Transcriptomic Analysis of Cardiomyocyte Extracellular Vesicles in Hypertrophic Cardiomyopathy Reveals Differential snoRNA Cargo.
Transcriptomic Analysis of Cardiomyocyte Extracellular Vesicles in Hypertrophic Cardiomyopathy Reveals Differential snoRNA Cargo.
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DOI：
10.1089/scd.2021.0202
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发表时间：
2021-12-15
影响因子：
4
通讯作者：
Smith JGW
中科院分区：
文献类型：
--
作者：
James V;Nizamudeen ZA;Lea D;Dottorini T;Holmes TL;Johnson BB;Arkill KP;Denning C;Smith JGW
肥厚型心肌病（HCM）的特征是左心室壁厚度增加，可导致心力衰竭和心源性猝死等毁灭性疾病。尽管进行了广泛的研究，但介导许多相关临床表现的机制仍然未知，需要人类模型。为了解决这一问题，从具有HCM相关突变（c.ACTC1G301A）的患者中产生了人诱导多能干细胞（hiPSC）系，并通过使用CRISPR/Cas9基因编辑技术校正突变来创建等基因对照。将心肌细胞（hiPSC-CM）与这些hiPSC区分开，并在基线和增加的收缩工作负荷（2 Hz电刺激）下进行分析。在24小时培养期后分离并表征释放的细胞外囊泡（EV），并对hiPSC-CM和释放的EV进行转录组学分析。细胞mRNA的转录组学分析表明，HCM突变引起已知HCM途径内的差异剪接，并破坏代谢途径。在增加收缩频率的分析显示代谢基因表达的进一步破坏，在HCM背景下具有累加效应。有趣的是，我们观察到HCM释放的EV内snoRNA货物的差异，当HCM hiPSC-CM经受增加的工作负荷时，这些差异特异性地改变。预测这些snoRNA在翻译后修饰和选择性剪接中具有作用，这些过程在HCM中受到差异调节。因此，本研究中鉴定的snoRNA可能揭示了对无法解释的HCM表型的机理性见解，并提供了未来作为HCM生物标志物或作为未来RNA靶向治疗靶点的潜在用途。
Hypertrophic cardiomyopathy (HCM) is characterized by increased left ventricular wall thickness that can lead to devastating conditions such as heart failure and sudden cardiac death. Despite extensive study, the mechanisms mediating many of the associated clinical manifestations remain unknown and human models are required. To address this, human-induced pluripotent stem cell (hiPSC) lines were generated from patients with a HCM-associated mutation (c.ACTC1G301A) and isogenic controls created by correcting the mutation using CRISPR/Cas9 gene editing technology. Cardiomyocytes (hiPSC-CMs) were differentiated from these hiPSCs and analyzed at baseline, and at increased contractile workload (2 Hz electrical stimulation). Released extracellular vesicles (EVs) were isolated and characterized after a 24-h culture period and transcriptomic analysis performed on both hiPSC-CMs and released EVs. Transcriptomic analysis of cellular mRNA showed the HCM mutation caused differential splicing within known HCM pathways, and disrupted metabolic pathways. Analysis at increasing contraction frequency showed further disruption of metabolic gene expression, with an additive effect in the HCM background. Intriguingly, we observed differences in snoRNA cargo within HCM released EVs that specifically altered when HCM hiPSC-CMs were subjected to increased workload. These snoRNAs were predicted to have roles in post-translational modifications and alternative splicing, processes differentially regulated in HCM. As such, the snoRNAs identified in this study may unveil mechanistic insight into unexplained HCM phenotypes and offer potential future use as HCM biomarkers or as targets in future RNA-targeting therapies.
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影响因子：
3.5
作者：
Good DJ;Kocher MA
通讯作者：
Kocher MA
影响因子：
3.7
作者：
Garcia NA;Ontoria-Oviedo I;González-King H;Diez-Juan A;Sepúlveda P
通讯作者：
Sepúlveda P
DOI：
10.1073/pnas.1707741114
发表时间：
2017-07-18
影响因子：
11.1
作者：
Ito, Kaoru;Patel, Parth N.;Seidman, J. G.
通讯作者：
Seidman, J. G.
心血管精密医学中的RNA疗法。
影响因子：
4
作者：
Laina A;Gatsiou A;Georgiopoulos G;Stamatelopoulos K;Stellos K
通讯作者：
Stellos K
影响因子：
3.7
作者：
Castle JC;Armour CD;Löwer M;Haynor D;Biery M;Bouzek H;Chen R;Jackson S;Johnson JM;Rohl CA;Raymond CK
通讯作者：
Raymond CK