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针对遗传性 BRCA1 女性的唯一乳腺癌预防策略 或 BRCA2 突变。虽然这种策略通常很有效，但它会显着影响生活质量，并且不会阻止 其他组织中发生的癌症（例如胰腺癌和前列腺癌）。这样，更好 迫切需要拦截策略。家长补助金的总体目标是表征最早的 乳腺癌高危人群乳腺组织的改变，并为乳腺癌的预防奠定基础 发现新的预防措施。我们与 Sam Aparicio 博士使用单细胞复制进行合作研究 数量分析已确定来自 BRCA1/2 的表面正常乳腺上皮细胞数量有所增加 携带拷贝数改变（CNA）的突变携带者。有趣的是，这些 CNA 属于 最常与早期 (DCIS) 和已形成 (IBC) 形式的乳腺癌相关，并且在之前就已存在 BRCA1/2 杂合性缺失 (LOH)，表明这些变化可能导致乳腺上皮细胞 细胞引发肿瘤的基因改变。该补充提案旨在检验总体假设 在 BRCA1 或 BRCA2 LOH 之前发生的基因改变使细胞能够在最终丧失 BRCA1 或 BRCA2 的两个拷贝，从而增加其细胞适应性和罹患恶性肿瘤的倾向 由同源重组缺陷引起的额外遗传改变的积累之后。我们 计划通过以下方式解决这一假设：1）通过确定单核苷酸来补充我们的 CNA 分析 使用 Tapestri 平台 (MissionBio) 观察癌前乳腺组织的变异和突变情况，这是一个 最先进的靶向单细胞 DNA 和蛋白质测序技术以及 2) 利用 3D 乳腺类器官 确定 CNA/SNV 在 BRCA1/2 缺失后生存中的功能作用以及是否需要额外的模型 下游的改变导致对肿瘤发展的敏感性。更好地了解多早 CNA/SNV 影响肿瘤发生，可以识别这些细胞的脆弱性，以便 在癌症进展之前消除它们，并制定检测异常细胞扩增的策略 作为监测癌前细胞增殖的手段。重要的是，实现该提案的目标将 使 Oliphant 博士能够进一步加强分析单细胞多组 DNA 所需的计算技能- 独立地分析 seq 和蛋白质表达数据集，并利用 3D 类器官模型来识别和 更有效地制定可行的治疗策略。它还将为 Oliphant 博士提供以下机会： 参与各种职业发展经验，包括参加会议和拓展 他的专业网络。总体而言，拟议项目的资金将确保奥利芬特博士扩大他的研究范围 科学和专业的培训，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