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Fen1 mutations result in autoimmunity, chronic inflammation and cancers

Abstract

Functional deficiency of the FEN1 gene has been suggested to cause genomic instability and cancer predisposition. We have identified a group of FEN1 mutations in human cancer specimens. Most of these mutations abrogated two of three nuclease activities of flap endonuclease 1 (FEN1). To demonstrate the etiological significance of these somatic mutations, we inbred a mouse line harboring the E160D mutation representing mutations identified in human cancers. Selective elimination of nuclease activities led to frequent spontaneous mutations and accumulation of incompletely digested DNA fragments in apoptotic cells. The mutant mice were predisposed to autoimmunity, chronic inflammation and cancers. The mutator phenotype results in the initiation of cancer, whereas chronic inflammation promotes the cancer progression. The current work exemplifies the approach of studying the mechanisms of individual polymorphisms and somatic mutations in cancer development, and may serve as a reference in developing new therapeutic regimens through the suppression of inflammatory responses.

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Figure 1: FEN1 mutations identified in human cancers have a segregated nuclease activity profile.
Figure 2: E160D cells are sensitive to DNA-damaging agents and show a strong mutator phenotype.
Figure 3: E160D mice accumulate apoptotic DNA in tissues as a result of retarded apoptotic DNA degradation and increased DNA damage–induced apoptosis.
Figure 4: E160D mice develop autoimmunity.
Figure 5: E160D mice are highly susceptible to cancers as a result of their mutator phenotype and chronic inflammation.

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Acknowledgements

We acknowledge G. Pfeifer, M. Lieber, W. Chen, K. Justus, L.D. Finger and S. Alas for their critical review of the manuscript. We thank R. Kolodner (University of California, San Diego) for kindly providing yeast strains, D. Zeng (City of Hope) for providing serum standard of antibodies to nuclear antigens and dsDNA, and the City of Hope DNA sequencing core facility for DNA sequencing analyses. This work was supported by a US National Institutes of Health grant R01CA073764 to B.H.S. and by the lung cancer program of City of Hope's Comprehensive Cancer Center.

Author information

Authors and Affiliations

Authors

Contributions

L.Z. designed and coordinated experiments for FEN1 mutation analysis in human mutation detection, conducted biochemical and mouse phenotype analyses, and contributed to manuscript preparation. H.F. and M.Z. conducted FEN1 mutation screening and genotyping and mouse anatomic analysis. M.L., J.Q. and W.T. constructed E160D Fen1 mutant mice and derived wild-type and Fen1-mutant MEF cells. P.S. conducted yeast genetic experiments. Q.H. conducted pathological and histological analyses. X.Z. and D.L. contributed to FEN1 mutation screening in human cancer specimens. K.K. supervised the FEN1 mutation screening and genotyping and histological analysis, and contributed to manuscript preparation. B.S. supervised the entire project, designed and coordinated most of the experiments in this study, and contributed to manuscript preparation.

Corresponding author

Correspondence to Binghui Shen.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

FEN1 mutation detection in human cancers. (PDF 30 kb)

Supplementary Fig. 2

Knock-in of E160D Fen1 mutant in mouse germline. (PDF 163 kb)

Supplementary Fig. 3

E160D mice develop subcutaneous inflammation, lymphoproliferative disorder, and extramedular hematopoiesis. (PDF 156 kb)

Supplementary Fig. 4

Analysis for loss of heterozygosity of lung tumors in heterozygous E160D mice. (PDF 98 kb)

Supplementary Table 1

FEN1 mutations identified in 12 major human cancers. (PDF 18 kb)

Supplementary Table 2

Cytokine profiles of WT normal and E160D inflammatory and adenoma lung tissues. (PDF 31 kb)

Supplementary Table 3

Oligonucleotide sequences and their applications in this study. (PDF 37 kb)

Supplementary Methods (PDF 21 kb)

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Zheng, L., Dai, H., Zhou, M. et al. Fen1 mutations result in autoimmunity, chronic inflammation and cancers. Nat Med 13, 812–819 (2007). https://doi.org/10.1038/nm1599

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