Cardiac transcriptome profiling of diabetic Akita mice using microarray and next generation sequencing

Abstract

Although diabetes mellitus (DM) causes cardiomyopathy and exacerbates heart failure, the underlying molecular mechanisms for diabetic cardiomyopathy/heart failure are poorly understood. Insulin2 mutant (Ins2+/-) Akita is a mouse model of T1DM, which manifests cardiac dysfunction. However, molecular changes at cardiac transcriptome level that lead to cardiomyopathy remain unclear. To understand the molecular changes in the heart of diabetic Akita mice, we profiled cardiac transcriptome of Ins2+/- Akita and Ins2+/+ control mice using next generation sequencing (NGS) and microarray, and determined the implications of differentially expressed genes on various heart failure signaling pathways using Ingenuity pathway (IPA) analysis. First, we validated hyperglycemia, increased cardiac fibrosis, and cardiac dysfunction in twelve-week male diabetic Akita. Then, we analyzed the transcriptome levels in the heart. NGS analyses on Akita heart revealed 137 differentially expressed transcripts, where Bone Morphogenic Protein-10 (BMP10) was the most upregulated and hairy and enhancer of split-related (HELT) was the most downregulated gene. Moreover, twelve long non-coding RNAs (lncRNAs) were upregulated. The microarray analyses on Akita heart showed 351 differentially expressed transcripts, where vomeronasal-1 receptor-180 (Vmn1r180) was the most upregulated and WD Repeat Domain 83 Opposite Strand (WDR83OS) was the most downregulated gene. Further, miR-101c and H19 lncRNA were upregulated but Neat1 lncRNA was downregulated in Akita heart. Eleven common genes were upregulated in Akita heart in both NGS and microarray analyses. IPA analyses revealed the role of these differentially expressed genes in key signaling pathways involved in diabetic cardiomyopathy. Our results provide a platform to initiate focused future studies by targeting these genes and/or non-coding RNAs, which are differentially expressed in Akita hearts and are involved in diabetic cardiomyopathy.

MiR-133a Mimic Alleviates T1DM-Induced Systolic Dysfunction in Akita: An MRI-Based Study

“…A report on Akita heart shows no hypertrophy and fibrosis, even with elevated cardiac levels of β-myosin heavy chain, a molecular marker of hypertrophy ( Basu et al, 2009 ). In contrast, other reports demonstrate increased cardiac hypertrophy ( Chavali et al, 2014 ; Nandi et al, 2016 ) and fibrosis ( Mishra et al, 2010b ; Kesherwani et al, 2017 ) in Akita. In streptozotocin-induced T1DM, another T1DM model system, shows increased cardiac hypertrophy ( Feng et al, 2010 ) and fibrosis ( Chen et al, 2014 ).…”

“…A speckle-tracking-based study reveals abnormal cardiac deformation in 12-week Akita ( Zhou et al, 2018 ). Hemodynamic study using pressure-volume loop shows reduced ± dp/dt and ejection fraction in 12- to 14-week Akita ( Kesherwani et al, 2017 ). Thus, there is controversy on cardiac dysfunction in Akita.…”

“…DM causes diabetic cardiomyopathy (DCM), a heart muscle disease independent of coronary artery disease, hypertension, or valvular disease, which leads to cardiac dysfunction ( Rubler et al, 1972 ). DCM and cardiac dysfunction can be evaluated by different methods including echocardiography ( Bugger et al, 2008 ; Basu et al, 2009 ; Mishra et al, 2010b ; Kesherwani et al, 2015 ; Negishi, 2018 ), pressure-volume loop ( Mishra et al, 2010a ; Kesherwani et al, 2017 ) and magnetic resonance imaging (MRI) ( Heijman et al, 2007 ; La Gerche et al, 2013 ). Echocardiography is a non-invasive method that utilizes ultrasound to measures mitral flow and cardiac dimension.…”

Analysis of macrophage transcriptome in atherogenesis

“…Результаты секвенирования с помощью ТСНП обладают высоким уровнем воспроизводимости. Кроме того, ТСНП не требует различных экспериментов по стандартизации таких параметров, как нелинейный эффект, обычный для микрочипов, из-за химического и оптического насыщения реакции гибридизации [26].…”

Downregulation of long non-coding RNA nuclear enriched abundant transcript 1 promotes cell proliferation and inhibits cell apoptosis by targeting miR-193a in myocardial ischemia/reperfusion injury

“…LncRNA nuclear enriched abundant transcript 1 (lnc-NEAT1) is transcribed by RNA polymerase II from a common promoter and widely expressed in mammalian cells, which functions as scaffolds of nuclear bodies [12][13][14]. Accumulating evidences have revealed that lnc-NEAT1 regulates cells activities through targeting multiple miRNAs or signaling pathways in several diseases including traumatic brain injury, diabetes mellitus as well as carcinomas [15][16][17]. Considering information about the underlying mechanism of lnc-NEAT1 in myocardial I/R injury is rarely known, we conducted this study to investigate the effect of lnc-NEAT1 on cell proliferation and apoptosis in myocardial I/R injury cells, and explore its target miRNAs.…”