疗初治患者（弥漫性）的单采和CAR-T产物 中线胶质瘤; NCT 04196413）与来自化疗治疗的那些相比具有改善的CAR-T适应性。 患者（骨肉瘤; NCT 04539366），随后使用单细胞RNAseq追踪持续性CAR-T 患者的转录谱。在目标3中，通过免疫调节培训，Ramakrishna博士将评估 是否可以通过调节CAR-T在化疗治疗的单采患者中的适应性来增强 通过cJun或伊曲替尼的分子途径。基于她之前大量的CAR-T研究和临床 Ramakrishna博士的经验，制定了一个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， 多维免疫分析的创新者;顾问委员会，大卫米克洛什博士，霍尔顿梅克尔博士， 和米歇尔·蒙杰博士;以及合作者，罗西·卡普兰博士和史蒂文·费尔德曼博士。在完成她的 与这个团队提出的计划，Ramakrishna博士将准备竞争R 01资金，并推出一个 转化研究计划，确定并克服儿童实体瘤患者的CAR-T局限性。