Defining epigenetic signaling to reshape pancreatic tumor microenvironment

定义表观遗传信号重塑胰腺肿瘤微环境

• 批准号: 10514158
• 负责人: Jiaqi Shi
• 金额: $ 46.26万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10514158
• 关键词: Activins; Animal Model; Biological Assay; CRISPR/Cas technology; Cancer Etiology; Cell Line; Cells; Cessation of life; ChIP-seq; Characteristics; Cytometry; Data; Development; Enhancers; Enzymes; Epigenetic Process; Failure; Fibroblasts; Future; Genetic; Genetic Transcription; Goals; Grant; Immune; Immunohistochemistry; Immunologics; Immunotherapy; Infiltration; Inflammatory; KRAS2 gene; Knock-out; Knowledge; Ligands; Light; Luciferases; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Modeling; Molecular; Mutation; Myeloid-derived suppressor cells; Neoadjuvant Therapy; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Preclinical Testing; Prognosis; Public Health; Regulation; Reporter; Research; Resistance; Role; Secondary to; Signal Transduction; T-Cell Activation; T-Lymphocyte; Technology; Testing; Transcriptional Regulation; Tumor-associated macrophages; Up-Regulation; Xenograft procedure; activin A; cancer cell; cancer genetics; cell behavior; clinical development; cytokine; effector T cell; epigenetic regulation; histone modification; improved; innovation; knock-down; loss of function mutation; macrophage; mouse model; neoplastic cell; novel; pancreas development; pancreatic cancer patients; pancreatic ductal adenocarcinoma cell; pancreatic neoplasm; preclinical development; protein expression; receptor; response; single-cell RNA sequencing; therapy resistant; three dimensional cell culture; three-dimensional modeling; transcription factor; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor progression; tumor-immune system interactions

PROJECT SUMMARY/ABSTRACT The 5-year survival of pancreatic cancer patients remains at 10% primarily due to the tumor resistance to standard chemo and immunotherapies. The profound immunosuppressive tumor microenvironment contributes to treatment resistance. Notably, 82% of the neoadjuvant-treated pancreatic cancers carry loss of function mutations for KMT2D, a histone modification enzyme. Epigenetics is known to impact cancer cell behavior. However, how tumor cell-intrinsic epigenetic alterations modulate the pancreatic tumor microenvironment remains elusive. The long-term goal is to develop new treatments to convert pancreatic cancer to an immunologically hot tumor and improve the efficacy of immunotherapy and patient survival. The overall objectives in this application are to 1) determine the impact of KMT2D signaling on the tumor microenvironment and 2) characterize the mechanisms by which KMT2D regulates activin A expression. The central hypothesis is that upregulation of activin A upon KMT2D loss in pancreatic cancer reprograms cancer- associated fibroblasts and immune cells to promote a pro-tumoral immunosuppressive microenvironment. The rationale for this project is that the knowledge of the mechanisms by which KMT2D regulates the tumor microenvironment will pave the way for future preclinical and clinical development of new strategies to treat pancreatic cancer. The central hypothesis will be tested by pursuing three specific aims: 1) Elucidate the impact of KMT2D signaling on the immune composition in the tumor microenvironment; 2) Define the regulation of heterogeneous cancer-associated fibroblasts by KMT2D signaling; and 3) Determine the molecular mechanisms of activin A regulation by KMT2D in pancreatic cancer. Under the first and second aims, we will use our established pancreatic cancer genetic mouse models with pancreas-specific inactivation of KMT2D to determine the impact of KMT2D signaling on the differentiation and activation of cancer- associated fibroblasts, macrophages, and T cells, and ligand-receptor interactions in the tumor microenvironment and the underlying mechanisms. Single-cell RNA sequencing technology, mass cytometry, and multiplex fluorescent immunohistochemistry will be used. For the third aim, we will characterize the interactions between KMT2D and co-factors and the mechanisms of KMT2D-mediated regulation of activin A transcription and enhancer activity using ChIP-seq, BruUV-seq, and functional luciferase reporter assays. The research proposed in this application is innovative because it focuses on a novel concept that epigenetic signaling from tumor cells remodels the tumor microenvironment and contributes to cancer progression and uses state-of-the-art animal models and approaches. The proposed research is significant because it will shed light on the mechanism of transcriptional regulation by KMT2D in pancreatic cancer and the effect of KMT2D signaling on pancreatic tumor microenvironment, which will set the stage for future preclinical and clinical development of new tumor microenvironment-targeting treatment strategies.

项目总结/摘要 胰腺癌患者的5年生存率保持在10%，主要是由于肿瘤对化疗的耐药性。 标准化疗和免疫疗法深刻的免疫抑制肿瘤微环境有助于 治疗抵抗。值得注意的是，82%的新治疗的胰腺癌携带功能丧失 KMT 2D是一种组蛋白修饰酶。已知表观遗传学会影响癌细胞的行为。 然而，肿瘤细胞内在的表观遗传改变如何调节胰腺肿瘤微环境， 仍然难以捉摸长期目标是开发新的治疗方法，将胰腺癌转化为 免疫热肿瘤和提高免疫治疗的疗效和患者的生存。整体 本申请的目的是1）确定KMT 2D信号传导对肿瘤的影响 微环境和2）表征KMT 2D调节激活素A表达的机制。的 中心假设是胰腺癌中KMT 2D缺失后激活素A的上调重新编程癌症- 相关的成纤维细胞和免疫细胞，以促进促肿瘤免疫抑制微环境。的 该项目的基本原理是KMT 2D调节肿瘤的机制的知识 微环境将为未来临床前和临床开发新的治疗策略铺平道路 胰腺癌中心假设将通过追求三个具体目标进行测试：1）阐明 KMT 2D信号传导对肿瘤微环境中免疫组成的影响; 2）定义KMT 2D信号传导对肿瘤微环境中免疫组成的影响。 通过KMT 2D信号传导调节异质性癌症相关成纤维细胞;和3）确定 胰腺癌中KMT 2D调节激活素A的分子机制在第一和第二 目的是利用我们建立的胰腺癌基因特异性失活的小鼠模型， 以确定KMT 2D信号传导对癌症的分化和活化的影响- 相关的成纤维细胞、巨噬细胞和T细胞，以及肿瘤中的配体-受体相互作用 微环境和潜在机制。单细胞RNA测序技术，质谱分析， 并将使用多重荧光免疫组织化学。对于第三个目标，我们将描述 KMT 2D与辅因子的相互作用及其调控激活素A的机制 使用ChIP-seq、BruUV-seq和功能性荧光素酶报告基因测定来分析转录和增强子活性。的 在这项申请中提出的研究是创新的，因为它集中在一个新的概念，表观遗传 来自肿瘤细胞的信号传导重塑肿瘤微环境并促进癌症进展， 使用最先进的动物模型和方法。这项研究很重要，因为它将使 KMT 2D在胰腺癌中的转录调控机制及其作用 胰腺肿瘤微环境的信号传导，这将为未来的临床前和临床 开发新的肿瘤微环境靶向治疗策略。