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/文摘