Myosins and MyomiR Network in Patients with Obstructive Hypertrophic Cardiomyopathy
Chiara Foglieni,
Maria Lombardi,
Davide Lazzeroni,
Riccardo Zerboni,
Edoardo Lazzarini,
Gloria Bertoli,
Annalinda Pisano,
Francesca Girolami,
Annapaola Andolfo,
Cinzia Magagnotti,
Giovanni Peretto,
Carmem L. Sartorio,
Iacopo Olivotto,
Giovanni La Canna,
Ottavio Alfieri,
Ornella E. Rimoldi,
Lucio Barile,
Giulia d’Amati,
Paolo G. Camici
Affiliations
Chiara Foglieni
Cardiovascular Research Center, IRCCS San Raffaele Hospital, 58, 20132 Milan, Italy
Maria Lombardi
Cardiovascular Research Center, IRCCS San Raffaele Hospital, 58, 20132 Milan, Italy
Davide Lazzeroni
Cardiovascular Research Center, IRCCS San Raffaele Hospital, 58, 20132 Milan, Italy
Riccardo Zerboni
Cardiovascular Research Center, IRCCS San Raffaele Hospital, 58, 20132 Milan, Italy
Edoardo Lazzarini
Laboratory for Cardiovascular Theranostics, Cardiocentro Ticino Institute, Ente Ospedaliero Cantonale and Faculty of Biomedical Sciences, Università Svizzera Italiana, 6900 Lugano, Switzerland
Gloria Bertoli
Institute of Molecular Bioimaging and Physiology, National Research Council (IBFM-CNR), 20054 Milan, Italy
Annalinda Pisano
Department of Radiological, Oncological and Pathological Sciences, Sapienza University of Rome, Policlinico Umberto I, 00185 Rome, Italy
Francesca Girolami
Cardiology Unit, Meyer Children’s Hospital, 50139 Florence, Italy
Annapaola Andolfo
ProMeFa, Proteomics and Metabolomics Facility, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
Cinzia Magagnotti
ProMeFa, Proteomics and Metabolomics Facility, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
Giovanni Peretto
IRCCS San Raffaele Scientific Institute, Vita-Salute University, 20132 Milan, Italy
Carmem L. Sartorio
Cardiovascular Research Center, IRCCS San Raffaele Hospital, 58, 20132 Milan, Italy
Iacopo Olivotto
Cardiomyopathy Unit, Careggi University Hospital, 50134 Florence, Italy
Giovanni La Canna
IRCCS Humanitas Clinical and Research Institute, 20089 Rozzano, Italy
Ottavio Alfieri
Cardiac Surgery Unit, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
Ornella E. Rimoldi
Institute of Molecular Bioimaging and Physiology, National Research Council (IBFM-CNR), 20054 Milan, Italy
Lucio Barile
Laboratory for Cardiovascular Theranostics, Cardiocentro Ticino Institute, Ente Ospedaliero Cantonale and Faculty of Biomedical Sciences, Università Svizzera Italiana, 6900 Lugano, Switzerland
Giulia d’Amati
Department of Radiological, Oncological and Pathological Sciences, Sapienza University of Rome, Policlinico Umberto I, 00185 Rome, Italy
Paolo G. Camici
Cardiovascular Research Center, IRCCS San Raffaele Hospital, 58, 20132 Milan, Italy
Hypertrophic cardiomyopathy (HCM) is the most common genetic cardiomyopathy. The molecular mechanisms determining HCM phenotypes are incompletely understood. Myocardial biopsies were obtained from a group of patients with obstructive HCM (n = 23) selected for surgical myectomy and from 9 unused donor hearts (controls). A subset of tissue-abundant myectomy samples from HCM (n = 10) and controls (n = 6) was submitted to laser-capture microdissection to isolate cardiomyocytes. We investigated the relationship among clinical phenotype, cardiac myosin proteins (MyHC6, MyHC7, and MyHC7b) measured by optimized label-free mass spectrometry, the relative genes (MYH7, MYH7B and MYLC2), and the MyomiR network (myosin-encoded microRNA (miRs) and long-noncoding RNAs (Mhrt)) measured using RNA sequencing and RT-qPCR. MyHC6 was lower in HCM vs. controls, whilst MyHC7, MyHC7b, and MyLC2 were comparable. MYH7, MYH7B, and MYLC2 were higher in HCM whilst MYH6, miR-208a, miR-208b, miR-499 were comparable in HCM and controls. These results are compatible with defective transcription by active genes in HCM. Mhrt and two miR-499-target genes, SOX6 and PTBP3, were upregulated in HCM. The presence of HCM-associated mutations correlated with PTBP3 in myectomies and with SOX6 in cardiomyocytes. Additionally, iPSC-derived cardiomyocytes, transiently transfected with either miR-208a or miR-499, demonstrated a time-dependent relationship between MyomiRs and myosin genes. The transfection end-stage pattern was at least in part similar to findings in HCM myectomies. These data support uncoupling between myosin protein/genes and a modulatory role for the myosin/MyomiR network in the HCM myocardium, possibly contributing to phenotypic diversity and providing putative therapeutic targets.
