Genetic Analysis of FAK kinase and scaffold functions in breast cancer

乳腺癌中 FAK 激酶和支架功能的遗传分析

• 批准号: 8907919
• 负责人: JUN-LIN GUAN
• 金额: $ 32.81万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8907919
• 关键词: Affect; Alleles; Biological; Breast; Breast Cancer therapy; Cell Adhesion; Cells; Development; Disease; Exhibits; Focal Adhesion Kinase 1; Genes; Genomics; Goals; Growth Factor Receptors; Health; Human; Incidence; Integrins; Knock-in Mouse; Knock-out; MMP14 gene; Maintenance; Malignant Neoplasms; Mammary Neoplasms; Mammary Tumorigenesis; Mammary gland; Mediating; Modeling; Molecular; Mouse Mammary Tumor Virus; Mus; Mutant Strains Mice; Mutation; Neoplasm Metastasis; Oncogenes; Phosphorylation; Phosphotransferases; Play; Protein Tyrosine Kinase; Regulation; Role; Signal Pathway; Signal Transduction; Signaling Protein; Stem cells; Surface; Tumor Suppressor Genes; Tumorigenicity; Woman; antitumor effect; base; cancer stem cell; epithelial to mesenchymal transition; genetic analysis; genetic approach; in vivo; insight; malignant breast neoplasm; mammary epithelium; migration; mortality; mouse model; mutant; neoplastic cell; novel; scaffold; self-renewal; targeted treatment; tumor growth; tumor initiation; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): Breast cancer exhibits enormous cellular, genomic and biological complexity wherein multiple gene aberrations act to drive tumorigenesis and cancer progression. The long term goal of the proposed studies is to understand the cellular and molecular mechanisms in the regulation of breast cancer development and metastasis. Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase that plays a major role in mediating signal transduction by integrins as well as growth factor receptors in the regulation of cell adhesion, migration, survival, proliferation and differentiation in a variety of cells. FAK has been implicated in human breast cancer as well as other malignancies. However, the cellular and molecular mechanisms by which FAK promotes mammary tumorigenesis in vivo are still not well understood. We showed recently that conditional knockout of FAK in the mammary epithelium suppressed mammary tumorigenesis and progression by affecting MaCSCs in a well characterized breast cancer mouse model. To investigate the unique role of FAK as both a tyrosine kinase and a scaffold in intracellular signaling in breast cancer, we have recently created two novel FAK knock-in mutant mice with the kinase-defective allele (FAK +/KD mice) or the P878/881A mutation allele (FAK +/PA mice) to disrupt its signaling through tyrosine kinase activity or specific scaffolding function for endophilin A2 phosphorylation by Src and regulation of MT1-MMP, respectively. Preliminary analyses of the PA mutant knockin mice using the MMTV-PyMT model (PA/PA-MT mice) revealed that FAK mediated endophilin A2 phosphorylation by Src plays an important role in mammary tumor growth and metastasis by promoting epithelial to mesenchymal transition (EMT) of mammary tumor cells, and by maintaining the content and tumorigenicity of MaCSCs. In addition, we identified a function of FAK in the maintenance of normal MaSCs and found that kinase-independent functions of FAK were able to promote self-renewal of MaSCs. Lastly, we also obtained preliminary results suggesting an important role of FAK in regulation of human MaCSCs, which is consistent with our more extensive findings in mouse models. Based on these preliminary and previous studies, we propose to 1). determine the mechanism of FAK scaffold function in mammary tumor growth and metastasis through regulation of EMT and MaCSCs, 2). examine the role of FAK kinase activity and dissect the downstream kinase-dependent and -independent signaling pathways in breast cancer development and progression, and 3). explore the strategies of targeting FAK scaffold and kinase functions in MaCSCs for breast cancer therapy. Together, these studies will provide significant insights into the molecular and cellular mechanisms of breast cancer that may contribute to novel therapies for this devastating disease.

描述（由申请人提供）：乳腺癌表现出巨大的细胞，基因组和生物学复杂性，其中多重基因畸变作用于驱动肿瘤发生和癌症的进展。拟议研究的长期目标是了解乳腺癌发育和转移调节的细胞和分子机制。局灶性粘附激酶（FAK）是一种细胞质酪氨酸激酶，它在整合素的信号转导以及生长因子受体在调节细胞粘附，迁移，生存，增殖和分化的各种细胞中起主要作用。 FAK与人类乳腺癌以及其他恶性肿瘤有关。然而，FAK在体内促进乳腺肿瘤发生的细胞和分子机制仍然不太了解。我们最近表明，在乳腺癌小鼠模型中影响MACSC，乳腺上皮中FAK的条件敲除抑制了乳腺肿瘤的发生和进展。为了调查FAK作为酪氨酸激酶和乳腺癌细胞内信号传导中的脚手座的独特作用，我们最近创建了两只新型的FAK敲击突变小鼠，其激酶缺陷等位基因（FAK +/KD小鼠）（FAK +/KD小鼠）或p878/881a突变（通过p878/881a突变）通过Tyros or Scaftil（Fak +/Pa）进行了挑战通过SRC和MT1-MMP的调节A2磷酸化。使用MMTV-PYMT模型（PA/PA/PA-MT小鼠）对PA突变型敲击蛋白小鼠进行初步分析表明，SRC通过SRC介导的内粒蛋白A2磷酸化介导的乳腺肿瘤生长和转移中的重要作用，通过促进上皮细胞（通过维护MESANDIG TAMOR）和TUMOR TUMOR和TUMOR TUMOR和TUMOR TUMOR和TUMOR TUMOR和TUMOR的乳腺症和转移作用。 MacScs。此外，我们确定了FAK在维持正常MASC中的功能，发现FAK的激酶无依赖性功能能够促进MASC的自我更新。最后，我们还获得了初步结果，这表明FAK在人类MacSC的调节中起着重要作用，这与我们在小鼠模型中更广泛的发现一致。基于这些初步和以前的研究，我们建议1）。通过调节EMT和MACSC，2）确定FAK支架功能在乳腺肿瘤生长和转移中的机制。检查FAK激酶活性的作用，并剖析下游激酶依赖性和非依赖性信号通路在乳腺癌发育和进展中以及3）。探索靶向MACSC中FAK支架和激酶功能的策略进行乳腺癌治疗。总之，这些研究将为乳腺癌的分子和细胞机制提供重大见解，这可能有助于这种毁灭性疾病的新疗法。