Adaptation of cancer cells from different entities to the MDM2 inhibitor nutlin-3 results in the emergence of p53-mutated multi-drug-resistant cancer cells

Michaelis, Martin; Rothweiler, Florian; Barth, Susanne; Činátl, Jindřich; Rikxoort, Marijke van; Löschmann, Nadine; Voges, Yvonne; Breitling, Rainer; Deimling, Andreas von; Rödel, Franz; Weber, Kristoffer; Fehse, Boris; Mack, Elisabeth; Stiewe, Thorsten; Doerr, Hans Wilhelm; Speidel, Daniel

doi:10.1038/cddis.2011.129

Cited by 162 publications

(227 citation statements)

References 40 publications

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“…Five unique TP53 mutations not found in the parental cell line were identified in subclones. In a separate study, six different cancer cell lines were exposed continuously to Nutlin‐3a for up to 14 passages 33. Here, 28 out of 35 Nutlin‐3a‐adapted sublines contained TP53 mutations.…”

Section: Discussionmentioning

confidence: 99%

Nutlin‐3a selects for cells harbouring TP53 mutations

Kucab

Hollstein

Arlt

et al. 2016

Intl Journal of Cancer

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TP53 mutations occur in half of all human tumours. Mutagen‐induced or spontaneous TP53 mutagenesis can be studied in vitro using the human TP53 knock‐in (Hupki) mouse embryo fibroblast (HUF) immortalisation assay (HIMA). TP53 mutations arise in up to 30% of mutagen‐treated, immortalised HUFs; however, mutants are not identified until TP53 sequence analysis following immortalisation (2–5 months) and much effort is expended maintaining TP53‐WT cultures. In order to improve the selectivity of the HIMA for HUFs harbouring TP53 mutations, we explored the use of Nutlin‐3a, an MDM2 inhibitor that leads to stabilisation and activation of wild‐type (WT) p53. First, we treated previously established immortal HUF lines carrying WT or mutated TP53 with Nutlin‐3a to examine the effect on cell growth and p53 activation. Nutlin‐3a induced the p53 pathway in TP53‐WT HUFs and inhibited cell growth, whereas most TP53‐mutated HUFs were resistant to Nutlin‐3a. We then assessed whether Nutlin‐3a treatment could discriminate between TP53‐WT and TP53‐mutated cells during the HIMA (n = 72 cultures). As immortal clones emerged from senescent cultures, each was treated with 10 µM Nutlin‐3a for 5 days and observed for sensitivity or resistance. TP53 was subsequently sequenced from all immortalised clones. We found that all Nutlin‐3a‐resistant clones harboured TP53 mutations, which were diverse in position and functional impact, while all but one of the Nutlin‐3a‐sensitive clones were TP53‐WT. These data suggest that including a Nutlin‐3a counter‐screen significantly improves the specificity and efficiency of the HIMA, whereby TP53‐mutated clones are selected prior to sequencing and TP53‐WT clones can be discarded.

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Section: Discussionmentioning

confidence: 99%

Nutlin‐3a selects for cells harbouring TP53 mutations

Kucab

Hollstein

Arlt

et al. 2016

Intl Journal of Cancer

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“…In particular, the p53 core domain has been the most studied both for its biological role in transcription process [96][97][98][99][100] and in cancer-related mutations. [101][102][103][104] MD conformational analyses were also performed for the N-terminal recognition α helix, [105][106][107][108] and for C-terminal fragments. 109,110 In order to study the entire monomeric protein behavior and its inter domain interactions, we have built a model for the full-length p53 (residues 1-393).…”

Section: Introductionmentioning

confidence: 99%

Molecular dynamics of the full-length p53 monomer

Chillemi¹,

Davidovich

D’Abramo³

et al. 2013

Cell Cycle

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The p53 protein is frequently mutated in a very large proportion of human tumors, where it seems to acquire gain-of-function activity that facilitates tumor onset and progression. A possible mechanism is the ability of mutant p53 proteins to physically interact with other proteins, including members of the same family, namely p63 and p73, inactivating their function. Assuming that this interaction might occurs at the level of the monomer, to investigate the molecular basis for this interaction, here, we sample the structural flexibility of the wild-type p53 monomeric protein. The results show a strong stability up to 850 ns in the DNA binding domain, with major flexibility in the N-terminal transactivations domains (TAD1 and TAD2) as well as in the C-terminal region (tetramerization domain). Several stable hydrogen bonds have been detected between N-terminal or C-terminal and DNA binding domain, and also between N-terminal and C-terminal. Essential dynamics analysis highlights strongly correlated movements involving TAD1 and the proline-rich region in the N-terminal domain, the tetramerization region in the C-terminal domain; Lys120 in the DNA binding region. The herein presented model is a starting point for further investigation of the whole protein tetramer as well as of its mutants

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“…Furthermore, a mutation that abrogates p53 transcriptional activity has been observed in cell lines after prolonged exposure to nutlin 3 treatment. 64,65 These data indicate that seeking the TP53 status in cohorts of patients before inclusion in clinical trials and after relapse could help strategize treatment plans for patients and could improve our understanding about these resistance-conferring mutations.…”

Section: Resultsmentioning

confidence: 99%

Activation of wild-type p53 by MDM2 inhibitors: a new strategy for lymphoma treatment

Pujals¹,

Favre²,

Gaulard³

et al. 2015

BLCTT

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Abstract:The tumor suppressor TP53 is frequently mutated or inactivated in human cancers. Mutations of TP53 are less common in lymphomas than in other tumors, but the protein is often inhibited by the overexpression of its main regulator -MDM2. In the past 10 years, major efforts have been made to develop drugs that can reactivate p53 and restore its functions. This review focuses on recent advances in the development of small inhibitors of MDM2, which are potentially relevant for the treatment of B-and T-cell lymphomas. We will describe the current state of development of these drugs and discuss their mechanism of action in these hematological malignancies.

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Adaptation of cancer cells from different entities to the MDM2 inhibitor nutlin-3 results in the emergence of p53-mutated multi-drug-resistant cancer cells

Cited by 162 publications

References 40 publications

Nutlin‐3a selects for cells harbouring TP53 mutations

Nutlin‐3a selects for cells harbouring TP53 mutations

Molecular dynamics of the full-length p53 monomer

Activation of wild-type p53 by MDM2 inhibitors: a new strategy for lymphoma treatment

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