The role of melanocyte precursors in zebrafish pigmentation disorders

描述（由申请人提供）：我的临床和研究培训长期以来一直面向职业生涯作为一个医生，科学家谁是能够弥合临床观察和基础科学方法之间的差距差距这些问题。这个应用程序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。项目概要：色素性黑素细胞的紊乱是许多人类疾病的原因，从白癜风的色素减退到恶性黑色素瘤的致命后果。了解参与神经嵴及其衍生物（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。在这个建议中，我计划使用斑马鱼来了解胚胎神经嵴和黑素细胞前体的正常发育中的扰动如何与成人色素沉着疾病如痣和黑色素瘤相关。我们已经开发出转基因BRAF;p53突变鱼，其在胚胎时形态正常，但在成年时具有严重的色素沉着破坏和黑色素瘤。基因表达分析表明BRAF;p53胚胎和肿瘤之间存在保守的基因特征，表明胚胎发生早期事件与肿瘤形成之间存在联系。在Specific Aim 1，1中，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中的起始事件相关的途径和分子。测试从该筛选中出现的命中物改变成体BRAF;p53表型的能力。初步数据的一个命中，MLT 013，表明它抑制色素条纹破坏以及黑色素瘤移植性。MLT 013可能通过内皮素受体阻断发挥作用。在目标2中，1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，为独立研究者未来的研究提供理论基础，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的了解，如痣和黑色素瘤，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，作为确定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.