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

Notch1 对乳腺癌增殖和粘附连接的非经典调控

基本信息

批准号：
10328889
负责人：
Matthew L Kutys
金额：
$ 24.9万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10328889
关键词：
3-Dimensional Actomyosin Adherens Junction Architecture Behavior Biochemical Biological Biological Models Biomimetics Boston Breast Breast Cancer Model Breast Cancer Patient Breast Cancer cell line Breast Cancer therapy Breast Epithelial Cells CRISPR/Cas technology Cancer Center Cancer Model Cell Proliferation Cell Shape Cell-Cell Adhesion Cells Collaborations Complex Contact Inhibition Crowding Cytoskeletal Modeling Cytoskeleton Development Disputes Duct (organ) structure Ductal Epithelium ERBB2 gene Endothelium Engineering Ensure Environment Epithelial Cell Proliferation Epithelial Cells Funding Future Genetic Transcription Goals Grant Growth Growth Factor Human Human Engineering In Vitro International Laboratory Study Leadership Link Malignant Neoplasms Mammary Duct Mammary Neoplasms Mammary Tumorigenesis Mechanics Mediating Mentors Microfabrication Microfluidics Modeling Molecular Molecular Genetics Morphogenesis Mutation NOTCH1 gene Noninfiltrating Intraductal Carcinoma Oncogenic Pathology Pathway interactions Patients Pattern Pharmaceutical Preparations Pharmacology Phase Phenotype Predisposition Pregnancy Process Puberty Receptor Signaling Regulation Research Research Training Role Shapes Signal Pathway Signal Transduction Study models System Testing Therapeutic Tissue Engineering Tissue Model Tissues Training Transcriptional Regulation Tumor Suppressor Proteins Tumor stage Universities Work Writing Xenograft Model base breast cancer progression cancer type career career development cell behavior clinical efficacy clinically relevant design disease heterogeneity efficacy testing genome-wide in vivo inhibitor insight interdisciplinary approach loss of function loss of function mutation malignant breast neoplasm mammary mammary epithelium mechanical stimulus mouse model notch protein novel prognostic value programs receptor response stem cell self renewal therapeutic target tool tumor tumor initiation tumor progression tumorigenesis

项目摘要

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.

项目概要人类乳腺肿瘤的全基因组测序揭示了显着的分子异质性疾病。然而，个性化乳腺癌治疗的进步需要更多的了解研究特定的基因改变如何影响作为发病和发展基础的细胞行为乳腺癌。我们最近发现了 Notch1 受体的一种新颖的、不依赖于转录的功能调节乳腺上皮增殖和粘附连接组织。本提案将利用一种跨学科方法，将 3D 组织工程人体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，并将建立先前的未描述的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 在乳腺粘附连接的转录独立调节中的域特异性作用皮质细胞骨架，由这种非典型 Notch1 信号控制的信号传导和增殖途径，并证明了转录独立的 NOTCH1 功能丧失对乳腺癌异种移植物的影响模型。在 R00 阶段，我们将在乳腺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过程中在粘附连接处激活（目标 2）。与此同时，我们将进一步利用我们的仿生乳腺导管模型探索两种主要复发性乳房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每个阶段的功效（目标 3）。拟议的研究将定义 NOTCH1 功能丧失突变对人类乳腺癌的影响，告知携带此类突变的患者的治疗靶点，并建立乳腺癌模型的新策略 3D 仿生文化中的进展和评估治疗。我将获得基于微流体的研究培训，体外组织工程，以及癌症信号传导、病理学和体内小鼠模型，同时同时通过资助写作、指导和领导力方面的培训来促进职业发展。我组建了一支出色的、互补的指导团队来帮助我实现我的研究和职业生涯目标：Christopher Chen博士，器官组织建模和细胞力学专家，将是我的主要导师 Andrea McClatchey 博士（麻省总医院癌症中心/哈佛大学），细胞骨架调控领域的国际领导者肿瘤发生、肿瘤抑制信号传导和癌症建模将是我的共同导师。机构波士顿大学生物设计中心提供的环境非常适合该提案，并且为生物学家、临床医生和工程师之间的科学讨论和合作提供机会，职业发展。总之，拟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，研究组织的潜在机制形态发生和肿瘤发生。