Abstract
High frequency of mutations seems to determine a higher occurrence of neoepitope formation and, thus, tumor immunogenicity. A somatic hypermutated status could thus act as a predictive biomarker of responsiveness to immunotherapy with recent immune checkpoint inhibitors. Among several factors involved in determining the hypermutated status, such as inactivating mutations in the DNA polymerases as well as exposure to external (cigarette smoke, UV radiation, chemicals) and endogenous (reactive oxygen species) mutagens, a defective DNA mismatch repair system may give rise to genetic instability and, particularly, to microsatellite instability (MSI). The occurrence of MSI has been associated with increased load of mutations and expression of abundant peptides that serve as neoantigens to elicit an immune response within a context of a favorable tumor microenvironment. Here we describe methodological strategies to investigate for the presence of the MSI phenotype in cancer samples, through a combination of molecular approaches performed on paraffin-embedded tissues.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cancer Genome Atlas Network (2012) Comprehensive molecular characterization of human colon and rectal cancer. Nature 487:330–337
Guinney J, Dienstmann R, Wang X et al (2015) The consensus molecular subtypes of colorectal cancer. Nat Med 21:1350–1356
Toth G, Gaspari Z, Jurka J (2000) Microsatellites in different eukaryotic genomes: survey and analysis. Genome Res 10:967–981
Schlotterer C (2000) Evolutionary dynamics of microsatellite DNA. Chromosoma 109:365–371
van Eyk CL, Richards RI (2012) Dynamic mutations: where are they now? Adv Exp Med Biol 769:55–77
Schmidt MH, Pearson CE (2016) Disease-associated repeat instability and mismatch repair. DNA Repair (Amst) 38:117–126
Dienstmann R, Vermeulen L, Guinney J et al (2017) Consensus molecular subtypes and the evolution of precision medicine in colorectal cancer. Nat Rev Cancer 17:79–92
Lengauer C, Kinzier KW, Volgestein B (1998) Genetic instabilities in human cancers. Nature 396:643–649
Muresu R, Sini MC, Cossu A et al (2002) Chromosomal abnormalities and microsatellite instability in sporadic endometrial cancer. Eur J Cancer 38:1802–1809
Jasin M (2000) Chromosome breaks and genomic instability. Cancer Invest 18:78–86
Sakofsky CJ, Malkova A (2017) Break induced replication in eukaryotes: mechanisms, functions, and consequences. Crit Rev Biochem Mol Biol 52:395–413
Diaz-Padilla I, Romero N, Amir E et al (2013) Mismatch repair status and clinical outcome in endometrial cancer: a systematic review and meta-analysis. Crit Rev Oncol Hematol 88:154–167
Richman S (2015) Deficient mismatch repair: read all about it. Int J Oncol 47:1189–1202
Bardi G, Pandis N, Schousboe K et al (1995) Near-diploid karyotypes with recurrent chromosome abnormalities characterize early-stage endometrial cancer. Cancer Genet Cytogenet 80:110–114
Giam M, Rancati G (2015) Aneuploidy and chromosomal instability in cancer: a jackpot to chaos. Cell Div 10:3
Palmieri G, Colombino M, Cossu A et al (2017) Genetic instability and increased mutational load: which diagnostic tool best direct patients with cancer to immunotherapy? J Transl Med 15:17
Chalmers ZR, Connelly CF, Fabrizio D et al (2017) Analysis of 100,000 human cancer genomes reveals the landscape of tumor mutational burden. Genome Med 9:34
Jamieson NB, Maker AV (2017) Gene-expression profiling to predict responsiveness to immunotherapy. Cancer Gene Ther 24:134–140
Le DT, Uram JN, Wang H et al (2015) PD-1 blockade in tumors with mismatch-repair deficiency. N Engl J Med 372:2509–2520
Dudley JC, Lin MT, Le DT et al (2016) Microsatellite instability as a biomarker for PD-1 blockade. Clin Cancer Res 22:813–820
Llosa NJ, Cruise M, Tam A et al (2015) The vigorous immune micro- environment of microsatellite instable colon cancer is balanced by multiple counter-inhibitory checkpoints. Cancer Discov 5:43–51
Fridman WH, Zitvogel L, Sautès-Fridman C et al (2017) The immune contexture in cancer prognosis and treatment. Nat Rev Clin Oncol 14:717–734
Shia J (2008) Immunohistochemistry versus microsatellite instability testing for screening colorectal cancer patients at risk for hereditary nonpolyposis colorectal cancer syndrome. Part I. The utility of immunohistochemistry. J Mol Diagn 10:293–300
Chang L, Chang M, Chang HM et al (2018) Microsatellite instability: a predictive biomarker for cancer immunotherapy. Appl Immunohistochem Mol Morphol 26:e15–e21
Colombino M, Cossu A, Manca A et al (2002) Prevalence and prognostic role of microsatellite instability in patients with rectal carcinoma. Ann Oncol 13:1447–1453
Palmieri G, Ascierto PA, Cossu A et al (2003) Assessment of genetic instability in melanocytic skin lesions through microsatellite analysis of benign nevi, dysplastic nevi, and primary melanomas along with their metastases. Melanoma Res 13:167–170
Colombino M, Cossu A, Arba A et al (2003) Microsatellite instability and mutation analysis among Southern Italian patients with colorectal carcinoma: detection of different alterations accounting for MLH1 and MSH2 inactivation in familial cases. Ann Oncol 14:1530–1536
Paliogiannis P, Attene F, Cossu A et al (2015) Impact of tissue type and content of neoplastic cells of samples on the quality of epidermal growth factor receptor mutation analysis among patients with lung adenocarcinoma. Mol Med Rep 12:187–191
Williams C, Ponten F, Moberg C et al (1999) A high frequency of sequence alterations is due to formalin fixation of archival specimens. Am J Pathol 155:1467–1471
Srinivasan M, Sedmak D, Jewell S (2002) Effect of fixatives and tissue processing on the content and integrity of nucleic acids. Am J Pathol 161:1961–1971
Apple S, Pucci R, Lowe AC et al (2011) The effect of delay in fixation, different fixatives, and duration of fixation in estrogen and progesterone receptor results in breast carcinoma. Am J Clin Pathol 135:592–598
Palomaki GE, McClain MR, Melillo S (2009) EGAPP supplementary evidence review: DNA testing strategies aimed at reducing morbidity and mortality from lynch syndrome. Genet Med 11:42–65
Pinol V, Castells A, Andreu M et al (2005) Accuracy of revised Bethesda guidelines, microsatellite instability, and immunohistochemistry for the identification of patients with hereditary nonpolyposis colorectal cancer. JAMA 293:1986–1994
Förster I, Brockmann M, Schildgen O et al (2018) Microsatellite instability testing in colorectal cancer using the QiaXcel advanced platform. BMC Cancer 18:484
Zhao H, Thienpont B, Yesilyurt BT et al (2014) Mismatch repair deficiency endows tumors with a unique mutation signature and sensitivity to DNA double-strand breaks. Elife 3:e02725
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
Acknowledgments
This work was partially supported by Associazione Italiana per la Ricerca sul Cancro (AIRC) “Programma di ricerca 5 per Mille 2018—Id.21073.”
Conflict of Interest: The authors have no conflict of interest to declare.
Author Contributions: GiP: Conception and design, acquisition of protocol data, drafting the manuscript. MiC, AM, GrP, MCS: Analysis and interpretation of molecular protocols, revising the manuscript. VD, AC: Analysis and interpretation of pathology aspects, revising the manuscript. MaC: Conception and design, contributed unpublished essential data or protocols, revising the manuscript.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Palmieri, G. et al. (2020). Genetic Instability Markers in Cancer. In: Thurin, M., Cesano, A., Marincola, F. (eds) Biomarkers for Immunotherapy of Cancer. Methods in Molecular Biology, vol 2055. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9773-2_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9773-2_6
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9772-5
Online ISBN: 978-1-4939-9773-2
eBook Packages: Springer Protocols