Inhibition of DYRK1a Enhances Cardiomyocyte Cycling After Myocardial Infarction.
Inhibition of DYRK1a Enhances Cardiomyocyte Cycling After Myocardial Infarction.
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DOI：
10.1161/circresaha.121.320005
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发表时间：
2022-04-29
影响因子：
20.1
通讯作者：
Wolf, Matthew J.
中科院分区：
文献类型：
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作者：
Young, Alexander;Bradley, Leigh A.;Farrar, Elizabeth;Bilcheck, Helen O.;Tkachenko, Svyatoslav;Saucerman, Jeffrey J.;Bekiranov, Stefan;Wolf, Matthew J.
双特异性酪氨酸磷酸化调节激酶1a（DYRK 1a）有助于控制细胞周期，包括心肌细胞。然而，抑制DYRK 1a对心肌梗死（MI）后心功能和心肌细胞循环的影响仍不清楚。我们使用αMHC-MerDreMer-Ki 67 p-RoxedCre：：Rox-Lox-tdTomato-eGFP（RLTG）（表示为αDKRC：：RLTG）和αMHC-Cre：：Fucci 2aR：：DYRK 1aflox/flox小鼠研究了DYRK 1a的药理学抑制和条件性基因消融对缺血-再灌注（I/R）MI中内源性心肌细胞循环和左心室（LV）收缩功能的影响。我们观察到，去氢骆驼蓬碱，一种DYRK 1a抑制剂，在I/R MI后两周改善了LV射血分数（LVEF）（分别为39.5 ± 1.6%和29.1 ± 1.6%，去氢骆驼蓬碱与安慰剂相比）。去氢骆驼蓬碱还增加了αDKRC：：RLTG小鼠I/R MI后的心肌细胞周期，分别为10.8 ± 1.5 vs 24.3 ± 2.6增强绿色荧光蛋白（eGFP）+心肌细胞，安慰剂vs去氢骆驼蓬碱，p=1.0x10−3）。在成年循环心肌细胞中表达诱导型白喉毒素的αDKRC：：DTA小鼠中，去氢骆驼蓬碱对LVEF的影响减弱。在αMHC-Cre：：Fucci 2aR：：DYRK 1aflox/flox（表示为DYRK 1a k/o）小鼠中，DYRK 1a的条件性心肌细胞特异性基因消融在基线时引起心肌细胞增生（每40倍视野分别为210 ± 28 vs. 126 ± 5个心肌细胞，DYRK 1a k/o vs对照，p = 1.7x10−2），与对照相比，心脏功能没有变化，或其他DYRK亚型表达的代偿性变化。I/R MI后，DYRK 1a k/o小鼠的LV功能改善（LVEF分别为41.8 ± 2.2%和26.4 ± 0.8%，DYRK 1a k/o与对照组相比，p = 3.7x10−2）。从I/R MI或假手术后的αMHC-Cre：：Fucci 2aR：：DYRK 1aflox/flox和αMHC-Cre：：Fucci 2aR小鼠中分离的心肌细胞的RNAseq鉴定出与α MHC-Cre：：Fucci 2aR相比，αMHC-Cre：：Fucci 2aR中有丝分裂细胞周期基因的富集。DYRK 1a的药理学抑制或心肌细胞特异性消融导致I/R MI后基线增生和改善心功能，细胞周期基因表达增加，表明DYRK 1a的抑制可作为治疗MI的治疗靶点。
Dual-specificity tyrosine phosphorylation-regulated kinase 1a (DYRK1a) contributes to the control of cycling cells, including cardiomyocytes. However, the effects of inhibition of DYRK1a on cardiac function and cycling cardiomyocytes after myocardial infarction (MI) remains unknown. We investigated the impacts of pharmacological inhibition and conditional genetic ablation of DYRK1a on endogenous cardiomyocyte cycling and left ventricular (LV) systolic function in ischemia-reperfusion (I/R) MI using αMHC-MerDreMer-Ki67p-RoxedCre::Rox-Lox-tdTomato-eGFP (RLTG) (denoted αDKRC::RLTG) and αMHC-Cre::Fucci2aR::DYRK1aflox/flox mice. We observed that harmine, an inhibitor of DYRK1a, improved LV Ejection Fraction (LVEF) (39.5 ± 1.6% and 29.1 ± 1.6%, harmine versus placebo, respectively), two weeks after I/R MI. Harmine also increased cardiomyocyte cycling after I/R MI in αDKRC::RLTG mice, 10.8 ± 1.5 verses 24.3 ± 2.6 enhanced Green Fluorescent Protein (eGFP)+ cardiomyocytes, placebo verses harmine, respectively, p=1.0x10−3). The effects of harmine on LVEF were attenuated in αDKRC::DTA mice that expressed an inducible diphtheria toxin in adult cycling cardiomyocytes. The conditional cardiomyocyte-specific genetic ablation of DYRK1a in αMHC-Cre::Fucci2aR::DYRK1aflox/flox (denoted DYRK1a k/o) mice caused cardiomyocyte hyperplasia at baseline (210 ± 28 vs. 126 ± 5 cardiomyocytes per 40x field, DYRK1a k/o versus controls, respectively, p = 1.7x10−2) without changes in cardiac function compared to controls, or compensatory changes in the expression of other DYRK isoforms. After I/R MI, DYRK1a k/o mice had improved LV function (LVEF 41.8 ± 2.2% and 26.4 ± 0.8%, DYRK1a k/o versus control, respectively, p = 3.7x10−2). RNAseq of cardiomyocytes isolated from αMHC-Cre::Fucci2aR::DYRK1aflox/flox and αMHC-Cre::Fucci2aR mice after I/R MI or Sham surgeries identified enrichment in mitotic cell cycle genes in αMHC-Cre::Fucci2aR::DYRK1aflox/flox compared to αMHC-Cre::Fucci2aR. The pharmacological inhibition or cardiomyocyte-specific ablation of DYRK1a caused baseline hyperplasia and improved cardiac function after I/R MI, with an increase in cell cycle gene expression, suggesting the inhibition of DYRK1a may serve as a therapeutic target to treat MI.