PAK Kinases Regulate Melanoma Chemoresistance and Metastasis

描述（由申请人提供）：黑色素瘤肿瘤在发展的早期阶段转移，并且众所周知对化疗药物具有抗性。我们最近利用系统水平的RNAi筛选方法来揭示黑色素瘤化疗耐药性的分子调节因子。这些研究确定RhoJ及其下游激酶PAK 1抑制黑色素瘤细胞中感知DNA损伤的途径。进一步的研究表明，RhoJ和PAK 1还通过调节肌动蛋白细胞骨架动力学来调节黑色素瘤侵袭。这些观察使我们假设PAK激酶磷酸化下游靶点，其抑制DNA损伤传感并促进黑色素瘤转移。在这个提议中，我们确定PAK 1如何抑制DNA损伤传感，并确定Pak 1是否抑制体内DNA损伤传感。进一步的研究将确定PAK激酶调节黑色素瘤细胞侵袭的靶点，并验证PAK激酶在体内调节黑色素瘤转移。在该提案的最终目的中，我们将检查PAK激酶是否在预后不良/化疗耐药的人黑色素瘤中被选择性激活，并确定PAK抑制剂是否可以使小鼠黑色素瘤对化疗药物敏感并抑制小鼠黑色素瘤转移。这些研究的完成将导致更有效的PAK抑制剂的设计，可用于治疗转移性黑色素瘤。

DESCRIPTION (provided by applicant): Melanoma tumors metastasize at an early stage of development and are notoriously resistant to chemotherapeutics. We recently utilized a systems-level RNAi screening approach to uncover the molecular regulators of melanoma chemoresistance. These studies determined that RhoJ and its downstream kinase PAK1 suppress pathways in melanoma cells that sense DNA damage. Additional studies revealed that RhoJ and PAK1 also regulate melanoma invasion by modulating actin cytoskeletal dynamics. These observations led us to hypothesize that PAK kinases phosphorylate downstream targets that suppress DNA damage sensing and promote melanoma metastasis. In this proposal, we determine how PAK1 suppresses DNA damage sensing and determine whether Pak1 suppresses DNA damage sensing in vivo. Additional studies will identify PAK kinase targets that modulate melanoma cell invasion and verify that PAK kinases regulate melanoma metastasis in vivo. In the final aim of the proposal, we will examine whether PAK kinases are selectively activated in poor prognosis/chemoresistant human melanomas and determine whether PAK inhibitors can both sensitize mouse melanomas to chemotherapeutics and inhibit murine melanoma metastasis. Completion of these studies will lead to the design of more effective PAK inhibitors that can be used to treat metastatic melanoma.