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 疾病的传播。尽管黑色素瘤形成的遗传基础已经得到很好的证实 黑色素瘤的转移性进展是由遗传事件还是表观遗传事件驱动的，目前尚不清楚 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筛查，确定了10个表观遗传学 调节剂是黑色素瘤转移的驱动因素。在这份提案中，我们将重点放在最热门的作品之一- UCHL5-是一种脱泛素酶，也是INO80染色质的哺乳动物特异性亚基 重塑综合体。利用体外和体内验证实验，我们已经验证了 UCHL5促进黑色素瘤侵袭和转移。重要的是，我们的初步研究 表明UCHL5涉及其脱泛素酶活性以及它与INO80在 宣扬侵略。此外，我们发现UCHL5可能调节重要的促转移 RhoA-LIMK-cofilin和Sit-Robo等信号通路通过直接调节 ARHGAP29和SLIT2。因此，这项建议的目的是研究 UCHL5在黑色素瘤发生中的作用及其对表观基因组重编程的调控 转移。在第一个目标中，我们提出了UCHL5介导的表观基因组的特征 使用人类黑色素瘤培养物结合动物模型和切割进行重新编程- 边缘表观基因组技术。我们将评估核小体位置的动力学和 用UCHL5重新编程超级增强子元件并评估INO80的贡献 YAP1，我们已经确定它是UCHL5的一个新的相互作用元件。在第二个目标中，我们将 研究UCHL5下游信号事件，重点研究ARHGAP29介导的调控 RhoA-LIMK-cofilin信号和肌动蛋白动力学的研究。在第三个目标中，我们建议研究 UCHL5抑制剂单独及与现行标准联合应用的治疗效果 转移性黑色素瘤的临床前转基因动物模型的CARE治疗。 总之，我们的建议不仅将为表观遗传驱动机制提供新的见解 黑色素瘤转移，但也提供了UCHL5抑制剂用于 转移性黑色素瘤的治疗策略。

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.