UCHL5 as a regulator and therapeutic target in metastatic melanoma

黑色素瘤是一个主要的世界健康问题，大多数死亡是由于转移性黑色素瘤。 疾病的传播。虽然黑色素瘤形成的遗传基础已经很好地建立 黑色素瘤的转移进展是否由遗传或表观遗传事件驱动， 不清楚假设表观遗传事件可能是转移的关键驱动力， 进展，我们进行了体内功能获得性筛选，确定了10个表观遗传 调节因子作为黑色素瘤转移的驱动因子。在这份提案中，我们关注的是热门之一- UCHL 5-是一种去泛素化酶，是Ino 80染色质的哺乳动物特异性亚基 重塑情结使用体外和体内验证实验，我们已经验证了 UCHL 5促进黑色素瘤侵袭和转移。重要的是，我们的初步研究 显示UCHL 5涉及其去泛素化酶活性以及其与Ino 80结合， 促进入侵。此外，我们发现UCHL 5可能调节重要的促转移因子， 信号通路如RhoA-LIMK-Cofilin和SLIT-ROBO信号通路通过直接调节 ARHGAP 29和SLIT 2。因此，本建议的目的是审查 UCHL 5及其调控表观基因组重编程在黑色素瘤中的作用 转移在第一个目标中，我们提出表征UCHL 5介导的表观基因组 使用人类黑色素瘤培养物结合动物模型和切割- 边缘表观基因组技术。我们将评估核小体位置的动态， 通过UCHL 5重编程超增强元件，并评估Ino 80的贡献 作为YAP 1，我们已经确定为UCHL 5的新相互作用物。第二个目标，我们将 研究UCHL 5下游的信号传导事件，重点关注ARHGAP 29介导的调控 RhoA-LIMK-Cofilin信号传导和肌动蛋白动力学。在第三个目标中，我们建议审查 UCHL 5抑制单独以及与目前的标准药物组合的治疗效用 在转移性黑色素瘤的临床前转基因动物模型中护理黑色素瘤疗法。 总之，我们的提案不仅将为表观遗传机制提供新的见解， 黑色素瘤转移，而且还提供了使用UCHL 5抑制剂作为 转移性黑素瘤的治疗策略。

Melanoma is a major world health problem with most deaths occurring due to metastatic spread of the disease. Although genetic basis of melanoma formation has been well established whether metastatic progression of melanoma is driven by genetic or epigenetic events remains unclear. With a hypothesis that epigenetic events may be a key driving force for metastatic progression, we performed an in vivo gain-of-function screen that identified 10 epigenetic regulators as drivers of melanoma metastasis. In this proposal, we focus on one of the top hits - UCHL5 - which is a deubiquitinase and a mammalian specific subunit of Ino80 chromatin remodeling complex. Using in vitro and in vivo validation experiments, we have verified role of UCHL5 in promoting melanoma invasion and metastasis. Importantly, our preliminary studies show that UCHL5 involves its deubiquitinase activity as well as its association with Ino80 in promotion of invasion. In addition, we find that UCHL5 may regulate important pro-metastatic pathways such as RhoA-LIMK-Cofilin and SLIT-ROBO signaling via direct regulation of ARHGAP29 and SLIT2 respectively. Therefore, the objective of this proposal is to examine the contribution of UCHL5 and its regulation of epigenome reprogramming in driving melanoma metastasis. In the first aim, we propose to characterize UCHL5 mediated epigenome reprogramming using human melanoma cultures in conjunction with animal models and cutting- edge epigenomic technologies. We will assess dynamics of nucleosome position and reprogramming of superenhancer elements by UCHL5 and assess contribution of Ino80 as well as YAP1, which we have identified as a novel interactor of UCHL5. In the second aim, we will study signaling events downstream of UCHL5 with a focus on ARHGAP29 mediated regulation of RhoA-LIMK-Cofilin signaling and actin dynamics. In the third aim, we propose to examine therapeutic utility of UCHL5 inhibition alone as well as in combination with current standard-of- care melanoma therapies in pre-clinical transgenic animal models of metastatic melanoma. Together, our proposal will not only provide new insights into the epigenetic mechanisms driving melanoma metastasis but also provide proof-of-concept evidence for use of UCHL5 inhibitors as a therapeutic strategy in metastatic melanoma.