Multi-Omics Analysis of the PDGF Response in Pulmonary Vascular Smooth Muscle Cells from Patients with Pulmonary Arterial Hypertension: Implication of the NMDAR

Abstract

Multi-Omics Analysis of the PDGF Response in Pulmonary Vascular Smooth Muscle Cells from Patients with Pulmonary Arterial Hypertension: Implication of the NMDAR

The N-methyl-D-aspartate receptor (NMDAR), expressed by PASMC, has been pointed as a novel therapeutic target in PAH, and is a new player involved in the PDGF-dependent proliferation of PASMC. Decoding the mechanisms of action of NMDAR antagonist is important to validate it as a therapeutic option in PAH.

Multi-omics approach was designed to study the transcriptional and translational response to PDGF, with or without NMDAR blockade, in PASMC of PAH patients. PASMC of PAH patients and controls in culture were stimulated with PDGF-BB 10 μM for 24 h and exposed to 100 μM MK-801, a noncompetitive NMDAR antagonist. The transcriptomic analysis was done on a Microarray Scanner DNA chip. Proteomic analysis was done using a high resolution Orbitrap mass spectrometer.

Transcriptomics revealed that PDGF induced the expression of genes involved in proliferation, migration and apoptosis resistance in PASMC. Exposure to MK-801 reversed these effects. Proteomics revealed 748 differentially expressed proteins between patient and control PASMC, and highlighted overexpression of a large number of members of the Rab protein family. Their expression was increased in patient PASMC exposed to PDGF and was decreased by MK-801 treatment. Consistent with transcriptomics, PDGF induced pro-proliferative proteins while MK-801 induced anti-proliferative proteins in PASMCs of PAH patients.

Multi-omics analysis showed that PDGF induced pro-proliferative gene and protein expression tended to normalize using NMDAR in PASMC. These results further support NMDARs as a therapeutic target for the treatment of PAHs.