Tropomyosin and tyrosine kinases in mechanics of cancer

原肌球蛋白和酪氨酸激酶在癌症机制中的作用

基本信息

批准号：
9247873
负责人：
MICHAEL Patrick SHEETZ
金额：
$ 30.37万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9247873
关键词：
Actin-Binding Protein Actinin Actins Address Adhesions Adhesives Affect Apoptosis Behavior Binding Binding Proteins Caliber Cell-Matrix Junction Cells Complex Contractile Proteins Cytoskeletal Proteins Dependence Development EGF gene Fibronectins Fostering Generations Goals Grant Growth Integrins Link Malignant Neoplasms Measures Mechanics Microfilaments Modification Molecular Molecular Weight Muscle Muscle Proteins Myosin ATPase Myosin Type II Natural regeneration Neoplasm Metastasis Pathway interactions Pattern Phenotype Phosphotransferases Play Process Protein Tyrosine Kinase Proteins Psychological reinforcement Recruitment Activity Role Sarcomeres Signal Transduction Site Surface Testing Time Tropomyosin Vinculin Work alpha Actinin cancer cell cell growth cell transformation cell type knock-down mechanotransduction nanofabrication paxillin polymerization public health relevance restoration sensor submicron tropomodulin tropomyosin kinase tumor growth

项目摘要

DESCRIPTION (provided by applicant): The major goal of our lab is to understand the molecular pathways involved in rigidity and force sensing at cell-matrix adhesions. In this grant we will explore how these pathways are altered in cancers related to depletion of tropomyosin and modifications of tyrosine kinases. Cancer cells typically show anchorage independence of growth and the cytoskeletal protein, tropomyosin (Tm), is depleted in many cancers. Further, restoration of normal Tm1 expression reverses the transformed phenotype. Recently our lab has shown that cell rigidity sensing depends upon local contraction units that displace matrix by 50-70nm and if a threshold force is exceeded rapidly, then rigid adhesions form. Local contraction units resemble muscle sarcomeres in size (~2 �, function, and composition (actin, myosin II, alpha-actinin, tropomodulin and tropomyosin). After knockdown of Tm1, the local contractions are dramatically altered and the cells no longer sense the rigidity of fibronectin-coated substrates. Similarly, the knockdown of tyrosine kinases alters both rigidity sensing and the pattern of contractions that are measured from displacements of 500nm diameter PDMS pillars. We propose now to follow the time course of force dependence and the concentration and dissipation of adhesion and contractile proteins at the pillars. This will tell us the order of binding and provide clues about the molecular steps involved in the cycles of contraction and release. We will then address the question of how the tyrosine kinases involved in rigidity sensing (AXL, ROR2 and EGF) interplay with the early adhesion complexes as a function of the force on the complexes. Since tropomyosin inhibits transformation and tumor growth, we will determine how tropomyosin depletion alters the pattern of adhesion maturation on soft surfaces that enables transformation. Thus, we will be able to better understand the mechanochemical basis of transformation.

描述（由适用提供）：我们实验室的主要目标是了解在细胞矩阵依从性下涉及刚性和力敏感性的分子途径。在这笔赠款中，我们将探讨与肿瘤耗尽和酪氨酸激酶修饰有关的癌症中这些途径的改变。癌细胞通常显示出生长的锚定独立性，并且在许多癌症中耗尽了细胞骨架蛋白，Tropomyosin（TM）。正常TM1表达的恢复会逆转转化的表型。最近，我们的实验室表明，细胞刚度感测取决于局部收缩单元，该单元将基质置于50-70nm，如果超过了阈值，则刚性粘附形式。局部收缩单位类似于大小的肌肉棒球（〜2�，功能和组成（肌动蛋白，肌球蛋白II，α-肌动蛋白，α-肌动蛋白，tropomodulin和tropomyosin）。敲除TM1后，局部收缩在动态上改变了局部收缩，并且不再感知细胞的固件。收缩的模式是从直径为500nm的PDMS支柱中测量的。结合并提供有关收缩和释放循环中涉及的分子步骤的线索。然后，我们将解决与早期粘合剂复合物相互作用的酪氨酸激酶如何与早期粘合剂复合物相互作用，这是对复合物上力的函数的函数。由于肌球蛋白抑制了转化和肿瘤的生长，因此我们将确定肌球蛋白耗竭如何改变在软表面上粘合剂成熟的模式，从而使转化。这，我们将能够更好地理解转换的机械化学基础。