The role of melanocyte precursors in zebrafish pigmentation disorders

描述（由申请人提供）：长期以来，我的临床和研究训练都是为了成为一名内科科学家，能够弥合临床观察和基础科学方法之间的差距，解决这些问题。这个申请是我在这方面成长的关键部分，并将确保我有必要的资源来实现这一目标。项目总结：色素黑素细胞的紊乱是导致许多人类疾病的原因，从白癜风的色素沉着到恶性黑色素瘤的致命后果。了解神经嵴及其衍生物（包括黑素细胞）发育的遗传途径是治疗这些谱系疾病的关键组成部分。在这个提案中，我计划使用斑马鱼来了解胚胎神经嵴和黑素细胞前体正常发育的扰动如何与成人色素沉着疾病（如痣和黑色素瘤）相关。我们开发了转基因BRAF；P53突变鱼胚胎形态正常，但成年后有严重的色素沉着破坏和黑色素瘤。基因表达分析表明BRAF之间存在保守的基因特征；P53胚胎和肿瘤，提示胚胎发生早期事件和肿瘤形成之间的联系。在Specific Aim 1中，我利用这些信息作为化学筛选的基础，以确定与预防黑色素瘤形成起始事件相关的途径和分子。从这个屏幕中出现的点击被测试其改变成人BRAF的能力；p53表现型。一种名为MLT013的药物的初步数据表明，它抑制了色素条纹的破坏以及黑色素瘤的可移植性。MLT013可能通过内皮素受体阻断起作用。在Aim 2中，我描述了确定内皮素是否与BRAF合作驱动黑色素瘤形成的实验。这些研究旨在揭示新的信号通路，为独立研究者的未来研究提供理论依据，并可能为痣和黑色素瘤患者提供治疗益处。摘要：我们目前对色素沉着相关疾病（如痣和黑色素瘤）的了解是有限的。斑马鱼已经对这些疾病的生物学产生了重要的见解，我计划研究这种鱼，作为识别治疗这些人类疾病的新途径和药物的工具。

DESCRIPTION (provided by applicant): My clinical and research training has long been geared towards a career as a physician-scientist who is able to bridge the gap between clinical observations and basic science approaches to these problems. This application is a critical part of my growth in this regard, and will ensure that I have the resources necessary to achieve this goal. PROJECT SUMMARY: Disorders of pigmented melanocytes are responsible for a number of human diseases, from the hypopigmentation of vitiligo to the fatal consequences of malignant melanoma. Understanding the genetic pathways that are involved in the development of the neural crest and its derivatives, including melanocytes, is a critical component of treating diseases of these lineages. In this proposal, I plan to use the zebrafish to understand how perturbations in the normal development of embryonic neural crest and melanocyte precursors is related to adult pigmentation disease such as nevi and melanoma. We have developed transgenic BRAF;p53 mutant fish that are morphologically normal as embryos yet have severe pigmentation disruption and melanoma as adults. Gene expression analyses demonstrates a conserved gene signature between BRAF;p53 embryos and tumors, suggesting a link between events early in embryogenesis and tumor formation. In Specific Aim 1,1 have utilized this information as the basis of a chemical screen to identify pathways and molecules that are relevant to preventing initiating events in melanoma formation. Hits which emerge from this screen are tested for their ability to alter the adult BRAF;p53 phenotype. Preliminary data of one hit, MLT013, demonstrates that it inhibits pigment stripe disruption as well as melanoma transplantability. MLT013 likely functions through endothelin receptor blockade. In Aim 2,1 describe experiments which determine whether endothelins cooperate with BRAF to drive melanoma formation. These studies aim to uncover novel signaling pathways that will provide the rationale for future studies as an independent investigator, and may offer therapeutic benefit to patients with nevi and melanoma. LAY SUMMARY: Our current understanding of diseases related to pigmentation, such as moles and melanoma, is limited. The zebrafish has yielded important insights into the biology of these diseases, and I plan to study this fish as a tool for identifying new pathways and drugs for the treatment of these human diseases.