Mechanisms of circuit failure and treatments in patient-derived neurons in autism

描述（由申请人提供）：缺乏有效的临床前模型来探索细胞机制，限制了严重自闭症的治疗发展。为了开发新的临床前模型和治疗方法，在患者来源的神经元中进行细胞表型和细胞对潜在药物反应的深入研究将是有利的。这项研究的长期目标是阐明严重自闭症病理的分子机制，这将作为开发创新治疗方法的基础，为这些目前治疗选择很少的患者提供帮助。我们最近发现了一种新的自闭症机制，涉及内体Na+/H+交换器（NHEs）。我们现在已经从一系列NHE6突变的患者身上产生了诱导多能干细胞系（iPSCs）。该应用程序的目的是阐明NHE6在人类轴突发育中的作用，并测试患者来源的神经元对可用的、基于机制的药物的反应，这些药物随后可能作为患者的治疗方法。NHE6调节核内体前体的外排。在我们对人类神经元的初步研究中，NHE6缺乏导致内体管腔过度酸化和轴突生长和分支缺陷。这种机制是偶然的，因为有许多众所周知的fda批准的靶向内体ph的药物。我们的中心假设是NHE6的缺乏由于极化膜添加减少而抑制轴突生长和分支。这项研究的基本原理是，它构成了开发新的治疗方法的关键的第一步

DESCRIPTION (provided by applicant): A lack of validated preclinical models for exploring cellular mechanisms has limited treatment development in severe autism. In order to develop new preclinical models and therapeutics, rig- orous studies of cell phenotypes and cellular responses to potential agents in patient-derived neurons will be advantageous. The long-term goal of this research is to elucidate the molecular mechanisms underlying the pathology of severe autism that will serve as the basis to develop innovative treatments for these patients with few current therapeutic options. We recently identi- fied a novel mechanism in autism involving endosomal Na+/H+ exchangers (NHEs). We have now generated induced pluripotent stem cell lines (iPSCs) from patients with a range of NHE6 mutations. The objective for this application is to elucidate the role of NHE6 in human axon de- velopment and to test the response of patient-derived neurons to available, mechanism-based agents that may subsequently serve as treatments in patients. NHE6 regulates the efflux of pro- tons from endosomes. In our preliminary studies in human neurons, NHE6 deficiency leads to over-acidification of the endosomal lumen and defects in axon growth and branching. This mechanism is fortuitous because there are a number of well-known FDA-approved agents that target intra-endosomal pH. Our central hypothesis is that deficiency of NHE6 inhibits axon growth and branching due to diminished polarized membrane addition. The rationale for this re- search is that it constitutes the first critical steps toward the development of new treatments for individuals with severe autism and related disorders caused by reductions in NHE6 expression and/or by defects in axonal growth or arborization. We have published studies on NHE6 function in mouse models, yet the transition to patient-derived tissues is warranted at this time. Screening of potential human therapeutic agents is best performed on human (and patient-derived) tis- sues. Three specific aims are proposed: (1) Determine the role of NHE6 in regulating endosome lumen acidity and endosome recycling. (2) Determine the extent to which the function of RAB10-associated SVs in axon growth is perturbed by NHE6 mutation in patient-derived neu- rons. (3) Determine the reversibility of axon growth and branching defects in patient-derived neurons by exogenous growth factors. The proposed research is innovative because we are studying a novel cellular mechanism in autism, namely, regulation of intra-endosomal pH in ax- on growth. This research is significant because it will lead to the development of critically need- ed preclinical models in patient-derived neurons and potential treatments for severe autism and related disorders, an urgent public health problem.