Abstract:

Objective To confirm whether PANoptosis is involved in the occurrence and development of non-syndromic cleft lip with or without cleft palate (NSCL/P) and identify key genetic variations and genes involved. Methods Firstly, a two-stage population was collected for genome-wide association study (GWAS) to determine the correlation between single nucleotide polymorphisms (SNPs) and NSCL/P. Next, the PANoptosis gene set was selected and SNPs were extracted to conduct quality control and gene-based association analysis in the stage Ⅰ population to screen candidate SNPs, and the stage Ⅱ samples were used as validation. The functional annotation of Haploreg, RegulomeDB, and ATAC sequencing combined with linkage disequilibrium-based clumping further identified the key functional SNP and risk gene. Finally, expression quantitative trait loci (eQTL) analysis, RNA sequencing, single-cell sequencing, and protein-protein interaction analysis were used to predict the regulatory effects of the locus and gene. Results rs71518324 was associated with the risk of NSCL/P (P_meta<0.001), and the surrounding region was enriched with a large number of active functional element markers. The eQTL effect of rs71518324 on PRKAR1B gene was significant (P<0.001). The expression of PRKAR1B in RNA sequencing generally increased over time. Five cell clusters were identified in single-cell RNA sequencing, with PRKAR1B mainly localized to the ectoderm. Conclusion rs71518324 in PRKAR1B, a PANoptosis related gene, is significantly associated with NSCL/P risk.

Key words: cleft lip, GWAS, PANoptosis