scholarly journals Assessment of mismatch repair protein expression by immunohistochemistry in endometrial carcinomas with clinicopathological correlation: A study from Indian Tertiary Cancer Care Centre

2020 ◽  
Vol 5 ◽  
pp. 101-107
Author(s):  
Anila Sharma ◽  
Meenakshi Kamboj ◽  
Ajit Panaych ◽  
Gurudutt Gupta ◽  
Sunil Pasricha ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mismatch Repair ◽  
Therapeutic Modality ◽  
Protein Loss ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Etiologic Heterogeneity ◽  
Study Population ◽  
Endometrial Carcinomas ◽  
Mmr Proteins

Objectives: Endometrial carcinomas (EC) are known to be histologically and biologically heterogeneous, and their recent molecular characterization has highlighted their etiologic heterogeneity. The aim of the present study was to analyze mutations in mismatch repair (MMR) proteins in ECs by immunohistochemistry (IHC), and correlates the data with their pathological parameters. Material and Methods: The expression of MMR proteins was analyzed using IHC in VENTANA BENCHMARK XT system, on formalin-fixed paraffin-embedded tumor tissue. The study population included 102 newly diagnosed cases of ECs over a duration of 2 years. Results: On histopathologic subtyping, 85.1% of cases were of Type 1 EC, 9.8% were Type 2 EC, and 4.9% were malignant mixed Mullerian tumors. On IHC for MMR protein expression, 22 of 102 cases (21.6%) showed loss of one or more protein, and mean age of patients with deficient MMR (dMMR) was 59.6 years. All of these dMMR cases were of endometrioid subtype, forming 25.3% of EEC. The combined loss of MLH1 and PMS2 was the most common abnormality detected (50% of dMMR). On pathological correlation, 54.5% of dMMR cases were found to be of higher grade (grade 2/3; P = 0.002) and 68.2% were higher stage tumors (T1b and above; P < 0.0001). The lymph-vascular invasion was seen in 50% of dMMR cases (4 of 8 cases). Conclusion: Detecting MMR protein loss in ECs by IHC is an efficient, relatively simple, and economical method. It needs to be routinely performed in all cases of ECs. Studies are still underway to utilize it as a therapeutic modality using immunotherapy.

scholarly journals PATH-08. EVALUATION OF MISMATCH REPAIR GENE EXPRESSION BY IMMUNOHISTOCHEMISTRY MAY DETECT EARLY PHASE OF MMR DEFICIENCY IN RECURRENT GLIOMAS

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii165-ii165
Author(s):  
Fumi Higuchi ◽  
Hadzki Matsuda ◽  
Takeo Uzuka ◽  
Phyo Kim ◽  
Keisuke Ueki
Keyword(s):  
Protein Expression ◽  
Mismatch Repair ◽  
Early Phase ◽  
Mismatch Repair Gene ◽  
Protein Loss ◽  
Primary Tumors ◽  
Recurrent Gliomas ◽  
Grade Ii ◽  
Mmr Deficiency ◽  
Mmr Proteins

Abstract BACKGROUND Mismatch Repair (MMR) Deficiency is common in recurrent gliomas from IDH-mutant and IDH-wild-type tumors. Emergence of MMR deficit is strongly associated with the chemotherapy using TMZ, and it is considered to be a key mechanism of acquiring resistance to TMZ. Several studies showed MMR protein loss detected by immunohistochemistry was well concordant with molecular genetics sequencing data, and sometimes even more reliable in detecting MMR functional deficit. Here we evaluated Mismatch Repair protein expression by immunohistrochemistry for the pairs of primary and recurrent gliomas. METHODS We investigated the expression of 4 MMR proteins (MLH1/MSH2/MSH6/PMS2) in 37 cases of paired primary-recurrent gliomas (7 grade II & III astrocytomas, 16 grade II & III oligodendrogliomas, and 14 glioblastomas). Clinical information including history of chemo-radiotherapy after primary surgery were collected retrospectively. RESULTS Except for one case of GBM, all primary tumors retained 4 MMR proteins. 10 of 37 recurrent tumors lost one or more MMR protein expression (4 GBM 3 astrocytoma, 3 oligodendroglioma), including one case in which primary tumor already had MSH2/MSH6 loss. 2 recurrent GBM lost MMR protein expression homogenously, while in other 8 tumors, the patterns of MMR protein loss were heterogenous. 8 of 10 tumors that lost MMR protein expression were recurrence after TMZ treatment. CONCLUSION Although the interpretation of the heterogenous loss of MMR protein expression is somewhat difficult, the immunohistochemical evaluation of MMR proteins appears to be feasible, and may be able to detect early phase of MMR deficiency and the rise of hypermutator phenotype.

Microsatellite instability evaluation: which test to use for endometrial cancer?

2021 ◽  
pp. jclinpath-2021-207723
Author(s):  
Paola Rafaniello-Raviele ◽  
Ilaria Betella ◽  
Alessandra Rappa ◽  
Davide Vacirca ◽  
Gianluca Tolva ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Microsatellite Instability ◽  
Automated System ◽  
Tissue Samples ◽  
Valuable Alternative ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed ◽  
Mmr Proteins

AimsAnalysis of microsatellite instability (MSI) is strongly recommended in endometrial cancer (EC) and colorectal cancer to screen for Lynch syndrome, to predict prognosis and to determine optimal treatment and follow-up. In a large monoinstitutional series of ECs, we evaluated the reliability and accuracy of Idylla assay, a rapid, fully automated system to detect MSI, and we compared its performance with two routine reference methods.MethodsWe evaluated MSI status in 174 formalin-fixed, paraffin-embedded EC tissue samples using immunohistochemistry (IHC) for mismatch repair (MMR) proteins and Idylla assay. Samples with discordant or equivocal results were analysed with a third technique, the Promega MSI kit.ResultsIdylla MSI assay and IHC were highly concordant (overall agreement: 154/170=90.59%, 95% CI 85.26% to 94.12%). However, in four samples, MMR-IHC staining was equivocal; moreover, 16 cases showed discordant results, that is, MMR deficient using IHC and microsatellite stable using Idylla. These 20 samples were reanalysed using the MSI-Promega kit, which showed the same results of Idylla assay in 18/20 cases (overall agreement: 90%, 95% CI 69.90% to 97.21%).ConclusionsOur results suggest that IHC is an efficient method to determine MMR status in ECs. However, the Idylla MSI assay is a rapid and reliable tool to define MSI status, and it could represent a valuable alternative to conventional MSI-PCR methods.

KIT Gene Mutations and Patterns of Protein Expression in Mucosal and Acral Melanoma

2012 ◽  
Vol 16 (2) ◽  
pp. 135-142 ◽  
Author(s):  
Suzan Abu-Abed ◽  
Nancy Pennell ◽  
Teresa Petrella ◽  
Frances Wright ◽  
Arun Seth ◽  
...  
Keyword(s):  
Protein Expression ◽  
Kinase Inhibitors ◽  
Case Series ◽  
Gene Mutations ◽  
Mucosal Melanoma ◽  
Acral Melanoma ◽  
Formalin Fixed Paraffin ◽  
Kit Gene ◽  
Formalin Fixed Paraffin Embedded ◽  
Optimized Conditions

Background: Recently characterized KIT (CD117) gene mutations have revealed new pathways involved in melanoma pathogenesis. In particular, certain subtypes harbor mutations similar to those observed in gastrointestinal stromal tumors, which are sensitive to treatment with tyrosine kinase inhibitors. Objective: The purpose of this study was to characterize KIT gene mutations and patterns of protein expression in mucosal and acral melanoma. Methods: Formalin-fixed, paraffin-embedded tissues were retrieved from our archives. Histologic assessment included routine hematoxylin-eosin stains and immunohistochemical staining for KIT. Genomic DNA was used for polymerase chain reaction-based amplification of exons 11 and 13. Results: We identified 59 acral and mucosal melanoma cases, of which 78% showed variable levels of KIT expression. Sequencing of exons 11 and 13 was completed on all cases, and 4 (6.8%) mutant cases were isolated. Conclusion: We successfully optimized conditions for the detection of KIT mutations and showed that 8.6% of mucosal and 4.2% of acral melanoma cases at our institution harbor KIT mutations; all mutant cases showed strong, diffuse KIT protein expression. Our case series represents the first Canadian study to characterize KIT gene mutations and patterns of protein expression in acral and mucosal melanoma.

scholarly journals Transcriptome Profiling Reveals New Insights into the Immune Microenvironment and Upregulation of Novel Biomarkers in Metastatic Uveal Melanoma

Cancers ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 2832 ◽  
Author(s):  
Yamini Krishna ◽  
Amelia Acha-Sagredo ◽  
Dorota Sabat-Pośpiech ◽  
Natalie Kipling ◽  
Kim Clarke ◽  
...  
Keyword(s):  
Protein Expression ◽  
Uveal Melanoma ◽  
Transcriptome Profiling ◽  
Epithelioid Cell ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Galectin 3 ◽  
Novel Biomarkers ◽  
And Control ◽  
Infiltrating Lymphocytes

Metastatic uveal melanoma (mUM) to the liver is incurable. Transcriptome profiling of 40 formalin-fixed paraffin-embedded mUM liver resections and 6 control liver specimens was undertaken. mUMs were assessed for morphology, nuclear BAP1 (nBAP1) expression, and their tumour microenvironments (TME) using an “immunoscore” (absent/altered/high) for tumour-infiltrating lymphocytes (TILs) and macrophages (TAMs). Transcriptomes were compared between mUM and control liver; intersegmental and intratumoural analyses were also undertaken. Most mUM were epithelioid cell-type (75%), amelanotic (55%), and nBAP1-ve (70%). They had intermediate (68%) or absent (15%) immunoscores for TILs and intermediate (53%) or high (45%) immunoscores for TAMs. M2-TAMs were dominant in the mUM-TME, with upregulated expression of ANXA1, CD74, CXCR4, MIF, STAT3, PLA2G6, and TGFB1. Compared to control liver, mUM showed significant (p < 0.01) upregulation of 10 genes: DUSP4, PRAME, CD44, IRF4/MUM1, BCL2, CD146/MCAM/MUC18, IGF1R, PNMA1, MFGE8/lactadherin, and LGALS3/Galectin-3. Protein expression of DUSP4, CD44, IRF4, BCL-2, CD146, and IGF1R was validated in all mUMs, whereas protein expression of PRAME was validated in 10% cases; LGALS3 stained TAMs, and MFGEF8 highlighted bile ducts only. Intersegmental mUMs show differing transcriptomes, whereas those within a single mUM were similar. Our results show that M2-TAMs dominate mUM-TME with upregulation of genes contributing to immunosuppression. mUM significantly overexpress genes with targetable signalling pathways, and yet these may differ between intersegmental lesions.

A Standardized Staining Protocol for L1CAM on Formalin-Fixed, Paraffin-Embedded Tissues Using Automated Platforms

2014 ◽  
Vol 29 (2) ◽  
pp. 180-183 ◽  
Author(s):  
Mina Fogel ◽  
Ayelet Harari ◽  
Elisabeth Müller-Holzner ◽  
Alain G. Zeimet ◽  
Gerhard Moldenhauer ◽  
...  
Keyword(s):  
Cell Adhesion Molecule ◽  
Staining Procedure ◽  
Type I ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Staining Protocol ◽  
Automated Platform ◽  
Endometrial Carcinomas ◽  
L1 Cell Adhesion Molecule ◽  
Formalin Fixed

The L1 cell adhesion molecule (L1CAM) is overexpressed in many human cancers and can serve as a biomarker for prognosis in most of these cancers (including type I endometrial carcinomas). Here we provide an optimized immunohistochemical staining procedure for a widely used automated platform (VENTANA™), which has recourse to commercially available primary antibody and detection reagents. In parallel, we optimized the staining on a semi-automated BioGenix (i6000) immunostainer. These protocols yield good stainings and should represent the basis for a reliable and standardized immunohistochemical detection of L1CAM in a variety of malignancies in different laboratories.

Loss of HLA class I and mismatch repair protein expression in sporadic endometrioid endometrial carcinomas

2012 ◽  
Vol 131 (8) ◽  
pp. 1828-1836 ◽  
Author(s):  
Renske A. de Jong ◽  
Annemarie Boerma ◽  
H. Marike Boezen ◽  
Marian J.E. Mourits ◽  
Harry Hollema ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mismatch Repair ◽  
Hla Class I ◽  
Class I ◽  
Repair Protein ◽  
Endometrial Carcinomas ◽  
Mismatch Repair Protein

scholarly journals Roles of Proliferation and Angiogenesis in Locally Aggressive Biologic Behavior of Ameloblastoma versus Ameloblastic Fibroma

2022 ◽  
Author(s):  
Amr Ibrahim ◽  
Emad Elqalshy ◽  
Ahmed El-Mohamadi ◽  
Kamal Abd El-Rahman ◽  
Magdy Alazzazi
Keyword(s):  
Protein Expression ◽  
Wide Distribution ◽  
Vascular Density ◽  
Image Analyzer ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Ameloblastic Fibroma ◽  
Formalin Fixed Paraffin Embedded ◽  
Embedded Blocks ◽  
The Mean

Background: The present study was carried out to evaluate the roles of proliferation and angiogenesis in locally aggressive biologic behavior of ameloblastoma versus ameloblastic fibroma; Methods: 30 formalin-fixed paraffin embedded blocks (15 cases of ameloblastoma &amp; 15 cases of ameloblastic fibroma) were used. To evaluate the proliferation, the tissue sections were stained with AgNORs stain. CD105 was used as immunohistochemical marker of angiogenesis. Quantitative evaluations of AgNORs were performed. The mean vascular density was evaluated as a measure for CD105 protein expression by using image analyzer computer system; Results: The mean number of AgNORs dots per nucleus was significantly higher in ameloblastoma as compared to ameloblastic fibroma. Also, the protein level of CD105 showed positive expression and wide distribution that the mean vascular density was significantly higher in ameloblastoma as compared to ameloblastic fibroma; Conclusion: Quantitative evaluation of AgNORs stain &amp; the mean vascular density utilizing CD105 protein expression may reflect a higher proliferative activity and a more locally aggressive biologic behavior of ameloblastoma when compared to ameloblastic fibroma, that other factors may be involved in biologic behavior of ameloblastic fibroma.

scholarly journals Detection of Mismatch Repair and Microsatellite Instability in Colorectal Cancer Patients

2020 ◽  
Vol 18 (2) ◽  
Author(s):  
Muhammad Ishaque Faizee
Keyword(s):  
Colorectal Cancer ◽  
Protein Expression ◽  
Microsatellite Instability ◽  
Mismatch Repair ◽  
Poor Response ◽  
Protein Product ◽  
Mismatch Repair Proteins ◽  
Altered Gene ◽  
Colorectal Cancer Patients ◽  
Mmr Proteins

Introduction:  Colorectal cancer is the third most common cancer worldwide. Microsatellite instability (MSI) contributes to be one of the main mechanisms in colorectal cancer. Individuals with MSI tumors have loss of expression of one or more Mismatch Repair proteins. MSI tumors have better survival rate than microsatellite stable (MSS) tumors, poor response to 5FU-based adjuvant chemotherapy and relatively successful immunotherapy in metastatic MSI tumors. Immunohistochemistry recognizes altered gene by recognizing loss of its protein product. Based on the presence or absence of Mismatch repair proteins, groups are classified into Mismatch repair proficient (MMR-p) and Mismatch repair deficient (MMR-d).  Aim:  To investigate the immunohistochemical profile of Mismatch repair proteins namely: hMLH1, hMSH2, hMSH6, and hPMS2 in surgically resected colorectal cancer specimens.  Materials and Method:  A total of 76 cases were selected from the Histopathology Department of HTAA to determine MMR protein expression status. Cases were either MMR-p or MMR-d. Results:  Of the specimens which were properly immunostained, seventeen out of seventy-six cases (22.37%) showed loss of one or more MMR proteins expression and thus were MMR-d. MLH1, MSH2, MSH6 and PMS2 protein expression was detected as 85.53% (65/76), 81.6% (62/76), 88.16% (67/76), and 76.32% (58/76), respectively. Conclusion: Mismatch repair proteins profile should be done using immunohistochemistry in  local laboratories on these selected cases before referring for the expensive molecular test.

scholarly journals Mismatch-Repair Protein Expression in High-Grade Gliomas: A Large Retrospective Multicenter Study

2020 ◽  
Vol 21 (18) ◽  
pp. 6716 ◽  
Author(s):  
Mario Caccese ◽  
Tamara Ius ◽  
Matteo Simonelli ◽  
Matteo Fassan ◽  
Daniela Cesselli ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mismatch Repair ◽  
Complete Loss ◽  
Molecular Characteristics ◽  
High Grade ◽  
Dna Mismatch ◽  
Anaplastic Gliomas ◽  
High Grade Gliomas ◽  
Mmr Proteins

Background: DNA mismatch repair (MMR) is a system for repairing errors in DNA replication. Cancer cells with MMR deficiency can have immunohistochemical loss of MMR protein expression leading to a hypermutable phenotype that may correlate with anti-PD1 efficacy. Scant data exist about immunohistochemical loss of MMR protein expression in high-grade gliomas (HGG). Materials and Methods: We performed a large multicenter retrospective study to investigate the frequency and the prognostic role of immunohistochemical loss of MMR protein expression in HGG patients; we nevertheless evaluated the association between this status and clinical or molecular characteristics. Immunohistochemical loss of MMR protein expression was recorded as partial or complete loss of at least 1 MMR protein. Results: We analyzed the expression of MMR proteins in tumor tissue of 355 consecutive patients. Partial and complete immunohistochemical loss of MMR proteins was found in 43/355 samples (12.1%) and among these, 15 cases (4.2%) showed a complete loss of at the least one MMR protein. Alteration of MSH2 expression was found in 55.8%, MSH6 in 46.5%, PMS2 in 34.9%, and MLH1 in 30.2%. Alteration of MMR protein expression was statistically more frequent in anaplastic gliomas, in recurrent disease, in patients treated with temozolomide, and in IDH-mut gliomas. Immunohistochemical loss of MMR proteins was not associated with survival, adjusting for clinically relevant confounders. Conclusions: MMR protein expression status did not affect survival in HGG patients. We identified clinical and molecular characteristics correlating with immunohistochemical loss of MMR proteins expression. A large study should be performed to analyze its predictive role of immune checkpoint inhibitor efficacy in these subgroups of patients.

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