Identification of Novel Gene Regulatory Networks for Dystrophin Protein in Vascular Smooth Muscle Cells by Single-Nuclear Transcriptome Analysis.
Identification of Novel Gene Regulatory Networks for Dystrophin Protein in Vascular Smooth Muscle Cells by Single-Nuclear Transcriptome Analysis.
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杜氏肌营养不良症是一种由营养不良蛋白突变引起的x连锁隐性疾病，可导致心力衰竭和呼吸衰竭。肌营养不良蛋白（Dystrophin， DMD）不仅在心肌细胞和骨骼肌细胞中表达，也在血管平滑肌细胞（VSMCs）中表达。据报道，DMD患者有低血压。单核RNA测序（snRNA-seq）是一种先进的技术，能够识别组织特异性细胞亚群的利基特异性基因程序。为了确定DMD突变是否会改变血压，我们比较了mdx小鼠（一种携带DMD基因无意义突变的DMD小鼠模型）和宽型对照小鼠的收缩压、舒张压和平均血压水平。我们发现mdx小鼠的收缩压、舒张压和平均血压明显低于对照组小鼠。为了了解DMD突变如何改变VSMCs的基因表达谱，我们分析了来自DMD突变小鼠（DMDmut）和对照组（Ctrl）小鼠肌核的snRNA-seq数据集。基因本体（Gene Ontology， GO）富集分析显示，DMDmut-VSMCs中最显著激活的通路涉及离子通道功能（钾通道活性、阳离子通道复合物和阳离子通道活性）。值得注意的是，我们发现DMDmut-VSMCs中KCNQ5和RYR2的表达显著上调，而最受抑制的途径是跨膜转运蛋白活性（如阴离子跨膜转运蛋白活性、无机阴离子跨膜转运蛋白活性、输入细胞和跨质膜输入）。此外，我们使用“scMetabolism”R包分析了京都基因与基因组百科全书（KEGG）中的代谢途径。DMDmut-VSMCs表现出丙酮酸代谢和核酸代谢失调。综上所述，通过snRNA-seq的应用，我们（首次）发现了DMD在VSMCs中上调KCNQ5基因表达的潜在分子调控，这有助于我们了解DMD患者低血压的发生机制。我们的研究可能为KCNQ5药物抑制的DMD患者全身性低血压的治疗干预提供新的可能性。
Duchenne muscular dystrophy is an X-linked recessive disease caused by mutations in dystrophin proteins that lead to heart failure and respiratory failure. Dystrophin (DMD) is not only expressed in cardiomyocytes and skeletal muscle cells, but also in vascular smooth muscle cells (VSMCs). Patients with DMD have been reported to have hypotension. Single nuclear RNA sequencing (snRNA-seq) is a state-of-the-art technology capable of identifying niche-specific gene programs of tissue-specific cell subpopulations. To determine whether DMD mutation alters blood pressure, we compared systolic, diastolic, and mean blood pressure levels in mdx mice (a mouse model of DMD carrying a nonsense mutation in DMD gene) and the wide-type control mice. We found that mdx mice showed significantly lower systolic, diastolic, and mean blood pressure than control mice. To understand how DMD mutation changes gene expression profiles from VSMCs, we analyzed an snRNA-seq dataset from the muscle nucleus of DMD mutant (DMDmut) mice and control (Ctrl) mice. Gene Ontology (GO) enrichment analysis revealed that the most significantly activated pathways in DMDmut-VSMCs are involved in ion channel function (potassium channel activity, cation channel complex, and cation channel activity). Notably, we discovered that the DMDmut-VSMCs showed significantly upregulated expression of KCNQ5 and RYR2, whereas the most suppressed pathways were transmembrane transporter activity (such as anion transmembrane transporter activity, inorganic anion transmembrane transporter activity, import into cell, and import across plasma membrane). Moreover, we analyzed metabolic pathways from the Kyoto Encyclopedia of Genes and Genomes (KEGG) using “scMetabolism” R package. DMDmut-VSMCs exhibited dysregulation of pyruvate metabolism and nuclear acid metabolism. In conclusion, via the application of snRNA-seq, we (for the first time) identify the potential molecular regulation by DMD in the upregulation of the expression of KCNQ5 genes in VSMCs, which helps us to understand the mechanism of hypotension in DMD patients. Our study potentially offers new possibilities for therapeutic interventions in systemic hypotension in DMD patients with pharmacological inhibition of KCNQ5.
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影响因子：
11.1
作者：
Krueger, Johanna;Schubert, Julian;Kegele, Josua;Labalme, Audrey;Mao, Miaomiao;Heighway, Jacqueline;Seebohm, Guiscard;Yan, Pu;Koko, Mahmoud;Aslan-Kara, Kezban;Caglayan, Hande;Steinhoff, Bernhard J.;Weber, Yvonne G.;Keo-Kosal, Pascale;Berkovic, Samuel F.;Hildebrand, Michael S.;Petrou, Steven;Krause, Roland;May, Patrick;Lesca, Gaetan;Maljevic, Snezana;Lerche, Holger
通讯作者：
Lerche, Holger
影响因子：
16.6
作者：
MacParland SA;Liu JC;Ma XZ;Innes BT;Bartczak AM;Gage BK;Manuel J;Khuu N;Echeverri J;Linares I;Gupta R;Cheng ML;Liu LY;Camat D;Chung SW;Seliga RK;Shao Z;Lee E;Ogawa S;Ogawa M;Wilson MD;Fish JE;Selzner M;Ghanekar A;Grant D;Greig P;Sapisochin G;Selzner N;Winegarden N;Adeyi O;Keller G;Bader GD;McGilvray ID
通讯作者：
McGilvray ID
影响因子：
8
作者：
Buddhe, Sujatha;Cripe, Linda;Olson, Aaron K.
通讯作者：
Olson, Aaron K.
影响因子：
4
作者：
Lopez, Jose R.;Uryash, Arkady;Adams, Jose A.
通讯作者：
Adams, Jose A.
DOI：
10.1152/ajpgi.00314.2013
发表时间：
2014-02-01
影响因子：
4.5
作者：
Alves, Gabriel A.;Silva, Luisa R.;Nouailhetas, Viviane L. A.
通讯作者：
Nouailhetas, Viviane L. A.