Tracking the evolution of breast cancer through single cell analyses of premalignant breast tissues from women at high risk for cancer development

通过对癌症高危女性的癌前乳腺组织进行单细胞分析来追踪乳腺癌的演变

• 批准号: 10683138
• 负责人: Joan Siefert Brugge
• 金额: $ 99.67万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10683138
• 关键词: BRCA mutations; BRCA1 Mutation; BRCA1 gene; BRCA2 Mutation; BRCA2 gene; Breast; Breast Cancer Prevention; Breast Cancer Risk Factor; Cell Lineage; Cell Proliferation; Cells; Cytometry; DNA Damage; Development; Evolution; Goals; High Risk Woman; High-Risk Cancer; Histology; Human; Image; Immunocompromised Host; Immunofluorescence Immunologic; In Situ; Inherited; Intervention; Malignant Neoplasms; Mammary Gland Parenchyma; Mammary Neoplasms; Masks; Mus; Mutation; Organoids; Periodicity; Population; Population Analysis; Premalignant Cell; Premalignant Change; Proliferation Marker; Quality of life; RNA; Structure; Surface; Susceptibility Gene; Technology; Tissues; Woman; brca gene; diagnostic strategy; high risk; high risk population; malignant breast neoplasm; mammary; mutant; mutation carrier; novel diagnostics; premalignant; prevent; programs; prophylactic; prophylactic mastectomy; psychologic; reconstitution; single cell analysis; single cell technology; single-cell RNA sequencing; tool; tumor; tumor progression; tumorigenesis

Abstract: The overarching objective of the proposed studies is to identify and characterize early premalignant changes in breast tissues from women that carry genetic alterations associated with a high risk of breast cancer to ultimately develop strategies to detect and prevent the development of breast cancer. To accomplish this goal, we have optimized three technologies to profile single breast mammary cells (MECs): (1) CyTOF mass cytometry to allow tracking in parallel of >30 cell lineage and proliferation markers, (2) single cell RNA sequencing to identify expression programs of cell populations enriched in mutation-carriers, and (3) multi-plex cyclic immunofluorescence imaging (CyCIF) to simultaneous image >50 markers in situ. These technologies make it possible to detect differences in small populations of cells that would be masked by bulk population analyses. To date, we have profiled breast tissues from over 30 women with wild-type or mutant BRCA1 or BRCA2 by CyTOF and have identified distinct, previously unrecognized subpopulations of cells that are enriched in breast tissues from BRCA1 and/or BRCA2 carriers. These enriched subpopulations may represent cells that are either directly on the path to malignancy or indirectly contribute to the development of cancer in these high-risk women. We have identified RNA signatures associated with these enriched subpopulations, which include surface markers to isolate them from breast tissue to investigate both these possibilities and to track them within breast tumors. The signatures associated with one of the enriched populations have provided clues as to the basis for their accumulation, as well as potential strategies to prevent their accumulation. Using CyCIF, we have been able to identify enriched subpopulations of cells in situ within breast tissues and track their association with aberrant histologies. We have also developed organoid cultures that maintain all of the major MEC lineages as well as the BRCA1/2-enriched populations, and that are able to reconstitute glandular structures in immunocompromised mice. We believe that these tools provide an unprecedented opportunity to track the development of human cancer. In the proposed studies, we will investigate whether and how the BRCA1/2+/mut-enriched subpopulations contribute to tumorigenesis in mutation carriers. We will also investigate the basis for the enrichment of these populations and the contribution of DNA damage to their enrichment. Later stage studies will focus on the development of strategies to interfere with tumor progression, and importantly to develop novel diagnostic strategies to inform on the timing of prophylactic interventions. In addition, we will examine tissues from women who carry mutations in other breast cancer predisposition genes to establish whether similar subpopulations are detected in other high-risk individuals. And lastly, we will examine the possibility that these cells represent cells-of-origin of sporadic breast tumors that arise more broadly in the population.

摘要： 拟议研究的总体目标是识别和表征早期癌前病变， 来自携带与乳腺癌高风险相关的基因改变的女性的乳腺组织， 制定检测和预防乳腺癌发展的策略。为了实现这一目标，我们 优化了三种技术来分析单个乳腺乳腺细胞（MEC）：（1）CyTOF质谱细胞术， 平行追踪>30种细胞谱系和增殖标志物，（2）单细胞RNA测序以鉴定 突变携带者中富集的细胞群的表达程序，和（3）多重循环免疫荧光 使用CyCIF成像（CyCIF）来同时原位成像>50个标记物。这些技术使我们有可能发现 小细胞群的差异会被大细胞群分析掩盖。到目前为止， 通过CyTOF分析来自30多名野生型或突变型BRCA 1或BRCA 2女性的乳腺组织， 从BRCA 1中鉴定出在乳腺组织中富集的不同的、以前未被识别的细胞亚群 和/或BRCA 2携带者。这些富集的亚群可以代表直接在向细胞转化的途径上的细胞。 恶性肿瘤或间接有助于这些高危妇女的癌症发展。我们已经识别出RNA 与这些富集亚群相关的特征，其中包括将它们与之分离的表面标记物 研究这些可能性，并在乳腺肿瘤中追踪它们。签名 与其中一个富集种群有关的化石也为它们的积累提供了线索， 作为防止其积累的潜在策略。使用CyCIF，我们已经能够识别富集的 乳腺组织内原位细胞亚群，并跟踪其与异常组织学的关联。我们有 还开发了维持所有主要MEC谱系以及BRCA 1/2富集的类器官培养物， 群体，并且能够在免疫受损小鼠中重建腺体结构。我们认为 这些工具为跟踪人类癌症的发展提供了前所未有的机会。拟议 研究中，我们将研究BRCA 1/2+/mut-enriched亚群是否以及如何有助于 突变携带者的肿瘤发生。我们还将调查这些人口富集的基础， DNA损伤对它们富集的贡献。后期研究将侧重于制定战略 干扰肿瘤进展，重要的是开发新的诊断策略， 预防性干预。此外，我们还将检查携带其他乳腺癌基因突变的妇女的组织。 癌症易感基因，以确定是否在其他高风险个体中检测到类似的亚群。 最后，我们将研究这些细胞是否代表散发性乳腺肿瘤的起源细胞， 在人群中更广泛地出现。