Mechanisms of resistance to imatinib mesylate in gastrointestinal stromal tumors and activity of the PKC412 inhibitor against imatinib-resistant mutants

doi:10.1053/j.gastro.2004.11.020

Gastroenterology

Volume 128, Issue 2, February 2005, Pages 270-279

https://doi.org/10.1053/j.gastro.2004.11.020 Get rights and content

Background & Aims: Resistance is a major challenge in the treatment of patients with gastrointestinal stromal tumors (GISTs). We investigated the mechanisms of resistance in patients with progressive GISTs with primary KIT mutations and the efficacy of the kinase inhibitor PKC412 for the inhibition of imatinib-resistant mutants. Methods: We performed a cytogenetic analysis and screened for mutations of the KIT and PDGFRA kinase domains in 26 resistant GISTs. KIT autophosphorylation status was assessed by Western immunoblotting. Imatinib-resistant GIST cells and Ba/F3 cells expressing these mutant proteins were tested for sensitivity to imatinib and PKC412. Results: Six distinct secondary mutations in KIT were detected in 12 progressive tumors, with V654A and T670I found to be recurrent. One progressive tumor showed acquired PDGFRA-D842V mutation. Amplification of KIT or KIT/PDGFRA was found in 2 patients. Eight of 10 progressive tumors available for analysis showed phosphorylated KIT. Two remaining progressive tumors lost KIT protein expression. GIST cells carrying KIT-del557-558/T670I or KIT-InsAY502-503/V654A mutations were resistant to imatinib, while PKC412 significantly inhibited autophosporylation of these mutants. Resistance to imatinib and sensitivity to PKC412 of KIT-T670I and PDGFRA-D842V mutants was confirmed using Ba/F3 cells. Conclusions: This study shows the high frequency of KIT/PDGFRA kinase domain mutations in patients with secondary resistance and defines genomic amplification of KIT/PDGFRA as an alternative cause of resistance to the drug. In a subset of patients, cancer cells lost their dependence on the targeted tyrosine kinase. Our findings show the sensitivity of the imatinib-resistant KIT-T670I and KIT-V654A and of PDGFRA-D842V mutants to PKC412.

Section snippets

Patients

Progressive tumors from 26 patients treated with imatinib in the Department of Oncology at University Hospital Leuven were evaluated. There were 20 men and 6 women, with a median age of 53 years (range, 37–77 years). Twenty-two of 26 patients had the primary tumor surgically removed. Chemotherapy and/or radiotherapy were applied in the advanced stage of the disease in 13 patients before treatment with imatinib. Criteria for inclusion in the imatinib study, study design, and procedures are

Results

Progressive tumors from 26 patients treated with imatinib were evaluated. Patient characteristics and clinical data are shown in Table 1. The median time from diagnosis to the proven malignancy of the disease was 48 weeks (range, 0–265 weeks), while the median time from diagnosis to imatinib treatment was 91 weeks (range, 6–304 weeks). Fifteen patients (57.6%) achieved partial remission and 10 patients (38.4%) showed stable disease during imatinib treatment, with an average duration of

Discussion

Preliminary studies described 2 categories of imatinib resistance: KIT-dependent or KIT-independent mechanisms.15 Based on our results, we conclude that reactivation of KIT is the most important mechanism for resistance. KIT was found to be phosphorylated (activated) in 8 of 10 progressive tumors that could be analyzed by Western blot during imatinib treatment. In 50% of these cases, reactivation of KIT was the consequence of secondary resistance mutations, while in the other 50% the cause for

References (28)

A.T. Van Oosterom et al.
Safety and efficacy of imatinib (STI571) in metastatic gastrointestinal tumorsa phase I study
Lancet
(2001)
B.J. Longley et al.
Classes of c-KIT activating mutationsproposed mechanisms of action and implications for disease classification and therapy
Leuk Res
(2001)
M. Debiec-Rychter et al.
Use of c-KIT/PDGFRA mutational analysis to predict the clinical response to imatinib in patients with advanced gastrointestinal stromal tumors entered on phase I and II studies of the EORTC Soft Tissue and Bone Sarcoma Group
Eur J Cancer
(2004)
Y. Ma et al.
The c-KIT mutation causing human mastocytosis is resistant to STI571 and other KIT kinase inhibitors; kinases with enzymatic site mutations show different inhibitor sensitivity profiles than wild-type kinases and those with regulatory-type mutations
Blood
(2002)
J. Cools et al.
PKC412 overcomes resistance to imatinib in a murine model of FIP1L1-PDGFRA-induced myeloproliferative disease
Cancer Cell
(2003)
E. Tamborini et al.
A new mutation in the KIT ATP pocket causes acquired resistance to imatinib in a gastrointestinal stromal tumor patient
Gastroenterology
(2004)
D. Fabbro et al.
Protein kinases as targets for anticancer agentsfrom inhibitors to useful drugs
Pharmacol Ther
(2002)
E. Weisberg et al.
Inhibition of mutant FLT3 receptors in leukemia cells by the small molecule tyrosine kinase inhibitor PKC412
Cancer Cell
(2002)
S. Hirota et al.
Familial gastrointestinal stromal tumors associated with dysphagia and novel type germline mutation of KIT gene
Gastroenterology
(2002)
J.J. Clark et al.
Variable sensitivity of FLT3 activation loop mutations to the small molecule tyrosine kinase inhibitor MLN518
Blood
(2004)

K. Kemmer et al.

KIT mutations are common in testicular seminomas

Am J Pathol

(2004)

S. Hirota et al.

Gain-of-function mutations of c-kit in human gastrointestinal stromal tumors

Science

(1998)

B.P. Rubin et al.

KIT activation is a ubiquitous feature of gastrointestinal stromal tumors

Cancer Res

(2001)

M.C. Heinrich et al.

PDGFRA activating mutations in gastrointestinal stromal tumors

Science

(2003)

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Retrospective study of the clinicopathological characteristics and prognostic factors of gastrointestinal stromal tumors in Chinese patients
2022, Annals of Diagnostic Pathology
The purpose of this study was to investigate the clinicopathological characteristics and prognostic factors of patients with gastrointestinal stromal tumors (GISTs) in mainland China. We retrospectively analyzed the clinicopathological characteristics and survival data of 149 patients with GISTs admitted to Shengjing Hospital of China Medical University from July 2011 to October 2017. The following details were collected from all patients: sex, age, symptoms, preoperative examination, pathology, surgical procedures, and follow-up data. Recurrence-free survival (RFS) and overall survival (OS) were used to assess survival outcomes. The Kaplan–Meier method was performed to draw survival curves and calculate the survival rate. The log-rank test was performed for univariate analysis, and the significant factors were included in multivariate analysis using a Cox proportional hazards model to determine prognostic factors. The 5-year RFS rate was 78.5 % and 5-year OS rate was 83.2 %. The univariate analysis showed that the following prognostic factors could significantly predict 5-year RFS and OS: tumor size, initial status, modified NIH classification, mitotic index, CD117 expression, Ki67 index, and surgical procedure (P < 0.05). The multivariate analysis showed that mitotic index, CD117, and Ki67 index were independent prognostic factors associated with 5-year RFS and 5-year OS. This study provides a reference for the clinicopathological characteristics and prognostic factors of patients with GISTs in mainland China, and the results suggest that focusing on immunohistochemical markers in clinical practice may be more reliable for the prediction of clinical outcomes.
A literature review and database of how the primary KIT/PDGFRA variant of a gastrointestinal stromal tumour predicts for sensitivity to imatinib
2022, Cancer Genetics
It is well recognized that the primary KIT or PDGFRA variant of a gastrointestinal stromal tumour (GIST) can predict sensitivity to imatinib. However, these data are currently spread across a wide range of publications and have not been collated as one reference. A broad-ranging literature search was therefore performed to assemble such a database which should help optimize imatinib-based management of GIST patients henceforth. Having excluded wild type GISTs and results for imatinib used as adjuvant therapy, 79 publications (dated August 2001 to March 2022) underwent data extraction. These data on imatinib sensitivity were either derived from in vitro studies, predicted by in silico analysis or based on in vivo clinical patient response. Data interpretation carried some caveats: there was a potential for replication of patient-derived data between older and new publications; only predicted protein sequences were presented; the criteria used to record clinical response were not uniform across all publications; and imatinib dosage could vary between different clinical publications. However, these data showed broad agreement of imatinib sensitivity amongst similar subtypes of KIT or PDGFRA variant. There was also agreement between in vivo versus in vitro/in silico derived sensitivity data for most variants when both data types were available.
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Supported by COST ACTION B19 “Molecular cytogenetics of solid tumors.” J.C. is a postdoctoral researcher from the FWO-Vlaanderen and is supported by grant SCIE2003-19 from the Belgian Federation Against Cancer. P.M. is supported by grant G.0507.04 from the FWO-Vlaanderen. B.W. is supported by a Marie Curie EC fellowship (contract HPMT-CT2001-00273).

This report presents research results of the Belgian program on Interuniversity Poles of Attraction initiated by the Belgian Sate, Prime Minister’s Office, Science Policy Programming. Its authors assume scientific responsibility.

¹: J.R. is an employee of Novartis Pharma AG, whose product (PKC412) is described in this report.

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Clinical-alimentary tractMechanisms of resistance to imatinib mesylate in gastrointestinal stromal tumors and activity of the PKC412 inhibitor against imatinib-resistant mutants

Section snippets

Patients

Results

Discussion

Lancet

Leuk Res

Eur J Cancer

Blood

Cancer Cell

Gastroenterology

Pharmacol Ther

Cancer Cell

Gastroenterology

Blood

Am J Pathol

Gain-of-function mutations of c-kit in human gastrointestinal stromal tumors

Science

KIT activation is a ubiquitous feature of gastrointestinal stromal tumors

Cancer Res

PDGFRA activating mutations in gastrointestinal stromal tumors

Science

Clinical-alimentary tract
Mechanisms of resistance to imatinib mesylate in gastrointestinal stromal tumors and activity of the PKC412 inhibitor against imatinib-resistant mutants