Dissecting the role of invadopodia-mediated degradation in metastasis

 描述(申请人提供)：转移是癌症患者死亡的主要原因。它是癌细胞从原发肿瘤扩散到远处器官的过程，这个过程需要降解基底膜和细胞外基质(ECM)的能力。肿瘤细胞通过形成称为侵袭足的膜突起来获得这种能力。它们是富含肌动蛋白的亚细胞结构，含有许多调节和适配蛋白，以招募金属蛋白酶，如MMP2和MMP9，MT1-MMPs，进行细胞外基质的降解。我们过去的工作表明，侵袭足在肿瘤转移中是必不可少的。鉴于这一想法，靶向肿瘤细胞中的内陷形成是一种潜在的抗转移治疗方法。过去抗转移治疗的研究主要集中在针对MMPs，因为它们在ECM降解中起直接作用。尽管基质金属蛋白酶抑制剂在临床前试验中显示了良好的效果，但由于广泛的抑制和毒性，它们在临床试验中失败了。靶向MMPs的另一种选择是靶向参与介体组装的蛋白质。在这项提议中，我的目标是通过进行免疫沉淀实验和质谱分析，更好地了解内侧足纲的组装以及蛋白酶是如何被招募到内侧足纲的。此外，还将进行降解分析、免疫细胞化学和体内研究，以了解已识别的蛋白质在介导性转移中的作用。

 DESCRIPTION (provided by applicant): Metastasis is the main cause of death in cancer patients. It is the spread of cancer cells from the primary tumor to distant organs and this process requires the ability to degrade the basement membrane and extracellular matrix (ECM). Tumor cells obtain this ability through the formation of membrane protrusions called invadopodia. They are actin-rich subcellular structures that contain a number of regulatory and adaptor proteins in order to recruit proteases, such as metalloproteinases, MMP2 and MMP9, MT1-MMP, to carry out ECM degradation. Our past work demonstrates that invadopodia are essential for tumor metastasis. Given this idea, targeting invadopodia formation in tumor cells is a potential anti-metastasis treatment. Past research on anti-metastasis therapy focused on targeting MMPs because of their direct role in ECM degradation. Although MMP inhibitors demonstrated promising results in preclinical trials, they failed in clinical trials due to broad spectrum of inhibition and toxicity. An alternative to targeting MMPs is to target proteins that ar involved in invadopodia assembly. In this proposal, I aim to better understand invadopodia assembly and how proteases are recruited to invadopodia by performing immunoprecipitation experiments followed by mass spectrometry. In addition, degradation assays, immunocytochemistry, and in vivo studies will be performed to understand the role of the identified protein in invadopodia-mediated metastasis.