Data_Sheet_1_Full-Length Transcriptome Sequencing: An Insight Into the Dog Model of Heart Failure.XLS

Heart failure (HF) leads to a progressive increase in morbidity and mortality rates. This study aimed to explore the transcriptional landscape during HF and identify differentially expressed transcripts (DETs) and alternative splicing events associated with HF. We generated a dog model of HF (n = 3) using right ventricular pacemaker implantation. We performed full-length transcriptome sequencing (based on nanopore platform) on the myocardial tissues and analyzed the transcripts using differential expression analysis and functional annotation methods [Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses]. Additionally, we estimated the expression of the selected genes by quantitative real-time PCR (qRT-PCR) and detected the proportion of immune cells using flow cytometry. We found that increased B-type natriuretic peptide reduced ejection fraction, and apparent clinical signs were observed in the dog model of HF. We identified 67,458 transcripts using full-length transcriptome sequencing. A total of 785 DETs were obtained from the HF and control groups. These DETs were mainly enriched in the immune responses, especially Th1, Th2, and Th17 cell differentiation processes. Furthermore, flow cytometry results revealed that the proportion of Th1 and Th17 cells increased in patients with HF compared to controls, while the proportion of Th2 cells decreased. Differentially expressed genes in the HF and control groups associated with Th1, Th2, and Th17 cell differentiation were quantified using qRT-PCR. We also identified variable splicing events of sarcomere genes (e.g., MYBPC3, TNNT2, TTN, FLNC, and TTNI3). In addition, we detected 4,892 transcription factors and 406 lncRNAs associated with HF. Our analysis based on full-length transcript sequencing provided an analysis perspective in a dog model of HF, which is valuable for molecular research in an increasingly relevant large animal model of HF.

心力衰竭（Heart Failure，HF）可导致发病率与死亡率进行性升高。本研究旨在探究心力衰竭过程中的转录组全景，并筛选与心力衰竭相关的差异表达转录本（Differentially Expressed Transcripts，DETs）以及可变剪接事件。本研究通过右心室起搏器植入术构建了3例心力衰竭犬模型。我们对心肌组织开展了基于纳米孔测序平台的全长转录组测序，并通过差异表达分析与功能注释方法[包括基因本体论（Gene Ontology，GO）和京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）富集分析]对转录本进行分析。此外，我们通过实时荧光定量PCR（Quantitative Real-time PCR，qRT-PCR）检测了筛选出的基因表达水平，并利用流式细胞术检测了免疫细胞的比例。研究发现，心力衰竭犬模型中B型钠尿肽水平升高且射血分数降低，并出现了明确的临床体征。通过全长转录组测序，我们共鉴定得到67458条转录本。心力衰竭组与对照组间共筛选得到785个差异表达转录本。这些差异表达转录本主要富集于免疫应答通路，尤其是Th1、Th2及Th17细胞分化过程。进一步的流式细胞术结果显示，与对照组相比，心力衰竭模型犬的Th1与Th17细胞比例升高，而Th2细胞比例降低。我们通过qRT-PCR对心力衰竭组与对照组中与Th1、Th2及Th17细胞分化相关的差异表达基因进行了定量验证。我们还鉴定得到了肌节基因的可变剪接事件，例如MYBPC3、TNNT2、TTN、FLNC及TTNI3。此外，我们还检测到4892个与心力衰竭相关的转录因子以及406个长链非编码RNA（long non-coding RNA，lncRNAs）。本研究基于全长转录组测序的分析为心力衰竭犬模型提供了新的研究视角，这对于日益受到关注的心力衰竭大动物模型的分子机制研究具有重要价值。