背景:神经发育障碍(NDD),如自闭症谱系障碍(ASD)和智力障碍(ID),是高度衰弱的儿童精神病。遗传因素被认为在NDD中起主要作用,涉及多个基因和基因组区域。虽然NDD相关基因的功能验证主要是使用小鼠模型进行的,小鼠和人类在脑结构和基因功能上的显著差异限制了小鼠模型在探索NDD潜在机制方面的有效性。因此,建立与人类在进化上更一致的替代动物模型是很重要的。
结果:在这项研究中,我们采用CRISPR/Cas9和体细胞核移植技术成功地产生了MIR137基因敲除的小型猪模型,它编码神经精神障碍相关的微小RNAmiR-137。MIR137(MIR137-/-)的纯合敲除有效抑制了成熟miR-137的表达,并导致死产或短命仔猪的出生。转录组学分析显示,MIR137-/-小型猪大脑中与神经发育和突触信号相关的基因发生了显着变化,反映人类ASD转录组数据的发现。与miR-137缺陷小鼠和人类诱导多能干细胞(hiPSC)衍生的神经元模型相比,在miR-137缺失后,小型猪模型在与人类相关的关键神经元基因中表现出更一致的变化.此外,一项比较分析确定了小型猪和hiPSC来源的神经元中与ASD和ID风险基因相关的差异表达基因。值得注意的是,人特异性miR-137靶标,例如CAMK2A,已知与认知障碍和NDD有关,MIR137-/-小型猪表现出失调。这些发现表明,小型猪中miR-137的丢失会影响对神经发育至关重要的基因。可能有助于NDD的发展。
结论:我们的研究强调了miR-137丢失对涉及MIR137-/-小型猪神经发育和相关疾病的关键基因的影响。它建立了小型猪模型作为研究神经发育障碍的有价值的工具,为人类研究中的潜在应用提供有价值的见解。
BACKGROUND: Neurodevelopmental disorders (NDD), such as autism spectrum disorders (ASD) and intellectual disorders (ID), are highly debilitating childhood psychiatric conditions. Genetic factors are recognized as playing a major role in NDD, with a multitude of genes and genomic regions implicated. While the functional validation of NDD-associated genes has predominantly been carried out using mouse models, the significant differences in brain structure and gene function between mice and humans have limited the effectiveness of mouse models in exploring the underlying mechanisms of NDD. Therefore, it is important to establish alternative animal models that are more evolutionarily aligned with humans.
RESULTS: In this study, we employed CRISPR/Cas9 and somatic cell nuclear transplantation technologies to successfully generate a knockout miniature pig model of the MIR137 gene, which encodes the neuropsychiatric disorder-associated microRNA miR-137. The homozygous knockout of MIR137 (MIR137-/-) effectively suppressed the expression of mature miR-137 and led to the birth of stillborn or short-lived piglets. Transcriptomic analysis revealed significant changes in genes associated with neurodevelopment and synaptic signaling in the brains of MIR137-/- miniature pig, mirroring findings from human ASD transcriptomic data. In comparison to miR-137-deficient mouse and human induced pluripotent stem cell (hiPSC)-derived neuron models, the miniature pig model exhibited more consistent changes in critical neuronal genes relevant to humans following the loss of miR-137. Furthermore, a comparative analysis identified differentially expressed genes associated with ASD and ID risk genes in both miniature pig and hiPSC-derived neurons. Notably, human-specific miR-137 targets, such as CAMK2A, known to be linked to cognitive impairments and NDD, exhibited dysregulation in MIR137-/- miniature pigs. These findings suggest that the loss of miR-137 in miniature pigs affects genes crucial for neurodevelopment, potentially contributing to the development of NDD.
CONCLUSIONS: Our study highlights the impact of miR-137 loss on critical genes involved in neurodevelopment and related disorders in MIR137-/- miniature pigs. It establishes the miniature pig model as a valuable tool for investigating neurodevelopmental disorders, providing valuable insights for potential applications in human research.