Flt3l gene-modified cDC1 in situ vaccination in NSCLC: mechanisms and therapeutic application

Flt3l 基因修饰的 cDC1 原位疫苗接种在 NSCLC 中的作用：机制和治疗应用

• 批准号: 10585591
• 负责人: Ramin Salehi-Rad
• 金额: --
• 依托单位: VA GREATER LOS ANGELES HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10585591
• 关键词: Address; Adverse event; Affect; Age; Antibodies; Antigen Presentation; Antigens; Apoptotic; Authorization documentation; Autologous; Automobile Driving; Bioinformatics; CCL21 gene; CD8-Positive T-Lymphocytes; CXCR6 gene; Cancer Etiology; Cancer Model; Cell Survival; Cell secretion; Cells; Cellular immunotherapy; Cessation of life; Clinical; Clinical Sciences; Clinical Trials; Combined Modality Therapy; Cross Presentation; Cytokine Gene; Data; Data Scientist; Dendritic Cell Vaccine; Dendritic Cells; Dependence; Development; Disease; Doctor of Philosophy; Engineering; Ethics; Evaluation; Evolution; FLT3 gene; FLT3 ligand; Flow Cytometry; Funding; Gene Expression Profiling; Gene Modified; Goals; Grant; Health system; Healthcare Systems; Immune; Immune checkpoint inhibitor; Immune response; Immunobiology; Immunofluorescence Immunologic; Immunologic Memory; Immunologic Sensitization; Immunology; Immunophenotyping; Immunosuppression; Immunotherapy; In Vitro; In complete remission; Incidence; Injections; Institution; International; K-Series Research Career Programs; Ligands; Los Angeles; Malignant neoplasm of lung; Manuscripts; Mediating; Mediator; Mentors; Molecular; Mus; Mutation; Natural Killer Cells; Nature; Non-Small-Cell Lung Carcinoma; PTPRC gene; Pathway interactions; Patients; Phenotype; Physicians; Population; Preparation; Proteins; Refractory; Research; Role; STK11 gene; Scientist; Spleen; Stable Disease; Survival Rate; T cell clonality; T cell response; T-Cell Activation; T-Lymphocyte; T-cell receptor repertoire; Therapeutic; Training; Translational Research; Tumor Antigens; Tumor Immunity; Tumor Promotion; Tumor-Infiltrating Lymphocytes; Vaccine Therapy; Vaccines; Veterans; Writing; anti-PD-1; antigen-specific T cells; authority; beta Chain Antigen T Cell Receptor; cancer immunotherapy; cancer infiltrating T cells; career development; checkpoint inhibition; chemokine; clinical translation; clinically relevant; congenic; cytokine; draining lymph node; exome sequencing; experience; immune resistance; improved; in situ vaccination; in vivo; innovation; interest; military veteran; mouse model; neoantigens; novel; novel strategies; novel vaccines; partial response; patient subsets; pembrolizumab; phase I trial; preclinical study; programmed cell death ligand 1; rational design; resistance mechanism; response; restoration; single-cell RNA sequencing; stem; synergism; tertiary lymphoid organ; trafficking; transcriptome sequencing; translational research program; tumor; tumor immunology; tumor microenvironment; vaccine evaluation

PROJECT SUMMARY/ABSTRACT Dr. Salehi-Rad is a Staff Pulmonologist at the VA Greater Los Angeles Healthcare System (VA GLAHS) with a clinical and research interest in lung cancer, the leading cause of cancer death among U.S. Veterans. In applying for the VA Career Development Award (CDA-2), Dr. Salehi-Rad’s goal is to establish an independent translational research program at the VA GLAHS, focused on improving our understanding of the immunopathogenesis of lung cancer for the development of novel approaches for cancer immunotherapy. He is supported by Steven Dubinett, MD (Primary-Mentor), a renowned VA Merit-funded physician-scientist and a leading expert in lung cancer, Antoni Ribas, MD, PhD (Co-Mentor), an internationally recognized authority in cancer immunology, and Paul Boutros, PhD (Co-mentor), a distinguished data scientist. Mentors were identified based on their complementary scientific expertise for the proposed research and their extensive experience in mentoring academic physician-scientists. Through UCLA Clinical and Translational Science Institute (CTSI), Dr. Salehi-Rad will have access to numerous career development seminars that address such topics as grant writing, manuscript preparation, and ethical research. He will also take graduate courses to obtain further training in immunology and bioinformatics. Dr. Salehi-Rad will have the full institutional support of both the VA and UCLA Health Systems to carry out his research. Dr. Salehi-Rad has established clinically relevant murine models of NSCLC with increased mutational burden and identified a novel targetable mechanism of resistance to immunotherapy in LKB1-deficient NSCLC. Utilizing these murine models, Dr. Salehi-Rad has shown that in situ vaccination (ISV) with elite antigen cross- presenting conventional type 1 DCs that are gene-modified to secrete FMS-like tyrosine kinase 3 ligand (FLT3L- cDC1), a cytokine that promotes DC viability and expansion, sensitize immune refractory NSCLC to immune checkpoint inhibition (ICI). In this proposal, Dr. Salehi-Rad aims to study the immune mechanisms of DC ISV. Aim 1.1 builds on preliminary in vitro data indicating enhanced viability of FLT3L-cDC1 compared to cDC1 and seeks to determine the molecular mechanisms that result in increased survival of FTL3L-cDC1. Aim 1.2 & 1.3 utilize various murine models to determine the vaccine and endogenous DC viability, antigen trafficking and antigen-specific T cell priming following DC ISV. Aim 2 of the proposal focuses on elucidating the immune determinants of response to DC ISV as monotherapy or as a combination therapy with ICI. Aim 2.1 & 2.2 combine single cell immunophenotyping by flow cytometry and single cell RNA-sequencing (scRNA-seq) with spatial analysis by multiplex immunofluorescence (MIF) to develop a comprehensive understanding of the local and systemic immune responses induced by DC ISV. Aim 2.3 utilizes antibody depletion studies to evaluate the dependency of DC ISV on T cells and natural killer cells. Aim 3 will determine the evolution of T cell repertoires by TCR-β CDR3 sequencing and tumor-neoantigen profiles by whole-exome sequencing (WES) to assess whether DC ISV induces the expansion of the TCR repertoire and promotes tumor immunoediting. Improved understanding of the nature of immunosuppression in NSCLC and the immunostimulatory mechanisms of DC ISV will represent a significant contribution to the field of lung cancer immunology and could facilitate the clinical translation of FLT3L-cDC1 ISV as an innovative therapeutic strategy for this devastating disease that affects many of our Veterans.

项目概要/摘要 Salehi-Rad 博士是 VA 大洛杉矶医疗系统 (VA GLAHS) 的一名肺科医生 对肺癌的临床和研究感兴趣，肺癌是美国退伍军人癌症死亡的主要原因。在 Salehi-Rad 博士申请 VA 职业发展奖 (CDA-2) 的目标是建立一个独立的 VA GLAHS 的转化研究项目，重点是提高我们对 肺癌的免疫发病机制，以开发癌症免疫治疗的新方法。他是 由史蒂文·杜比内特 (Steven Dubinett) 医学博士（主要导师）支持，他是一位著名的 VA 优秀资助医师兼科学家， 肺癌领域的领先专家 Antoni Ribas 博士（联合导师），国际公认的肺癌领域权威 癌症免疫学，以及杰出数据科学家 Paul Boutros 博士（共同导师）。导师已确定 基于他们对拟议研究的互补科学专业知识以及他们在以下方面的丰富经验 指导学术医师科学家。通过加州大学洛杉矶分校临床和转化科学研究所 (CTSI)，博士。 Salehi-Rad 将有机会参加众多职业发展研讨会，讨论资助等主题 写作、手稿准备和伦理研究。他还将参加研究生课程以获得进一步的培训 在免疫学和生物信息学领域。 Salehi-Rad 博士将获得 VA 和 UCLA 的全力机构支持 卫生系统开展他的研究。 Salehi-Rad 博士建立了临床相关的 NSCLC 小鼠模型，其中突变增加 负担，并确定了 LKB1 缺陷 NSCLC 中免疫治疗耐药的一种新的靶向机制。 Salehi-Rad 博士利用这些小鼠模型表明，原位疫苗接种 (ISV) 与精英抗原交叉 展示经过基因修饰的常规 1 型 DC，可分泌 FMS 样酪氨酸激酶 3 配体（FLT3L- cDC1)，一种促进 DC 活力和扩增的细胞因子，使免疫难治性 NSCLC 对免疫敏感 检查点抑制（ICI）。在这项提案中，Salehi-Rad 博士旨在研究 DC ISV 的免疫机制。 目标 1.1 基于初步体外数据，表明与 cDC1 相比，FLT3L-cDC1 的活力增强 旨在确定导致 FTL3L-cDC1 存活增加的分子机制。目标 1.2 和 1.3 利用各种小鼠模型来确定疫苗和内源性 DC 的活力、抗原运输和 DC ISV 后抗原特异性 T 细胞启动。该提案的目标 2 侧重于阐明免疫 DC ISV 作为单一疗法或与 ICI 联合疗法的反应的决定因素。目标 2.1 和 2.2 将流式细胞术的单细胞免疫表型分析与单细胞 RNA 测序 (scRNA-seq) 相结合 通过多重免疫荧光 (MIF) 进行空间分析，以全面了解局部 以及 DC ISV 诱导的全身免疫反应。目标 2.3 利用抗体耗竭研究来评估 DC ISV 对 T 细胞和自然杀伤细胞的依赖性。目标 3 将决定 T 细胞库的进化 通过 TCR-β CDR3 测序和全外显子组测序 (WES) 来评估肿瘤新抗原谱 DC ISV 是否会诱导 TCR 库的扩展并促进肿瘤免疫编辑。改进 了解 NSCLC 免疫抑制的本质和 DC 的免疫刺激机制 ISV 将代表对肺癌免疫学领域的重大贡献，并可以促进临床 将 FLT3L-cDC1 ISV 转化为这种破坏性疾病的创新治疗策略 我们的许多退伍军人。