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: 10817308
• 负责人: Joan Siefert Brugge
• 金额: $ 11.45万
• 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/10817308
• 关键词: 3-Dimensional; Address; BRCA mutations; BRCA1 Mutation; BRCA1 gene; BRCA2 Mutation; BRCA2 gene; Breast; Breast Cancer Prevention; Breast Epithelial Cells; Cells; Complement; DNA sequencing; Data Set; Defect; Development; Ensure; Evolution; Foundations; Funding; Goals; High Risk Woman; High-Risk Cancer; Individual; Inherited; Loss of Heterozygosity; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Malignant neoplasm of prostate; Mammary Gland Parenchyma; Modeling; Monitor; Mutation; Noninfiltrating Intraductal Carcinoma; Normal Cell; Organoids; Pathogenicity; Peptide Sequence Determination; Population; Predisposition; Premalignant Cell; Prevention Measures; Prevention approach; Prevention strategy; Quality of life; Research; Research Project Grants; Role; Single Nucleotide Polymorphism; Technology; Testing; Therapeutic; Tissues; Training; Woman; brca gene; career development; career networking; experience; fitness; high risk; homologous recombination; malignant breast neoplasm; multiple omics; mutant; mutation carrier; parent grant; premalignant; programs; prophylactic mastectomy; protein expression; single cell analysis; single cell technology; skills; symposium; tumor; tumor initiation; tumor progression; tumorigenesis

Prophylactic mastectomy is currently the only breast cancer prevention strategy for women with inherited BRCA1 or BRCA2 mutations. While often effective, this strategy significantly impacts quality of life and does not prevent the development of cancers arising in other tissues (e.g., pancreatic and prostate cancers). Thus, better interception strategies are urgently needed. The overall goal of the parent grant is to characterize the earliest alterations in breast tissues in individuals at high risk of developing breast cancer and to lay the groundwork for the discovery of new prevention measures. Our collaborative studies with Dr. Sam Aparicio using single-cell copy number analyses have identified expanded population of ostensibly normal breast epithelial cells from BRCA1/2 mutation carriers that carry copy number alterations (CNAs). Interestingly, these CNAs are among those that are most commonly associated with early (DCIS) and established (IBC) forms of breast cancer and are present prior to BRCA1/2 loss of heterozygosity (LOH), suggesting that these changes may predispose mammary epithelial cells to tumor-initiating genetic alterations. This supplement proposal aims to test the overarching hypothesis that the genetic alterations that occur prior to LOH of BRCA1 or BRCA2 allow cells to survive eventual loss of both copies of BRCA1 or BRCA2, thereby increasing their cellular fitness and predisposition to malignant cancer following accumulation of additional genetic alterations caused by defects in homologous recombination. We plan to address this hypothesis by 1) complementing our CNA analysis by determining the single nucleotide variant and mutational landscape of precancerous breast tissues using the Tapestri Platform (MissionBio), a state-of-the-art, targeted single cell DNA and protein sequencing technology and 2) utilizing 3D breast organoid models to determine the functional role of CNAs/SNVs in survival after loss of BRCA1/2 and whether additional alterations downstream result in sensitization to tumor development. A better understanding of how early CNAs/SNVs influence tumorigenesis can lead to the identification of vulnerabilities of these cells in order to eliminate them before cancer progresses, as well as to develop strategies to detect expansion of aberrant cells as a means of monitoring pre-malignant cell expansion. Importantly, accomplishing the goals of this proposal will enable Dr. Oliphant to further strengthen the computational skills needed to analyze single-cell multiomic DNA- seq and protein expression datasets independently, as well as leverage 3D organoid models to identify and develop actionable therapeutic strategies more effectively. It will also provide Dr. Oliphant with opportunities for engagement in various career development experiences including participating at conferences and expanding his professional network. Overall, the funding of the proposed project will ensure that Dr. Oliphant expands his scientific and professional training, while establishing the foundation for a viable independent academic research program.

预防性乳房切除术是目前遗传性BRCA1妇女唯一的乳腺癌预防策略 或BRCA2突变。虽然这种策略通常很有效，但它对生活质量有很大影响，并不能防止 发生在其他组织(如胰腺癌和前列腺癌)的癌症的发展。因此，更好地 迫切需要拦截策略。父母资助的总体目标是将最早的 乳腺癌高危人群乳腺组织的改变，为预防乳腺癌奠定基础 新的预防措施的发现。我们与Sam Aparicio博士使用单细胞复制进行的合作研究 数量分析已经确认了BRCA1/2表面正常的乳腺上皮细胞的扩大群体 携带拷贝数改变(CNA)的突变载体。有趣的是，这些CNA属于 最常与早期(DCIS)和已确诊(IBC)形式的乳腺癌有关，并优先出现 BRCA1/2杂合性缺失(LOH)，提示这些变化可能倾向于乳腺上皮 细胞到引发肿瘤的基因改变。这项补充建议旨在检验总体假设。 在BRCA1或BRCA2杂合性缺失之前发生的基因改变允许细胞在最终失去 BRCA1或BRCA2的两个副本，从而提高它们的细胞适应性和患恶性肿瘤的易感性 随着同源重组缺陷引起的额外基因改变的积累。我们 计划通过1)通过确定单核苷酸来补充我们的CNA分析来解决这一假设 使用Tapestri平台(MaskBio)研究癌前病变组织的变异和突变景观 最先进的、有针对性的单细胞DNA和蛋白质测序技术以及2)利用3D乳房器官 模型来确定CNAs/SNV在BRCA1/2缺失后存活中的功能作用以及是否有额外的 下游的改变导致对肿瘤发展的敏感化。更好地理解有多早 影响肿瘤发生的CNAS/SNV可以导致识别这些细胞的脆弱性，以便 在癌症进展之前消除它们，以及开发检测异常细胞扩张的策略 作为监测癌前细胞扩张的一种手段。重要的是，实现这项提案的目标将 使奥列芬特博士能够进一步加强分析单细胞多组DNA所需的计算技能-- 独立的SEQ和蛋白质表达数据集，以及利用3D有机模型来识别和 更有效地制定可行的治疗策略。它还将为奥列芬特博士提供机会 参与各种职业发展经历，包括参加会议和扩展 他的职业关系网。总体而言，拟议项目的资金将确保奥利芬特博士扩大他的 科学和专业培训，同时为可行的独立学术研究奠定基础 程序。

项目成果

Long-term culture, genetic manipulation and xenotransplantation of human normal and breast cancer organoids.

• DOI: 10.1038/s41596-020-00474-1
• 发表时间: 2021-04
• 期刊: Nature protocols
• 影响因子: 14.8
• 作者: Dekkers JF;van Vliet EJ;Sachs N;Rosenbluth JM;Kopper O;Rebel HG;Wehrens EJ;Piani C;Visvader JE;Verissimo CS;Boj SF;Brugge JS;Clevers H;Rios AC
• 通讯作者: Rios AC