Aberrations in the cGMP-PKG-Ca2+ pathway are implicated in cardiovascular complications of diverse etiologies, though involved molecular mechanisms are not understood. We performed RNA-Seq analysis to profile global changes in gene expression and exon splicing in Chagas disease (ChD) murine myocardium. Ingenuity-Pathway-Analysis of transcriptome dataset identified 26 differentially expressed genes associated with increased mobilization and cellular levels of Ca2+ in ChD hearts. Mixture-of-isoforms and Enrichr KEGG pathway analyses of the RNA-Seq datasets from ChD (this study) and diabetic (previous study) murine hearts identified alternative splicing (AS) in eleven genes (Arhgef10, Atp2b1, Atp2a3, Cacna1c, Itpr1, Mef2a, Mef2d, Pde2a, Plcb1, Plcb4, and Ppp1r12a) of the cGMP-PKG-Ca2+ pathway in diseased hearts. AS of these genes was validated by an exon exclusion-inclusion assay. Further, Arhgef10, Atp2b1, Mef2a, Mef2d, Plcb1, and Ppp1r12a genes consisted RBFOX2 (RNA-binding protein) binding-site clusters, determined by analyzing the RBFOX2 CLIP-Seq dataset. H9c2 rat heart cells transfected with Rbfox2 (vs. scrambled) siRNA confirmed that expression of Rbfox2 is essential for proper exon splicing of genes of the cGMP-PKG-Ca2+ pathway. We conclude that changes in gene expression may influence the Ca2+ mobilization pathway in ChD, and AS impacts the genes involved in cGMP/PKG/Ca2+ signaling pathway in ChD and diabetes. Our findings suggest that ChD patients with diabetes may be at increased risk of cardiomyopathy and heart failure and provide novel ways to restore cGMP-PKG regulated signaling networks via correcting splicing patterns of key factors using oligonucleotide-based therapies for the treatment of cardiovascular complications.

中文翻译：

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cGMP-PKG-Ca2 +信号通路的基因也可在心肌病中剪接：RBFOX2的作用。

尽管尚不清楚涉及的分子机制，但cGMP-PKG-Ca2 +途径的异常与多种病因的心血管并发症有关。我们进行了RNA-Seq分析，以分析Chagas病（ChD）鼠心肌中基因表达和外显子剪接的总体变化。转录组数据集的独创性-途径-分析鉴定了与ChD心脏中动员和细胞内Ca2 +水平升高相关的26个差异表达基因。来自ChD（本研究）和糖尿病（先前研究）的鼠心RNA-Seq数据集的同工型和Enrichr KEGG通路混合物的分析，在11种基因（Arhgef10，Atp2b1，Atp2a3，Cacna1c，Itpr1，患病心脏中cGMP-PKG-Ca2 +途径的Mef2a，Mef2d，Pde2a，Plcb1，Plcb4和Ppp1r12a）。这些基因的AS通过外显子排除-包含测定法验证。此外，Arhgef10，Atp2b1，Mef2a，Mef2d，Plcb1和Ppp1r12a基因由RBFOX2（RNA结合蛋白）结合位点簇组成，通过分析RBFOX2 CLIP-Seq数据集确定。用Rbfox2（相对于加扰）siRNA转染的H9c2大鼠心脏细胞证实，Rbfox2的表达对于cGMP-PKG-Ca2 +途径基因的正确外显子剪接至关重要。我们得出结论，基因表达的变化可能影响ChD中的Ca2 +动员途径，而AS影响ChD和糖尿病中cGMP / PKG / Ca2 +信号通路中涉及的基因。