Thiopurine-methyltransferase variants in inflammatory bowel disease: prevalence and toxicity in Brazilian patients.
OBJECTIVE
To analyze the prevalence of thiopurine-methyltransferase (TPMT) genotypes and their association with drug toxicity in inflammatory bowel disease (IBD) patients from southeastern Brazil.


METHODS
A total of 219 consecutive patients with IBD, of which 146 had Crohn's disease and 73 had ulcerative colitis, regularly seen at the outpatient unit of the Division of Gastroenterology at the University Hospital Pedro Ernesto of the State University of Rio de Janeiro, a tertiary referral center, were enrolled in this study from February 2009 to January 2011. We analyzed the presence of major TPMT genetic variants (TPMT 2, 3A, 3C) in IBD patients by means of a specific allele and RFLP-PCR. Genomic DNA was isolated from peripheral blood leukocytes by proteinase-K/Sodium Dodecyl Sulfate digestion and phenol-chloroform extraction. TPMT 2 (C238G), TPMT 3A (G460A/A719G), and TPMT 3C (A719G) genotypes were detected by real-time polymerase chain reaction followed by direct sequencing with specific primers. Clinical data were systematically recorded, and correlated with the genotype results.


RESULTS
The distribution of the selected TPMT gene polymorphism TPMT 2 (C238G), TPMT 3A (G460A/A719G), and TPMT 3C (A719G) genotypes was 3.6%, 5.4%, and 7.7% of the patients, respectively. Among the side effects recorded from patients taking azathioprine, 14 patients presented with pancreatitis and/or an elevation of pancreatic enzymes, while 6 patients had liver toxicity, and 2 patients exhibited myelosuppression/neutropenia. TPMT polymorphisms were detected in 37/219 patients (8 heterozygous for 2, 11 heterozygous for 3A, and 18 heterozygous for 3C). No homozygotic polymorphisms were found. Despite the prevalence of the TPMT 3C genotype, no differences among the genotype frequencies were significant. Although no association was detected regarding myelotoxicity or hepatotoxicity, a trend towards the elevation of pancreatic enzymes was observed for TPMT 2 and TPMT 3C genotypes.


CONCLUSIONS
The prevalence of TPMT genotypes was high among Brazilian patients. Variants genes 2 and 3C may be associated with azathioprine pancreatic toxicity in a IBD southeastern Brazilian population.
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Ana Teresa P
AT
Carvalho
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Barbara C
BC
Esberard
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Renata S B
RS
Fróes
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Davy C M
DC
Rapozo
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Ana B
AB
Grinman
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Tatiana A
TA
Simão
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Juliana C V C
JC
Santos
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Antonio José V
AJ
Carneiro
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Luis Felipe
LF
Ribeiro-Pinto
Ana Teresa P Carvalho, Barbara C Esberard, Renata S B Fróes, Ana B Grinman, Disciplina de Gastroenterologia e Endoscopia Digestiva, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20551-900, Brazil.
Heitor S P
HS
de Souza
D000328:Adult; D000483:Alleles; D001379:Azathioprine / Q000009:adverse effects; D001938:Brazil; D003093:Colitis, Ulcerative / Q000235:genetics*; D003424:Crohn Disease / Q000235:genetics*; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000235:genetics*; D007962:Leukocytes / Q000166:cytology / Q000378:metabolism; D008297:Male; D008780:Methyltransferases / Q000235:genetics*; D008875:Middle Aged; D010597:Pharmacogenetics; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D015995:Prevalence; D017422:Sequence Analysis, DNA
D008780:Methyltransferases; C022745:thiopurine methyltransferase; D001379:Azathioprine
Azathioprine; Drug toxicity; Inflammatory bowel disease; Pancreatitis; Thiopurine-methyl-transferase