cAMP signaling in melanoma pathogenesis: Evidence for multiple microdomains playi

描述（由申请人提供）：我在过去的十年里建立了cAMP微域研究的专业知识，特别是那些由新型蛋白可溶性腺苷酸环化酶（sAC）调节的微域。在我接受皮肤病学培训期间，我将我所掌握的技术和技能应用于对过度增殖角质细胞皮肤病的理解，并在最近的《皮肤病学调查杂志》上发表了一篇文章。在过去的一年里，我开始探索cAMP信号在黑色素瘤中的作用。我与纪念斯隆凯特琳癌症中心（Memorial Sloan Kettering Cancer Center， MSKCC）的黑色素瘤生物学专家Wolchok博士和Bastian博士进行了两次卓有成效的合作，并继续与sAC生物学专家Buck博士合作。Bastian博士和Buck博士将在我的培训期间担任我的导师，Wolchok博士将担任顾问。在接下来的五年里，我将有幸在世界一流的癌症研究机构MSKCC和领先的医学院威尔康奈尔医学中心（WCMC）接受培训，这两所大学都位于世界上最负盛名的研究角落之一（WCMC/MSKCC/Rockefeller），这使我能够获得前所未有的资源和机会。在接下来的五年里，我制定了一个计划，以进一步提高我在cAMP信号方面的专业知识，并发展我在黑色素瘤生物学方面的知识。我的短期计划是继续我的学业，以便成为一名皮肤科医生。我的长期目标是，在五年内，以独立调查员的身份获得R01基金。黑色素瘤是人类第六大常见癌症，也是最致命的癌症之一。一旦黑色素瘤扩散到皮肤以外，有效的医疗干预是有限的。虽然某些信号通路，如Ras/Raf/ERK通路，在控制黑色素瘤发病机制中有很好的作用，但最近的临床试验和基础科学报告表明，仅阻断该通路不足以治疗黑色素瘤。很明显，黑色素瘤有效治疗的发展依赖于对黑色素瘤信号复杂性的更好理解。cAMP与多种信号通路有关，这些信号通路是黑色素瘤发展和发病的关键，但在许多情况下，cAMP信号的来源和后果尚不清楚。最近，一类新的腺苷酸环化酶被称为可溶性腺苷酸环化酶（sAC），我参与了一系列新的小分子和基因抑制剂的开发，这些抑制剂能够区分sAC产生的和典型的g蛋白敏感跨膜腺苷酸环化酶（tmAC）产生的cAMP。在过去，我已经使用这些试剂建立了sAC在神经生长因子诱导的PC-12细胞Rap激活、netrin诱导的原代神经元生长锥趋化以及葡萄糖诱导的β细胞cAMP中的作用。在每一个案例中，cAMP的参与都是已知的，但缺乏一个合适的模型来解释它的作用。通过区分tmAC-与sac依赖的cAMP，已经开发出更连贯的模型。我已经将这些技术应用于黑色素瘤生物学，并发现当黑色素细胞获得细胞非典型性时，sAC蛋白在人类黑色素细胞增殖中迁移到细胞核中。在过去，我证明了核sAC激活cAMP反应性结合蛋白转录因子。我还发现sAC蛋白在黑色素瘤中上调，sAC抑制剂阻断黑素细胞生长。在这个提案中，我计划评估sAC和tmAC微域在黑色素瘤发病中的相对贡献。我预测，这条研究路线将导致黑色素瘤的新治疗靶点。

DESCRIPTION (provided by applicant): I have spent the last ten years building an expertise in the study of cAMP microdomains, specifically those regulated by the novel protein soluble adenylyl cyclase (sAC). During my training in dermatology, I applied the techniques and skills I have developed to the understanding of hyper proliferative keratinocyte skin disease, which led to a recent publication in the Journal of Investigative Dermatology. Over the past year, I began to explore the role of cAMP signaling in melanoma. I have developed two fruitful collaborations with experts in melanoma biology, Dr. Wolchok and Dr. Bastian at Memorial Sloan Kettering Cancer Center (MSKCC), and have continued my collaboration with an expert in sAC biology, Dr. Buck. Dr. Bastian and Dr. Buck will serve as my mentors during my training, and Dr. Wolchok will serve as an advisor. Over the next five years, I will have the privilege of being trained at both a world-class cancer research institution, MSKCC, and a leading medical college, Weill Cornell Medical Center (WCMC), both located at one of the most prestigious research corners in the world (WCMC/MSKCC/Rockefeller), proximity to which allows for unprecedented resources and exposure. Over the next five years, I have formulated a plan to further my expertise in cAMP signaling and develop my knowledge in melanoma biology. My short- term plan is to continue my education in order to become a physician scientist in dermatology. My long-term goal, in five years, is to secure R01 funding as an independent investigator. Melanoma is the sixth most common human cancer and one of the deadliest. Once melanoma spreads beyond the skin, effective medical interventions are limited. While certain signaling pathways, such as the Ras/Raf/ERK pathway, have well- established roles in controlling melanoma pathogenesis, recent clinical trials and basic science reports suggest that blocking this pathway alone is an insufficient treatment for melanoma. It has become apparent that the development of an effective treatment for melanoma relies on an improved understanding of the intricacies of melanoma signaling. cAMP has been implicated in a variety of signaling pathways that are key to the development and pathogenesis of melanoma, but in many cases the source and ramifications of cAMP signaling is poorly understood. A new class of adenylyl cyclases called soluble adenylyl cyclase (sAC) was recently characterized, and I was involved in developing a cadre of novel small molecule and genetic inhibitors capable of differentiating sAC-generated and the canonical G-protein sensitive transmembrane adenylyl cyclase (tmAC)-generated cAMP. In the past, I have used these reagents to establish the role of sAC in nerve growth factor-induced Rap activation in PC-12 cells, netrin-induced growth cone chemotaxis in primary neurons, and glucose-induced cAMP in beta cells. In each of these cases, involvement of cAMP was known, but a suitable model to explain its role was lacking. By differentiating tmAC- from sAC-dependent cAMP, more coherent models have been developed. I have applied these techniques to melanoma biology and have found that sAC protein migrates into the nucleus in human melanocytic proliferations as melanocytes acquire cellular atypical. In the past, I demonstrated that nuclear sAC activates the cAMP responsive binding protein transcription factor. I have also discovered that sAC protein is upregulated in melanoma, and inhibitors of sAC block melanocyte growth. In this proposal I plan to evaluate the relative contributions of sAC and tmAC microdomains in melanoma pathogenesis. I predict that this line of investigation will lead to novel therapeutic targets of melanoma.!