RESUMEN
We have recently shown that a temporary increase in sarcoplasmic reticulum (SR) cycling via adenovirus-mediated overexpression of sarcoplasmic reticulum ATPase (SERCA2) transiently improves relaxation and delays hypertrophic remodeling in a familial hypertrophic cardiomyopathy (FHC) caused by a mutation in the thin filament protein, tropomyosin (i.e., α-TmE180G or Tm180). In this study, we sought to permanently alter calcium fluxes via phospholamban (PLN) gene deletion in Tm180 mice in order to sustain long-term improvements in cardiac function and adverse cardiac remodeling/hypertrophy. While similar work has been done in FHCs resulting from mutations in thick myofilament proteins, no one has studied these effects in an FHC resulting from a thin filament protein mutation. Tm180 transgenic (TG) mice were crossbred with PLN knockout (KO) mice and four groups were studied in parallel: 1) non-TG (NTG), 2) Tm180, 3) PLNKO/NTG and 4) PLNKO/Tm180. Tm180 mice exhibit increased heart weight/body weight and hypertrophic gene markers compared to NTG mice, but levels in PLNKO/Tm180 mice were similar to NTG. Tm180 mice also displayed altered function as assessed via in situ pressure-volume analysis and echocardiography at 3-6 months and one year; however, altered function in Tm180 mice was rescued back to NTG levels in PLNKO/Tm180 mice. Collagen deposition, as assessed by Picrosirius Red staining, was increased in Tm180 mice but was similar in NTG and in PLNKO/Tm180 mice. Extracellular signal-regulated kinase (ERK1/2) phosphorylation increased in Tm180 mice while levels in PLNKO/Tm180 mice were similar to NTGs. The present study shows that by modulating SR calcium cycling, we were able to rescue many of the deleterious aspects of FHC caused by a mutation in the thin filament protein, Tm.
Asunto(s)
Biomarcadores/metabolismo , Proteínas de Unión al Calcio/deficiencia , Calcio/metabolismo , Cardiomiopatía Hipertrófica Familiar , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Tropomiosina/genética , Animales , Peso Corporal , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/uso terapéutico , Cardiomiopatía Hipertrófica Familiar/diagnóstico por imagen , Cardiomiopatía Hipertrófica Familiar/genética , Cardiomiopatía Hipertrófica Familiar/metabolismo , Cardiomiopatía Hipertrófica Familiar/fisiopatología , Cardiomiopatía Hipertrófica Familiar/terapia , Modelos Animales de Enfermedad , Ecocardiografía , Quinasas MAP Reguladas por Señal Extracelular/genética , Expresión Génica , Humanos , Ratones , Ratones Transgénicos , Mutación , Contracción Miocárdica/genética , Miocardio/citología , Miocardio/metabolismo , Tamaño de los Órganos , Fosforilación , Reacción en Cadena en Tiempo Real de la Polimerasa , Retículo Sarcoplasmático/genética , Retículo Sarcoplasmático/metabolismo , Tropomiosina/metabolismoRESUMEN
Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant genetic disorder linked to numerous mutations in the sarcomeric proteins. The clinical presentation of FHC is highly variable, but it is a major cause of sudden cardiac death in young adults with no specific treatments. We tested the hypothesis that early intervention in Ca(2+) regulation may prevent pathological hypertrophy and improve cardiac function in a FHC displaying increased myofilament sensitivity to Ca(2+) and diastolic dysfunction. A transgenic (TG) mouse model of FHC with a mutation in tropomyosin at position 180 was employed. Adenoviral-Serca2a (Ad.Ser) was injected into the left ventricle of 1-day-old non-transgenic (NTG) and TG mice. Ad.LacZ was injected as a control. Serca2a protein expression was significantly increased in NTG and TG hearts injected with Ad.Ser for up to 6 weeks. Compared to TG-Ad.LacZ hearts, the TG-Ad.Ser hearts showed improved whole heart morphology. Moreover, there was a significant decline in ANF and ß-MHC expression. Developed force in isolated papillary muscle from 2- to 3-week-old TG-Ad.Ser hearts was higher and the response to isoproterenol (ISO) improved compared to TG-Ad.LacZ muscles. In situ hemodynamic measurements showed that by 3 months the TG-Ad.Ser hearts also had a significantly improved response to ISO compared to TG-Ad.LacZ hearts. The present study strongly suggests that Serca2a expression should be considered as a potential target for gene therapy in FHC. Moreover, our data imply that development of FHC can be successfully delayed if therapies are started shortly after birth.
Asunto(s)
Cardiomiopatía Hipertrófica Familiar/fisiopatología , Cardiomiopatía Hipertrófica Familiar/terapia , Técnicas de Transferencia de Gen , Terapia Genética , Pruebas de Función Cardíaca , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/uso terapéutico , Remodelación Ventricular/fisiología , Citoesqueleto de Actina/efectos de los fármacos , Citoesqueleto de Actina/metabolismo , Adenoviridae/genética , Animales , Animales Recién Nacidos , Factor Natriurético Atrial/metabolismo , Proteínas de Unión al Calcio/metabolismo , Hemodinámica/efectos de los fármacos , Humanos , Inyecciones , Isoproterenol/farmacología , Ratones , Ratones Transgénicos , Contracción Miocárdica/efectos de los fármacos , Cadenas Pesadas de Miosina/metabolismo , Fosforilación/efectos de los fármacos , Isoformas de Proteínas/metabolismo , Conejos , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/genética , Remodelación Ventricular/efectos de los fármacosRESUMEN
Administration of ß-lactam antibiotics by extended infusion optimizes the pharmacodynamic properties and bactericidal activity of these agents resulting in a potential improvement in patient outcomes and reduction in drug expenditure. Consequently, a pharmacist-led piperacillin-tazobactam extended 4-hour infusion guideline was implemented hospital-wide at a 500-bed academic medical center. Each piperacillin-tazobactam infusion was prospectively monitored for 5 weeks to ensure accurate administration and identify barriers to guideline adherence. Overall, a total of 103 patients received 1215 doses of piperacillin-tazobactam by extended infusions. In all, 98% of the doses were administered at the correct extended infusion rate and 94% of the doses were given at the scheduled time. There were a total of 20 missed doses and 53 delayed doses, accounting for 2% and 4% of the total administered doses, respectively. The primary barrier to adherence was the patient not being on the unit at the time of the scheduled dose followed by the piperacillin-tazobactam dose not being available on the floor. While insufficient power prevented meaningful evaluation of clinical outcomes, we anticipate a conservative annual estimated cost savings of $108,529. Key elements contributing to our success included consistent pharmacy leadership, multidisciplinary involvement, thorough inservicing to health care professionals, hospital-wide implementation, and extensive quality assurance monitoring.