Abstract P3001: Hnrnpa2b1 An Rna-binding Protein That Orchestrates Smooth Muscle Cell Phenotype In Pulmonary Arterial Hypertension.

Abstract

Background: RNA-binding proteins (RPBs) are master orchestrators of gene expression regulation. RBPs regulate hundreds of transcripts at once by recognizing specific motifs. Characterizing RBP targets is critical to harvest their full therapeutic potential. However, such investigation has often been restricted to a few RBP targets, limiting our understanding of their function. In cancer, the RBP HNRNPA2B1 (A2B1) promotes the pro-proliferative / anti-apoptotic phenotype. The same phenotype in pulmonary arterial smooth muscle cells (PASMC) is responsible for the development of pulmonary arterial hypertension (PAH). However, A2B1 function has never been investigated in PAH. Methods: Through the integration of computational and experimental biology, we investigated the role of A2B1 in human PAH-PASMC. Bioinformatics and RNA-sequencing allowed us to investigate the transcriptome-wide function of A2B1, while RNA immunoprecipitation and A2B1 silencing experiments in vitro allowed us to decipher the intricate molecular mechanism at play. In addition, we performed a pre-clinical trial in the monocrotaline-induced pulmonary hypertension rat model to investigate the relevance of A2B1 inhibition in mitigating pulmonary hypertension severity. Results: In the present study, we found that A2B1 is up-regulated in human PAH-PASMC. We also show that A2B1 localization shifts in PAH-PASMC from cytoplasmic to nuclear mainly due to significantly higher expression of B1 isoform compared to A2 isoform of A2B1. Using bio-informatics, we identified three known motifs of A2B1 and all mRNAs carrying them. In PAH-PASMC, we demonstrated the complementary non-redundant function of A2B1 motifs as all motifs are implicated in different aspect of the cell cycle. In addition, we showed that in PAH-PASMC, A2B1 promotes the expression of its targets. A2B1 silencing in PAH-PASMC lead to a decrease of all tested mRNAs carrying an A2B1 motif, and an according decrease in proliferation and resistance to apoptosis. Finally, in vivo A2B1 inhibition in the lungs rescued pulmonary hypertension in rats. Conclusion: Through the integration of computational and experimental biology, our study revealed the role of A2B1 as a master orchestrator of the PAH-PASMC phenotype and its relevance as a therapeutic target in PAH.