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Serum antibodies against frameshift peptides in microsatellite unstable colorectal cancer patients with Lynch syndrome

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Abstract

High level microsatellite instability (MSI-H) occurs in about 15% of colorectal cancer (CRCs), either as sporadic cancers or in the context of hereditary non-polyposis cancer or Lynch syndrome. In MSI-H CRC, mismatch repair deficiency leads to insertion/deletion mutations at coding microsatellites and thus to the translation of frameshift peptides (FSPs). FSPs are potent inductors of T cell responses in vitro and in vivo. The present study aims at the identification of FSP-specific humoral immune responses in MSI-H CRC and Lynch syndrome. Sera from patients with history of MSI-H CRC (n = 69), healthy Lynch syndrome mutation carriers (n = 31) and healthy controls (n = 52) were analyzed for antibodies against FSPs using peptide ELISA. Reactivities were measured against FSPs derived from genes frequently mutated in MSI-H CRCs, AIM2, TGFBR2, CASP5, TAF1B, ZNF294, and MARCKS. Antibody reactivity against FSPs was significantly higher in MSI-H CRC patients than in healthy controls (P = 0.036, Mann–Whitney) and highest in patients with shortest interval between tumor resection and serum sampling. Humoral immune responses in patients were most frequently directed against FSPs derived from mutated TAF1B (11.6%, 8/69) and TGFBR2 (10.1%, 7/69). Low level FSP-specific antibodies were also detected in healthy mutation carriers. Our results show that antibody responses against FSPs are detectable in MSI-H CRC patients and healthy Lynch syndrome mutation carriers. Based on the high number of defined FSP antigens, measuring FSP-specific humoral immune responses is a highly promising tool for future diagnostic application in MSI-H cancer patients.

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Abbreviations

CRC:

Colorectal cancer

ELISA:

Enzyme-linked immunosorbent assay

HNPCC:

Hereditary non-polyposis cancer

MSI-H:

High level microsatellite instability

FSP:

Frameshift-derived peptide

MMR:

Mismatch repair

PBS:

Phosphate-buffered saline

References

  1. Ionov Y, Peinado MA, Malkhosyan S et al (1993) Ubiquitous somatic mutations in simple repeated sequences reveal a new mechanism for colonic carcinogenesis. Nature 363:558–561

    Article  CAS  PubMed  Google Scholar 

  2. Thibodeau SN, Bren G, Schaid D (1993) Microsatellite instability in cancer of the proximal colon. Science 260:816–819

    Article  CAS  PubMed  Google Scholar 

  3. Lynch HT, de la Chapelle A (2003) Hereditary colorectal cancer. N Engl J Med 348:919–932

    Article  CAS  PubMed  Google Scholar 

  4. Quehenberger F, Vasen HF, van Houwelingen HC (2005) Risk of colorectal and endometrial cancer for carriers of mutations of the hMLH1 and hMSH2 gene: correction for ascertainment. J Med Genet 42:491–496

    Article  CAS  PubMed  Google Scholar 

  5. Hampel H, Frankel WL, Martin E et al (2008) Feasibility of screening for Lynch syndrome among patients with colorectal cancer. J Clin Oncol 26:5783–5788

    Article  PubMed  Google Scholar 

  6. Watson P, Lin KM, Rodriguez-Bigas MA et al (1998) Colorectal carcinoma survival among hereditary nonpolyposis colorectal carcinoma family members. Cancer 83:259–266

    Article  CAS  PubMed  Google Scholar 

  7. Dolcetti R, Viel A, Doglioni C et al (1999) High prevalence of activated intraepithelial cytotoxic T lymphocytes and increased neoplastic cell apoptosis in colorectal carcinomas with microsatellite instability. Am J Pathol 154:1805–1813

    CAS  PubMed  Google Scholar 

  8. Buckowitz A, Knaebel HP, Benner A et al (2005) Microsatellite instability in colorectal cancer is associated with local lymphocyte infiltration and low frequency of distant metastases. Br J Cancer 92:1746–1753

    Article  CAS  PubMed  Google Scholar 

  9. Banerjea A, Bustin SA, Dorudi S (2005) The immunogenicity of colorectal cancers with high-degree microsatellite instability. World J Surg Oncol 3:26

    Article  PubMed  Google Scholar 

  10. Lynch HT, Drescher KM, de la Chapelle A (2008) Immunology and the Lynch syndrome. Gastroenterology 134:1246–1249

    Article  PubMed  Google Scholar 

  11. Saeterdal I, Bjorheim J, Lislerud K et al (2001) Frameshift-mutation-derived peptides as tumor-specific antigens in inherited and spontaneous colorectal cancer. Proc Natl Acad Sci U S A 98:13255–13260

    Article  CAS  PubMed  Google Scholar 

  12. Woerner SM, Benner A, Sutter C et al (2003) Pathogenesis of DNA repair-deficient cancers: a statistical meta-analysis of putative Real Common Target genes. Oncogene 22:2226–2235

    Article  CAS  PubMed  Google Scholar 

  13. Woerner SM, Kloor M, von Knebel Doeberitz M et al (2006) Microsatellite instability in the development of DNA mismatch repair deficient tumors. Cancer Biomark 2:69–86

    CAS  PubMed  Google Scholar 

  14. Schwitalle Y, Kloor M, Eiermann S et al (2008) Immune response against frameshift-induced neopeptides in HNPCC patients and healthy HNPCC mutation carriers. Gastroenterology 134:988–997

    Article  CAS  PubMed  Google Scholar 

  15. Reuschenbach M, von Knebel Doeberitz M, Wentzensen N (2009) A systematic review of humoral immune responses against tumor antigens. Cancer Immunol Immunother 58:1535–1544

    Article  CAS  PubMed  Google Scholar 

  16. Ishikawa T, Fujita T, Suzuki Y et al (2003) Tumor-specific immunological recognition of frameshift-mutated peptides in colon cancer with microsatellite instability. Cancer Res 63:5564–5572

    CAS  PubMed  Google Scholar 

  17. Steinitz M, Baraz L (2000) A rapid method for estimating the binding of ligands to ELISA microwells. J Immunol Methods 238:143–150

    Article  CAS  PubMed  Google Scholar 

  18. Reuschenbach M, Waterboer T, Wallin KL et al (2008) Characterization of humoral immune responses against p16, p53, HPV16 E6 and HPV16 E7 in patients with HPV-associated cancers. Int J Cancer 123:2626–2631

    Article  CAS  PubMed  Google Scholar 

  19. Korangy F, Ormandy LA, Bleck JS et al (2004) Spontaneous tumor-specific humoral and cellular immune responses to NY-ESO-1 in hepatocellular carcinoma. Clin Cancer Res 10:4332–4341

    Article  CAS  PubMed  Google Scholar 

  20. Jäger E, Stockert E, Zidianakis Z et al (1999) Humoral immune responses of cancer patients against “Cancer–Testis” antigen NY-ESO-1: correlation with clinical events. Int J Cancer 84:506–510

    Article  PubMed  Google Scholar 

  21. Shimada H, Takeda A, Arima M et al (2000) Serum p53 antibody is a useful tumor marker in superficial esophageal squamous cell carcinoma. Cancer 89:1677–1683

    Article  CAS  PubMed  Google Scholar 

  22. Lu H, Goodell V, Disis ML (2008) Humoral immunity directed against tumor-associated antigens as potential biomarkers for the early diagnosis of cancer. J Proteome Res 7:1388–1394

    Article  CAS  PubMed  Google Scholar 

  23. Zhang JY, Casiano CA, Peng XX et al (2003) Enhancement of antibody detection in cancer using panel of recombinant tumor-associated antigens. Cancer Epidemiol Biomarkers Prev 12:136–143

    CAS  PubMed  Google Scholar 

  24. Montgomery RB, Makary E, Schiffman K et al (2005) Endogenous anti-HER2 antibodies block HER2 phosphorylation and signaling through extracellular signal-regulated kinase. Cancer Res 65:650–656

    CAS  PubMed  Google Scholar 

  25. Speetjens FM, Kuppen PJ, Morreau H et al (2008) Immune response against frameshift-induced neopeptides in HNPCC patients and healthy HNPCC mutation carriers. Gastroenterology 135:711–712

    Article  PubMed  Google Scholar 

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Acknowledgments

This work was funded by a grant from the “Deutsche Krebshilfe” (grant number 106908).

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Authors and Affiliations

  1. Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 220, 69120, Heidelberg, Germany

    Miriam Reuschenbach, Matthias Kloor, Nicolas Wentzensen, Anja Germann, Yvette Garbe & Magnus von Knebel Doeberitz

  2. Group Cancer Early Detection, German Cancer Research Center (DKFZ) Heidelberg and Molecular Medicine Partnership Unit (MMPU), University Hospital Heidelberg, Heidelberg, Germany

    Miriam Reuschenbach, Matthias Kloor, Nicolas Wentzensen, Anja Germann, Yvette Garbe & Magnus von Knebel Doeberitz

  3. Medizinisch-Genetisches Zentrum, University of Munich, Munich, Germany

    Monika Morak & Elke Holinski-Feder

  4. Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Bethesda, MA, USA

    Nicolas Wentzensen

  5. Department of Surgery, University Hospital Heidelberg, Heidelberg, Germany

    Mirjam Tariverdian

  6. Institute of Clinical Chemistry, University Hospital Mannheim, Mannheim, Germany

    Peter Findeisen & Michael Neumaier

Authors
  1. Miriam Reuschenbach
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  2. Matthias Kloor
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  3. Monika Morak
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  4. Nicolas Wentzensen
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  10. Elke Holinski-Feder
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  11. Magnus von Knebel Doeberitz
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Corresponding author

Correspondence to Magnus von Knebel Doeberitz.

Additional information

Miriam Reuschenbach and Matthias Kloor contributed equally to this work.

Electronic supplementary material

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10689_2009_9307_MOESM1_ESM.pdf

Specific preabsorption of antibodies against FSPs. Representative serum reactive against TGFBR2(-1) and TGFBR2(-1)-C (A) and against TAF1B(-1) (B). ELISA results with different peptides as target antigens (A1-4; B1-3) after preincubation (blocking) of serum with different peptides at different concentrations (1–100 μg/ml). Preincubation of serum with TGFBR2(-1) and TGFBR2(-1)-C (A1, A3) and TAF1B(-1) (B1), decreases reactivity against the respective peptides while preincubation with other peptides does not show an effect. (PDF 89 kb)

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Reuschenbach, M., Kloor, M., Morak, M. et al. Serum antibodies against frameshift peptides in microsatellite unstable colorectal cancer patients with Lynch syndrome. Familial Cancer 9, 173–179 (2010). https://doi.org/10.1007/s10689-009-9307-z

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  • DOI: https://doi.org/10.1007/s10689-009-9307-z

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