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.

项目摘要/摘要 Salehi-Rad博士是VA大洛杉矶医疗保健系统（VA GLAHS）的肺科医生 伴有对肺癌的临床和研究兴趣，这是美国退伍军人癌症死亡的主要原因。在 申请VA职业发展奖（CDA-2），Salehi-Rad博士的目标是建立独立 VA Glah的翻译研究计划，重点是提高我们对 肺癌的免疫原理发生用于开发新方法的癌症免疫疗法。他是 由史蒂文·杜比尼特（Steven Dubinett），医学博士（小学生）的支持 肺癌领先的专家，马里兰州安东尼·里巴斯（Antoni Ribas） 癌症免疫学和杰出数据科学家的Paul Boutros博士（同事）。确定了导师 根据他们的完整科学专业知识，用于拟议的研究及其丰富的经验 指导学术医师科学家。通过UCLA临床和转化科学研究所（CTSI），博士 Salehi-Rad将可以访问众多职业发展中心，以解决诸如Grant之类的主题 写作，手稿准备和道德研究。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中。 Salehi-Rad博士将获得VA和UCLA的全部机构支持 卫生系统进行他的研究。 Salehi-Rad博士已经建立了NSCLC的临床相关鼠模型，并增加了突变 负担并确定了一种新型的可抵抗免疫疗法NSCLC免疫疗法的机制。 Salehi-Rad博士利用这些鼠模型表明，原位疫苗接种（ISV）具有精英抗原横断 呈现常规1型DC，这些DC经过基因修饰以分泌FMS样酪氨酸激酶3配体（FLT3L-- CDC1），一种促进直流生存力和膨胀的细胞因子，对免疫的敏感免疫难治性NSCLC 检查点抑制（ICI）。在此提案中，Salehi-Rad博士旨在研究DC ISV的免疫力学。 AIM 1.1建立在初步的体外数据上，表明与CDC1和CDC1和 试图确定导致FTL3L-CDC1存活增加的分子机制。目标1.2＆1.3 利用各种鼠模型来确定疫苗和内源性直流生存能力，抗原运输和 DC ISV之后的抗原特异性T细胞启动。提案的目标2着重阐明免疫 对DC ISV反应的决定因素作为单一疗法或与ICI的组合疗法。 AIM 2.1和2.2 通过流式细胞术和单细胞RNA-sequesing（SCRNA-SEQ）与单细胞免疫原化型结合在一起 通过多重免疫荧光（MIF）进行空间分析，以对局部有全面的了解 DC ISV诱导的全身免疫反应。 AIM 2.3利用抗体部署研究来评估 DC ISV对T细胞和天然杀伤细胞的依赖性。 AIM 3将确定T细胞库的演变 通过TCR-βCDR3测序和肿瘤神经抗原曲线，通过全外观测序（WES）评估 DC ISV是否诱导TCR库的扩张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将代表对肺癌免疫学领域的重要贡献，并可以促进临床 FLT3L-CDC1 ISV的翻译是这种毁灭性疾病的创新治疗策略 我们的许多退伍军人。