Supplementary file 1_Molecular basis of anti-CD36 alloimmunization: transcriptomic evidence linking intronic splice-site mutations to CD36 loss-of-function.docx

Allele-specific analysis confirmed mutant allele enrichment in aberrant transcripts (P = 0.024). Long-read sequencing showed that one patient only had two intronic mutations (c.430-1G>C and c.429 + 3insG). Indicating intronic mutation was significantly linked to type I CD36 deficiency.

Specifically, c.430-1G>C mutation induced aberrant exon 6 splicing, resulting in 17-bp pseudo-intron insertion or complete exon 6 skipping (28.6% or 30.3% skipped); the c.429 + 3insG mutation caused exon 5 skipping (38.3% skipped). Functional assays showed that the c.430-1G>C mutation led to complete loss of CD36 protein expression.</p>Conclusion<p>This study establishes that intronic mutations alone can also induce CD36 deficiency by impairing mRNA splicing, thereby extending the known mutational landscape of CD36 deficiency and reinforcing the necessity of screening intronic mutations in diagnostic and pathogenetic investigations.</p>