Mutant Shank3 macaque monkeys for neurobiological studies of ASD

项目摘要/摘要 对小鼠基因组进行基因修改的能力使生物医学研究发生了革命性的变化。然而， 它对我们理解大脑疾病的影响是有限的，部分原因是 啮齿动物和人类之间大脑的结构和生理学。最值得注意的是，前额叶皮质是 人类大脑最大和最发达的部分，也是病理过程的最佳候选者 在许多精神疾病中。然而，啮齿动物只有一个基本的前额叶皮质，因此受限于 表现出由该区域调节的复杂的认知功能。缺乏可预测的动物 模型现在被认为是开发有效的大脑疾病治疗方法的关键瓶颈之一。 非人灵长类动物与人类的亲缘关系比啮齿动物更密切，这一点在 他们的大脑发育、结构和生理。因此，人们越来越认识到，它们提供了一种 吸引人的模型来研究更高级的大脑功能和大脑疾病。高效节能技术的最新发展 CRISPR基因组编辑技术使直接操纵受精卵中的基因组成为可能，因此 将基因操作扩展到包括非人类灵长类在内的许多物种。 在过去的4年里，我们一直在与一个科学家团队合作，研究大脑认知和 中国科学院深圳高级技术研究院脑病研究所 用CRISPR/Cas9建立单基因ASD猕猴模型。我们现在已经成功地 产生了Shank3突变的食蟹猴。SHANK3是一种谷氨酸能突触后支架蛋白 对突触的发育和功能至关重要。人类Shank3基因杂合突变导致 费兰-麦克德米德综合征(PMS)，一种自闭症谱系障碍。对5位创始人的初步描述 SHANK3突变猴子表现出睡眠障碍、运动缺陷和重复行为增加，如 以及社交和学习障碍。对fMRI数据的无偏分析显示局部和全局都发生了变化 指示电路异常的连接模式。总而言之，这些结果与 人类自闭症和经前综合征的基因-电路-行为功能障碍。在此，我们建议，与我们的 SIAT的同事中国，以产生F1代的Shank3突变猴为初步验证 观察，以进一步描述行为和神经生理特征，并将突变精子带到美国 建立一个与自闭症研究社区分享的殖民地。

PROJECT SUMMARY/ABSTRACT The ability to genetically modify the mouse genome has revolutionized biomedical research. However, its impact on our understanding of brain disorders is limited partially due to the inherent differences in the structure and physiology of the brain between rodents and humans. Most notably, the prefrontal cortex is one of the largest and most developed portions of the human brain and a top candidate for pathological processes in many psychiatric disorders. Yet, rodents have only a rudimentary prefrontal cortex and are thus limited in exhibiting the complex cognitive functions that are mediated by this region. The lack of predictive animal models is now considered one of the key bottlenecks in developing effective treatments for brain disorders. Non-human primates are much more closely related to humans than are rodents, and this is reflected in their brain development, structure and physiology. Hence, it is increasingly recognized that they provide an attractive model to study higher brain function and brain disorders. The recent development of highly efficient CRISPR genome-editing technology made it feasible to directly manipulate the genome in zygotes, thus expanding genetic manipulations to many species including non-human primates. In the past 4 years, we have been collaborating with a team of scientists in the Brain Cognition and Brain Disease Institute, Shenzhen Institutes of Advanced Technology (SIAT), Chinese Academy of Sciences to use CRISPR/Cas9 to generate macaque monkey models of monogenic ASD. We have now successfully generated Shank3 mutant cynomolgus macaques. Shank3 is a glutamatergic postsynaptic scaffolding protein critical for synapse development and function. Heterozygous mutations of the Shank3 gene in humans lead to Phelan-McDermid syndrome (PMS), an autism spectrum disorder. Initial characterization of the 5 founder Shank3 mutant monkeys revealed sleep disturbances, motor deficits, and increased repetitive behaviors, as well as social and learning impairments. Unbiased analysis of fMRI data showed altered local and global connectivity patterns indicative of circuit abnormalities. Together, these results parallel some aspects of the gene-circuit-behavior dysfunction in human ASD and PMS. Here we propose, in collaboration with our colleagues in SIAT, China, to generate F1 generation of Shank3 mutant monkeys to validate initial observations, to further behavioral and neurophysiological characterization and to bring mutant sperms to US for establishing a colony for sharing with autism research community.