Autism spectrum disorder (ASD) has a strong genetic etiology. Germline mutation in the tumor suppressor gene PTEN is one of the best described monogenic risk cases for ASD. Animal modeling of cell-specific Pten loss or mutation has provided insight into how disruptions to the function of PTEN affect neurodevelopment, neurobiology, and social behavior. As such, there is a growing need to understand more about how various aspects of PTEN activity and cell-compartment-specific functions, contribute to certain neurological or behavior phenotypes. To understand more about the relationship between Pten localization and downstream effects on neurophenotypes, we generated the nuclear-predominant PtenY68H/+ mouse, which is identical to the genotype of some PTEN-ASD individuals. We subjected the PtenY68H/+ mouse to morphological and behavioral phenotyping, including the three-chamber sociability, open field, rotarod, and marble burying tests. We subsequently performed in vivo and in vitro cellular phenotyping and concluded the work with a transcriptomic survey of the PtenY68H/+ cortex, which profiled gene expression. We observe a significant increase in P-Akt downstream of canonical Pten signaling, macrocephaly, decreased sociability, decreased preference for novel social stimuli, increased repetitive behavior, and increased thigmotaxis in PtenY68H/+ six-week-old (P40) mice. In addition, we found significant microglial activation with increased expression of complement and neuroinflammatory proteins in vivo and in vitro accompanied by enhanced phagocytosis. These observations were subsequently validated with RNA-seq and qRT-PCR, which revealed overexpression of many genes involved in neuroinflammation and neuronal function, including oxytocin. Oxytocin transcript was fivefold overexpressed (P = 0.0018), and oxytocin protein was strongly overexpressed in the PtenY68H/+ hypothalamus. The nuclear-predominant PtenY68H/+ model has clarified that Pten dysfunction links to microglial pathology and this associates with increased Akt signaling. We also demonstrate that Pten dysfunction associates with changes in the oxytocin system, an important connection between a prominent ASD risk gene and a potent neuroendocrine regulator of social behavior. These cellular and molecular pathologies may related to the observed changes in social behavior. Ultimately, the findings from this work may reveal important biomarkers and/or novel therapeutic modalities that could be explored in individuals with germline mutations in PTEN with ASD.

中文翻译：

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种系核为主的 Pten 小鼠模型表现出社交和持久行为受损、小胶质细胞激活和催产素活性增加


自闭症谱系障碍（ASD）具有很强的遗传病因。肿瘤抑制基因 PTEN 的种系突变是自闭症谱系障碍（ASD）单基因风险案例中描述最充分的之一。细胞特异性 Pten 丢失或突变的动物模型为了解 PTEN 功能的破坏如何影响神经发育、神经生物学和社会行为提供了见解。因此，越来越需要更多地了解 PTEN 活性和细胞区室特异性功能的各个方面如何促进某些神经或行为表型。为了更多地了解 Pten 定位和下游对神经表型的影响之间的关系，我们培育了核优势型 PtenY68H/+ 小鼠，它与一些 PTEN-ASD 个体的基因型相同。我们对 PtenY68H/+ 小鼠进行形态学和行为表型分析，包括三室社交性、旷场、旋转和大理石埋藏测试。随后，我们进行了体内和体外细胞表型分析，并通过对 PtenY68H/+ 皮质进行转录组学调查来结束工作，该调查分析了基因表达。我们观察到 PtenY68H/+ 六周龄 (P40) 小鼠中典型 Pten 信号传导下游的 P-Akt 显着增加、巨头畸形、社交能力下降、对新社交刺激的偏好下降、重复行为增加和触动性增加。此外，我们发现小胶质细胞显着激活，体内和体外补体和神经炎症蛋白表达增加，并伴有吞噬作用增强。这些观察结果随后通过 RNA-seq 和 qRT-PCR 进行了验证，揭示了许多涉及神经炎症和神经元功能的基因的过度表达，包括催产素。 催产素转录物过表达五倍（P = 0.0018），催产素蛋白在 PtenY68H/+ 下丘脑中强烈过表达。核主导型 PtenY68H/+ 模型已阐明 Pten 功能障碍与小胶质细胞病理学相关，并且与 Akt 信号传导增强相关。我们还证明，Pten 功能障碍与催产素系统的变化有关，催产素系统是重要的 ASD 风险基因和社会行为的有效神经内分泌调节剂之间的重要联系。这些细胞和分子病理可能与观察到的社会行为变化有关。最终，这项工作的结果可能揭示重要的生物标志物和/或新的治疗方式，可以在患有 ASD 的 PTEN 种系突变个体中进行探索。