ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/020/SRR16293220/SRR16293220_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/022/SRR16293222/SRR16293222_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/024/SRR16293224/SRR16293224_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/016/SRR16293216/SRR16293216_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/015/SRR16293215/SRR16293215_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/016/SRR16293216/SRR16293216_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/019/SRR16293219/SRR16293219_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/023/SRR16293223/SRR16293223_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/025/SRR16293225/SRR16293225_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/026/SRR16293226/SRR16293226_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/022/SRR16293222/SRR16293222_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/021/SRR16293221/SRR16293221_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/015/SRR16293215/SRR16293215_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/017/SRR16293217/SRR16293217_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/026/SRR16293226/SRR16293226_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/025/SRR16293225/SRR16293225_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/024/SRR16293224/SRR16293224_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/019/SRR16293219/SRR16293219_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/018/SRR16293218/SRR16293218_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/020/SRR16293220/SRR16293220_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/021/SRR16293221/SRR16293221_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/023/SRR16293223/SRR16293223_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/017/SRR16293217/SRR16293217_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR162/018/SRR16293218/SRR16293218_2.fastq.gzprimaryOK200GenomicsInstitute of Pharmacology and Toxicology, Technical University of Munichhttps://www.ebi.ac.uk/ena/browser/view/PRJNA770440Homo sapiensxref:PubMed:35017523Abnormalities of ventricular action potential (AP) cause malignant cardiac arrhythmias and sudden cardiac death. Here, we sought to identify microRNAs (miRNAs) that regulate the human cardiac AP and asked whether their manipulation allows for therapeutic modulation of AP abnormalities. Quantitative analysis of the miRNA targetomes in human cardiac myocytes identified miR-365 as a primary miRNA to regulate repolarizing ion channels. AP recordings in patient-specific induced pluripotent stem cell-derived cardiac myocytes (iPSC-CMs) showed that elevation of miR-365 level significantly prolonged AP duration in hiPSC-CMs derived from a Short-QT syndrome (SQTS) patient, whereas specific inhibition of miR-365 normalized pathologically prolonged AP in Long-QT syndrome (LQTS) iPSC-CMs. Transcriptome analyses in human iPSC-CMs at bulk and single-cell level corroborated the key cardiac repolarizing channels as direct targets of miR-365, together with functionally synergistic regulation of additional AP-regulating genes by this miRNA. Whole-cell patch clamp experiments revealed miR-365-based regulation of repolarizing ionic currents. Finally, refractory period measurements in human myocardial slices substantiated the prolonging effect of miR-365 on AP duration also in adult human myocardial tissue. Our results delineate miR-365 to regulate human cardiac AP duration by targeting key factors of cardiac repolarization. Overall design: healthy hiPSC-CMs treated with mimic-365 and antimiR-365 were subjected to single cell RNA-seq. Untreated, antimiR-Ctrl and antimiR-365-treated LQT1-specific hiPSC-CMs were labeled with cell hashing oligos (HTO), multiplexed and subjected to single cell RNA-seq (10x Genomics).ENAcongenital myasthenia, HH, d230, Kv1.9, Fibroblast-Derived IPS Cell, Induced Pluripotent Stem Cell, IPS, hg., bar, Fibroblast Derived IPS Cells, dTAFII250, hiPSC, SQT2, IDOL, EfW1, IPS Cells, Hh, dmTAF[[II]]230, kvlqt1, human iPS cell line cell, anon-WO0134654.19, CMS, DmelCG4637, sqt2, dmTAF1, Taf230, Whole Transcriptome Shotgun Sequencing, Mir, MIR, congenital MG, TAF250, Miranda, Fibroblast-Derived IPS Cells, Taf200, dTAF[[II]]250, MONDOA, TFIID TAF250, cel, cell, Mrt, Taf1p, KVLQT1, myasthenia gravis congenital, mir, ATFB1, Human Induced Pluripotent Stem Cells, ATFB3, dTAF250, Myosin regulatory light chain-interacting protein, atfb3, kv1.9, bar-3, LQT, atfb1, bHLHe36, TAF, myasthenia gravis pseudoparalytica, Idol, 9430057C20Rik, LQT1, jlns1, l(3)hh, dTAF[[II]]230, TAF[[II]]250, Myosin regulatory light chain interacting protein, 6.3.2.-, lqt, TAF200, l(3)84Ab, RNA-seq, BG:DS00004.13, Kv7.1, TAFII-250, TAF250/230, kcna9, Cell, kcna8, CG4637, dTAF230, arf1gap, CG12249, AW228700, TAFII250, Fibroblast Derived Induced Pluripotent Stem Cells, p230, bar3, RWS, lqt1, TAF[[II]]250/230, TFIID, DmelCG12249, IPS Cell, JLNS1, Taf[[II]]250, familial limb-girdle myasthenia, TAF[[II]]230, l(3)neo56, l(3)neo57, KCNA8, KCNA9, rws, Mira, Fibroblast-Derived IPS, TAF[II]250, Inducible degrader of the LDL-receptor, WRS, CG17603, Fibroblast-Derived, TAF[[II]], k(v)7.1, DmelCG17603, Taf250, anon-WO0182946.19, SR3-5, Cells, Fibroblast-Derived Induced Pluripotent Stem Cells, MIRA, kv7.1, wrs, Human Induced Pluripotent Stem Cell, erb-Goldflam syndrome, TAF230, TAF1congenital myasthenia, HH, d230, Kv1.9, Fibroblast-Derived IPS Cell, Induced Pluripotent Stem Cell, IPS, hg., bar, Fibroblast Derived IPS Cells, dTAFII250, hiPSC, SQT2, IDOL, EfW1, IPS Cells, Hh, dmTAF[[II]]230, kvlqt1, human iPS cell line cell, anon-WO0134654.19, CMS, DmelCG4637, sqt2, dmTAF1, Taf230, Whole Transcriptome Shotgun Sequencing, Mir, MIR, congenital MG, TAF250, Miranda, Fibroblast-Derived IPS Cells, Taf200, dTAF[[II]]250, MONDOA, TFIID TAF250, cel, cell, Mrt, Taf1p, KVLQT1, myasthenia gravis congenital, mir, ATFB1, Human Induced Pluripotent Stem Cells, ATFB3, dTAF250, Myosin regulatory light chain-interacting protein, atfb3, kv1.9, bar-3, LQT, atfb1, bHLHe36, TAF, myasthenia gravis pseudoparalytica, Idol, 9430057C20Rik, LQT1, jlns1, l(3)hh, dTAF[[II]]230, TAF[[II]]250, Myosin regulatory light chain interacting protein, 6.3.2.-, lqt, TAF200, l(3)84Ab, RNA-seq, BG:DS00004.13, Kv7.1, TAFII-250, TAF250/230, kcna9, Cell, kcna8, CG4637, dTAF230, arf1gap, CG12249, AW228700, TAFII250, Fibroblast Derived Induced Pluripotent Stem Cells, p230, bar3, RWS, lqt1, TAF[[II]]250/230, TFIID, DmelCG12249, IPS Cell, JLNS1, Taf[[II]]250, familial limb-girdle myasthenia, TAF[[II]]230, l(3)neo56, l(3)neo57, KCNA8, KCNA9, rws, Mira, Fibroblast-Derived IPS, TAF[II]250, Inducible degrader of the LDL-receptor, WRS, CG17603, Fibroblast-Derived, TAF[[II]], k(v)7.1, DmelCG17603, Taf250, anon-WO0182946.19, SR3-5, Cells, Fibroblast-Derived Induced Pluripotent Stem Cells, MIRA, kv7.1, wrs, Human Induced Pluripotent Stem Cell, erb-Goldflam syndrome, TAF230, TAF1falsesingle cell RNA-seq in healthy and LQT1 hiPSC-CMs upon manipulation of miR-365single cell RNA-seq in healthy and LQT1 hiPSC-CMs upon manipulation of miR-3652025-09-242021-12-04PRJNA770440GSE185688960635017523