Novel Animal Models for Functional Analysis of Protein Phosphorylation in Breast Cancer Progression.

用于乳腺癌进展中蛋白质磷酸化功能分析的新型动物模型。

基本信息

批准号：
9022147
负责人：
ZONGHAN DAI
金额：
$ 16.64万
依托单位：
TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9022147
关键词：
Actins Affect Animal Model Biological Models Breast Cancer Cell Breast Cancer Patient Breast cancer metastasis Cancer Cell Growth Cell physiology Cytoskeleton Data Set Development Epidermal Growth Factor Epidermal Growth Factor Receptor Event Figs - dietary Gene Expression Gene Silencing Genes Genetic Recombination Goals Growth In Vitro Knock-in Link Malignant Neoplasms Mediating Metastatic breast cancer Metastatic malignant neoplasm to brain Modification Molecular Mutation Mutation Analysis Neoplasm Metastasis Phosphorylation Phosphotransferases Principal Investigator Prognostic Marker Protein Analysis Protein Tyrosine Kinase Proteins Public Health Research Role Signal Transduction Site System Technology Testing Therapeutic Time Translating Tyrosine Tyrosine Phosphorylation Xenograft procedure base cancer proteomics cost effective design gene replacement high throughput analysis improved in vivo innovation malignant breast neoplasm mimetics molecular targeted therapies mouse model mutant novel novel strategies overexpression personalized medicine phosphoproteomics prognostic programs public health relevance screening small hairpin RNA src-Family Kinases targeted treatment tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Recent advances in cancer proteomics have allowed for systemic identification of the phosphorylation events associated with cancer progression. However, validating and translating these large datasets from in vitro to in vivo systems remains a major challenge. The long-term goal of our research is to understand how the protein phosphorylation events induced by Abelson (ABL) tyrosine kinases contribute to breast cancer development. The objective of this exploratory study is to develop a streamlined model system for rapid in vivo analysis of the function of ABL-mediated protein phosphorylation in breast cancer growth and metastasis. We have recently developed a strategy to conditionally replace an endogenous gene expression with the expression of its mutant counterpart by using short hairpin RNA-based technology. We hypothesize that, by using this strategy combined with the recombination-mediated cassette exchange (RMCE) technology, a streamlined system can be developed to provide a fast, reversible, and cost-effective way to analyze the function of a given phosphorylation-deficient or -mimetic mutant in breast cancer development. We will test this hypothesis by defining the role of tyrosine phosphorylation of ABL interactor 1 (Abi1), an ABL substrate and key regulator of actin cytoskeleton remodeling, in breast cancer development. The rationale behind this research is that once this system is developed, we will be able to quickly analyze the function of a large set of ABL-mediated phosphorylation events obtained from cancer proteomic studies and identify those that are essential for breast cancer development for prognostic and therapeutic purposes. In specific aim 1, a set of conditional Abi1 gene silencing/re-expressing cassettes will be developed to analyze how ABL-mediated Abi1 tyrosine phosphorylation affects breast cancer cell function and signaling. In aim 2, how the ABL-mediated Abi1 phosphorylation contributes to breast cancer progression in vivo will be determined by using conditional gene replacement strategy combined with a brain metastasis mouse model recently developed in our lab. This project is innovative because it proposes a new strategy different from conventional gene knockin for in vivo protein mutation analysis. If successful, this line of research will enable a high-throughput function analysis of protein mutation in breast cancer development. Identifying ABL-mediated phosphorylation events essential for breast cancer development will not only help us to understand how ABL kinases contribute to breast cancer development, but also provide prognostic markers for selection of breast cancer patients who may benefit from targeted molecular therapy.

描述（由申请人提供）：癌症蛋白质组学的最新进展已经允许系统地鉴定与癌症进展相关的磷酸化事件，然而，从体外到体内系统验证和翻译这些大数据集仍然是一个重大挑战。我们研究的目标是了解 Abelson (ABL) 酪氨酸激酶诱导的蛋白质磷酸化事件如何促进乳腺癌的发展。这项探索性研究的目的是开发一个简化的模型系统。为了快速体内分析 ABL 介导的蛋白质磷酸化在乳腺癌生长和转移中的功能，我们最近开发了一种策略，通过使用基于短发夹 RNA 的技术，用其突变体对应物的表达有条件地替换内源基因表达。我们发现，通过使用这种策略与重组介导的盒式交换（RMCE）技术相结合，可以开发出一种简化的系统，以提供一种快速、可逆且经济有效的方法来分析给定的功能我们将通过定义 ABL 相互作用子 1 (Abi1) 的酪氨酸磷酸化在乳腺癌发展中的作用来检验这一假设，Abi1 是 ABL 底物和肌动蛋白细胞骨架重塑的关键调节因子。研究表明，一旦这个系统被开发出来，我们将能够快速分析从癌症蛋白质组学研究中获得的大量 ABL 介导的磷酸化事件的功能，并识别那些在特定目标 1 中，将开发一组条件 Abi1 基因沉默/重新表达盒，以分析 ABL 介导的 Abi1 酪氨酸磷酸化如何影响乳腺癌细胞功能和信号传导。在目标2中，将通过使用条件基因替换策略结合最近开发的脑转移小鼠模型来确定ABL介导的Abi1磷酸化如何促进体内乳腺癌进展我们实验室的这个项目是创新的，因为它提出了一种不同于传统基因敲入的体内蛋白质突变分析的新策略，如果成功，这一系列研究将能够对乳腺癌发展中的蛋白质突变进行高通量功能分析。 ABL 介导的对乳腺癌发展至关重要的磷酸化事件不仅有助于我们了解 ABL 激酶如何促进乳腺癌的发展，而且还为选择可能受益于靶向分子治疗的乳腺癌患者提供预后标志物。