Modeling Heterogeneity of Triple-Negative Breast Cancer Uncovers a Novel Combinatorial Treatment Overcoming Primary Drug Resistance

Lamballe, Fabienne; Ahmad, Fahmida; Vinik, Yaron; Castellanet, Olivier; Daian, Fabrice; Müller, Anna-Katharina; Köhler, Ulrike A.; Bailly, Anne-Laure; Josselin, Emmanuelle; Castellano, Rémy; Cayrou, Christelle; Charafe‐Jauffret, Emmanuelle; Mills, Gordon B.; Géli, Vincent; Borg, Jean-Paul; Lev, Sima; Maina, Flavio

doi:10.1101/2020.06.22.164418

Cited by 7 publications

(18 citation statements)

References 62 publications

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“…Cell viability assay was performed on MMTV-R26 Met MGT and human breast cancer cells as previously described [4]. Briefly, cells were plated in 96-well plates, and treated after 24hrs with either single or combined drugs at the indicated concentrations.…”

Section: Cell Viability Assaymentioning

confidence: 99%

“…Met cells were treated for 12hrs with vehicle, A1155463 (A11, 1µM), R547 (3µM), alone or in combination. After trypsinisation, cell suspension was then processed as previously described [4]. Three independent experiments were performed.…”

Section: Cell Cycle Analysis By Flow Cytometry Mmtv-r26mentioning

confidence: 99%

“…We have recently reported the generation of a rather unique mouse model in which a subtle increase in the wild-type MET RTK expression levels in the mammary gland (MMTV-R26 Met mice) leads to spontaneous tumour formation faithfully recapitulating TNBC features [4]. These include histological, molecular and signalling heterogeneity, as well as primary resistance to chemotherapy and approved targeted therapies [5,6].…”

Section: Introductionmentioning

confidence: 99%

“…These include histological, molecular and signalling heterogeneity, as well as primary resistance to chemotherapy and approved targeted therapies [5,6]. We further exploited the MMTV-R26 Met TNBC model to identify a highly effective therapeutic protocol based on the combined inhibition of the anti-apoptotic molecule BCL-XL and of the cell cycle checkpoint regulator WEE1 [4]. Cell cycle proteins are frequently overexpressed and/or overactivated in several cancer types including TNBC.…”

Section: Introductionmentioning

confidence: 99%

“…We provide evidence that this combinatorial targeting significantly reduces the levels of RTK and AKT signalling besides perturbing cell cycle and DNA repair. Met MGT cell lines were derived from independent MMTV-R26 Met tumours and established as previously described [4]. Cells were cultured in complete DMEM/F12 (Dulbecco's modified Eagle's media/F12, 1/1, ThermoFisher Scientific) medium (DMEM/F12, supplemented with 10% foetal bovine serum (FBS, ThermoFisher Scientific), penicillin-streptomycin (P/S, 100 U/ml/0.1 mg/ml, ThermoFisher Scientific), glutamine (2mM, ThermoFisher Scientific), glucose (0.25%, Sigma), insulin (10 µg/ml, Sigma), transferrin (10µg/ml, Sigma), sodium selenite (5ng/ml, Sigma), hydrocortisone (0.5 µg/ml, Sigma), EGF (20ng/ml, Roche), and HGF (10 ng/ml, Peprotech), at 37C° in a 5% CO 2 atmosphere.…”

Section: Introductionmentioning

confidence: 99%

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BCL-XL blockage in TNBC models confers vulnerability to inhibition of specific cell cycle regulators

Castellanet

Ahmad

Vinik

et al. 2021

Preprint

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Cell cycle regulators are frequently altered in Triple-Negative Breast Cancer (TNBC). Emerging agents targeting these signals offer the possibility to design new combinatorial therapies. However, preclinical models that recapitulate TNBC primary resistance and heterogeneity are essential to evaluate the potency of these combined treatments. Methods: Bioinformatic processing of human breast cancer datasets was used to analyse correlations between expression levels of cell cycle regulators and patient survival outcome. The MMTV-R26Met mouse model of TNBC resistance and heterogeneity was employed to analyse expression and targeting vulnerability of cell cycle regulators in the presence of BCL-XL blockage. Robustness of outcomes and selectivity was further explored using a panel of human breast cancer cells. Alterations of protein expression, phosphorylation, and/or cellular localisation were analysed by western blots, reverse phase protein array, and immunocytochemistry. Bioinformatics was performed to highlight drug's mechanisms of action. Results: We report that high expression levels of BCL-XL and specific cell cycle regulators correlate with poor survival outcomes of TNBC patients. Blockage of BCL-XL confers vulnerability to drugs targeting CDK1/2/4, but not FOXM1, CDK4/6, Aurora A and Aurora B, to all MMTV-R26Met and human TNBC cell lines tested. Mechanistically, we show that, co-targeting of BCL-XL and CDK1/2/4 synergistically inhibited cell growth by combinatorial depletion of survival and RTK/AKT signals, and concomitantly restoring FOXO3a tumour suppression actions. This was accompanied by an accumulation of DNA damage and consequently apoptosis. Conclusions: Our studies illustrate the possibility to exploit the vulnerability of TNBC cells to CDK1/2/4 inhibition by targeting BCL-XL. Moreover, they underline that specificity matters in targeting cell cycle regulators for combinatorial anticancer therapies.

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Section: Cell Viability Assaymentioning

confidence: 99%

Section: Cell Cycle Analysis By Flow Cytometry Mmtv-r26mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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BCL-XL blockage in TNBC models confers vulnerability to inhibition of specific cell cycle regulators

Castellanet

Ahmad

Vinik

et al. 2021

Preprint

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Walking Dead Tumor Cells for Targeted Drug Delivery Against Lung Metastasis of Triple‐Negative Breast Cancer

et al. 2022

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Lung metastasis is challenging in patients with triple‐negative breast cancer (TNBC). Surgery is always not available due to the dissemination of metastatic foci and most drugs are powerless because of poor retention at metastatic sites. TNBC cells generate an inflamed microenvironment and overexpress adhesive molecules to promote invasion and colonization. Herein, “walking dead” TNBC cells are developed through conjugating anti‐PD‐1 (programmed death protein 1 inhibitor) and doxorubicin (DOX)‐loaded liposomes onto cell corpses for temporal chemo‐immunotherapy against lung metastasis. The walking dead TNBC cells maintain plenary tumor antigens to conduct vaccination effects. Anti‐PD‐1 antibodies are conjugated to cell corpses via reduction‐activated linker, and DOX‐loaded liposomes are attached by maleimide–thiol coupling. This anchor strategy enables rapid release of anti‐PD‐1 upon reduction conditions while long‐lasting release of DOX at inflamed metastatic sites. The walking dead TNBC cells improve pulmonary accumulation and local retention of drugs, reprogram the lung microenvironment through damage‐associated molecular patterns (DAMPs) and PD‐1 blockade, and prolong overall survival of lung metastatic 4T1 and EMT6‐bearing mice. Taking advantage of the walking dead TNBC cells for pulmonary preferred delivery of chemotherapeutics and checkpoint inhibitors, this study suggests an alternative treatment option of chemo‐immunotherapy to augment the efficacy against lung metastasis.

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WEE1 Dependency and Pejorative Prognostic Value in Triple‐Negative Breast Cancer

Nonneville¹,

Finetti²,

Birnbaum³

et al. 2021

Advanced Science

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In their recent paper, [1] Lamballe et al. identify the combination treatment with BCL-XL and WEE1 inhibitors as a promising therapeutic approach in triple-negative breast cancer (TNBC). TNBC is the most aggressive molecular subtype of breast cancer, but patients with TNBC have less benefited from recognized molecular targets than patients with other subtypes. In the adjuvant setting, the only systemic treatment currently approved remains chemotherapy. [2] New systemic therapies are urgently needed. The Lamballe's study represents a promising new avenue for treatments targeting the cell cycle in TNBC. CDK4/6 inhibitors, which prevent phosphorylation of the RB tumor suppressor, thereby invoking cancer cell cycle arrest in G1, were recently approved for treatment of endocrine receptor-positive (ER+) breast cancers. But TNBC has been considered a poor candidate because of frequent loss of RB expression or high cyclin E expression, both of which being expected to confer resistance to CDK4/6 inhibitors. Moreover, many TNBC cell lines showed resistance to CDK4/6 inhibition in vitro and in vivo. [3] Recent works have pointed out that targeting other mitotic checkpoints in TNBC might help overcoming treatment resistance or synergizing drug effect. [4,5] The WEE1 G2 Checkpoint Kinase acts as a negative regulator of entry into mitosis (G2 to M transition) by protecting the nucleus from activated cyclin B1-complexed CDK1, and is thought to exert protumorigenic functions by securing a tolerable level of genomic instability, an intrinsic feature of cancer cells. [6] In order to reinforce and extend the Lamballe's results, we performed in silico analyses of WEE1 in large datasets of pre-clinical and clinical breast cancer samples.Using the genome-wide CRISPR screen of 808 cell lines derived from many cancer types from the Broad Institute, we found that WEE1 was essential for the viability of almost all cell lines, independently from their lineage.

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Modeling Heterogeneity of Triple-Negative Breast Cancer Uncovers a Novel Combinatorial Treatment Overcoming Primary Drug Resistance

Cited by 7 publications

References 62 publications

BCL-XL blockage in TNBC models confers vulnerability to inhibition of specific cell cycle regulators

BCL-XL blockage in TNBC models confers vulnerability to inhibition of specific cell cycle regulators

Walking Dead Tumor Cells for Targeted Drug Delivery Against Lung Metastasis of Triple‐Negative Breast Cancer

WEE1 Dependency and Pejorative Prognostic Value in Triple‐Negative Breast Cancer

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