Genetic Analysis of Breast Cancer Progression in Mice Using Inducible Transposition

使用诱导转座对小鼠乳腺癌进展进行遗传分析

• 批准号: 9922893
• 负责人: EDWARD J GUNTHER
• 金额: $ 35.16万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-16 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9922893
• 关键词: 3-Dimensional; Address; Advanced Malignant Neoplasm; Animal Model; Apoptosis; Automobile Driving; Benign; Breast; Breast Cancer Genetics; Breast Cancer Model; Candidate Disease Gene; Catalogs; Cell Proliferation; Cells; Clinical; Complement; Complex; Disease; Disease Progression; Disease Resistance; Disease remission; Doxycycline; Drug Targeting; Drug resistance; ERBB2 gene; Engineering; Enterobacteria phage P1 Cre recombinase; Estrogen Receptors; Event; Evolution; Fostering; Genes; Genetic; Genetic Screening; Genome; Goals; Growth; Hormonal; Human; Hyperplasia; Jumping Genes; Knowledge; Malignant - descriptor; Malignant Neoplasms; Mammary Neoplasms; Mammary Tumorigenesis; Mammary gland; Modeling; Monitor; Mouse Mammary Tumor Virus; Mus; Mutagenesis; Mutation; Oncogenes; Oncogenic; Organoids; Outcome; Pathway interactions; Pharmaceutical Preparations; Phenotype; Publishing; Recurrence; Refractory; Refractory Disease; Relapse; Reporting; Resistance; Scoring Method; Signal Transduction; Sleeping Beauty; Somatic Mutation; Sum; System; Testing; Tissues; Validation; WNT Signaling Pathway; Withdrawal; Work; advanced breast cancer; aggressive therapy; base; bioinformatics pipeline; breast cancer genomics; breast cancer progression; cancer genome; candidate validation; design; driver mutation; gene discovery; gene interaction; genetic analysis; genome database; genome-wide; imaging modality; imaging platform; in vivo; innovation; integration site; lapatinib; live cell imaging; malignant breast neoplasm; mouse model; neoplastic cell; novel; novel strategies; prospective; quantitative imaging; rare cancer; resistance gene; screening; targeted treatment; three dimensional cell culture; trait; tumor progression; validation studies

Project Summary Despite the impressive catalog of mutations amassed from sequencing human breast cancer genomes, most genomes decoded so far derive from early-stage disease, which likely leaves many genetic events driving disease progression undiscovered. Ongoing efforts to decode the more complex genomes from more advanced and drug-resistant breast cancers hold great promise, but the higher mutation burden in these cancers complicates the difficult task of distinguishing causative driver events from innocuous passengers events. Animal models that offer a genome-wide view of the mutations acquired during breast cancer progression can provide cross-species validation crucial for validating candidate driver genes. Drawing on the breast cancer genomics landscape and our own mouse modeling work, we formulated the hypothesis that breast cancer progression depends on driver mutations acquired in a disease stage- specific manner, obscured by continuously accruing passenger mutations. Based on this hypothesis, we developed the following long-term goal: discover novel drivers of breast cancer progression by performing genetic screens in mouse breast cancer models using timed mobilization of transposons (so-called “jumping genes”) at discrete disease stages. In unpublished preliminary studies, we engineered new mouse models for discovering and validating candidate mammary oncogenes. To enable oncogene discovery, we generated an inducible version of the Sleeping Beauty (SB) transposition system, which enables timed transposon mobilization in the mouse breast. We confirmed that this system provides an efficient cancer gene discovery platform by identifying known and novel oncogenes that cooperate with Wnt pathway activation to drive mammary tumorigenesis in vivo. To complement this high-throughput cancer gene discovery platform, we designed a novel strategy for efficiently validating candidate oncogenes and probing their mechanisms of action by monitoring the growth of mammary tissue fragments (organoids) grown in 3D culture. This strategy employs live-cell imaging for quantitative scoring of the oncogene-driven events that culminate in mammary cell overgrowth. We will address our hypothesis by completing three Specific Aims. In Aim 1, transposon-based gene discovery will be employed in the context of mouse models engineered to express known breast cancer- relevant oncogenes. Our goal is to identify novel candidate genes that drive the transition from mammary hyperplasia to focal mammary cancer. In Aim 2, delayed transposon mobilization will be initiated within established mammary cancers arising in the classic MMTV-Neu breast cancer model. Our goal is to identify candidate genes that drive resistance to Lapatinib, a clinically important drug that blocks Her2/Neu signaling. In Aim 3, we will optimize our live-cell imaging platform and apply it to test whether and how candidate cancer genes confer malignant capabilities to mammary cells grown in 3D culture.

项目摘要 尽管从人类乳腺癌基因组测序中积累了令人印象深刻的突变目录， 迄今为止解码的大多数基因组来自早期疾病，这可能会留下许多遗传事件 导致疾病进展未被发现。正在进行的努力，以解码更复杂的基因组，从更多的 晚期和耐药乳腺癌有很大的希望，但这些癌症中较高的突变负担， 癌症使区分致病的驾驶员事件和无害的乘客的困难任务复杂化 事件提供乳腺癌期间获得的突变的全基因组视图的动物模型 进展可以提供对验证候选驱动基因至关重要的跨物种验证。 利用乳腺癌基因组学的前景和我们自己的小鼠建模工作，我们制定了 乳腺癌进展取决于在疾病阶段获得的驱动突变的假设- 特定的方式，被不断积累的乘客突变所掩盖。基于这一假设，我们 制定了以下长期目标：通过执行，发现乳腺癌进展的新驱动因素。 使用转座子的定时动员（所谓的“跳跃”）在小鼠乳腺癌模型中进行遗传筛选 基因”）在不同的疾病阶段。 在未发表的初步研究中，我们设计了新的小鼠模型， 候选乳腺癌基因。为了发现癌基因，我们产生了一个诱导型的 睡美人（SB）转座系统，其使得能够在小鼠乳房中进行定时转座子动员。 我们证实，该系统提供了一个有效的癌症基因发现平台， 与Wnt通路活化协同驱动体内乳腺肿瘤发生的新癌基因。到 为了补充这个高通量癌症基因发现平台，我们设计了一种新的策略， 验证候选癌基因并通过监测乳腺癌的生长来探索其作用机制， 在3D培养中生长的组织碎片（类器官）。该策略采用活细胞成像进行定量 对导致乳腺细胞过度生长的癌基因驱动事件进行评分。 我们将通过完成三个具体目标来解决我们的假设。在目标1中，基于转座子的基因 这一发现将用于表达已知乳腺癌的小鼠模型， 相关癌基因。我们的目标是确定新的候选基因，驱动从乳腺癌的转变， 乳腺增生至局灶性乳腺癌。在目标2中，延迟转座子动员将在 在经典的MMTV-Neu乳腺癌模型中建立的乳腺癌。我们的目标是确定 候选基因驱动对拉帕替尼的耐药性，拉帕替尼是一种临床上重要的药物，可阻断Her 2/Neu信号传导。在 目标3，我们将优化我们的活细胞成像平台，并将其应用于测试候选癌症是否以及如何 基因赋予在3D培养中生长的乳腺细胞恶性能力。