Identification of vitamin B1 metabolism as a tumor-specific radiosensitizing pathway using a high-throughput colony formation screen

Tiwana, G S; Prevo, Remko; Buffa, Francesca M.; Yu, Sile; Ebner, Daniel; Howarth, Alison; Folkes, Lisa K.; Budwal, B; Chu, Kwun-Ye; Durrant, L.; Muschel, Ruth J.; McKenna, W. Gillies; Higgins, Geoff S.

doi:10.18632/oncotarget.3468

Cited by 35 publications

(33 citation statements)

References 25 publications

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“…To validate CDK1 as a tumor specific radiosensitization target identified in our siRNA kinome library screen [9], we tested the effect of the specific CDK1 inhibitor RO-3306 in three tumor lines and three normal lines that are commonly used for radiosensitivity studies [11,12]. The tumor lines were HeLa (used in the siRNA screen), the bladder transitional cell carcinoma line T24, and the head and neck squamous cell carcinoma SQ20B.…”

Section: Resultsmentioning

confidence: 99%

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CDK1 inhibition sensitizes normal cells to DNA damage in a cell cycle dependent manner

et al. 2018

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Cyclin-dependent kinase 1 (CDK1) orchestrates the transition from the G2 phase into mitosis and as cancer cells often display enhanced CDK1 activity, it has been proposed as a tumor specific anti-cancer target. Here we show that the effects of CDK1 inhibition are not restricted to tumor cells but can also reduce viability in non-cancer cells and sensitize them to radiation in a cell cycle dependent manner. Radiosensitization by the specific CDK1 inhibitor, RO-3306, was determined by colony formation assays in three tumor lines (HeLa, T24, SQ20B) and three non-cancer lines (HFL1, MRC-5, RPE). Initial results showed that CDK1 inhibition radiosensitized tumor cells, but did not sensitize normal fibroblasts and epithelial cells in colony formation assays despite effective inhibition of CDK1 signaling. Further investigation showed that normal cells were less sensitive to CDK1 inhibition because they remained predominantly in G1 for a prolonged period when plated in colony formation assays. In contrast, inhibiting CDK1 a day after plating, when the cells were going through G2/M phase, reduced their clonogenic survival both with and without radiation. Our finding that inhibition of CDK1 can damage normal cells in a cell cycle dependent manner indicates that targeting CDK1 in cancer patients may lead to toxicity in normal proliferating cells. Furthermore, our finding that cell cycle progression becomes easily stalled in non-cancer cells under normal culture conditions has general implications for testing anti-cancer agents in these cells.

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Section: Resultsmentioning

confidence: 99%

“…We recently screened a kinome siRNA library for novel radiosensitization targets using colony formation in HeLa cells after irradiation as an end point [9]. CDK1 was one of the potential radiosensitization targets identified in this screen.…”

Section: Introductionmentioning

confidence: 99%

CDK1 inhibition sensitizes normal cells to DNA damage in a cell cycle dependent manner

et al. 2018

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“…Owing to the complexity of the HR and NHEJ pathways, several cell-based systems have been developed to screening the novel DSB repair inhibitors. In these approaches, DSBs were mainly introduced by IR or rare cutting meganuclease I-SceI (39)(40)(41)(42), and the consequences of DSB repair were deduced by the results of neutral single-cell electrophoresis (39), colony formation assay (40), expressions of DSB repair-dependent fluorescent protein (41) or fluorescent staining of HR-and NHEJ-specific foci (42). Because NHEJ is the predominant DSB repair pathway, there is rationale in monitoring its activity to facilitate the screening of radiosensitizers.…”

Section: Discussionmentioning

confidence: 99%

A CRISPR/Cas9–Based Screening for Non-Homologous End Joining Inhibitors Reveals Ouabain and Penfluridol as Radiosensitizers

Shang

Chen

et al. 2018

Molecular Cancer Therapeutics

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Non-homologous end joining (NHEJ) is the major pathway responsible for the repair of ionizing radiation (IR)-induced DNA double-strand breaks (DSB), and correspondingly regulates the cellular response to IR. Identification of NHEJ inhibitors could substantially enhance the tumor radiosensitivity and improve the therapeutic efficiency of radiotherapy. In this study, we demonstrated a screening for NHEJ inhibitors using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and high-resolution melting (HRM) analysis. Because NHEJ is regarded as an error-prone mechanism, the NHEJ-mediated ligation of the site-specific DSB induced by Cas9 nuclease would eventually cause the mutation of the targeted sequence. Then, HRM analysis, a reliable and rapid assay for detecting sequence variation, was performed to evaluate the mutation efficiency of the targeted site. Validating analysis confirmed the NHEJ activities were positively correlated with the mutation frequencies. Next, an approved drug library containing 1,540 compounds was interrogated by using this screening strategy. Our results identified ouabain, a cardiotonic agent, and penfluridol, an antipsychotic agent, have the capacity to restrain NHEJ activity. Further experiments revealed the radiosensitizing effects of these compounds. Overall, we presented a cell-based screening for NHEJ inhibitors, which could promote the discovery of novel radiosensitizers.

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“…Because altered AKT/PI3K/mTOR signalling has been frequently reported to contribute to human disease, researchers have made efforts to develop small-molecule inhibitors for AKT, PI3K and mTOR. Although the central role of PTEN in glucose metabolism has been well defined [31,43,46], little is known about how PTEN affects lipid metabolism [32,38]. The following results show the metabolic alterations by PTEN dysregulation.…”

Section: Detecting One-hit Oncogenementioning

confidence: 72%

Oncogene inference optimization using constraint-based modelling incorporated with protein expression in normal and tumour tissues

Shu

et al. 2020

R. Soc. open sci.

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Cancer cells are known to exhibit unusual metabolic activity, and yet few metabolic cancer driver genes are known. Genetic alterations and epigenetic modifications of cancer cells result in the abnormal regulation of cellular metabolic pathways that are different when compared with normal cells. Such a metabolic reprogramming can be simulated using constraint-based modelling approaches towards predicting oncogenes. We introduced the tri-level optimization problem to use the metabolic reprogramming towards inferring oncogenes. The algorithm incorporated Recon 2.2 network with the Human Protein Atlas to reconstruct genome-scale metabolic network models of the tissue-specific cells at normal and cancer states, respectively. Such reconstructed models were applied to build the templates of the metabolic reprogramming between normal and cancer cell metabolism. The inference optimization problem was formulated to use the templates as a measure towards predicting oncogenes. The nested hybrid differential evolution algorithm was applied to solve the problem to overcome solving difficulty for transferring the inner optimization problem into the single one. Head and neck squamous cells were applied as a case study to evaluate the algorithm. We detected 13 of the top-ranked one-hit dysregulations and 17 of the top-ranked two-hit oncogenes with high similarity ratios to the templates. According to the literature survey, most inferred oncogenes are consistent with the observation in various tissues. Furthermore, the inferred oncogenes were highly connected with the TP53/AKT/IGF/MTOR signalling pathway through PTEN, which is one of the most frequently detected tumour suppressor genes in human cancer.

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Identification of vitamin B1 metabolism as a tumor-specific radiosensitizing pathway using a high-throughput colony formation screen

Cited by 35 publications

References 25 publications

CDK1 inhibition sensitizes normal cells to DNA damage in a cell cycle dependent manner

CDK1 inhibition sensitizes normal cells to DNA damage in a cell cycle dependent manner

A CRISPR/Cas9–Based Screening for Non-Homologous End Joining Inhibitors Reveals Ouabain and Penfluridol as Radiosensitizers

Oncogene inference optimization using constraint-based modelling incorporated with protein expression in normal and tumour tissues

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