Transcriptional signatures of participant-derived neural progenitor cells and neurons implicate altered Wnt signaling in Phelan-McDermid syndrome and autism
Transcriptional signatures of participant-derived neural progenitor cells and neurons implicate altered Wnt signaling in Phelan-McDermid syndrome and autism
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DOI：
10.1186/s13229-020-00355-0
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发表时间：
2020-06-19
期刊：
影响因子：
6.2
通讯作者：
Drapeau, Elodie
中科院分区：
文献类型：
--
作者：
Breen, Michael S.;Browne, Andrew;Drapeau, Elodie
phan - mcdermid综合征（PMS）是一种罕见的遗传性疾病，具有自闭症谱系障碍（ASD）、智力残疾和语言迟缓的高风险，由shank3基因22q13.3缺失或突变引起。迄今为止，尚不清楚经前综合征中shank3单倍性不足导致的分子和途径变化。揭示这些机制对于理解经前症候群的病理生物学以及最终开发新的治疗干预措施至关重要。方法通过对经前综合征患者（n= 7）及其未受影响兄弟姐妹（n= 6）的外周血样本进行重编程，建立基于人诱导多能干细胞（hiPSC）的经前综合征模型。对于每个参与者，最多生成三个hiPSC克隆并分化为诱导神经祖细胞（hiPSC- npc, n= 39）和诱导前脑神经元（hiPSC-神经元，n= 41）。全基因组rna测序应用于探索经前综合症先证者和未患病兄弟姐妹之间的转录差异。结果转录组分析发现，在PMS先显子和未受影响的兄弟姐妹（FDR < 5%）的细胞中，hipsc - npc中有391个差异表达基因（deg）， hipsc -神经元中有82个差异表达基因（deg）。PMS中低表达的基因与Wnt信号、胚胎发育和蛋白质翻译有关，而过表达的基因则与突触前和突触后密度基因、突触可塑性调节和g蛋白门控钾通道活性有关。基因共表达网络分析发现，hipsc神经元中有两个模块在PMS中过表达，涉及突触后信号传导和GDP结合，这两个模块都含有发育迟缓和智力残疾的遗传风险位点。最后，pms相关基因与其他ASD hiPSC转录组结果整合，并确定了几个趋同点，表明Wnt信号和细胞外基质发生了改变。考虑到这种情况的罕见性，我们无法在独立的生物样本中进行实验验证。此外，shank3缺失引起的功能和形态表型在这里没有被描述。结论这是经前症候群中分析的最大的人类神经样本。来自PMS患者的hipsc衍生神经细胞的全基因组rna测序揭示了hipsc - npc和hipsc -神经元之间共享和不同的转录特征，包括许多与ASD风险相关的基因，以及特定的神经生物学途径，包括Wnt途径。
Background Phelan-McDermid syndrome (PMS) is a rare genetic disorder with high risk of autism spectrum disorder (ASD), intellectual disability, and language delay, and is caused by 22q13.3 deletions or mutations in theSHANK3gene. To date, the molecular and pathway changes resulting fromSHANK3haploinsufficiency in PMS remain poorly understood. Uncovering these mechanisms is critical for understanding pathobiology of PMS and, ultimately, for the development of new therapeutic interventions. Methods We developed human-induced pluripotent stem cell (hiPSC)-based models of PMS by reprogramming peripheral blood samples from individuals with PMS (n= 7) and their unaffected siblings (n= 6). For each participant, up to three hiPSC clones were generated and differentiated into induced neural progenitor cells (hiPSC-NPCs;n= 39) and induced forebrain neurons (hiPSC-neurons;n= 41). Genome-wide RNA-sequencing was applied to explore transcriptional differences between PMS probands and unaffected siblings. Results Transcriptome analyses identified 391 differentially expressed genes (DEGs) in hiPSC-NPCs and 82 DEGs in hiPSC-neurons, when comparing cells from PMS probands and unaffected siblings (FDR < 5%). Genes under-expressed in PMS were implicated in Wnt signaling, embryonic development, and protein translation, while over-expressed genes were enriched for pre- and postsynaptic density genes, regulation of synaptic plasticity, and G-protein-gated potassium channel activity. Gene co-expression network analysis identified two modules in hiPSC-neurons that were over-expressed in PMS, implicating postsynaptic signaling and GDP binding, and both modules harbored a significant enrichment of genetic risk loci for developmental delay and intellectual disability. Finally, PMS-associated genes were integrated with other ASD hiPSC transcriptome findings and several points of convergence were identified, indicating altered Wnt signaling and extracellular matrix. Limitations Given the rarity of the condition, we could not carry out experimental validation in independent biological samples. In addition, functional and morphological phenotypes caused by loss ofSHANK3were not characterized here. Conclusions This is the largest human neural sample analyzed in PMS. Genome-wide RNA-sequencing in hiPSC-derived neural cells from individuals with PMS revealed both shared and distinct transcriptional signatures across hiPSC-NPCs and hiPSC-neurons, including many genes implicated in risk for ASD, as well as specific neurobiological pathways, including the Wnt pathway.