Full-Length Transcriptome Sequencing: An Insight Into the Dog Model of Heart Failure

Xiaoyan Liang; Zechen Bai; Feifei Wang; Yafan Han; Huaxin Sun; Jiasuoer Xiaokereti; Ling Zhang; Xianhui Zhou; Yanmei Lu; Baopeng Tang

doi:10.3389/fcvm.2021.712797

Full-Length Transcriptome Sequencing: An Insight Into the Dog Model of Heart Failure

Front Cardiovasc Med. 2021 Dec 16:8:712797. doi: 10.3389/fcvm.2021.712797. eCollection 2021.

Authors

Xiaoyan Liang^{1

2}, Zechen Bai³, Feifei Wang^{1

2

4}, Yafan Han^{1

2}, Huaxin Sun^{1

2}, Jiasuoer Xiaokereti^{1

2}, Ling Zhang², Xianhui Zhou^{1

2}, Yanmei Lu^{1

2}, Baopeng Tang^{1

2}

Affiliations

¹ Department of Pacing and Electrophysiology, The First Affiliated Hospital of Xinjiang Medical University, Ürümqi, China.
² Xinjiang Key Laboratory of Cardiac Electrophysiology and Cardiac Remodeling, The First Affiliated Hospital of Xinjiang Medical University, Ürümqi, China.
³ Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences (CAS), Shenzhen, China.
⁴ Xinjiang First Aid Center, People's Hospital of Xinjiang Uygur Autonomous Region, Ürümqi, China.

Abstract

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.

Keywords: full-length transcriptome sequencing; heart failure; helper T cell differentiation; target genes; transcript expression analysis.