Minigene analysis of intronic variants in common SPINK1 haplotypes associated with chronic pancreatitis

Kereszturi, Éva; Király, Orsolya; Sahin–Tóth, Miklós

doi:10.1136/gut.2008.164947

Cited by 65 publications

(59 citation statements)

References 27 publications

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“…The p.N34S variation had no effect on the secretion of SPINK1 protein from transfected cells and the trypsin inhibitory activity of the mutant protein was also unchanged (Kuwata et al 2002;Király et al 2007). The roles of co-segregating four intronic alterations have been excluded (Masamune et al 2007;Kereszturi et al 2009a). On the other hand, we have shown that the c.194+2T>C mutation affects the consensus splicing donor site, resulting in the skipping of exon 3 (Kume et al 2006).…”

Section: Spink1 Mutationsmentioning

confidence: 99%

Genetics of Pancreatitis: The 2014 Update

Masamune

2014

Tohoku J. Exp. Med.

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Chronic pancreatitis is a progressive inflammatory disease in which pancreatic secretory parenchyma is destroyed and replaced by fibrous tissue, eventually leading to malnutrition and diabetes. Alcohol is the leading cause in Western countries, but genetic factors are also implicated. Since the identification of mutations in the cationic trypsinogen (PRSS1) gene as a cause of hereditary pancreatitis in 1996, we have seen great progress in our understanding of the genetics of pancreatitis. It has been established that mutations in the genes related to the activation and inactivation of trypsin(ogen) such as PRSS1, serine protease inhibitor Kazal type 1 (SPINK1) and chymotrypsin C (CTRC) genes are associated with pancreatitis. In 2013, carboxypeptidase A1 (CPA1) was identified as a novel pancreatitis susceptibility gene. Endoplasmic reticulum stress in pancreatic acinar cells resulting from the mis-folding of mutated pancreatic enzymes has been shown to act as a novel mechanism underlying the susceptibility to pancreatitis. In Japan, the nationwide survey revealed 171 patients (96 males and 75 females) with hereditary pancreatitis in 59 families based on the European Registry of Hereditary Pancreatitis and Familial Pancreatic Cancer criteria. Because about 30% of families with hereditary pancreatitis do not carry mutations in any of the known pancreatitis susceptibility genes, other yet unidentified genes might be involved. Next generation sequencers can perform billions of sequencing reactions with a read length of 150-250 nucleotides. Comprehensive analysis using next generation sequencers will be a promising strategy to identify novel pancreatitis-associated genes and further clarify the pathogenesis of pancreatitis.

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Section: Spink1 Mutationsmentioning

confidence: 99%

Genetics of Pancreatitis: The 2014 Update

Masamune

2014

Tohoku J. Exp. Med.

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“…Our report suggests variable phenotypes of the homozygous IVS3+2T>C mutation-associated CP. This IVS3+2T>C mutation has been observed with an average allele frequency of 6.3% in Japanese patients with idiopathic, familial, and alcoholic CP (Kume et al 2005;Kereszturi et al 2009), but is not specific to Japanese. It was also found in two patients with tropical pancreatitis in Bangladesh (Rossi et al 2001) and Thailand (Snabboon et al 2006), four patients with CP in Europe (Witt et al 2000;Kalinin et al 2006), three patients with familial pancreatitis in the United States (Pfützer et al 2000), and one patient with pancreatitis in the United States (Keiles and kammesheidt 2006).…”

Section: Discussionmentioning

confidence: 88%

“…The p.N34S (c.101A>G) mutation has been found worldwide in CP patients and healthy controls, with an average allele frequency of 9.7% and 1%, respectively (Pfützer et al 2000;Witt et al 2000;Aoun et al 2008;Kereszturi et al 2009). The haplotype associated with the p.N34S mutation increases the risk of CP 11-fold on average, with higher risk observed in idiopathic and tropical cases than in alcoholic CP (Aoun et al 2008 and references therein).…”

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confidence: 99%

“…Indeed, gastric biopsies from carriers of the IVS3+2T>C mutation-associated haplotype revealed the predominant expression of a shorter SPINK1 mRNA lacking exon 3 (Kume et al 2006). This aberrant mRNA was predicted to code for a non-functional trypsin inhibitor, and trypsin inhibitor production was diminished in the minigene analysis (Kereszturi et al 2009). Thus, the IVS3+2T>C mutation results in functional loss of SPINK1 and disturbs the protease/antiprotease balance within the pancreas, leading to the development of pancreatitis.…”

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confidence: 99%

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Phenotypic Variability of the Homozygous IVS3+2T>C Mutation in the Serine Protease Inhibitor Kazal Type 1 (SPINK1) Gene in Patients with Chronic Pancreatitis

Ota

Masamune

Inui

et al. 2010

Tohoku J. Exp. Med.

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“…Even though p.N34S was presumed to cause a loss of function, the exact mechanism of action has never been identified, as no functional defect was demonstrated for p.N34S or any of the four intronic variants associated with this haplotype (5,7,19,20,24,26). In contrast, an unambiguous loss-of-function phenotype was evident for variant c.194ϩ2TϾC (28,35,39, and references therein), the second most frequent SPINK1 mutation that affects a splice site in intron-3 and causes exon skipping with markedly reduced SPINK1 mRNA expression (19,23). This intronic mutation is in complete linkage disequilibrium (i.e., found together) with the promoter variant c.-215GϾA.…”

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confidence: 99%

Functional significance of SPINK1 promoter variants in chronic pancreatitis

Derikx

Geisz

Kereszturi

et al. 2015

American Journal of Physiology-Gastrointestinal and Liver Physi

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Chronic pancreatitis is a progressive inflammatory disorder of the pancreas, which often develops as a result of genetic predisposition. Some of the most frequently identified risk factors affect the serine protease inhibitor Kazal type 1 ( SPINK1) gene, which encodes a trypsin inhibitor responsible for protecting the pancreas from premature trypsinogen activation. Recent genetic and functional studies indicated that promoter variants in the SPINK1 gene might contribute to disease risk in carriers. Here, we investigated the functional effects of 17 SPINK1 promoter variants using luciferase reporter gene expression assay in four different cell lines, including three pancreatic acinar cell lines (rat AR42J with or without dexamethasone-induced differentiation and mouse 266-6) and human embryonic kidney 293T cells. We found that most variants caused relatively small changes in promoter activity. Surprisingly, however, we observed significant variations in the effects of the promoter variants in the different cell lines. Only four variants exhibited consistently reduced promoter activity in all acinar cell lines, confirming previous reports that variants c.-108G>T, c.-142T>C, and c.-147A>G are risk factors for chronic pancreatitis and identifying c.-52G>T as a novel risk variant. In contrast, variant c.-215G>A, which is linked with the disease-associated splice-site mutation c.194+2T>C, caused increased promoter activity, which may mitigate the overall effect of the pathogenic haplotype. Our study lends further support to the notion that sequence evaluation of the SPINK1 promoter region in patients with chronic pancreatitis is justified as part of the etiological investigation.

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Minigene analysis of intronic variants in common SPINK1 haplotypes associated with chronic pancreatitis

Cited by 65 publications

References 27 publications

Genetics of Pancreatitis: The 2014 Update

Genetics of Pancreatitis: The 2014 Update

Phenotypic Variability of the Homozygous IVS3+2T>C Mutation in the Serine Protease Inhibitor Kazal Type 1 (SPINK1) Gene in Patients with Chronic Pancreatitis

Functional significance of SPINK1 promoter variants in chronic pancreatitis

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