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Molecular Medicine

, Volume 21, Issue 1, pp 479–486 | Cite as

Coding Microsatellite Frameshift Mutations Accumulate in Atherosclerotic Carotid Artery Lesions: Evaluation of 26 Cases and Literature Review

  • Carolin Kurz
  • Maani Hakimi
  • Matthias Kloor
  • Caspar Grond-Ginsbach
  • Marie-Luise Gross-Weissmann
  • Dittmar Böckler
  • Magnus von Knebel Doeberitz
  • Susanne Dihlmann
Research Article

Abstract

Somatic DNA alterations are known to occur in atherosclerotic carotid artery lesions; however, their significance is unknown. The accumulation of microsatellite mutations in coding DNA regions may reflect a deficiency of the DNA mismatch repair (MMR) system. Alternatively, accumulation of these coding microsatellite mutations may indicate that they contribute to the pathology. To discriminate between these two possibilities, we compared the mutation frequencies in coding microsatellites (likely functionally relevant) with those in noncoding microsatellites (likely neutral). Genomic DNA was isolated from carotid endarterectomy (CEA) specimens of 26 patients undergoing carotid surgery and from 15 nonatherosclerotic control arteries. Samples were analyzed by DNA fragment analysis for instability at three noncoding (BAT25, BAT26, CAT25) and five coding (AIM2, ACVR2, BAX, CASP5, TGFBR2) microsatellite loci, with proven validity for detection of microsatellite instability in neoplasms. We found an increased frequency of coding microsatellite mutations in CEA specimens compared with control specimens (34.6 versus 0%; p = 0.0013). Five CEA specimens exhibited more than one frameshift mutation, and ACVR2 and CASP5 were affected most frequently (5/26 and 6/26). Moreover, the rate of coding microsatellite alterations (15/130) differed significantly from that of noncoding alterations (0/78) in CEA specimens (p = 0.0013). In control arteries, no microsatellite alterations were observed, neither in coding nor in noncoding microsatellite loci. In conclusion, the specific accumulation of coding mutations suggests that these mutations play a role in the pathogenesis of atherosclerotic carotid lesions, since the absence of mutations in noncoding microsatellites argues against general microsatellite instability, reflecting MMR deficiency.

Notes

Acknowledgments

We thank Anja Spieler, Petra Hoefler and Heike Sartor for excellent technical assistance in tissue processing for histological analysis, immunohistochemistry and DNA extraction. We thank all patients and their relatives for making this study possible. This study was financially supported by a grant from the Koerber-Stiftung (young investigator grant) to Matthias Kloor. We acknowledge financial support by the Deutsche Forschungsgemeinschaft and Ruprecht-Karls-Universität Heidelberg within the funding program Open Access Publishing.

Supplementary material

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References

  1. 1.
    Libby P, Ridker PM, Hansson GK. (2009) Inflammation in atherosclerosis: from pathophysiology to practice. J. Am. Coll. Cardiol. 54:2129–38.CrossRefGoogle Scholar
  2. 2.
    Borghini A, et al. (2013) DNA modifications in atherosclerosis: from the past to the future. Atherosclerosis. 230:202–9.CrossRefGoogle Scholar
  3. 3.
    Cervelli T, et al. (2012) DNA damage and repair in atherosclerosis: current insights and future perspectives. Int. J. Mol. Sci. 13:16929–44.CrossRefGoogle Scholar
  4. 4.
    Lander ES, et al. (2001) Initial sequencing and analysis of the human genome. Nature. 409:860–921.CrossRefGoogle Scholar
  5. 5.
    Ellegren H. (2004) Microsatellites: simple sequences with complex evolution. Nat. Rev. Genet. 5:435–45.CrossRefGoogle Scholar
  6. 6.
    Gatchel JR, Zoghbi HY. (2005) Diseases of unstable repeat expansion: mechanisms and common principles. Nat. Rev. Genet. 6:743–55.CrossRefGoogle Scholar
  7. 7.
    Sammalkorpi H, et al. (2007) Background mutation frequency in microsatellite-unstable colorectal cancer. Cancer Res. 67:5691–8.CrossRefGoogle Scholar
  8. 8.
    Woerner SM, et al. (2006) Microsatellite instability in the development of DNA mismatch repair deficient tumors. Cancer Biomark. 2:69–86.CrossRefGoogle Scholar
  9. 9.
    Boland CR, et al. (1998) A National Cancer Institute Workshop on Microsatellite Instability for cancer detection and familial predisposition: development of international criteria for the determination of microsatellite instability in colorectal cancer. Cancer Res. 58:5248–57.PubMedGoogle Scholar
  10. 10.
    Woerner SM, et al. (2001) Systematic identification of genes with coding microsatellites mutated in DNA mismatch repair-deficient cancer cells. Int. J. Cancer. 93:12–9.CrossRefGoogle Scholar
  11. 11.
    Woerner SM, et al. (2003) Pathogenesis of DNA repair-deficient cancers: a statistical metaanalysis of putative Real Common Target genes. Oncogene. 22:2226–35.CrossRefGoogle Scholar
  12. 12.
    Woerner SM, et al. (2010) SelTarbase, a database of human mononucleotide-microsatellite mutations and their potential impact to tumorigenesis and immunology. Nucleic Acids Res. 38:D682–9.CrossRefGoogle Scholar
  13. 13.
    Kim TM, Laird PW, Park PJ. (2013) The landscape of microsatellite instability in colorectal and endometrial cancer genomes. Cell. 155:858–68.CrossRefGoogle Scholar
  14. 14.
    Biswas S, et al. (2008) Mutational inactivation of TGFBR2 in microsatellite unstable colon cancer arises from the cooperation of genomic instability and the clonal outgrowth of transforming growth factor beta resistant cells. Genes Chromosomes Cancer. 47:95–106.CrossRefGoogle Scholar
  15. 15.
    Findeisen P, et al. (2005) T25 repeat in the 3′ untranslated region of the CASP2 gene: a sensitive and specific marker for microsatellite instability in colorectal cancer. Cancer Res. 65:8072–8.CrossRefGoogle Scholar
  16. 16.
    Woerner SM, et al. (2005) Microsatellite instability of selective target genes in HNPCC-associated colon adenomas. Oncogene. 24:2525–35.CrossRefGoogle Scholar
  17. 17.
    Tsukano H, et al. (2010) The endoplasmic reticulum stress-C/EBP homologous protein pathway-mediated apoptosis in macrophages contributes to the instability of atherosclerotic plaques. Arterioscler. Thromb. Vasc. Biol. 30:1925–32.CrossRefGoogle Scholar
  18. 18.
    Dihlmann S, et al. (2014) Increased expression and activation of absent in melanoma 2 inflammasome components in lymphocytic infiltrates of abdominal aortic aneurysms. Mol. Med. 20:230–7.CrossRefGoogle Scholar
  19. 19.
    Hakimi M, et al. (2014) Inflammation-related induction of absent in melanoma 2 (AIM2) in vascular cells and atherosclerotic lesions suggests a role in vascular pathogenesis. J. Vasc. Surg. 59:794–803.CrossRefGoogle Scholar
  20. 20.
    Van Laer L, Dietz H, Loeys B. (2014) Loeys-Dietz syndrome. Adv. Exp. Med. Biol. 802:95–105.CrossRefGoogle Scholar
  21. 21.
    McCaffrey TA. (2009) TGF-beta signaling in atherosclerosis and restenosis. Front. Biosci. (Schol. Ed.). 2009:236–45.CrossRefGoogle Scholar
  22. 22.
    Hakimi M, et al. (2013) The expression of glycophorin A and osteoprotegerin is locally increased in carotid atherosclerotic lesions of symptomatic compared to asymptomatic patients. Int. J. Mol. Med. 32:331–8.CrossRefGoogle Scholar
  23. 23.
    Herpel E, et al. (2010) Quality management and accreditation of research tissue banks: experience of the National Center for Tumor Diseases (NCT) Heidelberg. Virchows Arch. 457:741–7.CrossRefGoogle Scholar
  24. 24.
    Clark KJ, et al. (2001) Microsatellite mutation of type II transforming growth factor-beta receptor is rare in atherosclerotic plaques. Arterioscler. Thromb. Vasc. Biol. 21:555–9.CrossRefGoogle Scholar
  25. 25.
    McCaffrey TA, et al. (1997) Genomic instability in the type II TGF-beta1 receptor gene in atherosclerotic and restenotic vascular cells. J. Clin. Invest. 100:2182–8.CrossRefGoogle Scholar
  26. 26.
    Woerner SM, et al. (2007) The putative tumor suppressor AIM2 is frequently affected by different genetic alterations in microsatellite unstable colon cancers. Genes Chromosomes Cancer. 46:1080–9.CrossRefGoogle Scholar
  27. 27.
    Sia EA, et al. (1997) Microsatellite instability in yeast: dependence on repeat unit size and DNA mismatch repair genes. Mol. Cell. Biol. 17:2851–8.CrossRefGoogle Scholar
  28. 28.
    Duval A, Hamelin R. (2002) Mutations at coding repeat sequences in mismatch repair-deficient human cancers: toward a new concept of target genes for instability. Cancer Res. 62:2447–54.PubMedGoogle Scholar
  29. 29.
    Schwartz SM, Murry CE. (1998) Proliferation and the monoclonal origins of atherosclerotic lesions. Annu. Rev. Med. 49:437–60.CrossRefGoogle Scholar
  30. 30.
    Bobik A. (2006) Transforming growth factor-betas and vascular disorders. Arterioscler. Thromb. Vasc. Biol. 26:1712–20.CrossRefGoogle Scholar
  31. 31.
    McCaffrey TA. (2000) TGF-betas and TGF-beta receptors in atherosclerosis. Cytokine Growth Factor Rev. 11:103–14.CrossRefGoogle Scholar
  32. 32.
    Dietz HC. (2007) 2006 Curt Stern Award Address: Marfan syndrome: from molecules to medicines. Am. J. Hum. Genet. 81:662–7.CrossRefGoogle Scholar
  33. 33.
    Hempen PM, et al. (2003) Evidence of selection for clones having genetic inactivation of the activin A type II receptor (ACVR2) gene in gastrointestinal cancers. Cancer Res. 63:994–9.PubMedGoogle Scholar
  34. 34.
    Yeager ME, et al. (2001) Microsatellite instability of endothelial cell growth and apoptosis genes within plexiform lesions in primary pulmonary hypertension. Circ. Res. 88:E2–11.CrossRefGoogle Scholar
  35. 35.
    Latz E, Xiao TS, Stutz A. (2013) Activation and regulation of the inflammasomes. Nat. Rev. Immunol. 13:397–411.CrossRefGoogle Scholar
  36. 36.
    Inafuku M, et al. (2004) Analysis of microsatellite instability and loss of heterozygosity in human aortic atherosclerotic lesions. Rinsho Byori. 52:961–5.PubMedGoogle Scholar
  37. 37.
    Hatzistamou J, et al. (1996) Loss of heterozygosity and microsatellite instability in human atherosclerotic plaques. Biochem. Biophys. Res. Commun. 225:186–90.CrossRefGoogle Scholar
  38. 38.
    Spandidos DA, et al. (1996) Microsatellite instability in human atherosclerotic plaques. Biochem. Biophys. Res. Commun. 220:137–40.CrossRefGoogle Scholar
  39. 39.
    Miniati P, et al. (2001) Loss of heterozygosity on chromosomes 1, 2, 8, 9 and 17 in cerebral atherosclerotic plaques. Int. J. Biol. Markers. 16:167–71.CrossRefGoogle Scholar
  40. 40.
    Flouris GA, et al. (2000) Loss of heterozygosity in DNA mismatch repair genes in human atherosclerotic plaques. Mol. Cell. Biol. Res. Commun. 4:62–5.CrossRefGoogle Scholar

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Authors and Affiliations

  • Carolin Kurz
    • 1
  • Maani Hakimi
    • 2
  • Matthias Kloor
    • 3
  • Caspar Grond-Ginsbach
    • 4
  • Marie-Luise Gross-Weissmann
    • 5
    • 6
  • Dittmar Böckler
    • 2
  • Magnus von Knebel Doeberitz
    • 3
  • Susanne Dihlmann
    • 2
  1. 1.Department of NeurologyTechnical University MunichMunichGermany
  2. 2.Department of Vascular and Endovascular SurgeryUniversity Hospital HeidelbergHeidelbergGermany
  3. 3.Applied Tumor Biology, Institute of PathologyUniversity Hospital HeidelbergHeidelbergGermany
  4. 4.Department of Neurology, Institute of PathologyUniversity Hospital HeidelbergHeidelbergGermany
  5. 5.General Pathology, Institute of PathologyUniversity Hospital HeidelbergHeidelbergGermany
  6. 6.Pathologie HeidelbergHeidelbergGermany

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