Background:Transcriptome sequencing can improve genetic diagnosis of Mendelian diseases but requires access to tissue expressing disease-relevant transcripts. We explored genetic testing of hypertrophic cardiomyopathy using transcriptome sequencing of patient-specific human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs). We also explored whether antisense oligonucleotides (AOs) could inhibit aberrant mRNA splicing in hiPSC-CMs.Methods:We derived hiPSC-CMs from patients with hypertrophic cardiomyopathy due to MYBPC3 splice-gain variants, or an unresolved genetic cause. We used transcriptome sequencing of hiPSC-CM RNA to identify pathogenic splicing and used AOs to inhibit this splicing.Results:Transcriptome sequencing of hiPSC-CMs confirmed aberrant splicing in 2 people with previously identified MYBPC3 splice-gain variants (c.1090+453C>T and c.1224-52G>A). In a patient with an unresolved genetic cause of hypertrophic cardiomyopathy following genome sequencing, transcriptome sequencing of hiPSC-CMs revealed diverse cryptic exon splicing due to an MYBPC3 c.1928-569G>T variant, and this was confirmed in cardiac tissue from an affected sibling. Antisense oligonucleotide treatment demonstrated almost complete inhibition of cryptic exon splicing in one patient-specific hiPSC-CM line.Conclusions:Transcriptome sequencing of patient specific hiPSC-CMs solved a previously undiagnosed genetic cause of hypertrophic cardiomyopathy and may be a useful adjunct approach to genetic testing. Antisense oligonucleotide inhibition of cryptic exon splicing is a potential future personalized therapeutic option.

中文翻译：

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肥厚型心肌病患者的转录测序揭示了 MYBPC3 中新的剪接变异体

背景：转录本测序可以改善孟德尔疾病的基因诊断，但需要获得表达疾病相关转录本的组织。我们使用患者特异性人诱导多能干细胞衍生心肌细胞 (hiPSC-CM) 的转录组测序探索了肥厚型心肌病的基因检测。我们还探讨了反义寡核苷酸（AOS）能否抑制的hiPSC-CM的异常mRNA剪接方法：我们得出的hiPSC-CMS患者肥厚型心肌病，由于MYBPC3剪接增益变异，或未解决的遗传原因。我们使用的hiPSC-CM RNA的转录组测序，以确定致病的剪接和使用的AO抑制这种splicing.Results：的hiPSC-CM的成绩单测序2人先前确定的证实异常拼接MYBPC3剪接变异体的增益（c.1090 + 453C> T 和 c.1224-52G>A)。在基因组测序后肥厚性心肌病遗传原因未解决的患者中，hiPSC -CM 的转录组测序揭示了由于MYBPC3导致的多种隐秘外显子剪接c.1928-569G>T 变异，这在来自受影响同胞的心脏组织中得到证实。反义寡核苷酸治疗表明，在一种患者特异性 hiPSC-CM 系中几乎完全抑制了隐秘外显子剪接。隐性外显子剪接的反义寡核苷酸抑制是未来潜在的个性化治疗选择。