Investigating the role of pericytes as regulators of breast tissue homeostasis and cancer.

研究周细胞作为乳腺组织稳态和癌症调节剂的作用。

• 批准号: 10435520
• 负责人: Kevin Nee
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10435520
• 关键词: 3-Dimensional; Adipocytes; Affect; Alleles; Attention; BRCA1 Mutation; BRCA1 gene; Basal Cell; Biological Assay; Biological Markers; Blood Vessels; Breast; Breast Cancer Risk Factor; Breast Epithelial Cells; Cancer Etiology; Cartilage; Cell Proliferation; Cells; Clinical Treatment; Connective Tissue; DNA Repair Enzymes; Data; Disease; Endothelium; Epithelial; Epithelial Cells; Fatty acid glycerol esters; Fibroblasts; Flow Cytometry; Genetic Transcription; Germ-Line Mutation; Goals; Growth; Heterogeneity; Heterozygote; Homeostasis; Human; Hydrogels; Immune; In Vitro; Individual; Malignant Neoplasms; Mammary Gland Parenchyma; Mammospheres; Mesenchymal; Mesenchymal Differentiation; Mesenchymal Stem Cells; Modeling; Molecular; Morphogenesis; Mus; Mutate; Mutation; NGFR Protein; Nerve Growth Factors; Oncogenic; Operative Surgical Procedures; Organoids; Paracrine Communication; Patients; Pericytes; Play; Population; Predisposition; Premalignant Change; Preneoplastic Change; Prevention strategy; Primary Cancer Prevention; Production; Prognosis; Recombinants; Resolution; Risk; Role; Sampling; Stromal Cells; System; Testing; Tissues; Woman; Work; aggressive breast cancer; base; bone; breast tumorigenesis; cancer initiation; cancer risk; cell type; cohort; defined contribution; high risk; human tissue; humanized mouse; in vivo; insight; malignant breast neoplasm; mammary; mammary epithelium; mammary gland development; mouse model; multidimensional data; mutant; mutation carrier; neoplastic; novel; paracrine; premalignant; progenitor; screening; single-cell RNA sequencing; stem; stem cell differentiation; stem cells; stromal progenitor; targeted treatment; tissue culture; transplant model; tumor; tumor initiation; tumor microenvironment; tumor progression; tumorigenic

PROJECT SUMMARY Despite advancements in clinical treatment and screening, breast cancer remains the second most deadly and most frequent cause of cancer in women. BRCA1 related cancer is associated with dramatically increased risk of cancer occurrence. Most BRCA1 related breast cancers will be ‘triple-negative’, with no targeted therapies, poor prognosis, and decreased overall survival. Pre-neoplastic changes in BRCA1 mutation carrier epithelium have been well studied, as well has the tumor microenvironment. However, BRCA1 germline mutations are likely to affect all cells in the breast, not just the epithelium. Less well studied, and the focus of this proposal, are the pre-neoplastic changes that occur in the stromal cells of the breast microenvironment, and their contributions to cancer initiation. The overarching hypothesis of this proposal is that the breast microenvironment is distorted by BRCA1 mutations and contribute to cancer initiation. A persistent challenge in assaying the stroma of the breast microenvironment is the diversity and heterogeneity of this tissue compartment. To overcome this challenge, our lab has performed single cell RNA sequencing (scRNAseq) of 81,023 stromal and epithelial cells from a cohort of high-risk BRCA1 mutation carrier and control patient surgical breast samples. scRNAseq has the resolution to identify specific cell populations in the breast microenvironment and how they change in the preneoplastic microenvironment. Using this highly multidimensional data, this proposal aims to 1) describe altered paracrine interactions in the pre-neoplastic breast and to 2) identify how microenvironment composition may be distorted in BRCA1 mutation carrier breasts. Using scRNAseq and in vitro organoid models, preliminary work has defined a pro-proliferative paracrine interaction of pericytes with basal epithelium. Additionally, this study uses scRNAseq data to develop a novel flow cytometry strategy to specifically isolate pericytes to demonstrate that pericytes have multipotent mesenchymal potential. This study proposes that these multipotent pericytes may serve as the stromal mesenchymal stem cell progenitor. To assess how these interactions contribute to breast cancer initiation, this study proposes two in vivo humanized mouse models that allow growth of oncogenic patient breast epithelium with defined patient-derived stromal populations. If successful, this study will provide insights into paracrine interactions that have been previously undefined but may have roles in breast cancer initiation. Furthermore, identification of a stromal progenitor may yield new biomarkers and targets for high-risk breast cancer.

项目总结 尽管在临床治疗和筛查方面取得了进步，但乳腺癌仍位居第二。 致命的，也是女性癌症最常见的原因。BRCA1相关癌症与戏剧性 增加癌症发生的风险。大多数与BRCA1相关的乳腺癌将是“三阴性”的，没有针对性的 治疗方法，预后差，总存活率下降。BRCA1突变携带者的癌前改变 对上皮细胞以及肿瘤微环境已有较好的研究。然而，BRCA1生殖系 突变可能会影响乳房的所有细胞，而不仅仅是上皮细胞。较少的研究，以及这一点的重点 是发生在乳腺微环境的间质细胞中的癌前改变，以及它们的 对癌症启动的贡献。这一建议的首要假设是乳房微环境 被BRCA1突变扭曲，并导致癌症的发生。 分析乳房微环境间质的一个长期挑战是多样性和 这个组织隔间的异质性。为了克服这一挑战，我们的实验室进行了单细胞RNA 高危BRCA1突变携带者队列中81,023个基质和上皮细胞的测序(ScRNAseq) 并控制患者的手术乳房样本。ScRNAseq具有识别特定细胞群的分辨率 乳房微环境及其在癌前微环境中的变化。高度利用这一点 多维数据，这项建议旨在1)描述癌前病变中旁分泌相互作用的改变 2)确定BRCA1突变携带者乳房的微环境成分可能会如何扭曲。 利用scRNAseq和体外器官模型，初步工作定义了一种促增殖的旁分泌 周细胞与基底上皮细胞的相互作用。此外，本研究使用scRNAseq数据开发了一种新的 特异性分离周细胞以证明周细胞具有多能性的流式细胞术策略 间充质潜能。本研究认为，这些多能周细胞可能作为基质细胞。 间充质干细胞的祖细胞。为了评估这些相互作用如何促进乳腺癌的发生，这项研究 研究提出了两种体内人源化小鼠模型，允许致癌患者乳腺上皮生长 具有定义的患者来源的基质种群。如果成功，这项研究将为旁分泌提供洞察力 以前没有定义但可能在乳腺癌发生中起作用的相互作用。此外， 对基质前体细胞的鉴定可能会为高危乳腺癌产生新的生物标志物和靶点。