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作为一名内科科学家的职业生涯，他能够弥合临床观察和基础科学方法之间的差距，以解决这些问题。这份申请是我在这方面成长的关键部分，它将确保我拥有实现这一目标所需的资源。项目摘要：色素黑素细胞的紊乱是许多人类疾病的罪魁祸首，从白癜风的色素减退到恶性黑色素瘤的致命后果。了解涉及神经脊及其衍生物(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。在这项提议中，我计划使用斑马鱼来了解胚胎神经脊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)有关。我们已经开发出转基因BRAF；p53突变鱼，它们在胚胎时形态正常，但成年后有严重的色素破坏和黑色素瘤。基因表达分析表明，BRAF；P53胚胎和肿瘤之间存在保守的基因信号，这表明胚胎发生早期事件与肿瘤形成之间存在联系。在特定的目标1中，1利用这些信息作为化学筛选的基础，以确定与防止黑色素瘤形成中的起始事件相关的途径和分子。从这个屏幕上出现的点击是为了测试它们改变成人BRAF；P53表型的能力。一个Hit，MLT013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可能通过阻断内皮素受体发挥作用。在目标2中，1描述了确定内皮素是否与BRAF协同推动黑色素瘤形成的实验。这些研究旨在发现新的信号通路，为作为独立调查者的未来研究提供理论基础，并可能为色素痣和黑色素瘤患者提供治疗益处。LAY摘要：我们目前对色素沉着相关疾病的了解有限，如葡萄胎和黑色素瘤。斑马鱼已经对这些疾病的生物学产生了重要的见解，我计划将这种鱼作为一种工具来研究，以确定治疗这些人类疾病的新途径和药物。

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.