The polymorphism analysis for CD36 among platelet donors

Area of Science:

• Genetics and Molecular Biology
• Immunology
• Biochemistry

Background:

• CD36 deficiency, characterized by defects in CD36 on platelets and/or monocytes, has incompletely understood correlations between molecular and protein expression levels.
• Investigating these correlations is crucial for understanding the pathophysiology of CD36 deficiency.

Purpose of the Study:

• To determine the polymorphisms of the CD36 gene, its RNA levels, and CD36 protein expression on platelets and in plasma.
• To correlate genetic variations with molecular and protein expression changes in CD36 deficiency.

Main Methods:

• Sanger sequencing for CD36 gene polymorphisms.
• Bioinformatic analysis using HotMuSiC, CUPSAT, SAAFEC-SEQ, and FoldX.
• Quantitative PCR (qPCR), flow cytometry, and ELISA for RNA and protein level analysis.

Main Results:

• Identified c.1228_1239delATTGTGCCTATT as the most frequent mutation (allele frequency 0.0072).
• Confirmed 5 mutations in the extracellular domain, with four (c.284T>C, c.512A>G, c.572C>T, c.869T>C) predicted to deleteriously impact CD36 protein stability.
• Observed significantly lower mean fluorescence intensity (MFI) of platelet CD36 expression in mutation carriers and deficient individuals.
• Found significantly lower soluble CD36 (sCD36) levels in type II deficiency and lower CD36 RNA levels in platelets of type II individuals compared to controls.

Conclusions:

• RNA level alterations may underlie type II CD36 deficiency.
• The prevalent mutation c.1228_1239delATTGTGCCTATT was identified in the Kunming cohort.
• Soluble CD36 (sCD36) shows potential as a biomarker for assessing immune reaction risk in CD36-deficient individuals, warranting further investigation.