Human Hereditary Cardiomyopathy Shares a Genetic Substrate With Bicuspid Aortic Valve.

The complex genetics underlying human cardiac disease is evidenced by its heterogenous manifestation, multigenic basis, and sporadic occurrence. These features have hampered disease modeling and mechanistic understanding. Here, we show that 2 structural cardiac diseases, left ventricular noncompaction (LVNC) and bicuspid aortic valve, can be caused by a set of inherited heterozygous gene mutations affecting the NOTCH ligand regulator MIB1 (MINDBOMB1) and cosegregating genes. We used CRISPR-Cas9 gene editing to generate mice harboring a nonsense or a missense MIB1 mutation that are both found in LVNC families. We also generated mice separately carrying these MIB1 mutations plus 5 additional cosegregating variants in the ASXL3, APCDD1, TMX3, CEP192, and BCL7A genes identified in these LVNC families by whole exome sequencing. Histological, developmental, and functional analyses of these mouse models were carried out by echocardiography and cardiac magnetic resonance imaging, together with gene expression profiling by RNA sequencing of both selected engineered mouse models and human induced pluripotent stem cell-derived cardiomyocytes. Potential biochemical interactions were assayed in vitro by coimmunoprecipitation and Western blot. Mice homozygous for the MIB1 nonsense mutation did not survive, and the mutation caused LVNC only in heteroallelic combination with a conditional allele inactivated in the myocardium. The heterozygous MIB1 missense allele leads to bicuspid aortic valve in a NOTCH-sensitized genetic background. These data suggest that development of LVNC is influenced by genetic modifiers present in affected families, whereas valve defects are highly sensitive to NOTCH haploinsufficiency. Whole exome sequencing of LVNC families revealed single-nucleotide gene variants of ASXL3, APCDD1, TMX3, CEP192, and BCL7A cosegregating with the MIB1 mutations and LVNC. In experiments with mice harboring the orthologous variants on the corresponding Mib1 backgrounds, triple heterozygous Mib1 Apcdd1 Asxl3 mice showed LVNC, whereas quadruple heterozygous Mib1 Cep192 Tmx3;Bcl7a mice developed bicuspid aortic valve and other valve-associated defects. Biochemical analysis suggested interactions between CEP192, BCL7A, and NOTCH. Gene expression profiling of mutant mouse hearts and human induced pluripotent stem cell-derived cardiomyocytes revealed increased cardiomyocyte proliferation and defective morphological and metabolic maturation. These findings reveal a shared genetic substrate underlying LVNC and bicuspid aortic valve in which MIB1-NOTCH variants plays a crucial role in heterozygous combination with cosegregating genetic modifiers.This study was supported by grants PID2019-104776RB-I00 and PID2020-120326RB-I00, CB16/11/00399 (CIBER CV) financed by MCIN/AEI/10.13039/501100011033, a grant from the Fundación BBVA (Ref. BIO14_298), and a grant from Fundació La Marató de TV3 (Ref. 20153431) to J.L.d.l.P. M.S.-A. was supported by a PhD contract from the Severo Ochoa Predoctor-al Program (SVP-2014-068723) of the MCIN/AEI/10.13039/501100011033. J.R.G.-B. was supported by SEC/FEC-INV-BAS 21/021. A.R. was funded by grants from MCIN (PID2021123925OB-I00), TerCel (RD16/0011/0024), AGAUR (2017-SGR-899), and Fundació La Marató de TV3 (201534-30). J.M.P.-P. was supported by RTI2018-095410-B-I00 (MCIN) and PY2000443 (Junta de Andalucía). B.I. was supported by the European Commission (H2020-HEALTH grant No. 945118) and by MCIN (PID2019-107332RB-I00). DO’R was sup-ported by the Medical Research Council (MC-A658-5QEB0) and KAMcG by the British Heart Foundation (RG/19/6/34387, RE/18/4/34215). The cost of this publication was supported in part with funds from the European Regional Devel-opment Fund. The Centro Nacional de Investigaciones Cardiovasculares is sup-ported by the ISCIII, the MCIN, and the Pro Centro Nacional de Investigaciones Cardiovasculares Foundation and is a Severo Ochoa Center of Excellence (grant CEX2020001041-S) financed by MCIN/AEI/10.13039/501100011033. For the purpose of open access, the authors have applied a CC BY public copyright license to any Author Accepted Manuscript version arising.S

Modeling MYBPC3-and ACTC1-related Hypertrophic Cardiomyopathy and LeftVentricular Non-Compaction in mice

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de Lectura: 25-02-2022Esta Tesis tiene embargado el acceso al texto completo hasta el 25-12-2023La compactación ventricular es un proceso fundamental en el desarrollo del corazón en el que se forma la pared muscular gruesa y uniforme presente en los ventrículos del corazón adulto. Este proceso, que tiene lugar durante la gestación y etapa perinatal, está regulado por numerosas vías de señalización y factores de transcripción que controlan la proliferación y maduración del miocardio. Durante la compactación, el miocardio trabecular (que favorece el suministro de oxígeno y nutrientes al miocardio durante el desarrollo temprano del corazón) se integran en la pared ventricular. Los defectos en este proceso dan lugar a la miocardiopatía no compactada (LVNC), caracterizada por la presencia de trabéculas en el corazón adulto y que puede cursar asintomáticamente o requerir un trasplante por insuficiencia cardíaca. Está enfermedad puede darse de manera aislada o de forma simultánea con otras cardiomiopatías, como la hipertrofia cardíaca (HCM). La HCM es la cardiomiopatía más frecuente y se caracteriza por un engrosamiento de las paredes y/o el septo ventricular. Aunque la LVNC y HCM son dos enfermedades diferentes que se desarrollan de manera distinta, pueden coexistir afectando a un mismo paciente y compartiendo una causa genética común. Este es el caso de mutaciones identificadas en los genes MYBPC3 y ACTC1 en pacientes afectados con LVNC y HCM. MYBPC3 codifica la proteína de unión a MIOSINA C, una proteína sarcomérica que regula la contracción del sarcómero cardíaco. En contra, ACTC1 codifica la ACTINA cardíaca, la isoforma principal de esta proteína en el corazón. Con el objetivo de identificar los mecanismos que producen HCM y LVNC a partir de mutaciones individuales en genes del sarcómero, hemos usado la herramienta de edición genética CRISPR-Cas9 para generar tres modelos murinos portadores de tres mutaciones diferentes (truncadoras y no truncadoras) en MYBPC3. Además, hemos modelado en el genoma murino una cuarta mutación no truncadora identificada en ACTC1. Hemos podido establecer que los ratones portadores de variantes truncadoras de Mybpc3 en homocigosis presentan un fenotipo fetal de hipertrabeculación transitorio que, en estadios posnatales, evoluciona a hipertrofia cardíaca. Hemos analizado el perfil transcripcional de estos dos fenotipos y, además, hemos determinado como contribuye el linaje celular del miocardio compacto (Hey2+) al desarrollo de ambos fenotipos. Hemos establecido también que la variante no truncadora Mybp3 V176M da lugar, en homocigosis, a un fenotipo de hipertrofia cardíaca menos severo. Por último, hemos determinado que los ratones heterocigotos para la variante Actc1 E101K/+ muestran alteraciones en el sistema de conducción cardíaco características del crecimiento biauricular tras someterlos a estrés cardíacoLa financiación del proyecto provino los siguientes proyectos adjudicados al Dr. José Luis de la Pompa: SAF2016-78370-R, PID2019-104776RB-I00 y CB16/11/00399 (CIBER CV) por parte del Ministerio de Ciencia e Innovación (MCIN) y de la de la Agencia Estatal de Investigación (AEI) 10.13039/501100011033Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC

Myocardial Notch1-Rbpj deletion does not affect NOTCH signaling, heart development or function.

During vertebrate cardiac development NOTCH signaling activity in the endocardium is essential for the crosstalk between endocardium and myocardium that initiates ventricular trabeculation and valve primordium formation. This crosstalk leads later to the maturation and compaction of the ventricular chambers and the morphogenesis of the cardiac valves, and its alteration may lead to disease. Although endocardial NOTCH signaling has been shown to be crucial for heart development, its physiological role in the myocardium has not been clearly established. Here we have used mouse genetics to evaluate the role of NOTCH in myocardial development. We have inactivated the unique and ubiquitous NOTCH effector RBPJ in early cardiomyocytes progenitors, and examined its consequences in cardiac development and function. Our results show that mice with Tnnt2-Cre-mediated myocardial-specific deletion of Rbpj develop to term, with homozygous mutant animals showing normal expression of cardiac development markers, and normal adult heart function. Similar observations have been obtained after Notch1 deletion with Tnnt2-Cre. We have also deleted Rbpj in both myocardial and endocardial progenitor cells, using the Nkx2.5-Cre driver, resulting in ventricular septal defect (VSD), double outlet right ventricle (DORV), and bicuspid aortic valve (BAV), due to NOTCH signaling abrogation in the endocardium of cardiac valves. Our data demonstrate that NOTCH-RBPJ inactivation in the myocardium does not affect heart development or adult cardiac function

Disfunción ventricular sistólica, un nuevo marcador de enfermedad coronaria en pacientes con estenosis aórtica sin infarto de miocardio previo

La determinación de factores clínicos predictivos de la presencia de enfermedad coronaria podría obviar la necesidad de realizar una coronariografía en pacientes seleccionados con estenosis aórtica severa. Se estudió a 315 pacientes (68 ± 8 años) sin infarto previo con estenosis aórtica severa a los que se realizó coronariografía. En el análisis univariable, resultaron significativas la edad (p = 0,001), la dislipemia (p = 0,003), la angina (p = 0,018), el gradiente aórtico (p = 0,001) y la fracción de eyección (FE) reducida (p = 0,006). En el análisis multivariable las variables asociadas de forma independiente a la lesión coronaria fueron la edad (odds ratio [OR] = 1,079, p = 0,01), la FE < 40% (OR = 2,685, p = 0,02), la angina (OR = 2,518, p = 0,04) y la dislipemia (OR = 2,34, p = 0,008). La disfunción ventricular se correlaciona de forma independiente con la presencia de lesiones coronaria

Comunicación multidimensional. Derechos humanos, internet, redes sociales y periodismo digital (Tomo I)

Comunicación Multidimensional, Tomo 1, reúne textos en los que se aborda la importancia de la comunicación en temas de derechos humanos, una mirada que va más allá de la democratización de la palabra; evidencia el empoderamiento de colectivos y comunidades en torno a temáticas competentes en sus contextos para que los individuos trabajen en conjunto y en favor de los suyos; hace una crítica de la visión colonizadora — vigente aún en nuestras modernidades— y enfatiza en la lucha de las minorías. Así mismo muestra el quehacer periodístico desde los entornos virtuales y las redes sociales, un campo que se posiciona con mayor frecuencia y cotidianidad. Pone sobre la mesa el cuestionamiento la información que se consume desde las virtualidades, y propone generar contenidos verificados y contrastados

A Human Hereditary Cardiomyopathy Shares a Genetic Substrate With Bicuspid Aortic Valve

Background:The complex genetics underlying human cardiac disease is evidenced by its heterogenous manifestation, multigenic basis, and sporadic occurrence. These features have hampered disease modeling and mechanistic understanding. Here, we show that 2 structural cardiac diseases, left ventricular noncompaction (LVNC) and bicuspid aortic valve, can be caused by a set of inherited heterozygous gene mutations affecting the NOTCH ligand regulator MIB1 (MINDBOMB1) and cosegregating genes. Methods:We used CRISPR-Cas9 gene editing to generate mice harboring a nonsense or a missense MIB1 mutation that are both found in LVNC families. We also generated mice separately carrying these MIB1 mutations plus 5 additional cosegregating variants in the ASXL3, APCDD1, TMX3, CEP192, and BCL7A genes identified in these LVNC families by whole exome sequencing. Histological, developmental, and functional analyses of these mouse models were carried out by echocardiography and cardiac magnetic resonance imaging, together with gene expression profiling by RNA sequencing of both selected engineered mouse models and human induced pluripotent stem cell-derived cardiomyocytes. Potential biochemical interactions were assayed in vitro by coimmunoprecipitation and Western blot. Results:Mice homozygous for the MIB1 nonsense mutation did not survive, and the mutation caused LVNC only in heteroallelic combination with a conditional allele inactivated in the myocardium. The heterozygous MIB1 missense allele leads to bicuspid aortic valve in a NOTCH-sensitized genetic background. These data suggest that development of LVNC is influenced by genetic modifiers present in affected families, whereas valve defects are highly sensitive to NOTCH haploinsufficiency. Whole exome sequencing of LVNC families revealed single-nucleotide gene variants of ASXL3, APCDD1, TMX3, CEP192, and BCL7A cosegregating with the MIB1 mutations and LVNC. In experiments with mice harboring the orthologous variants on the corresponding Mib1 backgrounds, triple heterozygous Mib1 Apcdd1 Asxl3 mice showed LVNC, whereas quadruple heterozygous Mib1 Cep192 Tmx3;Bcl7a mice developed bicuspid aortic valve and other valve-associated defects. Biochemical analysis suggested interactions between CEP192, BCL7A, and NOTCH. Gene expression profiling of mutant mouse hearts and human induced pluripotent stem cell-derived cardiomyocytes revealed increased cardiomyocyte proliferation and defective morphological and metabolic maturation. Conclusions:These findings reveal a shared genetic substrate underlying LVNC and bicuspid aortic valve in which MIB1-NOTCH variants plays a crucial role in heterozygous combination with cosegregating genetic modifiers