Mutation of Threonine 766 in the Epidermal Growth Factor Receptor Reveals a Hotspot for Resistance Formation against Selective Tyrosine Kinase Inhibitors

Blencke, Stephanie; Ullrich, Axel; Daub, Henrik

doi:10.1074/jbc.m211158200

Cited by 105 publications

(72 citation statements)

References 26 publications

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“…29 Introduction of bulkier hydrophobic side chains at the Thr-790 position fully preserved the cellular kinase activity of the EGFR in the presence of selective kinase inhibitors, indicating potential mechanisms of molecular resistance formation as previously found for BCR-Abl at T315I. Previous in vitro study showed that mutation of T790M in the EGFR revealed a hotspot for resistance formation against gefitinib, 30 also in vivo.…”

Section: Discussionmentioning

confidence: 98%

EGFR and erbB2 mutation status in Japanese lung cancer patients

Sasaki

Shimizu

Endo

et al. 2005

Intl Journal of Cancer

152

111

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Much evidence has accumulated that the epidermal growth factor receptor (EGFR) and its family members are strongly implicated in the development and progression of lung cancers. Somatic mutations of the EGFR gene were found in about 25-40% of Japanese lung cancer patients. More recently, erbB2 mutations are found in about 4% of European-derived lung cancer patients. We have investigated EGFR and erbB2 mutation status in 95 surgically treated nonsmall cell lung cancer (NSCLC) cases from Nagoya City University Hospital. Seventy-five adenocarcinoma cases were included. The presence or absence of EGFR and ernB2 mutations of kinase domains were analyzed by reverse transcription polymerase chain reaction (RT-PCR) amplifications and direct sequences. We have also investigated erbB2 mutation status in 27 surgically treated NSCLC cases followed by treatment with gefitinib from Kinki-chuo Chest Medical Center. EGFR mutations (CTGfiCGG; L858R) were found from 14 of 95 lung cancer patients. We also detected the deletion 1a-type mutations from 9 patients and deletion 4-type mutations from 6 patients in exon 19. In exon 20, 4 mutations including 2 novel mutations were found. Total EGFR mutations were present in 35 patients (36.8%). These mutation statuses were significantly correlated with gender (women 73.3% vs. men 20%, p < 0.0001), smoking status (never smoker 69.4% vs. smoker 16.9%, p < 0.0001), pathologic subtypes (adenocarcinoma 45.1% vs. nonadenocarcinoma 12.5%, p 5 0.0089) and differentiation status of the lung cancers (well 51% vs. moderately or poorly 18.4%, p 5 0.0021). On the other hand, erbB2 mutation was only found from 1 of 95 patients, at exon 20. This patient was female and a never smoker with adenocarcinoma. This 12 nucleotide insertion mutation (2324-2325 ins ATACGTGATGGC) was located in the exon 20 at kinase domain (775-776 ins YVMA). There was no erbB2 mutation in 27 gefitinib-treated NSCLC patients. In total, we have found only 1 erbB2 mutation from 122 (0.8%) Japanese NSCLC patients. There was a significantly higher erbB2 positive (21/31) ratio in EGFR mutant patients (13/25, 52.0%) compared to EGFR wild-type patients (10/ 62, 16.1%; p 5 0.0247). The NSCLC specimen with erbB2 mutation showed 11 immunoreactivity. The EGFR mutation status might correlate with the clinicopathologic features related to good response to gefitinib, such as gender, smoking history and pathologic subtypes of lung cancers. However, erbB2 mutation is rare from Japanese lung cancer and is of limited value for molecular target therapy. ' 2005 Wiley-Liss, Inc.Key words: EGFR; lung cancer; mutations; erbB2; Japanese Lung cancer is a major cause of death from malignant diseases, due to its high incidence, malignant behavior and lack of major advancements in treatment strategy.1 Lung cancer was the leading indication for respiratory surgery (42.2%) in 1998 in Japan.2 More than 15,000 patients underwent surgical operations at Japanese institutions in 1998. 2 The clinical behavior of the lung cancer is largely associated with its stage. ...

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Section: Discussionmentioning

confidence: 98%

EGFR and erbB2 mutation status in Japanese lung cancer patients

Sasaki

Shimizu

Endo

et al. 2005

Intl Journal of Cancer

152

111

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“…COS-7 cells were transiently transfected as previously described (15). Epidermal growth factor (EGF) stimulation of starved HeLa cells was carried out as described (18).…”

Section: Methodsmentioning

confidence: 99%

Proteome-wide Identification of Cellular Targets Affected by Bisindolylmaleimide-type Protein Kinase C Inhibitors

Brehmer

Godl

Zech

et al. 2004

Molecular & Cellular Proteomics

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Bisindolylmaleimide compounds such as GF109203X are potent inhibitors of protein kinase C (PKC) activity. Although bisindolylmaleimides are not entirely selective for PKC and are known to inhibit a few other protein kinases, these reagents have been extensively used to study the functional roles of PKC family enzymes in cellular signal transduction for more than a decade. Here, we establish a proteomics approach to gain further insights into the cellular effects of this compound class. Functional immobilization of suitable bisindolylmaleimide analogues in combination with the specific purification of cellular binding proteins by affinity chromatography led to the identification of several known and previously unknown enzyme targets. Subsequent in vitro binding and activity assays confirmed the protein kinases Ste20-related kinase and cyclin-dependent kinase 2 (CDK2) and the non-protein kinases adenosine kinase and quinone reductase type 2 as novel targets of bisindolylmaleimide inhibitors. As observed specifically for CDK2, minor chemical variation of the ligand by immobilizing the closely related bisindolylmaleimides III, VIII, and X dramatically affected target binding. These observed changes in affinity correlated with both the measured IC 50 values for in vitro CDK2 inhibition and results from molecular docking into the CDK2 crystal structure. Moreover, the conditions for affinity purification could be adapted in a way that immobilized bisindolylmaleimide III selectively interacted with either PKC␣ or ribosomal S6 protein kinase 1 only after activation of these kinases. Thus, we have established an efficient technique for the rapid identification of cellular bisindolylmaleimide targets and further demonstrate the comparative selectivity profiling of closely related kinase inhibitors within a cellular proteome. Molecular & Cellular Proteomics 3:490 -500, 2004.

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“…Intriguingly, it was initially generated in laboratory studies when findings from imatinib-resistant Bcr-Abl were projected to EGFR (Blencke et al, 2003). However, current clinical data suggest a low frequency of this alteration in NSCLC and the incidence of EGFR-T790M appears to be independent from treatment, though subpopulations of corresponding cells may be enriched during TKI regimen (Pao et al, 2005a, Toyooka et al, 2005Inukai et al, 2006).…”

Section: Mechanisms Of Resistance and Clinical Relapsementioning

confidence: 99%

EGFR kinase domain mutations – functional impact and relevance for lung cancer therapy

2007

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In 2004 remarkable clinical responses in non-small-cell lung cancer (NSCLC) patients treated with the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor gefitinib were reported to correlate with the presence of certain somatic EGFR kinase domain mutations in tumors. Since then, a surge of enthusiasm has been encountered in the field of molecular and clinical oncology. Beyond the promise of a tailored medicine, questions about the molecular mechanisms underlying the observed effects have arisen. In vitro analysis of NSCLC cells with endogenous EGFR mutations, recombinant expression of EGFR variants by transfection of several cell lines and the generation of transgenic mice expressing mutant EGFR were applied to study the impact of these genetic alterations on cellular signaling and cell fate. This review outlines the current mechanistic knowledge derived from such studies and discusses the relevance of EGFR kinase domain mutations for EGFR-directed therapies, including monoclonal antibodies.

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Mutation of Threonine 766 in the Epidermal Growth Factor Receptor Reveals a Hotspot for Resistance Formation against Selective Tyrosine Kinase Inhibitors

Cited by 105 publications

References 26 publications

EGFR and erbB2 mutation status in Japanese lung cancer patients

EGFR and erbB2 mutation status in Japanese lung cancer patients

Proteome-wide Identification of Cellular Targets Affected by Bisindolylmaleimide-type Protein Kinase C Inhibitors

EGFR kinase domain mutations – functional impact and relevance for lung cancer therapy

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