Elucidating fibroblast/immune crosstalk and plasticity in pancreatic ductal adenocarcinoma

阐明胰腺导管腺癌中的成纤维细胞/免疫串扰和可塑性

• 批准号: 10338112
• 负责人: Jennifer Susan Thalappillil
• 金额: $ 3.95万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2023-02-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10338112
• 关键词: 3-Dimensional; Affect; Biology; Bone Marrow; Cancer Etiology; Cancer Survivor; Cell Culture Techniques; Cells; Cessation of life; Characteristics; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Data; Desmoplastic; Development; Diagnosis; Diphtheria Toxin; Disease; Disease Progression; Engineering; Extracellular Matrix; Fibroblasts; Ganciclovir; Genes; Genetically Engineered Mouse; Goals; Heterogeneity; Immune; Immunosuppression; In Vitro; Individual; Inflammatory; Knockout Mice; Laboratories; Lesion; Macrophage Activation; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Modeling; Morphology; Mus; Organoids; Pancreas; Pancreatic Ductal Adenocarcinoma; Pancreatic duct; Patient-Focused Outcomes; Patients; Penetration; Pharmaceutical Preparations; Phenotype; Play; Population; Reporter; Role; Signal Transduction; Smooth Muscle Actin Staining Method; Spatial Distribution; Stromal Cells; Suicide; System; Testing; Therapeutic; Time; Treatment Efficacy; Tumor-associated macrophages; United States; Work; base; cancer cell; cancer subtypes; cell type; conditional knockout; cytokine; ds-DNA; effective therapy; improved; in vivo; innovation; macrophage; men; mouse model; neoplastic cell; novel; novel strategies; novel therapeutic intervention; organoid transplantation; pancreatic cancer model; pancreatic cancer patients; pancreatic neoplasm; pancreatic stellate cell; promoter; survival outcome; therapeutic target; tumor; tumor initiation; tumor progression; tumorigenesis

Project Summary Pancreatic ductal adenocarcinoma (PDA) is a deadly malignancy, and only 8% of patients diagnosed with this cancer survive 5 years after diagnosis. The poor outcomes of patients with PDA are attributed to the advanced stage of disease at the time of diagnosis and the lack of effective therapies, highlighting the need for new approaches. PDA is characterized by an abundance of extracellular matrix (ECM) around the cancer cells, referred to as a desmoplastic reaction. The desmoplasia provides a niche for different types of stromal cells, including immune cells and cancer-associated fibroblasts (CAFs), which are derived primarily from pancreatic stellate cells (PSCs). PDA CAFs have recently been found to display heterogeneity with distinct phenotypes and spatial distributions: myofibroblastic CAFs (myCAFs) express high levels of α-smooth muscle actin (αSMA) and are in direct physical contact with cancer cells, while inflammatory CAFs (iCAFs) are farther away from tumor cells and secrete inflammatory cytokines. Although this finding has provided opportunities for new therapeutic approaches, their interaction with other cell types within the immunosuppressive, tumor-promoting microenvironment of PDA requires further study. Tumor-associated macrophages (TAMs) are prominent in PDA tumors and are known to have an immunosuppressive function, making them an attractive therapeutic target. However, the mechanisms through which TAMs signal to and impact the biology of iCAFs and myCAFs remains to be explored. This study aims to understand the crosstalk between macrophages and the two CAF subtypes and how these cell populations fluctuate during PDA progression. My preliminary studies show that PSCs promote the proliferation and alternative activation of macrophages in vitro. In addition, in a genetically engineered mouse model (GEMM) of pancreatic cancer, PanIN lesions (precursors of PDA) display an abundance of both αSMA+ fibroblasts and alternatively activated macrophages in close proximity to each other. These results suggest that the fibroblasts and macrophages signal to each other, contributing to the CAF and macrophage phenotypes in PDA. To identify factors responsible for such crosstalk, I will use an innovative, three- dimensional triple co-culture system that combines pancreatic ductal organoids, PSCs, and macrophages. To better understand how the CAF subtypes impact tumor progression and macrophage phenotype in vivo, I will engineer iCAF- and myCAF-conditional knockout mice and perturb each subtype in an orthotopically transplanted organoid mouse model. Ultimately, the identification of iCAF/myCAF and macrophage dynamics and interactions will illuminate the role these cells play in the disease, opening avenues for new and much- needed therapeutic targets.

项目摘要 胰腺导管腺癌（PDA）是一种致命的恶性肿瘤，只有8%的患者被诊断患有这种疾病。 癌症在诊断后存活5年。PDA患者的不良结局归因于晚期 在诊断时疾病的阶段和缺乏有效的治疗，突出了新的需要， 接近。PDA的特征在于癌细胞周围存在丰富的细胞外基质（ECM）， 称为促纤维增生反应。结缔组织增生为不同类型的基质细胞提供了一个小生境， 包括免疫细胞和癌症相关成纤维细胞（CAF），它们主要来源于胰腺癌。 星状细胞（PSC）。最近发现PDA CAF显示具有不同表型的异质性， 空间分布：肌纤维母细胞CAFs（myCAFs）表达高水平的α-平滑肌肌动蛋白（αSMA）， 与癌细胞直接物理接触，而炎性CAF（iCAF）远离肿瘤 细胞并分泌炎性细胞因子。尽管这一发现为新的治疗方法提供了机会， 方法，它们与免疫抑制、肿瘤促进和免疫调节中的其他细胞类型的相互作用， PDA的微环境需要进一步研究。肿瘤相关巨噬细胞（TAM）在PDA中很突出 已知它们具有免疫抑制功能，使它们成为有吸引力的治疗靶点。 然而，TAM向iCAF和myCAF发出信号并影响其生物学的机制仍然存在 有待探索本研究旨在了解巨噬细胞和两种CAF亚型之间的串扰 以及这些细胞群在PDA进展过程中如何波动。我的初步研究表明PSC 促进体外巨噬细胞的增殖和交替激活。此外，在基因 在胰腺癌的工程小鼠模型（GEMM）中，PanIN病变（PDA的前体）显示了胰腺癌的早期转移。 αSMA+成纤维细胞和交替激活的巨噬细胞的丰度彼此非常接近。 这些结果表明，成纤维细胞和巨噬细胞相互传递信号，促成CAF， 巨噬细胞表型。为了确定这种串扰的因素，我将使用一个创新的，三- 三维三元共培养系统，其组合了胰腺导管类器官、PSC和巨噬细胞。到 为了更好地了解CAF亚型如何影响体内肿瘤进展和巨噬细胞表型，我将 改造iCAF和myCAF条件性敲除小鼠，并在原位干扰 移植类器官小鼠模型。最终，iCAF/myCAF和巨噬细胞动力学的鉴定 和相互作用将阐明这些细胞在疾病中所起的作用，为新的和更多的- 需要的治疗目标。