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DNA C-circles are specific and quantifiable markers of alternative-lengthening-of-telomeres activity

Nature Biotechnology volume 27, pages 1181–1185 (2009)Cite this article

Abstract

Alternative lengthening of telomeres (ALT)1 is likely to be an important target for anticancer treatment as ∼10% of cancers depend on this telomere maintenance mechanism for continued growth2, and inhibition of ALT can cause cellular senescence3. However, no ALT inhibitors have been developed for therapeutic use because of the lack of a suitable ALT activity assay and of known ALT-specific target molecules. Here we show that partially single-stranded telomeric (CCCTAA)n DNA circles (C-circles) are ALT specific. We provide an assay that is rapidly and linearly responsive to ALT activity and that is suitable for screening for ALT inhibitors. We detect C-circles in blood from ALT+ osteosarcoma patients, suggesting that the C-circle assay (CC assay) may have clinical utility for diagnosis and management of ALT+ tumors.

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Figure 1: Detection of C-circles in ALT+ cells by C-circle (CC) assay.
Figure 2: CC assay is specific for ALT.
Figure 3: CC assay is rapidly responsive to ALT activity changes.
Figure 4: C-circles are increased in blood from ALT+ osteosarcoma patients.

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Acknowledgements

We thank Tumour Bank (Children's Hospital Westmead, Australia) for patient blood specimens, kConFab (Australia) for LFS-05 skin sample, P.R. Potts (University of Texas Southwestern) for MMS21 antibody, M.A. Cerone (Breakthrough Breast Cancer Research Centre Institute of Cancer Research) for C3-c16 cells, P. Crowe and R. Das Gupta (Prince of Wales Hospital) for DOS16 cells, W.-Q. Jiang and K. Perrem (Children's Medical Research Institute) for IIICF/c-10, 11, 16 and 17 cells and GM847/hTERT-3 and GM847/hTERT-6 cells, respectively, D. Spector (Cold Spring Harbor Laboratory) for EYFP-SP100 construct and L. Naldini (San Raffaele Telethon Institute for Gene Therapy) for pMD2-VSVG envelope vector, and A. Muntoni for critical review of this manuscript.

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  1. Children's Medical Research Institute, University of Sydney, New South Wales, Australia

    Jeremy D Henson, Ying Cao, Lily I Huschtscha, Andy C Chang, Amy Y M Au, Hilda A Pickett & Roger R Reddel

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  1. Jeremy D Henson
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  2. Ying Cao
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  3. Lily I Huschtscha
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  6. Hilda A Pickett
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  7. Roger R Reddel
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Contributions

J.D.H. conceived and developed the assay and wrote the manuscript. L.I.H. established the LFS-05F-24 cell line. Experiments were performed by J.D.H., Y.C., L.I.H., A.C.C., A.Y.M.A. and H.A.P. R.R.R. was responsible for all aspects of the project and edited the manuscript.

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Correspondence to Roger R Reddel.

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Henson, J., Cao, Y., Huschtscha, L. et al. DNA C-circles are specific and quantifiable markers of alternative-lengthening-of-telomeres activity. Nat Biotechnol 27, 1181–1185 (2009). https://doi.org/10.1038/nbt.1587

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