Pharmacological intervention to restore connectivity deficits of neuronal networks derived from ASD patient iPSC with a TSC2 mutation.
Pharmacological intervention to restore connectivity deficits of neuronal networks derived from ASD patient iPSC with a TSC2 mutation.
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DOI：
10.1186/s13229-020-00391-w
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发表时间：
2020-10-19
期刊：
影响因子：
6.2
通讯作者：
Harwood AJ
中科院分区：
文献类型：
--
作者：
Alsaqati M;Heine VM;Harwood AJ
多发性硬化症（TSC）是一种罕见的遗传性多系统性疾病，由TSC 1或TSC 2基因的常染色体显性突变引起。其特征在于雷帕霉素复合物1（mTORC 1）通路的机械靶点的过度活化，并具有严重的神经发育和神经学成分，包括自闭症，智力残疾和癫痫。在人类和啮齿类动物模型中，TSC蛋白的丢失导致神经元过度兴奋和突触功能障碍，尽管这些变化对发育中的中枢神经系统的后果目前尚不清楚。在这里，我们应用基于多电极阵列的测定来研究TSC 2损失对使用自闭症谱系障碍（ASD）患者来源的iPSC的神经元网络活动的影响。我们研究了神经元爆发的时间同步和整个网络电极之间的空间连接。我们发现，ASD患者来源的神经元与TSC 2的功能丧失，除了拥有神经元活动过度，发展功能失调的神经元网络与减少同步神经元爆发和较低的空间连接。这些网络功能的缺陷与抑制性GABA信号传导和谷氨酸信号传导的基因表达升高相关，表明突触兴奋性信号传导的潜在异常。mTORC 1活性在蛋白激酶、mTOR、AMP依赖性蛋白激酶1（AMPK）和Unc-51样自噬激活激酶1（ULK 1）的稳态三联体中发挥作用，其协调合成代谢细胞生长和分解代谢自噬的相互作用，同时平衡能量和营养稳态。mTOR抑制剂雷帕霉素抑制神经元过度活跃，但不增加同步网络活动，而AMPK的激活恢复了网络活动的某些方面。相比之下，ULK 1激活剂林恩-1604增加了网络行为，缩短了网络突发长度，并减少了不相关尖峰的数量。尽管在多个独立的iPSC培养物中观察到稳健且一致的表型，但结果是基于一名患者。具有不同TSC 2突变的患者之间可能存在更微妙的差异，或者其基因组内多基因背景的差异。这可能会影响TSC 2患者之间的网络缺陷或药理学反应的严重程度。我们的观察结果表明，与TSC 2突变的ASD患者相关的体外神经元网络的网络连接性降低，这可能是由于抑制性突触处GABA信号传导增加而引起的兴奋性/抑制性失衡。通过激活ULK 1可以有效地抑制这种异常。
Tuberous sclerosis complex (TSC) is a rare genetic multisystemic disorder resulting from autosomal dominant mutations in the TSC1 or TSC2 genes. It is characterised by hyperactivation of the mechanistic target of rapamycin complex 1 (mTORC1) pathway and has severe neurodevelopmental and neurological components including autism, intellectual disability and epilepsy. In human and rodent models, loss of the TSC proteins causes neuronal hyperexcitability and synaptic dysfunction, although the consequences of these changes for the developing central nervous system are currently unclear. Here we apply multi-electrode array-based assays to study the effects of TSC2 loss on neuronal network activity using autism spectrum disorder (ASD) patient-derived iPSCs. We examine both temporal synchronisation of neuronal bursting and spatial connectivity between electrodes across the network. We find that ASD patient-derived neurons with a functional loss of TSC2, in addition to possessing neuronal hyperactivity, develop a dysfunctional neuronal network with reduced synchronisation of neuronal bursting and lower spatial connectivity. These deficits of network function are associated with elevated expression of genes for inhibitory GABA signalling and glutamate signalling, indicating a potential abnormality of synaptic inhibitory–excitatory signalling. mTORC1 activity functions within a homeostatic triad of protein kinases, mTOR, AMP-dependent protein Kinase 1 (AMPK) and Unc-51 like Autophagy Activating Kinase 1 (ULK1) that orchestrate the interplay of anabolic cell growth and catabolic autophagy while balancing energy and nutrient homeostasis. The mTOR inhibitor rapamycin suppresses neuronal hyperactivity, but does not increase synchronised network activity, whereas activation of AMPK restores some aspects of network activity. In contrast, the ULK1 activator, LYN-1604, increases the network behaviour, shortens the network burst lengths and reduces the number of uncorrelated spikes. Although a robust and consistent phenotype is observed across multiple independent iPSC cultures, the results are based on one patient. There may be more subtle differences between patients with different TSC2 mutations or differences of polygenic background within their genomes. This may affect the severity of the network deficit or the pharmacological response between TSC2 patients. Our observations suggest that there is a reduction in the network connectivity of the in vitro neuronal network associated with ASD patients with TSC2 mutation, which may arise via an excitatory/inhibitory imbalance due to increased GABA-signalling at inhibitory synapses. This abnormality can be effectively suppressed via activation of ULK1.
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AMP激活的蛋白激酶对ULK1（HATG1）的磷酸化将能量传感连接到线粒体。
DOI：
10.1126/science.1196371
发表时间：
2011-01-28
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Egan DF;Shackelford DB;Mihaylova MM;Gelino S;Kohnz RA;Mair W;Vasquez DS;Joshi A;Gwinn DM;Taylor R;Asara JM;Fitzpatrick J;Dillin A;Viollet B;Kundu M;Hansen M;Shaw RJ
通讯作者：
Shaw RJ
影响因子：
21.3
作者：
通讯作者：
--
影响因子：
5.3
作者：
Nyfeler, Beat;Bergman, Philip;Murphy, Leon O.
通讯作者：
Murphy, Leon O.
影响因子：
6.2
作者：
Martin, Pauline;Wagh, Vilas;Ramesh, Vijaya
通讯作者：
Ramesh, Vijaya
影响因子：
64.5
作者：
Inoki, Ken;Ouyang, Hongjiao;Guan, Kun-Liang
通讯作者：
Guan, Kun-Liang