In vitro chemosensitivity of freshly explanted tumor cells to pemetrexed is correlated with target gene expression

Hanauske, Axel-Rainer; Eismann, Ulrike; Oberschmidt, Olaf; Pospisil, Heike; Hoffmann, Steve; Hanauske-Abel, Hartmut M.; Ma, Doreen; Chen, Victor; Paoletti, Paolo; Niyikiza, Clet

doi:10.1007/s10637-007-9060-9

Cited by 78 publications

(60 citation statements)

References 20 publications

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“…Sensitivity to pemetrexed was also associated with mRNA levels of genes involved in the mechanism of action of this drug in freshly explanted human tumor specimens. In particular, low TS mRNA, as well as reduced levels of GARFT and DHFR mRNA, significantly correlated with chemosensitivity to pemetrexed [34]. Several other studies showed that the expression of TS protein was correlated with outcome in NSCLC specimens from patients treated with pemetrexed-based chemotherapy, particularly in nonsquamous carcinomas [35,36].…”

Section: Pharmacogenetics Of Pemetrexedmentioning

confidence: 89%

On the pharmacogenetics of non-small cell lung cancer treatment

Santarpía

Rolfo

Peters

et al. 2016

Expert Opinion on Drug Metabolism & Toxicology

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Introduction. Despite many clinical efforts, non-small-cell lung cancer (NSCLC) has a dismal 5-year survival rate of 16%, and high incidence of recurrence. The success of biologically targeted agents, as well as the activity of well-established chemotherapeutic regimens, has been limited by inherited/ acquired resistance, and biomarkers to adapt the prescription of anticancer drugs to patients' features are urgently warranted. Areas covered. In oncology, pharmacogenetics should provide the way to select patients who may benefit from a specific therapy that best match the individual and tumor genetic profile, thus allowing maximum activity and minimal toxicity. The present review summarizes the main findings on NSCLC pharmacogenetics, critically reappraising the most important studies on polymorphisms correlated with outcome of pemetrexed and EGFR-inhibitors, and provides perspective on clinical application of genomic tests for treatment decision-making. Expert Opinion. A major challenge in NSCLC is the identification of subgroups of diseases/patients that will truly benefit from specific treatments. Ideally, convenient and minimally invasive tests to decipher biomarkers of chemosensitivity/resistance and toxicity should be developed alongside novel anticancer treatments. Integration with the latest generation of whole-genome analyses and liquid biopsies as well as prospective validation in large cohorts of patients will overcome the limitations of the traditional pharmacogenetic approaches. ARTICLE HISTORY

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Section: Pharmacogenetics Of Pemetrexedmentioning

confidence: 89%

On the pharmacogenetics of non-small cell lung cancer treatment

Santarpía

Rolfo

Peters

et al. 2016

Expert Opinion on Drug Metabolism & Toxicology

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“…To evaluate the possible role of breast cancer resistance protein (BCRP) and multidrug-related proteins (MRPs) expression on pemetrexed sensitivity, as suggested in a study on freshly explanted human tumor specimens (Hanauske et al, 2007), we performed Western blot analysis of BCRP, MRP1, MRP2, MRP3, MRP4, and MRP5 expression in the panel of NSCLC cells. Total lysates were prepared in buffer containing 50 mM Tris, pH 7.6, 20% (v/v) glycerol, 5 mM dithiothreitol, 0.5% (v/v) Nonidet P-40, and 4.0% (v/v) of a protease inhibitor cocktail.…”

Section: Erlotinib and Pemetrexed In Nsclc 1291mentioning

confidence: 99%

Molecular Mechanisms Underlying the Synergistic Interaction of Erlotinib, an Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor, with the Multitargeted Antifolate Pemetrexed in Non-Small-Cell Lung Cancer Cells

Giovannetti

Lemos²,

Tekle³

et al. 2008

Mol Pharmacol

141

123

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Because the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor erlotinib and the multitargeted antifolate pemetrexed are registered in the treatment of second-line nonsmall-cell lung cancer (NSCLC), empirical combinations of these drugs are being tested. This study investigated molecular mechanisms underlying their combination in six NSCLC cell lines. Cells were characterized by heterogeneous expression of pemetrexed determinants, including thymidylate synthase (TS) and dihydrofolate reductase (DHFR), and mutations potentially affecting chemosensitivity. Pharmacological interaction was studied using the combination index (CI) method, whereas cell cycle, apoptosis induction, and EGFR, extracellular signalregulated kinases 1 and 2, and Akt phosphorylation were studied by flow cytometry, fluorescence microscopy, and enzyme-linked immunosorbent assays. Reverse-transcriptase polymerase chain reaction (RT-PCR), Western blot, and activity assays were performed to assess whether erlotinib influenced TS. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium assays demonstrated that EGFR and k-Ras mutations were related to erlotinib sensitivity, whereas TS and DHFR expression were related to pemetrexed sensitivity. Synergistic cytotoxicity was found in all cells, most pronounced with pemetrexed ϩ erlotinib (24 h) 3 erlotinib (48 h) sequence (CI, 0.09 -0.40), which was associated with a significant induction of apoptosis. Pemetrexed increased EGFR phosphorylation and reduced Akt phosphorylation, which was additionally reduced by drug combination (Ϫ70.6% in H1650). Erlotinib significantly reduced TS expression and activity, possibly via E2F-1 reduction, as detected by RT-PCR and Western blot, and the combination decreased TS in situ activity in all cells. Erlotinib and pemetrexed showed a strong synergism in NSCLC cells, regardless of their genetic characteristics. Induction of apoptosis, modulation of EGFR and Akt phosphorylation, and changes in the expression of critical genes involved in pemetrexed activity contribute to this synergistic interaction and support the clinical investigation of these markers.Non-small-cell lung cancer (NSCLC) is the leading cause of cancer-related deaths in the Western world. Chemotherapy represents the backbone of treatment of advanced NSCLC, which represents more than 50% of cases diagnosed. Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org. doi:10.1124/mol.107.042382.ABBREVIATIONS: NSCLC, non-small-cell lung cancer; 5-FU, 5-fluorouracil; AI, apoptotic index; BCRP, breast cancer resistance protein; CI, combination index; DHFR, dihydrofolate reductase; EGFR, epidermal growth factor receptor; FA, fraction affected; FPGS, folyl-polyglutamate synthetase; GARFT, glycinamide ribonucleotide formyltransferase; MRPs, multidrug-related protein; PI3K, phosphatidylinositide 3-kinase; RFC, reduced folate carrier; TKI, tyrosine-kinase inhibitor; TS, thymidylate synthase; LY294002, (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one ...

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“…Therefore, most studies have focused on pemetrexed effects on TS. TS mRNA levels were inversely correlated with pemetrexed activity in different tumor cells (19,20), whereas other studies suggested a correlation between high levels of TS protein expression and reduced sensitivity to pemetrexed in colon and lung cancer cells (21,22). Furthermore, TS mRNA and protein expression were predictive of responsiveness in patients with advanced breast cancer treated with pemetrexed alone and in NSCLC patients treated with pemetrexed/gemcitabine neoadjuvant therapy, respectively (23).…”

Section: Introductionmentioning

confidence: 98%

Thymidylate Synthase and Excision Repair Cross-Complementing Group-1 as Predictors of Responsiveness in Mesothelioma Patients Treated with Pemetrexed/Carboplatin

Zucali

Giovannetti

Destro

et al. 2011

Clinical Cancer Research

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Purpose: The pemetrexed/platinum agent combination represents the standard of care in first-line treatment for malignant pleural mesothelioma (MPM). However, there are no established indicators of responsiveness that can be used to optimize the treatment. This retrospective study aimed to assess the role of excision repair cross-complementing group-1 (ERCC1) and thymidylate synthase (TS) in tumors, and correlate expression levels and polymorphisms of these key determinants of drug activity with the outcome of MPM patients treated with carboplatin/pemetrexed in first-line setting.Experimental design: Analysis of TS and ERCC1 polymorphisms, mRNA and protein expression was done by PCR and immunohistochemistry [with the H-score (histologic score)] in tumor specimens from 126 MPM patients, including 99 carboplatin-/pemetrexed-treated patients.Results: A significant correlation between low TS protein expression and disease control (DC) to carboplatin/pemetrexed therapy (P ¼ 0.027), longer progression-free survival (PFS; P ¼ 0.017), and longer overall survival (OS; P ¼ 0.022) was found when patients were categorized according to median H-score. However, patients with the higher tertile of TS mRNA expression correlated with higher risk of developing progressive disease (P ¼ 0.022), shorter PFS (P < 0.001), and shorter OS (P < 0.001). At multivariate analysis, the higher tertile of TS mRNA level and TS H-score confirmed their independent prognostic role for DC, PFS, and OS. No significant associations were found among ERCC1 protein expression, TS and ERCC1 polymorphisms, and clinical outcome.Conclusions: In our series of carboplatin-/pemetrexed-treated MPM patients, low TS protein and mRNA levels were significantly associated to DC, improved PFS, and OS. Prospective trials for the validation of the prognostic/predictive role of TS in MPM patients treated with pemetrexed-based regimens are warranted.

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In vitro chemosensitivity of freshly explanted tumor cells to pemetrexed is correlated with target gene expression

Abstract: These results form a rational basis for the design of clinical trials to evaluate the expression of these enzymes as predictors for treatment outcome.

Cited by 78 publications

References 20 publications

On the pharmacogenetics of non-small cell lung cancer treatment

On the pharmacogenetics of non-small cell lung cancer treatment

Molecular Mechanisms Underlying the Synergistic Interaction of Erlotinib, an Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor, with the Multitargeted Antifolate Pemetrexed in Non-Small-Cell Lung Cancer Cells

Thymidylate Synthase and Excision Repair Cross-Complementing Group-1 as Predictors of Responsiveness in Mesothelioma Patients Treated with Pemetrexed/Carboplatin

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