Functional, structural and molecular characterization of a new mitral valve prolapse animal model: the FLNA-P637Q KI rat

Abstract

Abstract Introduction Mitral Valve Prolapse (MVP) affects 3% of the population and is characterized by a heterogeneous mitral leaflet remodeling. The pathophysiological mechanisms involved in MVP development are not fully understood, the only therapeutic option remains the surgical valve replacement. We previously identified FLNA as the first gene causing MVP and recently generated a unique knock-in rat model for the FLNA-P637Q mutation that now paves the road to study the molecular mechanisms involved in MVP development. Purpose The aim of our study was to characterize the morphological, functional and molecular expression of the valvular disease in our unique KI rat model. Methods 5 wild-type (WT) and 10 KI rats were evaluated at 3, 6 and 13 weeks. Comprehensive 2D echocardiography was performed to determine valve function and morphology. 3D quantitative analysis of the mitral valve (MV) remodelling was done using micro computed tomography (microCT). MV tissue composition was analysed based on histological and immunohistochemistry. Transcriptomic comparison was performed using RNA-sequencing approach. Results Based on the qualitative echocardiographic assessment of the valve, a high genotype-phenotype concordance was established for WT and KI animals (100%, 93% and 100% matching for each time points). The anterior leaflet was longer in KI comparatively to WT rats (+12 to +14% increase at all time points (p<0.01)). Increased lengths corroborated the increased leaflets volume assessed by microCT analysis (+20 to +58% in KI compared to WT all time points (p<0.05)). Histological and immunohistological analyses (leaflet's thickening, hypercellularity, proteoglycans accumulation without calcification) pointed out towards a myxomatous valve disease. The differential gene expression profile established by RNAseq analysis revealed that inflammation, epithelial cell migration or mechanical transduction pathways were specifically activated in KI valves. Genes such as Itgb2 (+1.30x), Ccl12 (+2.44x), Ccl2 (+1.79x), Ccl28 (+1.53x), Ccl7 (+2.95x), S100a8 (+8.40x) or S100a9 (+2.67x) were significantly upregulated in the GO:0060326 “cell chemotaxis”, p=2.31x10–5. In the GO:0043542 “endothelial cell migration”, p=1.59x10–6, Klf4 (+1.31x) and Tgfbr1 (+1.21x) were upregulated. Genes part of the GO:0048771 “tissue remodelling” (p=5.52x10–5) were also found upregulated. Conclusion These results establish that our unique KI FLNA-P637Q rat develops a myxomatous MV dystrophy comparable to the one described in MVP patients and thus constitutes a pertinent model to study the pathophysiological molecular mechanisms associated with MVP development. Our results point to molecular pathways including inflammation and epithelial activation, which constitute potential therapeutic targets. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Connect Talent