中国科学院昆明动物研究所机构知识库

脑疾病是世界范围的主要公共健康问题之一。阿尔兹海默氏病（俗称老年痴呆，Alzheimer’s disease，AD）、帕金森氏病（Parkinson’s disease，PD）和自闭症谱系障碍（Autism spectrum disorders，ASD）是发生率较高的脑疾病，且目前尚无治疗的有效手段。同时，这三种脑疾病都受到遗传因素的影响，已报道很多与疾病发生相关的易感基因，如与AD发病相关的APP、ABCA7和PSEN等，与PD发病相关的PINK1、LRRK2和GBA等，以及与ASD发病相关的WAC、ADNP 和ANKRD11等。另外，传统的模式动物，如线虫、果蝇、斑马鱼和啮齿类动物的大脑与人类大脑差异巨大，用这些动物建立的脑疾病模型存在非常大的局限性。相比之下，非人灵长类 (non-human primate, NHP) 无论在遗传还是大脑结构和功能上都与人类相近，是建立脑疾病模型的理想动物。目前，主要有三种NHP脑疾病模型：诱导损伤模型、基因工程模型和自然发生模型。诱导损伤模型建模快，但很难完全模拟疾病的发生机制；基因工程模型可以较好地模拟疾病发生的遗传机制并验证基因功能，是脑疾病研究中一个非常重要的工具。但是基因操作NHP模型的成功率很低、周期很长且成本很高。自然发生的NHP疾病模型对疾病的研究非常有价值，因为它排除了人为因素的影响。但是用传统的手段很难在猴群中大规模筛选自然发生的疾病动物。本研究利用分子倒位探针（molecular inversion probe，MIP）技术在猕猴群体中大规模扫描AD、PD和ASD易感基因，以期建立自然发生的疾病动物模型。通过文献汇总我们共选取了63个基因，包括12个AD易感基因，11个PD易感基因和40个ASD易感基因。我们设计了2,928个分子倒位单针来捕获这些基因的编码区序列并通过高通量测序筛选天然发生的基因功能缺失突变。我们共计扫描了2233只动物（648只食蟹猴和1585只恒河猴），发现了129个个体携带的71个功能缺失突变（AD: 33个个体的14个突变；PD：21个个体的15个突变；ASD：75个个体的42个突变）。我们通过独立采样和Sanger测序对大多数突变进行了验证，并对部分突变进行了mRNA水平的验证。研究结果将为建立非人灵长类脑疾病的自然发病模型提供重要的基础数据。 

Brain diseases are among the major worldwide public health concerns. Alzheimer’s disease (AD), Parkinson’s disease (PD) and Autism Spectrum Disorder (ASD) are of the common brain diseases with high prevalence, and no effective treatments have been developed for these disorders effectively. Genetic components play an important role in these three brain disorders and many risk genes have been reported, for examples, APP, ABCA7 and PSEN1 for AD; PINK1, LRRK2 and GBA for PD, and WAC, ADNPand ANKRD11 for ASD. Although a wide range of model organisms (worm, fruit fly, zebrafish and rodent) have been used in brain disease studies, their contributions were rather limited owing to the large differences between the brains of these species and humans. In contrast, non-human primates (NHPs) are closely related with humans in view of gene sequences and brain structure and function, and they serve as the ideal animal model for human brain disorders.There are three types of NHP disease models: the induced-damage model, the genetically-engineered model and the naturally-occurred model. The induced-damage model can be constructed quickly, but can not completely mimic the etiology of human brain diseases. The genetically manipulated model can reflect disease mechanism and be used to study gene function, serving as an important tool for investigating brain diseases. However, the low success rate and high cost of the NHP transgenic model prevent its application. In contrast, the naturally-occurred model is highly valuable for brain disease study because it is free from artificial disturbance. Previously, it was difficult to scan a large monkey colony for generating the naturally-occurred disease model due to technical limitations. Here we use (Molecular inversion probe (MIP) to conduct large-scale screening of susceptibility genes for AD, PD and ASD in large monkey populations .Based on our current understanding of disease susceptibility genes, we chose 63 genes including 12 AD genes, 11 PD genes and 40 ASD genes. We designed 2,928 MIPs to capture the coding sequences of these genes in order to identify loss-of-functional mutations in these gene coding regions. Totally, we have screened 2,243 monkeys (648 rhesus macaques and 1,585 cynomolgus monkeys）, and we found 71 loss-of-function mutations in 129 monkeys (AD:14 mutations in 33 monkeys, PD: 15 mutations in 21monkeys, and ASD: 42 mutations in 75 monkeys). Most of the mutations have been validated by independent sampling and Sanger sequencing, and part of the mutations were validated at mRNA level. The data generated in this study is valuable for the establishment of naturally-occurred NHP models of brain diseases.