Heart RNA-seq of therapeutic RBM20 antisense oligonucleotide (ASO) treatment in a mouse model of heart failure with preserved ejection fraction (HFpEF)

We report the heart RNA-sequencing of control animals and animals of a mouse model of heart failure treated or not with RBM20 antisense oligonucleotide. This study aims to investigate the therapeutic effect of antisense oligonucleotide (ASO)-mediated downregulation of the cardiac splice factor RBM20 in a mouse model of heart failure with preserved ejection fraction. 8-week old wild type and titin N2B KO mice with increased wall stiffness were subcutaneously injected with 50 mg/kg/week RBM20 ASO for 8 weeks. RNA from 4 left ventricles per group were isolated using TRIzol followed by a clean-up with the QIAGEN RNA micro kit. The cDNA libraries were generated using the Illumina TruSeq Stranded mRNA LT Sample Prep Kit and 2x150 paired end sequenced on HiSeq4000. The RNA-Sequencing analysis confirms that RBM20 ASO treatment increased the expression of compliant titin isoforms and improved impaired ventricular filling and cardiac function. RNA-seq confirmed RBM20 dependent isoform changes of known targets such as Titin, Ldb3, Camk2d, and Ank3, and served as a sensitive indicator of potential side effects, largely limited to genes related to the immune response.本研究报道了对照组动物，以及心力衰竭小鼠模型动物经RBM20反义寡核苷酸（RBM20 antisense oligonucleotide）处理或未处理的心脏RNA测序数据。本研究旨在探究反义寡核苷酸（antisense oligonucleotide，ASO）介导的心脏剪接因子RBM20下调，在射血分数保留型心力衰竭小鼠模型中的治疗效果。选取8周龄、心肌壁硬度升高的野生型小鼠及肌联蛋白（titin, TTN）N2B基因敲除（KO）小鼠，每周皮下注射50 mg/kg的RBM20 ASO，持续8周。每组取4个左心室样本，采用TRIzol试剂提取总RNA，随后使用QIAGEN RNA微量提取试剂盒进行RNA纯化。采用Illumina TruSeq Stranded mRNA LT样本制备试剂盒构建cDNA文库，并在HiSeq4000测序平台上进行2×150 bp双端测序。RNA测序分析结果证实，RBM20 ASO处理可上调顺应性肌联蛋白亚型的表达，并改善受损的心室充盈功能与心脏整体功能。RNA测序结果证实了RBM20依赖的已知靶基因（包括肌联蛋白、Ldb3、Camk2d及Ank3）的剪接亚型变化，同时可作为潜在不良反应的敏感检测指标，其不良反应主要局限于与免疫应答相关的基因。