A KRAS hot spot mutation correlates with MLH1 methylation in endometrial carcinomas with microsatellite instability: A potential triage tool for Lynch syndrome evaluation

e17119 Background: Universal tumor testing for defective DNA mismatch repair (MMR) is recommended for all women diagnosed with endometrial cancer (EC) to identify those with underlying Lynch syndrome (LS). However, since its implementation in 2013, the effectiveness of this screening method on identifying individuals with LS across the population has not been well studied. The aim of this study was to evaluate outcomes of MMR immunohistochemistry (IHC), MLH1 methylation, and microsatellite instability (MSI) analysis among EC patients. Methods: We conducted a complete systematic search of online databases PubMed, Embase, Medline, and Cochrane Library between 1990-2018. A DerSimonian–Laird random-effects model meta-analysis was utilized to estimate the weighted prevalence of LS diagnoses. Results: The comprehensive search produced 3,427 publications. 29 peer-review studies met the inclusion criteria. 6,649 EC patients were identified, 206 (3%) were confirmed to have LS following positive universal tumor molecular screening.5,917 patients underwent tumor IHC, 28% had abnormal staining. 3,140 patients underwent MSI analysis, 31% had MSI instability. Among EC patients with deficient IHC staining or positive MSI analysis, the weighted prevalence of LS was 15% and 19% respectively. 1159 patients exhibited loss of MLH1 staining, 143 (13.7%) were found to be MLH1 methylation negative, 32 demonstrated a germline MLH1 mutation (2.8% of all MLH1 absent staining; 22.4% of all MLH1 methylation negative). 43% of EC patients diagnosed with LS via tumor typing would have been missed by family history-based screening alone. Conclusions: Despite widespread implementation of universal tumor testing in EC, data regarding results have previously been limited. For the first time, this study provides large-scale predictive values that will help practitioners evaluate abnormal results in the context of LS and aid in patient counseling. [Table: see text]

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Microsatellite Instability and Epigenetic Inactivation of MLH1 and Outcome of Patients With Endometrial Carcinomas of the Endometrioid Type

Journal of Clinical Oncology ◽

10.1200/jco.2006.08.2107 ◽

2007 ◽

Vol 25 (15) ◽

pp. 2042-2048 ◽

Cited By ~ 134

Author(s):

Israel Zighelboim ◽

Paul J. Goodfellow ◽

Feng Gao ◽

Randall K. Gibb ◽

Matthew A. Powell ◽

...

Keyword(s):

Endometrial Cancer ◽

Microsatellite Instability ◽

Restriction Analysis ◽

Methylation Status ◽

Disease Free Survival ◽

Survival Outcomes ◽

Gynecology And Obstetrics ◽

Endometrial Carcinomas ◽

The Impact ◽

Mlh1 Methylation

Purpose Most studies of microsatellite instability (MSI) and outcomes in endometrial cancer patients have included varied histologic subtypes. Nonetheless, MSI occurs almost exclusively in endometrioid tumors. The impact of MSI on outcomes in patients with endometrial cancer is controversial. We sought to determine whether MSI and MLH1 methylation are associated with clinicopathologic variables and survival outcomes in a large series of patients with endometrial carcinomas of the endometrioid type. Patients and Methods Tumor samples, blood, and clinicopathologic data were prospectively collected and analyzed for 446 patients with endometrioid carcinomas. MSI was determined using five National Cancer Institute (NCI) consensus panel markers, and the methylation status of the MLH1 promoter was determined by combined bisulfite restriction analysis (COBRA). Associations with clinicopathologic variables and survival outcomes were evaluated. Results MSI was identified in 147 cases (33%). MSI was associated with higher International Federation of Gynecology and Obstetrics (FIGO) grade (P < .0001). MSI+ tumors without MLH1 methylation were associated with younger age (P < .001). MSI was not associated with overall survival (OS; hazard ratio [HR], 1.011; 95% CI, 0.688 to 1.484; P = .96) or disease-free survival (DFS; HR 0.951; 95% CI, 0.554 to 1.635; P = .86). The combined MSI/MLH1 methylation status (treating MSI− as the reference) did not predict OS (MSI+/MLH1-U: HR, 0.62; 95% CI, 0.27 to 1.44; P = .26; MSI+/MLH1-M: HR, 0.95; 95% CI, 0.62 to 1.46; P = .82) or DFS (MSI+/MLH1-U: HR, 0.51; 95% CI, 0.22 to 1.19; P = .12; MSI+/MLH1-M: HR, 0.93; 95% CI, 0.62 to 1.40; P = .72). Conclusion MSI is not associated with survival in patients with endometrioid endometrial cancer.

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Combined Microsatellite Instability, MLH1 Methylation Analysis, and Immunohistochemistry for Lynch Syndrome Screening in Endometrial Cancers From GOG210: An NRG Oncology and Gynecologic Oncology Group Study

Journal of Clinical Oncology ◽

10.1200/jco.2015.63.9518 ◽

2015 ◽

Vol 33 (36) ◽

pp. 4301-4308 ◽

Cited By ~ 96

Author(s):

Paul J. Goodfellow ◽

Caroline C. Billingsley ◽

Heather A. Lankes ◽

Shamshad Ali ◽

David E. Cohn ◽

...

Keyword(s):

Family History ◽

Lynch Syndrome ◽

Microsatellite Instability ◽

Gynecologic Oncology ◽

Gynecologic Oncology Group ◽

Methylation Analysis ◽

Screening Practice ◽

Mutation Carriers ◽

Number Of Patients ◽

Mlh1 Methylation

Purpose The best screening practice for Lynch syndrome (LS) in endometrial cancer (EC) remains unknown. We sought to determine whether tumor microsatellite instability (MSI) typing along with immunohistochemistry (IHC) and MLH1 methylation analysis can help identify women with LS. Patients and Methods ECs from GOG210 patients were assessed for MSI, MLH1 methylation, and mismatch repair (MMR) protein expression. Each tumor was classified as having normal MMR, defective MMR associated with MLH1 methylation, or probable MMR mutation (ie, defective MMR but no methylation). Cancer family history and demographic and clinical features were compared for the three groups. Lynch mutation testing was performed for a subset of women. Results Analysis of 1,002 ECs suggested possible MMR mutation in 11.8% of tumors. The number of patients with a family history suggestive of LS was highest among women whose tumors were classified as probable MMR mutation (P = .001). Lynch mutations were identified in 41% of patient cases classified as probable mutation (21 of 51 tested). One of the MSH6 Lynch mutations was identified in a patient whose tumor had intact MSH6 expression. Age at diagnosis was younger for mutation carriers than noncarriers (54.3 v 62.3 years; P < .01), with five carriers diagnosed at age > 60 years. Conclusion Combined MSI, methylation, and IHC analysis may prove useful in Lynch screening in EC. Twenty-four percent of mutation carriers presented with ECs at age > 60 years, and one carrier had an MSI-positive tumor with no IHC defect. Restricting Lynch testing to women diagnosed at age < 60 years or to women with IHC defects could result in missing a substantial fraction of genetic disease.

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TUMOUR PATHOLOGY PREDICTS MICROSATELLITE INSTABILITY IN A POPULATION-BASED SERIES OF COLORECTAL CANCER CASES

Clinical & Investigative Medicine ◽

10.25011/cim.v31i4.4807 ◽

2008 ◽

Vol 31 (4) ◽

pp. 12

Author(s):

A J Hyde ◽

D Fontaine ◽

R C Green ◽

M Simms ◽

P S Parfrey ◽

...

Keyword(s):

Colorectal Cancer ◽

Lynch Syndrome ◽

Microsatellite Instability ◽

Population Based ◽

Pathological Features ◽

Predictive Tool ◽

Entire Cohort ◽

Dna Mismatch ◽

Bethesda Guidelines ◽

Revised Bethesda Guidelines

Background: Lynch Syndrome is an autosomal dominant trait that accounts forapproximately 3% of all cases of colorectal cancer (CRC). It is caused by mutations in DNA mismatch repair (MMR) genes, most commonly MLH1 or MSH2. These MMR defects cause high levels of microsatellite instability (MSI-H) in the tumours. MSI testing of all CRCs to identify potential Lynch Syndrome cases is not practical, so the Bethesda Guidelines, which use clinical and pathological features, were created to identify those tumours most likely to be MSI-H^1. In 2007 Jenkins et. al. created MsPath, a tool based on the pathological features described in the rarely used 3^rd Bethesda criterion, to improve prediction of MSI-H tumours among CRC cases diagnosed before age 60 years^2. Methods: We collected a population-based cohort of 716 CRC cases diagnosed before age 75 years in Newfoundland. For each of these cases we collected family history, performed MSI analysis, and scored a number of pathological features for the purpose of evaluating the accuracy of the Bethesda Criteria and MsPath at predicting MSI-H tumours. Results: Our work validates the MsPath tool in the Newfoundland population for the same age group used to create the tool. We found it identified MSI-H cases with a sensitivity of 95% and specificity of 35% in our population of CRCcases diagnosed before age 60 years (n=290). We also tested this tool on our older population of CRCcases, diagnosed at ages 60 to 74 years (n=426). We found it to be at least as predictive in this population,with a sensitivity of 95% and a specificity of 42%. We then used our entire cohort (N=716) to compare MsPath with the other Bethesda criteria.Bethesda criteria 1, 2, 4 and 5 together predicted MSI-H cases with a sensitivity of 67% and a specificity of 51%. MsPath was better at identifying these cases, with a sensitivity of 95% and a specificity of 39%. Conclusions: We conclude that MsPath can be extended to include patients diagnosed with CRC before age 75 years. As well, we have found that MsPath is a better predictive tool than the Revised Bethesda Guidelines for identifying MSI-H cases within a population-based setting of colorectal cancer. References: 1. Umar, A. et. al. J Natl Cancer Inst 2004;96:261-8 2.Jenkins, M.A. et. al. Gastroenterology 2007;133:48-56

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High rates of discordance between BRAF and MLH1 methylation tests in CRC patients over 70 years old in a Lynch syndrome universal screening program

10.26226/morressier.57c5383cd462b80296c9b7c3 ◽

2016 ◽

Author(s):

Tomer Adar

Keyword(s):

Lynch Syndrome ◽

Universal Screening ◽

Screening Program ◽

Mlh1 Methylation

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Idylla MSI test as a new tool for microsatellite instability detection in sebaceous tumours and keratoacanthomas

Journal of Clinical Pathology ◽

10.1136/jclinpath-2021-207606 ◽

2021 ◽

pp. jclinpath-2021-207606

Author(s):

Loëtitia Favre ◽

Ruiqian Chen ◽

Yaëlle Bellahsen-Harrar ◽

Nicolas Ortonne ◽

Anaïs Pujals

Keyword(s):

Lynch Syndrome ◽

Microsatellite Instability ◽

Sensitivity And Specificity ◽

Molecular Techniques ◽

Molecular Methods ◽

Specific Method ◽

Skin Tumours ◽

Automated Method ◽

Mmr Deficiency ◽

Mmr Proteins

AimSebaceous tumours and keratoacanthomas can be associated with mismatch repair (MMR) deficiency and thus microsatellite instability (MSI). In such tumours, MSI phenotype could be an argument to search for an underlying Muir-Torre syndrome (MTS). MTS has been recognised as a variant of Lynch syndrome, characterised by a deficiency of the MMR proteins. In Lynch syndrome, the sensitivity and specificity of the techniques used to detect MSI is well described, which is not the case for skin tumours. In our hands, immunohistochemistry is a sensitive and specific method to detect MMR deficiency in those tumours. Contrasting with tumours of Lynch spectrum, sensitivity and specificity of molecular methods has not been extensively studied. This study aimed at evaluating two molecular methods to detect MSI phenotype in MTS associated tumours: a commonly used pentaplex PCR using Bethesda markers and the fully automated method using the Idylla MSI assay.MethodsA comparison between PCR, and Idylla was performed on 39 DNA extracted from cutaneous tumours. Immunohistochemistry was used as the gold standard to calculate sensitivity and specificity of both molecular techniques.ResultsConcordant results were found in 32 cases (82%) with pentaplex PCR and in 36 cases (92%) with Idylla. The sensitivity of pentaplex PCR to detect MSI phenotype was 76% whereas Idylla sensitivity was 90%.ConclusionIdylla is more performant than PCR, for the detection of MSI in MTS-associated tumours and is a reliable additional technique to help detecting MTS in these tumours.

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Variable Expression of MSH6 in Endometrial Carcinomas With Intact Mismatch Repair and With MLH1 Loss Due to MLH1 Methylation

International Journal of Gynecological Pathology ◽

10.1097/pgp.0000000000000655 ◽

2019 ◽

Vol 39 (6) ◽

pp. 507-513 ◽

Cited By ~ 1

Author(s):

Nidhi Tandon ◽

Courtney Hudgens ◽

Bryan Fellman ◽

Michael T. Tetzlaff ◽

Russell R. Broaddus

Keyword(s):

Mismatch Repair ◽

Variable Expression ◽

Endometrial Carcinomas ◽

Mlh1 Methylation

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Clinicopathologic features of endometrial carcinomas from patients with Lynch syndrome, Lynch-like syndrome and intact DNA mismatch repair: A systematic review and meta-analysis

Gynecologic Oncology ◽

10.1016/j.ygyno.2019.04.547 ◽

2019 ◽

Vol 154 ◽

pp. 236

Author(s):

S. Gordhandas ◽

R.M. Kahn ◽

B. Maddy ◽

B. Baltich Nelson ◽

P.J. Christos ◽

...

Keyword(s):

Systematic Review ◽

Lynch Syndrome ◽

Mismatch Repair ◽

Dna Mismatch Repair ◽

Meta Analysis ◽

Clinicopathologic Features ◽

Dna Mismatch ◽

Endometrial Carcinomas

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Understanding Inherited Risk in Unselected Newly Diagnosed Patients With Endometrial Cancer

JCO Precision Oncology ◽

10.1200/po.18.00338 ◽

2019 ◽

pp. 1-15

Author(s):

Karen A. Cadoo ◽

Diana L. Mandelker ◽

Semanti Mukherjee ◽

Carolyn Stewart ◽

Deborah DeLair ◽

...

Keyword(s):

Endometrial Cancer ◽

Next Generation Sequencing ◽

Lynch Syndrome ◽

Microsatellite Instability ◽

Mismatch Repair ◽

Cancer Predisposition ◽

Next Generation ◽

Germline Variants ◽

Predisposition Genes ◽

Generation Sequencing

PURPOSE Mutations in DNA mismatch repair genes and PTEN, diagnostic of Lynch and Cowden syndromes, respectively, represent the only established inherited predisposition genes in endometrial cancer to date. The prevalence of other cancer predisposition genes remains unclear. We determined the prevalence of pathogenic germline variants in unselected patients with endometrial cancer scheduled for surgical consultation. PATIENTS AND METHODS Patients prospectively consented (April 2016 to May 2017) to an institutional review board–approved protocol of tumor-normal sequencing via a custom next-generation sequencing panel—the Memorial Sloan Kettering–Integrated Mutation Profiling of Actionable Cancer Targets—that yielded germline results for more than 75 cancer predisposition genes. Tumors were assessed for microsatellite instability. Per institutional standards, all tumors underwent Lynch syndrome screening via immunohistochemistry (IHC) for mismatch repair proteins. RESULTS Of 156 patients who consented to germline genetic testing, 118 (76%) had stage I disease. In 104 patients (67%), tumors were endometrioid, and 60 (58%) of those tumors were grade 1. Twenty-four pathogenic germline variants were identified in 22 patients (14%): seven (4.5%) had highly penetrant cancer syndromes and 15 (9.6%) had variants in low-penetrance, moderate-penetrance, or recessive genes. Of these, five (21%) were in Lynch syndrome genes (two MSH6, two PMS2, and one MLH1). All five tumors had concordant IHC staining; two (40%) were definitively microsatellite instability–high by next-generation sequencing. One patient had a known BRCA1 mutation, and one had an SMARCA4 deletion. The remaining 17 variants (71%) were incremental findings in low- and moderate-penetrance variants or genes associated with recessive disease. CONCLUSION In unselected patients with predominantly low-risk, early-stage endometrial cancer, germline multigene panel testing identified cancer predisposition gene variants in 14%. This finding may have implications for future cancer screening and risk-reduction recommendations. Universal IHC screening for Lynch syndrome successfully identifies the majority (71%) of high-penetrance germline mutations.

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