Clinical Cancer Research Molecular Diagnostics in Cancer Therapeutic Development: Fulfilling the Promise of Personalized Medicine Translational Cancer Medicine 2008: Cancer Clinical Trials and Personalized Medicine
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Cell Growth & Differentiation
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lecomte, T.
Right arrow Articles by Loriot, M.-A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lecomte, T.
Right arrow Articles by Loriot, M.-A.
Clinical Cancer Research Vol. 12, 3050-3056, May 15, 2006
© 2006 American Association for Cancer Research

Imaging, Diagnosis, Prognosis

Glutathione S-Transferase P1 Polymorphism (Ile105Val) Predicts Cumulative Neuropathy in Patients Receiving Oxaliplatin-Based Chemotherapy

Thierry Lecomte1,3, Bruno Landi1, Philippe Beaune2,3, Pierre Laurent-Puig2,3 and Marie-Anne Loriot2,3

Authors' Affiliations: Departments of 1 Gastroenterology and 2 Biochemistry, Assistance Publique-Hôpitaux de Paris, European Georges Pompidou Hospital; and 3 Université Paris-Descartes, Faculté de Médecine, Institut National de la Sante et de la Recherche Medicale UMRS 775, Paris, France

Requests for reprints: Pierre Laurent-Puig, U775 Bases Moléculaires de la Réponse aux Xénobiotiques, 45 rue des Saints Pères, 75006 Paris, France. Phone: 33-1-428-62081; Fax: 33-1-428-62072; E-mail: pierre.laurent-puig{at}biomedicale.univ-paris5.fr.


   Abstract
Top
 Abstract
Patients and Methods
 Results
 Discussion
 References
 
Purpose: Glutathione S-transferases (GST) are xenobiotic metabolizing enzymes involved in the detoxification of a variety of chemotherapeutic drugs, including platinum derivatives. Genetic polymorphisms of GSTs have been associated with enzyme activity variations. Thus, a study was done to investigate the relationship between GST polymorphisms and oxaliplatin-related cumulative neuropathy in gastrointestinal cancer patients treated with oxaliplatin-based chemotherapy.

Experimental Design: Ninety patients were included. Clinical neurologic evaluation was done at baseline and before each cycle of treatment. We determined genetic variants for GSTP1 exon 5 (Ile105Val), GSTP1 exon 6 (Ala114Val), GSTM1 (homozygous deletion), and GSTT1 (homozygous deletion). We conducted analyses in a subgroup of 64 patients receiving a minimal cumulative dose of 500 mg/m2 of oxaliplatin to examine whether the GST polymorphisms are associated with oxaliplatin-related cumulative neuropathy.

Results: Among patients receiving a minimal cumulative dose of 500 mg/m2 of oxaliplatin, 15 patients showed clinically evident oxaliplatin-related cumulative neuropathy scored grade 3 according to an oxaliplatin-specific scale. The oxaliplatin-related cumulative neuropathy scored grade 3 was significantly more frequent in patients homozygous for the GSTP1 105Ile allele than in patients homozygous or heterozygous for the GSTP1 105Val allele (odds ratio, 5.75; 95% confidence interval, 1.08-30.74; P = 0.02). No association was found with respect to any of the GSTM1, GSTT1, or GSTP1 exon 6 genotypes.

Conclusions: The results of the current study suggest that the 105Val allele variant of the GSTP1 gene at exon 5 confers a significantly decreased risk of developing severe oxaliplatin-related cumulative neuropathy.

Pharmacogenetics is becoming an increasingly important field in the study of cancer chemotherapy treatment. Genetic factors affecting the metabolism and transport of drugs partly explain interindividual variability in drug response, both in terms of efficacy and safety. Polymorphisms in genes encoding specific drug-metabolizing enzymes can result in individuals in the general population being characterized as low, rapid, or even ultrarapid metabolizers. Phenotyping and genotyping tests are now available to determine and predict the metabolic status of an individual and, thus, enabling the evaluation of risk of treatment failure or toxicity.

Currently, oxaliplatin is one of the most frequently used chemotherapy agent used in the palliative treatment of gastrointestinal cancers and, more recently, in the adjuvant treatment of colorectal cancer. The dose-limiting side effect of oxaliplatin is a cumulative peripheral neuropathy usually observed after 4 to 6 months of treatment (1–3). The oxaliplatin-induced cumulative neuropathy is often a cause for dose reduction or treatment discontinuation. Approximately 10% to 18% of patients develop oxaliplatin-induced cumulative neuropathy progressively when a cumulative dose of 800 mg/m2 is reached (4). Because oxaliplatin-induced neurotoxicity causes significant discomfort, alters patient quality of life, and may be accompanied by significant disability, effort could be made to optimize oxaliplatin-based chemotherapy.

Glutathione S-transferases (GST) are a multigene family of enzymes (cytosolic and membrane-bound), which catalyze the conjugation of glutathione to electrophilic xenobiotics to inactivate them and facilitate their excretion from the body (5). GSTs play an important role in the metabolism of potentially genotoxic compounds by preventing of DNA damage and adduct formation. The cytosolic isoenzymes are divided at least into five major classes ({alpha}, µ, {pi}, {theta}, {zeta}) among which genetic polymorphisms have been detected in the genes encoding GSTM3, GSTM1 (µ class), GSTT1 ({theta} class), GSTP1 ({pi} class), and GSTZ1 (GST {zeta} class; refs. 6–8). Among them, the GSTM1, GSTT1, and GSTP1 genotypes have been extensively studied for their potential modulating role in individual susceptibility to cancer or drug response (8–10). Growing evidence indicates that GST enzymes determine cytotoxicity of a variety of chemotherapeutic drugs (11–18). The isoenzyme GSTP1 participates in the detoxification of platinum derivatives, especially cisplatin, and is an important mediator of both intrinsic and acquired resistance to platinum. In addition to GSTP1, the activity of GSTM1 and GSTT1 could be involved in the response to treatment (19). However, the role of these enzymes specifically in the oxaliplatin pathway remains unknown.

Genetic variations of GSTs have been associated with changes in enzyme activity. Four common polymorphisms in the GSTP1, GSTT1, and GSTM1 genes either decrease or abolish GST enzyme activity. GSTP1 is a polymorphic gene located on chromosome 11 with two single-nucleotide substitutions in exon 5 (A313G) and exon 6 (C341T) that give rise to Ile105Val and Ala114Val amino acid substitutions, respectively (6). These amino acid substitutions seem to be within the GSTP1 active site and lead to the alteration of substrate affinity (5, 6, 20, 21).

By contrast, inherited homozygous deletion of GSTT1 or GSTM1 gene leads to a complete absence of enzymatic activity. To date, in spite of the biochemical evidence that the GSTs mediate inactivation of platinum drugs, the association between polymorphisms in the GST genes and the neurotoxicity of these compounds remains to be explored. This prompted us to evaluate the different GSTP1, GSTT1, and GSTM1 genetic polymorphisms as predictive factors for the oxaliplatin-induced neuropathy in 64 patients receiving a minimal cumulative dose of 500 mg/m2 of oxaliplatin.


   Patients and Methods
Top
 Abstract
 Patients and Methods
Results
 Discussion
 References
 
Patients. From January 2002 to September 2004, 90 adult patients (52 men, 38 women; mean age, 64 years; range, 24-84 years) with gastrointestinal solid tumors who received an oxaliplatin-based chemotherapy were included in this retrospective analysis. This study was conducted in one medical center. All patients entered in this study signed an informed consent form for blood sample collection to establish the clinical significance of genetic polymorphisms in gastrointestinal cancer. Because the aim of this study was to determine whether genetic polymorphisms may be predictive of cumulative neuropathy in patients receiving oxaliplatin-based chemotherapy, we enrolled only patients who received a minimal cumulative dose of 500 mg/m2 of oxaliplatin. The patients with neuropathy or who received platinum-based chemotherapy before oxaliplatin-based chemotherapy were excluded. Among the 90 patients who participated in this study, 64 patients received a minimal cumulative dose of 500 mg/m2 oxaliplatin and were eligible for the analysis. In the group of nonenrolled patients receiving lesser than 500 mg/m2, no grade 3 neuropathy was observed. The characteristics of eligible patients are summarized in Table 1 . All tumors were classified as adenocarcinoma. Fifty-two patients with stage IV colorectal cancer received palliative oxaliplatin-based chemotherapy as first-line in 39 cases and as second-line in 13 cases. The palliative treatments were semimonthly regimens of 5-fluorouracil, leucovorin, and oxaliplatin previously described: FOLFOX4 regimen (oxaliplatin 85 mg/m2), n = 38; FOLFOX7 regimen (oxaliplatin 130 mg/m2), n = 7; FOLFOX6 regimen (oxaliplatin 100 mg/m2), n = 6 (22–24). One patient received oxaliplatin combined with raltitrexed as previously described (TOMOX; 25). Six patients with stage III colorectal cancer received adjuvant oxaliplatin-based chemotherapy (FOLFOX4 regimen). Six patients with other gastrointestinal cancer received palliative oxaliplatin-based chemotherapy. The treatments as first-line chemotherapy were as follows: oxaliplatin combined with gemcitabine as previously described (GEMOX), n = 4; FOLFOX4 regimen, n = 2 (26). Physical examination and a full blood count were done before each cycle. The same chemotherapy regimen was maintained until disease progression or severe toxicity. Cumulative oxaliplatin-induced neuropathy was scored according to the oxaliplatin-specific scale reported by Caussanel et al. (27). On this oxaliplatin-specific scale, the grade of cumulative neuropathy depends on the duration and intensity of symptoms (grade 1: paresthesia, dysesthesia of short duration; grade 2: paresthesia, dysesthesia persisting between cycles; grade 3: paresthesia, dysesthesia causing functional impairment).


View this table:
[in this window]
[in a new window]
  		Table 1. Clinical and pathologic characteristics of patients (n = 64)

 
DNA extraction. The genomic DNA was purified from peripheral lymphocytes using a commercial kit (QiAmp Blood DNA kit, Qiagen, Courtaboeuf, France) and stored at –20°C until use.

PCR. The DNA samples (10-50 ng/µL) were analyzed using the PCR. All the PCR analyses were done using a reaction volume of 25 µL containing 2 µL DNA, 2.5 µL of 10x PCR buffer (GeneAmp, Applied Biosystems, Courtaboeuf, France), MgCl2 (GeneAmp, Applied Biosystems) at 1.5 mmol/L final concentrations, 200 µmol/L deoxynucleotide triphosphates (ABgene, Courtaboeuf, France), 400 nmol/L of each primer (Genset, Paris, France) and 1.5 IU Taq polymerase (AmpliTaq DNA Polymerase, Applied Biosystems). The PCR were conducted in a 9700 thermal cycler (Applied Biosystems). The amplification products (5 µL) underwent electrophoresis on 1.5% agarose gel (Invitrogen, Cergy Pontoise, France) and staining with ethidium bromide.

GSTM1, GSTT1, and exon 5 GSTP1 genotyping. The GSTM1, GSTT1, and GSTP1 genotypes were determined by a slightly modified method previously described (21, 28). This method allows the simultaneous genotyping of all three GSTs in the same tube. Briefly, a multiplex PCR was conducted using primer pairs specific to each GST gene (28). The fragment lengths of the PCR products were 480 bp for GSTT1, 215 bp for GSTM1, and 176 bp for GSTP1. The presence of the GSTM1 or GSTT1 gene generated positive PCR signals, whereas the deletion did not. The results were validated using the GSTP1 PCR fragment as an internal amplification control. The PCR product was then subjected to restriction fragment analysis using BsmAI enzyme (New England Biolabs, Saint Quentin en Yvelines, France). Only the 176 bp GSTP1 fragment was cut into 91 and 85 bp fragments when the GSTP1 105Val allele was present and not cut when the GSTP1 105Ile allele was present. The digestion products were revealed in a polyacrylamide (8%) gel electrophoresis.

Genotyping of exon 6 GSTP1 polymorphism. The polymorphism Ala114Val located in exon 6 was detected by use of a 5' nuclease allelic discrimination assay (ABI PRISM 7700 Sequence Detection System; Applied Biosystems, Foster City, CA). The SNP (National Center for Biotechnology Information Reference rs1799811) was identified based on the TaqMan Drug Metabolism Genotyping Assay (commercially available, Applied Biosystems), in the conditions recommended by the manufacturer.

Statistical analysis. The primary end point was the development of a grade 3 cumulative neuropathy to oxaliplatin. For quantitative variables, the cutoff level chosen was the median value. For qualitative variables, patients were grouped according to the presence or absence of each variable. The Mann-Whitney test was used to compare the means of two independent groups. GSTP1, GSTT1, and GSTM1 polymorphisms were analyzed separately to evaluate the association between the polymorphisms and cumulative neuropathy to oxaliplatin. The {chi}2 test was used to compare the observed genotype distributions with those expected by the Hardy-Weinberg equilibrium. {chi}2 or Fisher's two-tailed exact test was used to determine the relationship between cumulative neuropathy to oxaliplatin with the GSTT1, GSTP1, and GSTM1 genotypes. Furthermore, the two GSTP1 genotypes were combined to define four haplotypes. GSTP1*A has Ile at codon 105 and Ala at codon 114. GSTP1*B has Val at codon 105 and Ala at codon 114. GSTP1*C has both Val at codon 105 and 114, and GSTP1*D has Ile at position 105 and Val at position 114. We grouped the patients homozygous for *A/*A and tested them against the others. Associations were quantified by calculating odds ratios (OR) with profile likehood-based 95% confidence intervals (95% CI). The calculation of ORs and 95% CIs was adjusted for potential confounding factors (i.e., age, gender, and treatment protocol). Second, multiple logistic regression analysis was applied to adjust ORs for these potential confounding factors. The probability of survival without grade 3 neuropathy during oxaliplatin-based chemotherapy was estimated using the methods of Kaplan and Meier (29). The comparison of survival without grade 3 neuropathy during oxaliplatin-based chemotherapy for patients with or without GSTP1 105Val allele was done by log-rank test. The log-rank test was stratified according to the treatment protocol (i.e., FOLFOX4 regimen versus other regimens). All statistical tests were two-sided, and P < 0.05 was used to indicate statistical significance.


   Results
Top
 Abstract
 Patients and Methods
 Results
Discussion
 References
 
In the group of eligible patients, a total of 680 cycles was administered (median, 10 per patient); the median dose-intensity of oxaliplatin was 43 mg/m2/wk (range, 33-65), whereas the median cumulative dose of oxaliplatin was 850 mg/m2 (range, 510-2,205). Among them, maximal cumulative peripheral neuropathy per patient was grade 3 in 23% (15 patients), grade 2 in 27% (17 patients), and grade 1 in 42% (27 patients) of patients, respectively. Forty-two patients (45%) received at least cumulative dose of 800 mg/m2 oxaliplatin, 19 patients (30%) at least 1,000 mg/m2, and 9 patients (14%) at least 1,200 mg/m2. The probability of survival without grade 3 neuropathy during oxaliplatin-based chemotherapy was 0.83 (95% CI, 0.69-0.90) and 0.53 (95% CI, 0.28-0.73) at 20 and 40 weeks of treatment, respectively. The mean dose of oxaliplatin was 876 mg/m2 (range, 580-1,275) in the group of patients who developed grade 3 cumulative peripheral neuropathy and 916 mg/m2 (range, 510-2,205) in those without (Mann-Whitney test, P = 0.9). There are no significant differences between the group of patients who developed grade 3 cumulative peripheral neuropathy to oxaliplatin and those without in terms of age, gender, and chemotherapy regimen (Table 2 ).


View this table:
[in this window]
[in a new window]
  		Table 2. Distribution of characteristics of study participants according to oxaliplatin-related cumulative neuropathy scored grade 3

 
Genotype data for the GSTP1 exon 5 and GSTT1 polymorphisms were available for all 64 eligible patients, whereas data were available for 63 patients for the GSTP1 exon 6 and GSTM1 polymorphisms (Table 3 ). The distribution of the exon 5 GSTP1 genotypes was 61% for the Ile/Ile genotype (39 patients; 95% CI, 49-72%), 31% for the Ile/Val genotype (20 patients; 95% CI, 21-43%), and 8% for the Val/Val genotype (5 patients; 95% CI, 3-17%). The above distribution is in close agreement with that predicted by the Hardy-Weinberg equilibrium ({chi}2 = 1.07, P = 0.3). Our observed allele frequency for GSTP1 105Val allele was 0.23 (30 of 128) and was similar to previous reports on allele frequencies for healthy Caucasians and for colorectal cancer patients (5, 30). The distribution of the exon 6 GSTP1 genotype was 90% for the Ala/Ala genotype (57 patients; 95% CI, 80-96%) and 10% for the Ala/Val genotype (6 patients; 95% CI, 4-20%). The above distribution is in close agreement with that predicted by the Hardy-Weinberg equilibrium ({chi}2 = 0.16, P = 0.7). The observed frequency for GSTP1 114Val was 0.047 (6 of 126) and was similar to previous reports on allele frequencies for healthy Caucasians and for colorectal cancer patients (5, 30). When we combined these two genotypes, 38 (62.3%) patients were *A/*A homozygous, 18 were *A/*B heterozygous (29.5%), 1 was *B/*B homozygous (1.7%), and 4 were *B/*C heterozygous (6.5%). The haplotypes cannot be unambiguously deduced in two patients because they were heterozygous for both 105 and 114 polymorphisms and they are possibly *A/*C or *B/*D. The frequencies of the GSTT1 and GSTM1 homozygous gene deletions were 0.23 (i.e., 15 of 64 patients had homozygous deletions of GSTT1) and 0.57 (i.e., 36 of 63 patients had homozygous deletions of GSTM1), respectively. Eleven patients (17.5%) showed a homozygous deletion of both genes.


View this table:
[in this window]
[in a new window]
  		Table 3. Distribution of GSTM1, GSTT1, and GSTP1 genotypes and GSTP1 haplotypes

 
Grade 3 cumulative peripheral neuropathy was significantly more common in patients with Ile/Ile GSTP1 exon 5 genotype (13 of 39 patients, 33%) compared with patients with Ile/Val or Val/Val GSTP1 exon 5 genotype (2 of 25 patients, 8%; {chi}2 = 5.45, P = 0.02; Table 4 ). As a result, patients who were genotyped as Ile/Ile GSTP1 exon 5 had a higher risk of developing a grade 3 cumulative peripheral neuropathy to oxaliplatin than did those with other GSTP1 exon 5 genotypes (unadjusted OR, 5.75; 95% CI, 1.08-30.74; P = 0.02).


View this table:
[in this window]
[in a new window]
  		Table 4. Distribution of GSTM1, GSTT1, GSTP1 exons 5 and 6 genotypes and GSTP1 haplotypes based on analysis of a dichotomous trait (with and without grade 3 cumulative neuropathy to oxaliplatin)

 
Our sample size of 63 patients combined with the distribution of GSTP1 haplotypes over the four categories did not allow a reasonable, matched assessment of the association of GSTP1 haplotypes with risk of neuropathy grade 3. However, we compared patients homozygous for the *A/*A haplotype to the others. The ORs for GSTP1*A/GSTP1*A category compared with the non-GSTP1*A/GSTP1*A category was 5.3 (P =0.03). The same significant association was observed when we adjusted our analysis by age (dichotomized on the median age of all the patients), gender, and chemotherapy regimen (FOLFOX4 regimen versus other regimens; Table 5 ). When logistic regression analysis was applied, it was found that GSTP1*A/GSTP1*A haplotype was significantly associated with the risk of developing a grade 3 cumulative peripheral neuropathy to oxaliplatin (adjusted OR, 5.54; 95% CI, 1.1-28.02; P = 0.03; Table 6 ). The mean dose of oxaliplatin was 921 mg/m2 (range, 510-1,530) in the group of patients with GSTP1*A/GSTP1*A haplotype versus 883 mg/m2 (range, 510-2,205) in those with other combinations of GSTP1 haplotypes (Mann-Whitney test, P = 0.24).


View this table:
[in this window]
[in a new window]
  		Table 5. ORs for GSTP1 haplotypes adjusted by age, gender, and chemotherapy regimen

 

View this table:
[in this window]
[in a new window]
  		Table 6. Adjusted ORs evaluated by logistic regression

 
The patients with non-GSTP1*A/GSTP1*A haplotypes had a significantly better survival without grade 3 neuropathy during oxaliplatin-based chemotherapy than the ones with GSTP1*A/GSTP1*A haplotype (Fig. 1 ). The 30 weeks probability of survival without grade 3 neuropathy during oxaliplatin-based chemotherapy for patients with GSTP1*A/GSTP1*A haplotype was 0.42 (95% CI, 0.11-0.64), compared with 0.92 (95%CI, 0.64-0.97) for patients without GSTP1*A/GSTP1*A haplotype (P = 0.048 by the log-rank test stratified according to the chemotherapy regimen).


Figure 1
View larger version (11K):
[in this window]
[in a new window]
  		Fig. 1. Probability of survival without grade 3 neuropathy during oxaliplatin-based chemotherapy by GSTP1 haplotypes. P is calculated by log-rank adjusted according to the chemotherapy regimen.

 

   Discussion
Top
 Abstract
 Patients and Methods
 Results
 Discussion
References
 
In the present study, we investigated whether the determination of common polymorphisms in GSTP1, GSTT1, and GSTM1 genes in gastrointestinal cancer patients who received oxaliplatin-based chemotherapy could be used to predict the occurrence of oxaliplatin-related cumulative neuropathy. Our results suggest that a relevant polymorphism of GSTP1 gene can modulate the risk of severe neuropathy among gastrointestinal cancer patients who received oxaliplatin-based chemotherapy. This is the first study reporting an association between a GSTP1 genetic polymorphism and oxaliplatin-related cumulative neuropathy.

Neurotoxicity is the main and dose-limiting toxicity of oxaliplatin, with two distinct syndromes. First, a unique syndrome of acute neurosensory toxicity is generally triggered and exacerbated by exposure to cold (31). This acute neurotoxicity of oxaliplatin is associated with abnormal function of voltage-gated potassium channels in peripheral nerves and characterized by peripheral-nerve hyperexcitability (32). Second, long-term administration of oxaliplatin produces a sensory neuropathy, with loss of sensation and dysesthesias in the distal extremities. Development of sensory neuropathy is correlated with the cumulative dose of oxaliplatin and is most commonly seen in patients who have received total doses ≥540 mg/m2 (33). For instance, in one randomized study, grade 3 neurotoxicity was observed in 18% of patients, and the estimated incidence of severe neuropathy, with a dose of 85 mg/m2 per cycle, was 10% after nine cycles, 25% after 12 cycles, and 50% after 14 cycles (1). In our study, the incidence of grade 3 oxaliplatin-related neuropathy was similar to previous reports.

On cessation of drug, the chronic neurotoxicity recovers in the majority of patients within 4 to 6 months and will completely resolve in ~40% of patients by 6 to 8 months (34, 35).

The mechanism of platinum-induced neurotoxicity has been proposed to involve the accumulation of platinum within the peripheral nerve system (36, 37). The major site of damage is likely the dorsal root ganglia, consistent with the platinum accumulation studies; it results in axonopathy of peripheral nerves, especially in the large myelinated fibers responsible for sensory nerve conduction (37, 38).

Oxaliplatin-induced chronic neuropathy is a growing, relevant clinical problem because, actually, use of oxaliplatin is expanding (a) in metastatic colorectal cancer as well as in the adjuvant setting, as indicated by the results of MOSAIC trial and (b) in several other cancers (1–3). Only one previous unpublished report has studied the association between GSTP1 and GSTM1 genotypes and oxaliplatin-related cumulative neuropathy. In a series of patients with advanced colorectal cancer treated with an oxaliplatin-based chemotherapy, McLeod et al. (39) did not find an association between GSTP1 and GSTM1 genotypes and the severity of oxaliplatin-related cumulative neuropathy. The analysis limited to the patients who received an oxliplatin minimal cumulative dose of 500 mg/m2 is likely more efficient to evidence the role of GSTP1 genotypes in the occurrence of a severe neuropathy, because under this dose the frequency of neuropathy is very low.

In our study, the GSTP1 105Ile allele was significantly more frequent in patients with grade 3 cumulative peripheral neuropathy. GSTP1 belongs to a multigene family of enzymes that catalyze the conjugation of glutathione to electrophilic molecules to inactivate them and facilitate their excretion (40).

Previous studies revealed that the Ile105Val substitution modify the substrate affinity of the GSTP1 enzyme. Individuals homozygous for the GSTP1 105Val genotype had an altered catalytic activity depending on substrates compared with individuals homozygous for the GSTP1 105Ile allele (5, 41). In particular, there is in vitro evidence that GSTP1 105Val allele alone or in combination with the GSTP1 114Val allele are much more active than the wild-type alleles against cisplatin and carboplatin (42). Thus, the increase rate of grade 3 neuropathy in patients homozygous for GSTP1 *A/*A haplotype could be explained by the diminished capacity of detoxifying oxaliplatin. The involvement of GSTP1 in cellular apoptosis could be another hypothesis to explain the protective effect of the variant genotype. Indeed, the role of GSTP1 in cellular defense was recently proposed as a consequence of its ability to interact with c-Jun NH2-terminal kinase (JNK). Adler et al. (43) have shown that GSTP1 is an inhibitor of the stress-inducible JNK and has also a role in regulating of the constitutive expression of target genes of the JNK signaling pathway, in particular when containing an activator protein-1 binding site. The absence of GSTP1 protein in GSTP1/P2–/– mice results in increased constitutive JNK activity leading to the up-regulation of specific downstream genes implicated in antioxidant cellular response (44–46). The GSTP-null mice are therefore highly resistant to the hepatotoxic effects of acetaminophen and we hypothesized that the patients carrying GSTP1 105Val allele are similarly protected against cumulative peripheral neuropathy to oxaliplatin (45). Recently, Wang et al. (47) have shown that d-JNKI-1, a JNK inhibitor, potentiates the ototoxic effects of cisplatin, suggesting that this pathway is important in protecting against toxicity of this compound. In cells treated with cisplatin, the activation of JNK is necessary for caspase activation and apoptosis induction. It has also been showed that cisplatin induces activator protein-1 DNA binding activity (48). Altogether, the following hypothesis can be put forward: The GSTP1 105Ile protein could enhance oxaliplatin neurotoxicity through its inhibition of JNK, whereas the GSTP1 105Val protein allows a higher activity of JNK, inducing the expression of genes involved in cellular defense and, thus, protecting the cells against platinum toxicity.

As the use of oxaliplatin becomes established in the treatment of colorectal cancer both as palliative or adjuvant chemotherapy, it is important to determine the predictive factors of neurotoxicity to this agent. Our study had several limitations. First, we evaluated only a relatively small number of patients. Second, patients received different oxaliplatin-based chemotherapies. Therefore, our data should be considered as preliminary results and more large studies with identical oxaliplatin-based chemotherapy are need to confirm that the GSTP1 genotype may be a powerful predictor of cumulative neuropathy to oxaliplatin-based chemotherapy. This finding could be provide the opportunity for safer chemotherapy for patients with gastrointestinal cancer requiring oxaliplatin-based treatment, and raises the possibility of being able to individualize the oxaliplatin dose by the identification of patients requiring monitoring or adjustment. Thus, this genotyping may be useful to prevent or limit the oxaliplatin-induced cumulative neuropathy.


   Acknowledgments
 
We thank the "Ligue Nationale de Lutte Contre le Cancer" and the Assistance Publique-Hôpitaux de Paris (CRC 02036P021011) for their financial support.


   Footnotes
 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Note: T. Lecomte and B. Landi contributed equally to this work.

Received 9/22/05; revised 3/ 5/06; accepted 3/14/06.


   References
Top
 Abstract
 Patients and Methods
 Results
 Discussion
 References
 

  1. de Gramont A, Figer A, Seymour M, et al. Leucovorin and fluorouracil with or without oxaliplatin as first-line treatment in advanced colorectal cancer. J Clin Oncol 2000;18:2938–47.[Abstract/Free Full Text]
  2. Goldberg RM, Sargent DJ, Morton RF, et al. A randomized controlled trial of fluorouracil plus leucovorin, irinotecan, and oxaliplatin combinations in patients with previously untreated metastatic colorectal cancer. J Clin Oncol 2004;22:23–30.[Abstract/Free Full Text]
  3. Andre T, Boni C, Mounedji-Boudiaf L, et al. Oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment for colon cancer. N Engl J Med 2004;350:2343–51.[Abstract/Free Full Text]
  4. Culy CR, Clemett D, Wiseman LR. Oxaliplatin. A review of its pharmacological properties and clinical efficacy in metastatic colorectal cancer and its potential in other malignancies. Drugs 2000;60:895–924.[CrossRef][Medline]
  5. Watson MA, Stewart RK, Smith GB, Massey TE, Bell DA. Human glutathione S-transferase P1 polymorphisms: relationship to lung tissue enzyme activity and population frequency distribution. Carcinogenesis 1998;19:275–80.[Abstract/Free Full Text]
  6. Ali-Osman F, Akande O, Antoun G, Mao JX, Buolamwini J. Molecular cloning, characterization, and expression in Escherichia coli of full-length cDNAs of three human glutathione S-transferase {pi} gene variants. Evidence for differential catalytic activity of the encoded proteins. J Biol Chem 1997;272:10004–12.[Abstract/Free Full Text]
  7. Blackburn AC, Woollatt E, Sutherland GR, Board PG. Characterization and chromosome location of the gene GSTZ1 encoding the human {zeta} class glutathione transferase and maleylacetoacetate isomerase. Cytogenet Cell Genet 1998;83:109–14.[CrossRef][Medline]
  8. Ketterer B, Harris JM, Talaska G, et al. The human glutathione S-transferase supergene family, its polymorphism, and its effects on susceptibility to lung cancer. Environ Health Perspect 1992;98:87–94.[Medline]
  9. Hirvonen A. Polymorphisms of xenobiotic-metabolizing enzymes and susceptibility to cancer. Environ Health Perspect 1999;107:37–47.[Medline]
  10. Stoehlmacher J, Park DJ, Zhang W, et al. Association between glutathione S-transferase P1, T1, and M1 genetic polymorphism and survival of patients with metastatic colorectal cancer. J Natl Cancer Inst 2002;94:936–42.[Abstract/Free Full Text]
  11. Cullen KJ, Newkirk KA, Schumaker LM, Aldosari N, Rone JD, Haddad BR. Glutathione S-transferase {pi} amplification is associated with cisplatin resistance in head and neck squamous cell carcinoma cell lines and primary tumors. Cancer Res 2003;63:8097–102.[Abstract/Free Full Text]
  12. Hoban PR, Robson CN, Davies SM, et al. Reduced topoisomerase II and elevated {alpha} class glutathione S-transferase expression in a multidrug resistant CHO cell line highly cross-resistant to mitomycin C. Biochem Pharmacol 1992;43:685–93.[CrossRef][Medline]
  13. Chao CC, Huang YT, Ma CM, Chou WY, Lin-Chao S. Overexpression of glutathione S-transferase and elevation of thiol pools in a multidrug-resistant human colon cancer cell line. Mol Pharmacol 1992;41:69–75.[Abstract]
  14. Wang AL, Tew KD. Increased glutathione-S-transferase activity in a cell line with acquired resistance to nitrogen mustards. Cancer Treat Rep 1985;69:677–82.[Medline]
  15. Buller AL, Clapper ML, Tew KD. Glutathione S-transferases in nitrogen mustard-resistant and -sensitive cell lines. Mol Pharmacol 1987;31:575–8.[Abstract]
  16. Robson CN, Lewis AD, Wolf CR, et al. Reduced levels of drug-induced DNA cross-linking in nitrogen mustard-resistant Chinese hamster ovary cells expressing elevated glutathione S-transferase activity. Cancer Res 1987;47:6022–7.[Abstract/Free Full Text]
  17. McGown AT, Fox BW. A proposed mechanism of resistance to cyclophosphamide and phosphoramide mustard in a Yoshida cell line in vitro. Cancer Chemother Pharmacol 1986;17:223–6.[CrossRef][Medline]
  18. Evans CG, Bodell WJ, Tokuda K, Doane-Setzer P, Smith MT. Glutathione and related enzymes in rat brain tumor cell resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea and nitrogen mustard. Cancer Res 1987;47:2525–30.[Abstract/Free Full Text]
  19. Hengstler JG, Arand M, Herrero ME, Oesch F. Polymorphisms of N-acetyltransferases, glutathione S-transferases, microsomal epoxide hydrolase and sulfotransferases: influence on cancer susceptibility. Recent Results Cancer Res 1998;154:47–85.[Medline]
  20. Reinemer P, Dirr HW, Ladenstein R, et al. Three-dimensional structure of class {pi} glutathione S-transferase from human placenta in complex with S-hexylglutathione at 2.8 A resolution. J Mol Biol 1992;227:214–26.[CrossRef][Medline]
  21. Zimniak P, Nanduri B, Pikula S, et al. Naturally occurring human glutathione S-transferase GSTP1-1 isoforms with isoleucine and valine in position 104 differ in enzymic properties. Eur J Biochem 1994;224:893–9.[Medline]
  22. Andre T, Bensmaine MA, Louvet C, et al. Multicenter phase II study of bimonthly high-dose leucovorin, fluorouracil infusion, and oxaliplatin for metastatic colorectal cancer resistant to the same leucovorin and fluorouracil regimen. J Clin Oncol 1999;17:3560–8.[Abstract/Free Full Text]
  23. Maindrault-Goebel F, Louvet C, Andre T, et al. Oxaliplatin added to the simplified bimonthly leucovorin and 5-fluorouracil regimen as second-line therapy for metastatic colorectal cancer (FOLFOX6). GERCOR. Eur J Cancer 1999;35:1338–42.[CrossRef][Medline]
  24. Maindrault-Goebel F, de Gramont A, Louvet C, et al. High-dose intensity oxaliplatin added to the simplified bimonthly leucovorin and 5-fluorouracil regimen as second-line therapy for metastatic colorectal cancer (FOLFOX 7). Eur J Cancer 2001;37:1000–5.[CrossRef][Medline]
  25. Cascinu S, Graziano F, Ferrau F, et al. Raltitrexed plus oxaliplatin (TOMOX) as first-line chemotherapy for metastatic colorectal cancer. A phase II study of the Italian Group for the Study of Gastrointestinal Tract Carcinomas (GISCAD). Ann Oncol 2002;13:716–20.[Abstract/Free Full Text]
  26. Louvet C, Andre T, Lledo G, et al. Gemcitabine combined with oxaliplatin in advanced pancreatic adenocarcinoma: final results of a GERCOR multicenter phase II study. J Clin Oncol 2002;20:1512–8.[Abstract/Free Full Text]
  27. Caussanel JP, Levi F, Brienza S, et al. Phase I trial of 5-day continuous venous infusion of oxaliplatin at circadian rhythm-modulated rate compared with constant rate. J Natl Cancer Inst 1990;82:1046–50.[Abstract/Free Full Text]
  28. Nedelcheva Kristensen V, Andersen TI, Erikstein B, et al. Single tube multiplex polymerase chain reaction genotype analysis of GSTM1, GSTT1 and GSTP1: relation of genotypes to TP53 tumor status and clinicopathological variables in breast cancer patients. Pharmacogenetics 1998;8:441–7.[CrossRef][Medline]
  29. Kaplan EL, Meier P. Non parametric estimation from incomplete observation. J Am Stat Assoc 1958;53:457–81.[CrossRef]
  30. Welfare M, Monesola Adeokun A, Bassendine MF, Daly AK. Polymorphisms in GSTP1, GSTM1, and GSTT1 and susceptibility to colorectal cancer. Cancer Epidemiol Biomarkers Prev 1999;8:289–92.[Abstract/Free Full Text]
  31. Wilson RH, Lehky T, Thomas RR, Quinn MG, Floeter MK, Grem JL. Acute oxaliplatin-induced peripheral nerve hyperexcitability. J Clin Oncol 2002;20:1767–74.[Abstract/Free Full Text]
  32. Grolleau F, Gamelin L, Boisdron-Celle M, Lapied B, Pelhate M, Gamelin E. A possible explanation for a neurotoxic effect of the anticancer agent oxaliplatin on neuronal voltage-gated sodium channels. J Neurophysiol 2001;85:2293–7.[Abstract/Free Full Text]
  33. Cersosimo RJ. Oxaliplatin-associated neuropathy: a review. Ann Pharmacother 2005;39:128–35.[Abstract/Free Full Text]
  34. Brienza S, Vignoud J, Itzahki M, Krikorian A. Oxaliplatin (L-OHP): global safety in 682 patients [abstract]. Proc Am Soc Clin Oncol 1995;14:209.
  35. Maindrault-Goebel F, Tournigand C, Andre T, et al. Oxaliplatin reintroduction in patients previously treated with leucovorin, fluorouracil and oxaliplatin for metastatic colorectal cancer. Ann Oncol 2004;15:1210–4.[Abstract/Free Full Text]
  36. Cavaletti G, Tredici G, Marmiroli P, Petruccioli MG, Barajon I, Fabbrica D. Morphometric study of the sensory neuron and peripheral nerve changes induced by chronic cisplatin (DDP) administration in rats. Acta Neuropathol (Berl) 1992;84:364–71.[Medline]
  37. Gregg RW, Molepo JM, Monpetit VJ, et al. Cisplatin neurotoxicity: the relationship between dosage, time, and platinum concentration in neurologic tissues, and morphologic evidence of toxicity. J Clin Oncol 1992;10:795–803.[Abstract/Free Full Text]
  38. Luo FR, Wyrick SD, Chaney SG. Comparative neurotoxicity of oxaliplatin, ormaplatin, and their biotransformation products utilizing a rat dorsal root ganglia in vitro explant culture model. Cancer Chemother Pharmacol 1999;44:29–38.[CrossRef][Medline]
  39. McLeod HL, Sargent DJ, Marsh S, et al. Pharmacogenetic analysis of systemic toxicity and response after 5-fluorouracil (5FU)/CPT-11, 5FU/oxaliplatin (oxal), or CPT-11/oxal therapy for advanced colorectal cancer (CRC): results from an intergroup trial [Abstract]. Proc Am Soc Clin Oncol 2003;22:253.
  40. Henderson CJ, McLaren AW, Moffat GJ, Bacon EJ, Wolf CR. {pi}-Class glutathione S-transferase: regulation and function. Chem Biol Interact 1998;111–2:69–82.
  41. Srivastava SK, Singhal SS, Hu X, Awasthi YC, Zimniak P, Singh SV. Differential catalytic efficiency of allelic variants of human glutathione S-transferase {pi} in catalyzing the glutathione conjugation of thiotepa. Arch Biochem Biophys 1999;366:89–94.[CrossRef][Medline]
  42. Ishimoto TM, Ali-Osman F. Allelic variants of the human glutathione S-transferase P1 gene confer differential cytoprotection against anticancer agents in Escherichia coli. Pharmacogenetics 2002;12:543–53.[CrossRef][Medline]
  43. Adler V, Yin Z, Fuchs SY, et al. Regulation of JNK signaling by GSTp. EMBO J 1999;18:1321–34.[CrossRef][Medline]
  44. Kitteringham NR, Powell H, Jenkins RE, et al. Protein expression profiling of glutathione S-transferase {pi} null mice as a strategy to identify potential markers of resistance to paracetamol-induced toxicity in the liver. Proteomics 2003;3:191–207.[CrossRef][Medline]
  45. Henderson CJ, Wolf CR, Kitteringham N, Powell H, Otto D, Park BK. Increased resistance to acetaminophen hepatotoxicity in mice lacking glutathione S-transferase {pi}. Proc Natl Acad Sci U S A 2000;97:12741–5.[Abstract/Free Full Text]
  46. Elsby R, Kitteringham NR, Goldring CE, et al. Increased constitutive c-Jun N-terminal kinase signaling in mice lacking glutathione S-transferase {pi}. J Biol Chem 2003;278:22243–9.[Abstract/Free Full Text]
  47. Wang J, Ladrech S, Pujol R, Brabet P, Van De Water TR, Puel JL. Caspase inhibitors, but not c-Jun NH2-terminal kinase inhibitor treatment, prevent cisplatin-induced hearing loss. Cancer Res 2004;64:9217–24.[Abstract/Free Full Text]
  48. Sanchez-Perez I, Perona R. Lack of c-Jun activity increases survival to cisplatin. FEBS Lett 1999;453:151–8.[CrossRef][Medline]



This article has been cited by other articles:


Home page
 Cancer Res.Home page
A. M. Moyer, O. E. Salavaggione, T.-Y. Wu, I. Moon, B. W. Eckloff, M. A.T. Hildebrandt, D. J. Schaid, E. D. Wieben, and R. M. Weinshilboum
Glutathione S-Transferase P1: Gene Sequence Variation and Functional Genomic Studies
Cancer Res., June 15, 2008; 68(12): 4791 - 4801.
[Abstract] [Full Text] [PDF]

Home page
 J. Mol. Diagn.Home page
A. Buchard, J. J. Sanchez, K. Dalhoff, and N. Morling
Multiplex PCR Detection of GSTM1, GSTT1, and GSTP1 Gene Variants: Simultaneously Detecting GSTM1 and GSTT1 Gene Copy Number and the Allelic Status of the GSTP1 Ile105Val Genetic Variant
J. Mol. Diagn., November 1, 2007; 9(5): 612 - 617.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
L. Gamelin, O. Capitain, A. Morel, A. Dumont, S. Traore, L. B. Anne, S. Gilles, M. Boisdron-Celle, and E. Gamelin
Predictive Factors of Oxaliplatin Neurotoxicity: The Involvement of the Oxalate Outcome Pathway
Clin. Cancer Res., November 1, 2007; 13(21): 6359 - 6368.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
A. Ruzzo, F. Graziano, F. Loupakis, E. Rulli, E. Canestrari, D. Santini, V. Catalano, R. Ficarelli, P. Maltese, R. Bisonni, et al.
Pharmacogenetic Profiling in Patients With Advanced Colorectal Cancer Treated With First-Line FOLFOX-4 Chemotherapy
J. Clin. Oncol., April 1, 2007; 25(10): 1247 - 1254.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
J. Oldenburg, S. M. Kraggerud, M. Cvancarova, R. A. Lothe, and S. D. Fossa
Cisplatin-Induced Long-Term Hearing Impairment Is Associated With Specific Glutathione S-Transferase Genotypes in Testicular Cancer Survivors
J. Clin. Oncol., February 20, 2007; 25(6): 708 - 714.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lecomte, T.
Right arrow Articles by Loriot, M.-A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lecomte, T.
Right arrow Articles by Loriot, M.-A.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Cell Growth & Differentiation