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

Colony formation is the gold standard assay for determining reproductive cell death after radiation treatment, since effects on proliferation often do not reflect survival. We have developed a high-throughput radiosensitivity screening method based on clonogenicity and screened a siRNA library again...

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Bibliographic Details
Published in:Oncotarget Vol. 6; no. 8; p. 5978
Main Authors: Tiwana, Gaganpreet S, Prevo, Remko, Buffa, Francesca M, Yu, Sheng, Ebner, Daniel V, Howarth, Alison, Folkes, Lisa K, Budwal, Balam, Chu, Kwun-Ye, Durrant, Lisa, Muschel, Ruth J, McKenna, W Gillies, Higgins, Geoff S
Format: Journal Article
Language:English
Published: United States 20.03.2015
Subjects:
Cell Line, Tumor
Colony-Forming Units Assay
DNA Damage
HCT116 Cells
HeLa Cells
High-Throughput Screening Assays
Humans
Neoplasms - metabolism
Neoplasms - radiotherapy
Pyrithiamine - pharmacology
Radiation Tolerance
Radiation-Sensitizing Agents - pharmacology
Thiamin Pyrophosphokinase - metabolism
Thiamine - metabolism
Transfection
thiamine
tumor radiosensitivity
TPK1
high-throughput screening
ISSN:1949-2553, 1949-2553
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Summary:Colony formation is the gold standard assay for determining reproductive cell death after radiation treatment, since effects on proliferation often do not reflect survival. We have developed a high-throughput radiosensitivity screening method based on clonogenicity and screened a siRNA library against kinases. Thiamine pyrophosphokinase-1 (TPK1), a key component of Vitamin B1/thiamine metabolism, was identified as a target for radiosensitization. TPK1 knockdown caused significant radiosensitization in cancer but not normal tissue cell lines. Other means of blocking this pathway, knockdown of thiamine transporter-1 (THTR1) or treatment with the thiamine analogue pyrithiamine hydrobromide (PyrH) caused significant tumor specific radiosensitization. There was persistent DNA damage in cells irradiated after TPK1 and THTR1 knockdown or PyrH treatment. Thus this screen allowed the identification of thiamine metabolism as a novel radiosensitization target that affects DNA repair. Short-term modulation of thiamine metabolism could be a clinically exploitable strategy to achieve tumor specific radiosensitization.
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ISSN:1949-2553
1949-2553
DOI:10.18632/oncotarget.3468