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.

项目总结/摘要 Salehi-Rad博士是VA大洛杉矶医疗保健系统（VA GLAHS）的一名工作人员肺病学家 肺癌是美国退伍军人癌症死亡的主要原因，他对肺癌有临床和研究兴趣。在 申请VA职业发展奖（CDA-2），Salehi-Rad博士的目标是建立一个独立的 翻译研究计划在VA GLAHS，重点是提高我们的理解， 肺癌的免疫发病机制，用于开发癌症免疫治疗的新方法。他是 由Steven Dubinett博士（初级导师）支持，他是著名的VA Merit资助的医生科学家， Antoni Ribas，MD，PhD（Co-Mentor），国际公认的肺癌领域权威 癌症免疫学和Paul Boutros博士（共同导师），杰出的数据科学家。导师已确定 基于他们对拟议研究的互补性科学专业知识和他们在以下方面的广泛经验， 指导学术物理学家和科学家。通过UCLA临床和转化科学研究所（CTSI），博士。 Salehi-Rad将有机会参加许多职业发展研讨会，这些研讨会涉及赠款等主题 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上。Salehi-Rad博士将得到VA和UCLA的全力支持 卫生系统进行他的研究。 Salehi-Rad博士已经建立了临床相关的NSCLC小鼠模型， 负荷，并确定了LKB 1缺陷型NSCLC免疫治疗耐药的新靶向机制。 利用这些小鼠模型，Salehi-Rad博士已经表明，用精英抗原交叉免疫的原位疫苗接种（ISV）， 呈现经基因修饰以分泌FMS样酪氨酸激酶3配体（FLT 3L-1）的常规1型DC。 cDC 1），一种促进DC存活和扩增的细胞因子，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。 目的1.1建立在初步体外数据的基础上，这些数据表明FLT 3L-cDC 1与cDC 1相比活力增强， 旨在确定导致FTL 3L-cDC 1存活增加的分子机制。目标1.2和1.3 利用各种鼠模型来确定疫苗和内源性DC活力、抗原运输和 DC ISV后抗原特异性T细胞引发。该提案的目标2侧重于阐明免疫 作为单药治疗或作为与ICI的联合治疗的DC ISV应答的决定因素。目标2.1和2.2 通过流式细胞术和单细胞RNA测序（scRNA-seq）进行联合收割机单细胞免疫表型分析， 通过多重免疫荧光（MIF）进行空间分析，以全面了解当地 和DC ISV诱导的全身免疫应答。目的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将确定T细胞库的演变 通过TCR-β CDR 3测序和通过全外显子组测序（WES）的肿瘤-新抗原谱来评估 DC ISV是否诱导TCR库的扩增并促进肿瘤免疫编辑。改进 了解NSCLC中免疫抑制的性质和DC的免疫刺激机制 ISV将对肺癌免疫学领域做出重大贡献，并有助于临床 FLT 3L-cDC 1 ISV的翻译作为这种破坏性疾病的创新治疗策略， 我们的许多退伍军人。