The Contribution of Melanocyte-like Cells to Atrial Function and Development

描述（由申请人提供）：我们最近在人和小鼠的肺静脉和心房中描述了一种新的黑色素细胞样细胞群。在发育和成熟的心脏中，黑色素细胞样细胞以一种不同于任何已知细胞谱系的模式被发现，并且在经常引起临床心房心律失常的解剖区域被发现。这些细胞表达一种独特的转录特征，与心房肌细胞或真皮黑色素细胞不同。有趣的是，分离的小鼠黑色素细胞样细胞可电兴奋并产生心房肌细胞样动作电位。我们发现，人类和小鼠心脏黑色素细胞都表达的多巴铬变异体酶（Dct）的遗传缺失，揭示了这些细胞中动作电位延长和后去极化的病理状态。此外，成熟的Dct敲除小鼠保留了黑色素细胞样细胞，心脏结构正常，但心房心律失常的易感性增加。虽然在心脏中有黑色素细胞样细胞的野生型小鼠在基线时没有增加心房心律失常，但与心脏中缺乏黑色素细胞样细胞的c-kit突变小鼠相比，在使用毒蕈碱受体激动剂carbachol时，它们确实有更多的心房心律失常。此外，用活性氧清除剂治疗Dct基因敲除小鼠时，心房心律失常的发生率更低。因此，黑素细胞样细胞可能对通常诱发临床心房心律失常的应激增加（即自主神经刺激或活性氧）做出反应，从而导致心房心律失常。尽管我们最初的特征，在正常的生理和病理生理状态下，黑素细胞样细胞的功能仍然不清楚。此外，虽然我们有一些证据表明黑素细胞样细胞可兴奋并可能影响心律失常的发生；这些细胞的潜在电生理特征需要进一步研究，以了解它们对心律失常的潜在贡献。因此，我们提出了一系列体外和体内实验，使用基因工程小鼠模型来表征这些细胞的细胞电生理，并确定它们对心房心律失常的贡献。建议的具体目标包括：1)阐明分离小鼠黑素细胞样细胞电兴奋性的电压依赖性电流以及黑素细胞样细胞对心房心律失常触发的直接贡献；2)确定自主神经刺激对ct阳性黑素细胞样细胞兴奋性的影响及其对心房心律失常的贡献；3)评估活性氧对ct阳性黑色素细胞样细胞兴奋性的影响及其对心房心律失常的影响；4)研究黑色素细胞样细胞在正常心脏中的作用。我们将获得的关于黑素细胞样细胞的基本生物学知识可能为我们对心房电生理学的理解开辟新的途径，并有可能为心房心律失常的发病机制提供范式转变的见解。

DESCRIPTION (provided by applicant): We recently described a novel population of melanocyte-like cells in the pulmonary veins and atria of humans and mice. In the developing and mature heart, melanocyte-like cells are found in a pattern unlike that of any currently known cell lineage and are found in anatomic regions that often give rise to clinical atrial arrhythmia triggers. These cells express a unique transcription signature that is distinct from that of atrial myocytes or dermal melanocytes. Interestingly, isolated murine melanocyte-like cells are electrically excitable and generate atrial myocyte-like action potentials. We have found that genetic deletion of the enzyme dopachrome tautomerase (Dct), which is expressed by both human and murine cardiac melanocytes, unmasks a pathological state with action potential prolongation and afterdepolarizations in these cells. Furthermore, mature Dct knockout mice retain melanocyte-like cells and have structurally normal hearts, yet display increased susceptibility to atrial arrhythmias. While wild-type mice with melanocyte-like cells in their hearts do not have increased atrial arrhythmias at baseline, they do have more atrial arrhythmias when challenged with the muscarinic agonist carbachol compared to c-kit mutant mice that lack melanocyte-like cells in their hearts. In addition, Dct knockout mice have fewer atrial arrhythmias when treated with reactive oxygen species scavengers. Hence, melanocyte-like cells may contribute to atrial arrhythmias in response to increased stresses (i.e. autonomic stimulation or reactive oxygen species) that commonly induce clinical atrial arrhythmias. Despite our initial characterization, the function of melanocyte-like cells during normal physiologic and pathophysiologic states remains obscure. Furthermore, while we have some evidence melanocyte-like cells are excitable and may influence arrhythmogenesis; the underlying electrophysiologic characteristics of these cells require further investigation to understand their potential contribution to arrhythmias. Therefore, we are proposing a series of in vitro and in vivo experiments using genetically engineered mouse models to characterize the cellular electrophysiology of these cells and determine their contribution to atrial arrhythmias. The specific aims proposed include: 1) elucidating the voltage-dependent currents underlying the electrical excitability of isolated murine melanocyte-like cells and the direct contribution of melanocyte-like to atrial arrhythmia triggers, 2) determining the effects of autonomic stimulation upon Dct-positive melanocyte-like cellular excitability and their contribution atrial arrhythmias, 3) assessing the effects of reactive oxygen species upon the excitability of Dct-positive melanocyte-like cells and their influence upon atrial arrhythmias, and 4) investigating the role of melanocyte-like cells in the normal heart. The knowledge we will gain about the basic biology of melanocyte-like cells is likely to open new avenues in our understanding of atrial electrophysiology, with the potential for paradigm shifting insights into the pathogenesis of atrial arrhythmias.