Statistics from Altmetric.com
Predisposition to hereditary pancreatitis has been associated with mutations in three genes: protease, serine, 1 (PRSS1), which codes for cationic trypsinogen,1 cystic fibrosis transmembrane conductance regulator (CFTR),2 and serine protease inhibitor Kazal type 1 (SPINK1).3
We have identified a novel PRSS1 mutation in seven subjects with chronic pancreatitis (CP) from three generations of an Italian family. The index patient was a 57 year old man with CP referred to our hospital for ductal adenocarcinoma of the pancreatic head. Eleven relatives were examined, and an uncle, also with CP, had died in an accident.
Congenital malformations and alcoholic, biliary, obstructive, and autoimmune pancreatitis were ruled out. Eleven subjects gave their written consent to the study.
The cystic fibrosis assay (CF-OLA; Applied Biosystems, California, USA) was used to look for 31 frequent CFTR mutations in all subjects. The five exons of the PRSS1 gene were sequenced with the oligonucleotides described by Nishimori and colleagues.4 The four SPINK1 exons were investigated by denaturant gradient gel electrophoresis (DGGE). No CFTR or SPINK1 mutations were found although subject III-8 (with CP) carried the N1303K mutation in heterozygosis in the cystic fibrosis gene.
The PRSS1 exon 2 sequence of the index patient revealed a T>C change at nucleotide 116 (c.116 T>C) causing a valine to alanine substitution at codon 39 (V39A). This mutation was present in another six subjects with CP, diagnosed from exocrine insufficiency and computer tomography and magnetic resonance imaging demonstrations of typical ductal alterations and parenchymal calcifications. Two of these patients were also diabetic. In a further two patients, the genetic analysis was not performed, but CP was confirmed by clinical and morphological findings. The remaining four subjects had a normal pancreas and did not carry the V39A mutation (fig 1).
The lod score calculated for the association between V39A and CP was z = 3.0 at θ = 0.0. This mutation was not found in a DGGE investigation of 130 patients with sporadic CP.
Mean age of the patients was 47.22 (±13.64) years (median 54 (range 25–60)). Mean age at onset was 30.0 (±7.35) years (median 32 (range 19–40)) whereas in patients displaying other PRSS1 mutations, onset was typically during childhood or adolescence.5,6
An acute attack requiring hospitalisation formed the clinical overture in six of the nine CP patients. The other three (III-4, III-5 and IV-2) presented morphological and functional evidence of CP at the time of the study but were asymptomatic. It is clear therefore that damage to the pancreas may occur prior to the clinical onset of CP.
In hereditary CP, the mechanism of the R122H mutation has been elucidated.1 This substitution removes a hydrolysis start site and makes both trypsin and trypsinogen autolysis resistant. A similar mechanism has been proposed for the N29I mutation which alters protein conformation and masks the R122 site.7
Valine 39 is evolutionarily conserved in the trypsinogen gene of all terrestrial vertebrates8 and would thus seem of importance in the protein’s structure and function. As V39 is only 10 amino acids distant from N29, its replacement by alanine may result in abnormal conformation of the peptide and mask arginine 122 against enzymatic degradation. Further work is needed to define the mechanism and confirm this interpretation.
In conclusion, the presence of the V39A mutation in seven of the CP patients, its absence in their healthy relatives, the 3.0 lod score, and the strong evolutionary conservation of V39, all indicate that the novel mutation is the cause of CP in this family.
We would like to thank Professors JP Neoptolemos and DC Whitcomb for their valuable assistance and Mr J Iliffe. This work was supported by Compagnia di San Paolo and Regione Piemonte.
Conflict of interest: None declared.
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.