A NOVEL TRANSLATIONAL MODEL OF AUTISUM SPECTRUM DISORDER

描述（由申请人提供）：最近的报告表明，自闭症谱系障碍（ASD）的发病率急剧上升，这在全球范围内造成了毁灭性的医疗和经济影响。复杂的遗传关联和缺乏复制人类症状谱的转译模型严重阻碍了治疗开发和干预的努力。在这个提案中，我们将利用令人兴奋的新的基因靶向技术以及全基因组关联研究，这些研究已经确定了SHANK3作为主要的疾病风险因子，从而推动了ASD研究的新时代。shRNA和ZFN策略的发展操纵遗传结构为产生单倍体不足或完全敲除疾病危险因素的新型恒河猴模型带来了巨大的希望。本提案的目的包括两种有针对性的方法，用于开发ASD转基因模型的探索阶段；1)用shRNA基因敲低SHANK3, 2)用ZFN基因敲低或敲除SHANK3。此外，神经祖细胞（npc）将从上述ESCs中提取，用于评估SHANK3对神经元功能的影响。总的来说，该提案的结果将为SHANK3调节发育过程和由此产生的神经活动提供关键的早期基准。更广泛地说，除了ASD之外，这笔拨款的目的可能对人类隐性遗传疾病有非常广泛的影响。值得注意的是，我们的团队拥有成功实施上述目标所必需的广泛专业知识，最重要的是，我们已经获得了初步数据，强烈暗示了成功的结果。有了这个探索性奖项的支持，我们有信心必要的基础可以建立起来，从而实现创建未来ASD转化模型的里程碑式努力。虽然这项拨款的目标是高风险的，但我们坚信，最近科学技术的进步是及时的，可以启动一种新的方法来模拟复杂的人类疾病。

DESCRIPTION (provided by applicant): Recent reports have indicated a dramatic rise in the incidence of Autism spectrum disorders (ASD), which has devastating medical and financial impact worldwide. Complex genetic associations and the lack translational models that replicate the spectrum of human symptoms have significantly impeded efforts for therapeutic development and intervention. In this proposal, we will capitalize on exciting new genetic targeting technologies as well as genome-wide association studies, which have pinpointed SHANK3 as a leading disease risk factor prompting a new era in ASD research. The development of shRNA and ZFN strategies to manipulate genetic structure holds great promise for the generation of novel rhesus models with haploinsufficency or complete knockout of disease risk factors. The aims of this proposal involve two targeted approaches for the exploratory stage in the path to develop an ASD transgenic model; 1) genetic knockdown of SHANK3 with shRNA and 2) knockdown or knockout of SHANK3 with ZFN. Additionally, neural progenitor cells (NPCs) will be derived from ESCs listed above for evaluation of SHANK3 on neuronal functions. Collectively, the results yielded from this proposal will provide a critical and early benchmark for SHANK3 modulation on developmental processes and resulting neurological activity. More generally, the aims of this grant could have very broad implications for human recessive genetic diseases in addition to ASD. Notably, our team has extensive expertise necessary for successful implementation of the goals outlined above and, most importantly, have already obtained preliminary data strongly suggestive of a successful outcome. With the support of this exploratory award, we are confident that the necessary groundwork can be established such that the milestone effort to create a future ASD translational model will be achieved. Although the goals of this grant are high-risk, we strongly believe that the recent advancements in scientific technology are timely for the launching of a new approach to modeling complex human diseases.