Non-canonical Notch1 regulation of proliferation and adherens junctions in breast cancer

项目摘要 人类乳腺肿瘤的全基因组测序揭示了乳腺癌组织中显著的分子异质性。 疾病然而，个性化乳腺癌治疗的进步需要更好的理解， 特定的遗传改变如何促进细胞行为，这些细胞行为是疾病发生和发展的基础。 乳腺癌我们最近发现了Notch1受体的一种新的、不依赖于转录的功能 在调节乳腺上皮细胞增殖和粘附连接组织中的作用。该提案将利用 一种跨学科的方法，将3D组织工程人类乳腺导管平台与 分子和遗传工具来剖析癌症增殖信号通路，并将建立一个以前的 在乳腺癌中的肿瘤抑制Notch1通路。在K99阶段（目标1），我们将确定 Notch1在乳腺粘连连接的转录非依赖性调节中的结构域特异性作用， 皮质细胞骨架，由这种非经典Notch1信号传导控制的信号传导和增殖途径， 并证明了非转录依赖性NOTCH1功能丧失在乳腺癌异种移植物中的作用 模型在R00阶段，我们将在乳腺癌的背景下构建肿瘤抑制Notch1功能， 接触抑制增殖，并确定Notch1的分子机制和机制， 在乳腺组织生长期间在粘附连接处活化（Aim 2）。与此同时，我们将进一步利用 仿生乳腺导管模型探索两种主要复发乳腺的不同形态发生表型 癌症突变，并测试临床活性药物在肿瘤进展的每个阶段的疗效（目标3）。 这项拟议的研究将确定NOTCH1功能丧失突变在人类乳腺癌中的作用， 在携带此类突变的患者中的治疗靶点，并建立一种新的乳腺癌模型策略 在3D仿生培养物中进行治疗和评估。我将获得微流体方面的研究培训， 体外组织工程，以及癌症信号，病理学和体内小鼠建模，而 同时通过赠款编写、指导和领导能力方面的培训，加强职业发展。我 我组建了一个特殊的，互补的指导团队，以帮助我实现我的研究和职业生涯 目标：克里斯托弗·陈博士，器官型组织建模和细胞力学方面的专家，将是我的主要导师 和Andrea McClatchey博士（MGH癌症中心/哈佛），细胞骨架调节的国际领导者， 肿瘤发生，肿瘤抑制信号，和癌症建模，将是我的共同导师。体制 波士顿大学生物设计中心提供的环境非常适合这项提议， 为生物学家、临床医生和工程师之间的科学讨论和合作提供了机会， 职业发展。总之，建议的学习和职业发展培训将确保我实现我的目标。 目标是建立一个成功的，独立资助的实验室，研究组织的基本机制， 形态发生和肿瘤发生。

Project Summary Genome-wide sequencing of human breast tumors has revealed the remarkable molecular heterogeneity of the disease. However, the advancement of personalized breast cancer therapies requires a greater understanding of how specific genetic alterations contribute to the cellular behaviors that underlie the onset and development of breast cancer. We have recently identified a novel, transcription-independent function of the Notch1 receptor in the regulation of mammary epithelial proliferation and adherens junction organization. This proposal will utilize an interdisciplinary approach that combines a 3D tissue engineered human mammary duct platform with molecular and genetic tools to dissect cancer proliferative signaling pathways and will establish a previously undescribed, tumor suppressive Notch1 pathway in breast cancer. During the K99 phase (Aim 1), we will identify domain-specific roles of Notch1 in the transcription-independent regulation of mammary adherens junctions and cortical cytoskeleton, the signaling and proliferative pathways controlled by this non-canonical Notch1 signaling, and demonstrate the effects of transcription-independent NOTCH1 loss-of-function in breast cancer xenograft models. During the R00 phase, we will frame tumor suppressive Notch1 function in the context of mammary contact inhibition of proliferation and identify the molecular mechanisms and mechanics by which Notch1 is activated at adherens junctions during mammary tissue growth (Aim 2). In parallel, we will further leverage our biomimetic mammary duct model to explore to the distinct morphogenic phenotypes of two major recurring breast cancer mutations and test the efficacy of clinically active drugs at each stage of their tumor progression (Aim 3). The proposed research will define effects of NOTCH1 loss-of-function mutations in human breast cancer, inform therapeutic targets in patients harboring such mutations, and establish a new strategy to model breast cancer progression and assess therapies in 3D biomimetic cultures. I will gain research training in microfluidic-based, in vitro tissue engineering, as well as cancer signaling, pathology, and in vivo mouse modeling, while simultaneously enhancing career development through training in grant writing, mentoring, and leadership. I have assembled an exceptional, complementary mentoring team to help me achieve my research and career goals: Dr. Christopher Chen, expert in organotypic tissue modeling and cell mechanics, will be my primary mentor and Dr. Andrea McClatchey (MGH Cancer Center/Harvard), an international leader in cytoskeletal regulation of tumorigenesis, tumor suppressor signaling, and cancer modeling, will be my co-mentor. The institutional environment provided by the Biological Design Center at Boston University is ideally suited for this proposal and offers opportunities for scientific discussion, collaboration between biologists, clinicians, and engineers, and career development. Together, the proposed studies and career development training will ensure I achieve my goal of establishing a successful, independently-funded laboratory studying underlying mechanisms of tissue morphogenesis and tumorigenesis.