Microvascular preservation & cardiomyocyte hyperplasia underlie adaptive right ventricle development in congenital heart disease-pulmonary arterial hypertension

Right ventricular (RV) failure is the primary cause of death among patients with pulmonary arterial hypertension (PAH). Patients with congenital heart disease-associated PAH (CHD-PAH) demonstrate improved outcomes compared to patients with other forms of PAH, which is related to the maintenance of an adaptively hypertrophied RV. In an ovine model of CHD-PAH, we aimed to elucidate the cellular, microvascular, and transcriptional adaptations to congenital pressure overload that support RV function. Fetal surgery was performed on late gestation lambs to insert an aortopulmonary graft, leading to a persistent congenital left-right shunt and RV pressure load. At 3 days and 4-6 weeks of life, shunt RV myocardial structure, growth mechanisms, and transcriptomes were compared to age-matched control and unoperated fetal RV. At 4-6 weeks of age, shunt lambs demonstrate significant RV enlargement (shunt 37.1±2.9g vs control 15.9±1.0g, p<0.001) but maintain stable microvascular density (fetal 3.0±0.6 vs shunt 2.9±0.3 vs control 3.1±0.6 capillaries per 1000 µm3, p>0.05). Shunt RV cardiomyocytes are significantly more numerous and smaller by cross-sectional area than age-matched controls (shunt 73.3±5.5µm2 vs control 99.2±4.9µm2, p=0.013). At 3 days, shunt RV cardiomyocytes show evidence of increased proliferative capacity and ongoing hyperplasia compared to controls. RNA sequencing analyses reveal a distinct transcriptomic profile in shunt RV consistent with a delay in terminal differentiation and metabolic adaptations to support adaptive function. This study provides novel insights into the roles of microvascular preservation and cardiomyocyte hyperplasia in the development of adaptive RV hypertrophy in CHD-PAH. RNA sequencing was performed on snap frozen tissue obtained from the right ventricle free wall of fetal lambs, 4-6 week old control lambs, and 4-6 week old shunt lambs. Shunt animals had previously undergone a fetal surgery to insert an aortopulmonary graft that introduced a congenital left-to-right shunt and pressure load on the right ventricle.