MerTK regulation of the PTTG and RPE phagocytosis

描述(申请人提供)：我的长期目标是阐明MerTK对视网膜色素上皮(RPE)细胞吞噬作用的细胞和分子机制。MerTK是一种受体型蛋白酪氨酸激酶，属于家族。MerTK基因敲除小鼠发生自身免疫性疾病，成人视网膜色素变性(RP)，其特征是巨噬细胞和RPE细胞分别吞噬凋亡的淋巴细胞和耗尽的视网膜外节(OS)。在皇家外科学院(RCS)大鼠中观察到并深入研究了由于RPE吞噬失败而导致的光感受器退化，在该模型中，RPE细胞携带MerTK零突变。MerTK为空也会导致人类RP。体内和体外研究表明，MerTK受体参与了OS的摄取。然而，MerTK调节RPE吞噬功能的分子机制还不是很清楚。我们分析了MerTK突变体RPE的基因表达谱，并对受影响的基因进行了功能研究。其中，MerTK突变显著上调了PTTG，并在MerTK-/-RPE中敲除了PTTG的一个拷贝，部分阻止了体内光感受器的退化。我们将选择并重点研究MerTK介导的基因调控，以研究其在吞噬调节中的功能作用。在这个方案中，我们计划研究MerTK是如何调节PTTG的，以及PTTG是否影响RPE的吞噬功能。我们还将通过ERG或OKR测量来研究低PTTG对中央光感受器结构的保护是否与功能恢复相对应。我们的研究将有助于进一步了解MerTK调节RPE功能的分子机制。公共卫生相关性：视网膜色素变性是一组遗传性视网膜退行性疾病，全球患病率为1：3000，是遗传性失明的主要原因。RP是由一组无关基因突变引起的，MerTK就是其中之一。我们的实验旨在通过研究MerTK调控下的一个候选基因来阐明MerTK调控RPE吞噬功能的分子机制，而MerTK调控RPE功能可能会反过来影响RPE功能。这些研究的数据有望为我们提供对RPE功能的新认识和理解，这将使我们能够开发和实施新的治疗方法来治疗由RPE功能障碍引起的RP。

DESCRIPTION (provided by applicant): My long-term goal is to elucidate the cellular and molecular mechanism by which MerTK exerts its effects on phagocytosis of the retina pigmental epithelium (RPE) cell. MerTK is a receptor-type protein tyrosine kinase, belonging to the TAM family. MerTK knockout mice develop autoimmune disease, retinitis pigmentosa (RP) in adults with characterization of defective phagocytosis of the apoptotic lymphocytes and spent retinal outer segments (OS) by macrophage and RPE cells, respectively. Photoreceptor degeneration caused by a failure of the RPE phagocytosis has been observed and intensively studied in the Royal College of Surgeons (RCS) rat, a model in which the RPE cells carry a MerTK null mutation. MerTK null also causes human RP. Both in vivo and in vitro studies showed that the MerTK receptor participated during OS ingestion. However, the molecular mechanism on how the MerTK regulate RPE phagocytosis is still not very clear. We have analyzed gene expression profile in MerTK mutant RPE and performed functional studies on the affected genes. Of those, the PTTG was dramatically upregulated by MerTK mutation and knockout one copy of PTTG in the MerTK-/- RPE partially prevented photoreceptor degeneration in vivo. We will select and focus on MerTK mediated gene regulation to study its functional role in regulation of phagocytosis. In this proposal, we project to study how the MerTK regulate PTTG and whether the PTTG affect RPE phagocytosis. We will also investigate whether structural preservation of the central photoreceptor by lower PTTG is corresponding to function recovery with ERG or OKR measurements. Our investigation will aid in one step forward to understand the molecular mechanism on the MerTK regulation of RPE functions. PUBLIC HEALTH RELEVANCE: Retinitis pigmentosa (RP) is a group of inherited retinal degenerative diseases with a worldwide prevalence of 1:3000 and a leading cause of inherited blindness. RP is caused by mutation in a group of unrelated genes, one of these is MerTK. Our experiments in this proposal aim to elucidate the molecular mechanism of the MerTK regulation on RPE phagocytosis through studies of one candidate gene under MerTK regulation, which may in turn affect RPE function. The data from these studies is expected to provide us the new knowledge and understanding of the RPE function, which will allow us to develop and implement new therapies for treatment of RP caused by RPE dysfunction.