Attenuated cell-cycle division protein 2 and elevated mitotic roles of polo-like kinase 1 characterize deficient myoblast fusion in peripheral arterial disease.
Attenuated cell-cycle division protein 2 and elevated mitotic roles of polo-like kinase 1 characterize deficient myoblast fusion in peripheral arterial disease.
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DOI：
10.1016/j.bbrc.2022.03.161
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发表时间：
2022-06-18
影响因子：
3.1
通讯作者：
Sachdev, Ulka
中科院分区：
文献类型：
--
作者：
Ferrari, Ricardo;Cong, Guangzhi;Chattopadhyay, Ansuman;Xie, B.;Assaf, E.;Morder, K.;Calderon, Michael J.;Watkins, Simon C.;Sachdev, Ulka
我们认为PAD中MuSC衍生的成肌细胞具有转录组学差异，可以突出缺血诱导的肌病的潜在原因。比较了灌注和缺血的人成肌细胞的分化能力。在mRNA的下一代测序之后，进行不稳定性途径分析（IPA）以用于经典途径富集。进行活细胞成像和免疫荧光以基于IPA的见解确定肌细胞融合指数和蛋白质表达，特别是关于细胞周期调节剂，包括细胞分裂周期蛋白2（CDC 2）和polo样激酶1（PLK 1）。缺血性成肌细胞形成减弱的肌管，表明融合减少。此外，来自缺血节段的成肌细胞在与PLK 1（上调）和G2/M DNA损伤检查点调节（下调）相关的经典途径中显示出显著差异。用BI 2536抑制PLK 1在24小时内不影响任何组中的细胞活力，但在PAD成肌细胞中更显著地阻止融合。此外，PLK 1抑制降低了灌注细胞中的检查点蛋白CDC 2的表达，但对缺血细胞没有影响。来自缺血肌肉的分化成肌细胞在基因表达方面具有显着差异，包括对DNA损伤检查点调节和细胞周期进程至关重要的基因表达。DNA损伤检查点失调可能导致PAD中的肌病。
We propose that MuSC-derived myoblasts in PAD have transcriptomic differences that can highlight underlying causes of ischemia-induced myopathy. Differentiation capacity among perfused and ischemic human myoblasts was compared. Following next generation sequencing of mRNA, Ingenuity Pathway Analysis (IPA) was performed for canonical pathway enrichment. Live cell imaging and immunofluorescence were performed to determine myocyte fusion index and protein expression based on insights from IPA, specifically concerning cell cycle regulators including cell-division cycle protein 2 (CDC2) and polo-like kinase 1 (PLK1). Ischemic myoblasts formed attenuated myotubes indicative of reduced fusion. Additionally, myoblasts from ischemic segments showed significant differences in canonical pathways associated with PLK1 (upregulated) and G2/M DNA damage checkpoint regulation (downregulated). PLK1 inhibition with BI2536 did not affect cell viability in any group over 24 hours but deterred fusion more significantly in PAD myoblasts. Furthermore, PLK1 inhibition reduced the expression of checkpoint protein CDC2 in perfused but not ischemic cells. Differentiating myoblasts derived from ischemic muscle have significant differences in gene expression including those essential to DNA-damage checkpoint regulation and cell cycle progress. DNA-damage checkpoint dysregulation may contribute to myopathy in PAD.
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DOI：
10.1083/jcb.150.5.1085
发表时间：
2000-09-04
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lee JY;Qu-Petersen Z;Cao B;Kimura S;Jankowski R;Cummins J;Usas A;Gates C;Robbins P;Wernig A;Huard J
通讯作者：
Huard J
影响因子：
3.3
作者：
Kajiguchi, Masahiro;Kondo, Takahisa;Murohara, Toyoaki
通讯作者：
Murohara, Toyoaki
影响因子：
5.8
作者：
Dumont, Nicolas A.;Bentzinger, C. Florian;Rudnicki, Michael A.
通讯作者：
Rudnicki, Michael A.
DOI：
10.1016/j.ejvs.2012.04.024
发表时间：
2012-08-01
影响因子：
5.7
作者：
Gasparini, M.;Sabovic, M.;Pisot, R.
通讯作者：
Pisot, R.
影响因子：
16.6
作者：
Montaudon, Elodie;Nikitorowicz-Buniak, Joanna;Marangoni, Elisabetta
通讯作者：
Marangoni, Elisabetta