MALDI-TOF mass spectrometry screening of cholelithiasis risk markers in the gene of HNF1alpha

In recent years MALDI-TOF MS gained importance for high-throughput DNA analysis. In the present study this technique was used for the pathogenetic analysis of gallstone disease. The intestinal apical sodium-dependent bile acid transporter (ASBT) shows a genetic association with gallstone disease. ASBT has 3 binding sites in its 5′UTR for hepatocyte nuclear factor 1alpha (HNF1alpha). We hypothesized that genetic alterations in the HNF1alpha gene could influence ASBT expression. The gene HNF1alpha was sequenced in 46 Stuttgart random samples, composed of 16 controls and 30 gallstone patients. Subsequently, two independent cohorts (Stuttgart: 67 gallstones carriers, 109 controls, Leutkirch: 112 gallstone carriers, 99 controls) were screened by MALDI-TOF MS. The subjects were further divided by gender and weight. 24 known polymorphisms and two novel SNPs in the 3′UTR of HNF1alpha were detected (c.*220G>A and c.*1151G>A). After gender-specific sub-division of the pooled cohorts, 4 SNPs resulted in significant differences between male gallstone carriers and male controls (Stuttgart/Leutkirch: rs2255531 OR = 2.78; p = 0.006, rs1169288 OR = 2.13; p = 0.032, rs7310409 OR = 2.34; p = 0.025 and rs1169294 OR = 2.13; p = 0.031). Two novel variants in the 3′UTR of HNF1alpha were detected and four SNPs of HNF1alpha show a significant association to cholelithiasis in male gallstone patients. This article is part of a Special Section entitled: Understanding genome regulation and genetic diversity by mass spectrometry.

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► MALDI-TOF MS gained in importance for high-throughput DNA analysis in the last years.
► The HNF1alpha gene was genotyped by MALDI-TOF MS in two independent cohorts.
► 24 known and two novel SNPs were detected in the gene.
► 4 SNPs resulted in significant associated with cholelithiasis in male gallstone patients.