Functionalization of CD36 cardiovascular disease and expression associated variants by interdisciplinary high...

Area of Science:

• Genetics
• Molecular Biology
• Cardiovascular Research

Background:

• CD36 is a platelet glycoprotein involved in oxidized low-density lipoprotein (oxLDL) binding and platelet activation.
• Genome-wide association studies link CD36 gene variants to platelet count, volume, lipid levels, and cardiovascular disease (CVD) risk.
• Platelet CD36 expression is associated with oxLDL response and thrombo-embolism risk, but causative variants remain elusive.

Purpose of the Study:

• To identify and functionally validate genetic variants that regulate platelet CD36 expression.
• To investigate the impact of these variants on CD36 transcription and platelet function.
• To assess the clinical relevance of identified variants using large-scale genetic data.

Main Methods:

• Screening of 81 expression quantitative trait loci (eQTL) single nucleotide polymorphisms (SNPs) using Massively Parallel Reporter Assays (MPRA).
• Analysis of MPRA data with a novel Bayesian statistical method.
• Validation of transcriptional regulation using luciferase assays, electromobility shift assays, and CRISPR/Cas9 gene editing in cell lines.
• Assessment of clinical relevance using the PhenoScanner tool.

Main Results:

• Ten eQTLs showed significant allele-specific transcription shifts, located upstream of CD36 transcriptional start sites.
• rs2366739 and rs1194196 variants were confirmed to alter transcriptional regulation and DNA:protein binding.
• CRISPR/Cas9-mediated deletion of the regulatory locus upregulated CD36 transcription.
• PhenoScanner analysis reinforced the clinical relevance of the identified variants.

Conclusions:

• A novel genetic variant regulating CD36 expression and subsequent platelet function has been identified.
• This study establishes a generalizable method for functional testing of genetic variants.
• Findings support potential applications in patient management and precision therapy development for cardiovascular diseases.