Fibrocytes: A novel cell population promoting obesity-induced breast tumor desmoplasia

纤维细胞：促进肥胖诱导的乳腺肿瘤结缔组织形成的新型细胞群

基本信息

批准号：
10317094
负责人：
Genevra Marie Kuziel
金额：
$ 3.52万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10317094
关键词：
Adenocarcinoma Adipocytes Adipose tissue Adoptive Transfer Biological Assay Body Weight decreased Bone Marrow Bone Marrow Cells Breast Breast Cancer Model Breast Cancer Patient CD34 gene Cancer Patient Cell Lineage Cells Characteristics Chronic Collagen Communication Data Deposition Desmoplastic Development Diagnosis Diet Disease Environment Epidemic Estrogen Receptor alpha Extracellular Matrix Fatty acid glycerol esters Fibroblasts Fibrosis Flow Cytometry Goals Green Fluorescent Proteins Growth High Fat Diet Hyperplasia ITGAM gene Immunofluorescence Immunologic In Vitro Incidence Inflammation Inflammatory Knowledge Lead Link Malignant Neoplasms Mammary Duct Mammary Neoplasms Mammary gland Menopausal Status Mentorship Mission Modeling Mus Myelogenous Myeloid Progenitor Cells Myofibroblast Neoplasm Transplantation Obese Mice Obesity Obesity associated cancer Pathology Population Postmenopause Prognosis Public Health Research Research Personnel Resistance Risk Factors Role Science Sirius Red F3B Smooth Muscle Actin Staining Method Stains Testing Thinness Tissues Training Tumor Weights Woman Work Xenograft procedure base biomarker panel cancer therapy cancer type career cell type infiltrating duct carcinoma innovation macrophage malignant breast neoplasm mammary monocyte mouse model neoplastic cell novel novel therapeutics obese patients recruit response skills targeted treatment therapeutic target therapy resistant tumor tumor growth tumor progression weight loss intervention wound healing

项目摘要

PROJECT ABSTRACT Obesity rates are rising worldwide. Obesity is a significant risk factor for development of breast cancer, and obese breast cancer patients have a worse prognosis. Breast tumors from obese patients have increased desmoplasia, with significantly more smooth muscle actin (SMA) positive cells, a marker for myofibroblasts, within tumors. Obesity induces a state of chronic, macrophage-driven inflammation. It is not understood how inflammation within the obese breast contributes to adipose tissue fibrosis and tumor desmoplasia. In obesity, myeloid progenitor cells are significantly increased within bone marrow. Fibrocytes, a myeloid lineage cell type with characteristics of both macrophages and myofibroblasts, are derived from myeloid progenitor cells and are increased in chronic inflammatory and fibrotic diseases. The long-term goal is to understand how the systemic inflammatory conditions of obesity contribute to fibrotic tumors. The rationale is that tumor fibrosis is associated with more aggressive tumors, and understanding this inflammatory microenvironment will lead to discoveries of much needed therapeutic targets for obese cancer patients. The overall objective of this proposal is to identify how obesity increases fibrocytes to enhance collagen deposition within tumors, and how weight loss intervention reverses these effects of obesity. The central hypothesis is that obesity reversibly increases myeloid progenitor cells and fibrocytes within the bone marrow and mammary gland, respectively, leading to an increase in collagen deposition within tumors. This hypothesis will be tested in three specific aims: 1) To identify how obesity-induced fibrocytes contribute to mammary tumor stroma; 2) To elucidate how obesity-induced fibrocytes enhance fibrosis during tumor growth; 3) To examine how weight loss decreases fibrocytes within the normal mammary gland and mammary tumors, resulting in reduced fibrosis. A diet-induced mouse model of obesity and trp53-/- mammary tumor cell xenografts will be utilized to test these aims. Myeloid progenitor cells and monocytes/ macrophages in bone marrow and mammary tumors will be quantified from lean, obese, and weight loss mice using FACS and flow cytometry. Colony formation from FACS isolated cells will be used to quantify mammary tumor fibrocytes. Adoptive transfer using FACS isolated GFP-expressing bone marrow cells from obese mice will be used to examine cell populations responsible for tumor fibrosis. Picrosirius red staining multiplexed with immunofluorescence will be used to examine collagen deposition in mammary tumors and localize fibrocytes. This approach is innovative because fibrocytes have not been studied in obesity, and the mechanisms underlying obesity-induced tumor desmoplasia are not well understood. Identifying how obesity regulates fibrocytes and tumor fibrosis is important to develop novel therapies to treat the more aggressive tumors observed in obese cancer patients. Beyond new technical knowledge, the training described provides opportunities to enhance science communication, networking, and mentorship skills within the collaborative research environment at UW- Madison. The training plan will help the applicant work toward a career as an independent cancer researcher.

项目摘要肥胖率在全球范围内上升。肥胖是乳腺癌发展的重要危险因素，肥胖的乳腺癌患者预后较差。肥胖患者的乳腺肿瘤增加了脱木质，具有更平滑肌肉肌动蛋白（SMA）阳性细胞，是肌纤维细胞的标志物，在肿瘤内。肥胖会诱导慢性巨噬细胞驱动的炎症状态。不明白如何肥胖乳房内的炎症有助于脂肪组织纤维化和肿瘤脱发。在肥胖中，骨髓内的髓样祖细胞显着增加。纤维细胞，髓样谱系细胞类型具有巨噬细胞和肌纤维细胞的特征，源自髓样祖细胞，是慢性炎症和纤维化疾病的增加。长期目标是了解系统性肥胖的炎症条件有助于纤维化肿瘤。理由是肿瘤纤维化是相关的有了更具侵略性的肿瘤，了解这种炎症性微环境将导致发现肥胖癌症患者急需的治疗靶标。该提议的总体目的是确定肥胖如何增加纤维细胞以增强肿瘤内的胶原蛋白沉积，以及如何减肥干预逆转肥胖的影响。中心假设是肥胖可逆地增加髓样祖细胞分别在骨髓和乳腺内的细胞和纤维细胞，导致胶原蛋白增加肿瘤内沉积。该假设将以三个特定目的进行检验：1）确定肥胖诱导的如何纤维细胞有助于乳腺肿瘤基质； 2）阐明肥胖诱导的纤维细胞如何增强纤维化在肿瘤生长期间； 3）检查体重减轻如何减少正常乳腺内的纤维细胞和乳腺肿瘤，导致纤维化降低。饮食引起的肥胖小鼠模型和TRP53 - / - 乳腺肿瘤细胞异种移植物将用于测试这些目标。髓样祖细胞和单核细胞/ 骨髓和乳腺肿瘤中的巨噬细胞将从瘦肉，肥胖和减肥小鼠中量化使用FACS和流式细胞仪。 FACS分离细胞的菌落形成将用于定量乳腺肿瘤纤维细胞。使用FACS分离的表达GFP的骨髓细胞的收养转移来自肥胖小鼠将用于检查负责肿瘤纤维化的细胞群。 Picrosirius红色染色与免疫荧光将用于检查乳腺肿瘤中的胶原蛋白沉积并定位纤维细胞。这种方法具有创新性，因为纤维细胞尚未在肥胖症中进行研究，并且机制是基础的肥胖引起的肿瘤脱木质尚不清楚。确定肥胖如何调节纤维细胞和肿瘤纤维化对于开发新型疗法以治疗肥胖症中观察到的更具侵略性的肿瘤很重要癌症患者。除了新的技术知识之外，所描述的培训还提供了增强的机会科学沟通，网络和指导技能在UW- 麦迪逊。培训计划将帮助申请人致力于成为独立癌症研究员的职业。