Elisabeth Sivy Nginamau

• Dongre, Harsh Nitin; Elnour, Rammah; Tornaas, Stian; Fromreide, Siren; Thomsen, Liv Cecilie Vestrheim & Kolseth, Ingrid Moss [Show all 11 contributors for this article] (2023). TP53 mutation and human papilloma virus status as independent prognostic factors in a Norwegian cohort of vulva squamous cell carcinoma. Acta Obstetricia et Gynecologica Scandinavica. ISSN 0001-6349. 103(1). doi: 10.1111/aogs.14689. Full text in Research Archive Show summary

  Introduction: Vulva squamous cell carcinoma (VSCC) develops through two separate molecular pathways—one involving high-risk human papilloma virus infection (HPV-associated), and the other without HPV infection (HPV-independent) often involving TP53 mutation. HPV-associated VSCC generally has a better progression-free survival than HPV-independent VSCC. The aim of this study was to determine TP53 mutation status using immunohistochemistry, compare different methods of HPV detection and correlate both with survival in a retrospective cohort of 123 patients with VSCC. Material and methods: Immunohistochemistry for p53, Ki67 and p16INK4A (a surrogate marker for HPV infection) was performed on formalin-fixed paraffin-embedded tissues from a cohort of surgically treated VSCC patients to identify molecular subtypes of VSCC. Presence of HPV infection was detected by HPV DNA PCR and HPV mRNA in situ hybridization (ISH). The Pearson chi-square test and multivariable Cox regression model were used to investigate the association of different parameters with progression-free survival and disease-specific survival (DSS), and Kaplan–Meier curves were used to show the association of different parameters with survival. Results: The results of p53 and p16INK4A immunohistochemistry confirmed three VSCC subtypes associated with different prognosis. The TP53 mutation status was identified as an independent prognostic factor of worse progression-free survival (p = 0.024) after adjustment for FIGO stage. p16INK4A immunohistochemistry, mRNA ISH, and DNA PCR had excellent concordance in terms of HPV detection. According to the multivariable Cox regression model, the presence of hrHPV mRNA correlated significantly with increased progression-free survival (p = 0.040) and DSS (p = 0.045), after adjustment for other confounders. Conclusions: p53 and p16INK4A immunohistochemistry stratify VSCC cohort into three subtypes with TP53mutated patients having the worst prognosis. The detection of hrHPV mRNA by ISH was an independent predictor of increased survival. Thus, the combined detection of p53 and HPV mRNA might improve risk stratification in VSCC.

• Mohamed, Nuha Mohamed Gaafar; Osman, Tarig Al-Hadi; Elsheikh, Mariam; Ahmed, Israa Abdul Rhman Hassaballah Alhaj; Dongre, Harsh Nitin & Fromreide, Siren [Show all 10 contributors for this article] (2022). Epithelial PD-L1 expression at tumor front predicts overall survival in a cohort of oral squamous cell carcinomas from Sudan. Clinical and Experimental Dental Research. p. 1–11. doi: 10.1002/cre2.666. Full text in Research Archive Show summary

  Background We recently described the tumor immune microenvironment (TIME) in oral squamous cell carcinomas (OSCC) from Sudan by assessing the core of the lesions. However, the invasive tumor front (ITF) is the most active part of OSCC lesions; thus, TIME should also be characterized at the ITF in this patient cohort. Objectives We aimed to evaluate patterns of immune cell infiltration at the ITF in a cohort of OSCC patients from Sudan previously investigated at the tumor center and their association with clinicopathological parameters. Methods This study was performed on a prospective cohort of 22 OSCC patients attending Khartoum Dental Teaching Hospital with a median follow-up of 48 months. Inflammatory infiltrate densities of CD4-, CD8-, FoxP3-, CD20-, CD66b-, M1 (CD80/CD68)-, M2 (CD163/CD68)-, and PD-L1-positive cells were assessed at the ITF by immunohistochemistry, followed by digital quantitative analysis at the stromal and epithelial compartments separately. Histopathological parameters such as the worst pattern of invasion, differentiation, and tumor budding (TB) were also assessed. Correlations between clinicopathological parameters and survival analysis were investigated using SPSS. Results All inflammatory cell subsets investigated were found to be higher in the stromal compartment as compared to the epithelial one, except for the PD-L1+ subset. Stromal infiltration with the CD8+ cell subset was associated with low TB. Kaplan–Meier analyses identified higher epithelial and stromal CD4+ cell subsets. The presence of PD-L1 was found to be associated with unfavorable overall survival. Further, Cox's regression analysis using an age- and tumor-stage-adjusted model identified epithelial PD-L1 expression at the ITF as the only independent prognosticator. Conclusions Epithelial PD-L1 expression at the ITF was found to be an independent prognostic biomarker for OSCC in a cohort of Sudanese patients.

• Mohamed, Nazar; Litlekalsøy, Jorunn; Ahmed, Israa Abdul Rhman; Martinsen, Einar Marius Hjellestad; Furriol, Jessica & Javier-Lopez, Ruben [Show all 15 contributors for this article] (2021). Analysis of salivary mycobiome in a cohort of oral squamous cell carcinoma patients from Sudan identifies higher salivary carriage of malassezia as an independent and favorable predictor of overall survival. Frontiers in Cellular and Infection Microbiology. 11, p. 1–15. doi: 10.3389/fcimb.2021.673465. Full text in Research Archive Show summary

  Background: Microbial dysbiosis and microbiome-induced inflammation have emerged as important factors in oral squamous cell carcinoma (OSCC) tumorigenesis during the last two decades. However, the “rare biosphere” of the oral microbiome, including fungi, has been sparsely investigated. This study aimed to characterize the salivary mycobiome in a prospective Sudanese cohort of OSCC patients and to explore patterns of diversities associated with overall survival (OS). Materials and Methods: Unstimulated saliva samples (n = 72) were collected from patients diagnosed with OSCC (n = 59) and from non-OSCC control volunteers (n = 13). DNA was extracted using a combined enzymatic–mechanical extraction protocol. The salivary mycobiome was assessed using a next-generation sequencing (NGS)-based methodology by amplifying the ITS2 region. The impact of the abundance of different fungal genera on the survival of OSCC patients was analyzed using Kaplan–Meier and Cox regression survival analyses (SPPS). Results: Sixteen genera were identified exclusively in the saliva of OSCC patients. Candida, Malassezia, Saccharomyces, Aspergillus, and Cyberlindnera were the most relatively abundant fungal genera in both groups and showed higher abundance in OSCC patients. Kaplan–Meier survival analysis showed higher salivary carriage of the Candida genus significantly associated with poor OS of OSCC patients (Breslow test: p = 0.043). In contrast, the higher salivary carriage of Malassezia showed a significant association with favorable OS in OSCC patients (Breslow test: p = 0.039). The Cox proportional hazards multiple regression model was applied to adjust the salivary carriage of both Candida and Malassezia according to age (p = 0.029) and identified the genus Malassezia as an independent predictor of OS (hazard ratio = 0.383, 95% CI = 0.16–0.93, p = 0.03). Conclusion: The fungal compositional patterns in saliva from OSCC patients were different from those of individuals without OSCC. The fungal genus Malassezia was identified as a putative prognostic biomarker and therapeutic target for OSCC.

• Rajthala, Saroj; Dongre, Harsh; Parajuli, Himalaya; Min, Anjie; Nginamau, Elisabeth Sivy & Kvalheim, Arild [Show all 10 contributors for this article] (2021). Combined in situ hybridization and immunohistochemistry on archival tissues reveals stromal microRNA-204 as prognostic biomarker for oral squamous cell carcinoma. Cancers. 13(6). doi: 10.3390/cancers13061307. Full text in Research Archive Show summary

  Micro-RNAs (miRs) are emerging as important players in carcinogenesis. Their stromal expression has been less investigated in part due to lack of methods to accurately differentiate between tumor compartments. This study aimed to establish a robust method for dual visualization of miR and protein (pan-cytokeratin) by combining chromogen-based in situ hybridization (ISH) and immunohistochemistry (IHC), and to apply it to investigate stromal expression of miR204 as a putative prognostic biomarker in oral squamous cell carcinoma (OSCC). Four different combinations of methods were tested and ImageJ and Aperio ImageScope were used to quantify miR expression. All four dual ISH-IHC methods tested were comparable to single ISH in terms of positive pixel area percentage or integrated optical density of miRs staining. Based on technical simplicity, one of the methods was chosen for further investigation of miR204 on a cohort of human papilloma virus (HPV)-negative primary OSCC (n = 169). MiR204 stromal expression at tumor front predicted recurrence-free survival (p = 0.032) and overall survival (p = 0.036). Multivariate Cox regression further confirmed it as an independent prognostic biomarker in OSCC. This study provides a methodological platform for integrative biomarker studies based on simultaneous detection and quantification of miRs and/or protein and reveals stromal miR204 as a prognostic biomarker in OSCC.

• Mohamed, Nazar; Goor, Rens van de; El-Sheikh, Mariam; Elrayah, Osman; Osman, Tarig & Nginamau, Elisabeth Sivy [Show all 10 contributors for this article] (2021). Feasibility of a portable electronic nose for detection of oral squamous cell carcinoma in sudan. Healthcare. 9(5), p. 1–11. doi: 10.3390/healthcare9050534. Full text in Research Archive Show summary

  Background: Oral squamous cell carcinoma (OSCC) is increasing at an alarming rate particularly in low-income countries. This urges for research into noninvasive, user-friendly diagnostic tools that can be used in limited-resource settings. This study aims to test and validate the feasibility of e-nose technology for detecting OSCC in the limited-resource settings of the Sudanese population. Methods: Two e-nose devices (Aeonose™, eNose Company, Zutphen, The Netherlands) were used to collect breath samples from OSCC (n = 49) and control (n = 35) patients. Patients were divided into a training group for building an artificial neural network (ANN) model and a blinded control group for model validation. The Statistical Package for the Social Sciences (SPSS) software was used for the analysis of baseline characteristics and regression. Aethena proprietary software was used for data analysis using artificial neural networks based on patterns of volatile organic compounds. Results: A diagnostic accuracy of 81% was observed, with 88% sensitivity and 71% specificity. Conclusions: This study demonstrates that e-nose is an efficient tool for OSCC detection in limited-resource settings, where it offers a valuable cost-effective strategy to tackle the burden posed by OSCC.

• Mohamed, Nuha Mohamed Gaafar; Osman, Tarig Al-Hadi; Ahmed, Israa A.H.; Elsheikh, Mariam; Dongre, Harsh Nitin & Jacobsen, Martha Rolland [Show all 12 contributors for this article] (2021). Characterization of immune cell infiltrate in tumor stroma and epithelial compartments in oral squamous cell carcinomas of Sudanese patients. Clinical and Experimental Dental Research. p. 1–11. doi: 10.1002/cre2.501. Full text in Research Archive Show summary

  Background Tumor immune infiltrate has been explored in oral squamous cell carcinoma (OSCC), but studies on simultaneous characterization of multiple immune cell subtypes separately in stromal and intraepithelial tumor compartments are limited. Objectives We aimed to investigate the immune cell infiltrate in OSCC by using immunohistochemistry (IHC) for a panel of inflammatory cells in stromal and epithelial tumor compartments for a better characterization of the tumors. Methods Thirty-six OSCC lesions and nine normal oral mucosa (NOM) samples from patients attending Khartoum Dental Teaching Hospital, Sudan were investigated for presence of tumor infiltrating lymphocytes, tumor-associated macrophages, tumor-associated neutrophils, and PD-L1 positive cells in the inflammatory infiltrate by single and double IHC. Digital quantitative analysis (Aperio Technologies Inc.) was performed separately for stromal and epithelial compartments. Results OSCC cases displayed a higher inflammatory infiltrate in the associated stroma, but not in the epithelial compartment when compared to NOM. The immunosuppressive type of inflammatory infiltrate, that is, T regulatory cells (FoxP3+ cells) was identified to be significantly higher in the epithelial compartment of tumors with advanced clinical state. An immunoscore developed by combining intraepithelial FoxP3+ and CD4+ cells was found significantly higher in lesions from elderly patients, localized at toombak dipping-related sites, poorly differentiated OSCCs, or with loco-regional lymph node spreading. Conclusions Despite heavy immune cell infiltration in tumor-associated stroma, the majority of OSCCs in this cohort displayed a low intraepithelial immune infiltration. An immunoscore based on combined CD4 and FoxP3 intraepithelial expression may serve as an indicator of advanced tumor progression and should be further investigated for its use as potential prognostic biomarker in OSCC.

• Bjerkli, Inger-Heidi; Hadler-Olsen, Elin Synnøve; Nginamau, Elisabeth Sivy; Laurvik, Helene; Søland, Tine Merete & Costea, Daniela Elena [Show all 8 contributors for this article] (2020). A combined histo-score based on tumor differentiation and lymphocytic infiltrate is a robust prognostic marker for mobile tongue cancer. Virchows Archiv. ISSN 0945-6317. 477(477), p. 865–872. doi: 10.1007/s00428-020-02875-9. Full text in Research Archive Show summary

  We wanted to evaluate the prognostic value of common histopathological variables in a large cohort of patients with cancer in the mobile tongue as such information can be important for treatment stratification of the individual patient, especially for patients with low-stage disease. In addition, we wanted to investigate whether an alternative scoring model with fewer options would compromise the prognostic value. One hundred fifty patients with oral tongue squamous cell carcinomas that were treated in curative intent and with available HE-stained tumor sections were included. We reclassified all tumors and performed univariate and multivariate survival analyses of histopathological and clinical variables. For the complete cohort, lymph node status, grade of differentiation, perineural infiltration, and lymphocytic infiltration were independent prognosticators. In the low-stage disease group, independent prognostic factors were tumor size, grade of differentiation, and lymphocytic infiltrate. For patients with low-stage disease, a histo-score combining the scores for tumor differentiation and lymphocytic infiltrate identified a group of patients with particularly low survival, as patients with moderately or poorly differentiated tumors and little lymphocytic infiltrate had a less favorable 5-year survival outcome than patients in the high-stage disease group. This study shows that a histo-score combining tumor differentiation and lymphocytic infiltration should be given special consideration in treatment planning. Our results also illustrate that many variables can be scored with fewer options than previously suggested to increase their reproducibility, and still maintain their prognostic value.

• Bjerkli, Inger-Heidi; Laurvik, Helene; Nginamau, Elisabeth Sivy; Søland, Tine Merete; Costea, Daniela Elena & Hov, Håkon [Show all 9 contributors for this article] (2020). Tumor budding score predicts lymph node status in oral tongue squamous cell carcinoma and should be included in the pathology report. PLOS ONE. 15:e0239783(9), p. 1–13. doi: 10.1371/journal.pone.0239783. Full text in Research Archive Show summary

  Background The majority of oral cavity cancers arise in the oral tongue. The aim of this study was to evaluate the prognostic value of tumor budding in oral tongue squamous cell carcinoma, both as a separate variable and in combination with depth of invasion. We also assessed the prognostic impact of the 8th edition of the American Joint Committee on Cancer’s TNM classification (TNM8), where depth of invasion (DOI) supplements diameter in the tumor size (T) categorization. Methods Patients diagnosed with primary oral tongue squamous cell carcinoma were evaluated retrospectively. Spearman bivariate correlation analyses with bootstrapping were used to identify correlation between variables. Prognostic value of clinical and histopathological variables was assessed by Log rank and Cox regression analyses with bootstrapping using 5-year disease specific survival as outcome. The significance level for the hypothesis test was 0.05. Results One-hundred and fifty patients had available material for microscopic evaluation on Hematoxylin and Eosin-stained slides and were included in the analyses. Reclassification of tumors according to TNM8 caused a shift towards a higher T status compared to the previous classification. The tumor budding score was associated with lymph node metastases where 23% of the patients with low-budding tumors had lymph node metastases, compared with 43% of those with high-budding tumors. T-status, lymph node status, tumor budding, depth of invasion, and the combined tumor budding/depth of invasion score were all significantly associated with survival in univariate analyses. In multivariate analyses only N-status was an independent prognosticator of survival. Conclusion Reclassification according to TNM8 shifted many tumors to a higher T-status, and also increased the prognostic value of the T-status. This supports the implementation of depth of invasion to the T-categorization in TNM8. Tumor budding correlated with lymph node metastases and survival. Therefore, information on tumor budding can aid clinicians in treatment planning and should be included in pathology reports of oral tongue squamous cell carcinomas.

• Steigen, Sonja Eriksson; Søland, Tine Merete; Nginamau, Elisabeth Sivy; Laurvik, Helene; Costea, Daniela Elena & Johannessen, Anne Christine [Show all 10 contributors for this article] (2020). Grading of oral squamous cell carcinomas – Intra and interrater agreeability: Simpler is better? Journal of Oral Pathology & Medicine. ISSN 0904-2512. 49, p. 630–635. doi: 10.1111/jop.12990. Full text in Research Archive Show summary

  Background: Numerous studies have been presented on histological grading of oral squamous cell carcinomas (OSCC) for predicting survival, but uncertainty of their usefulness rises due to discordances of results. A scoring system should be robust and well validated, and intra‐ and interrater agreement can be used as a tool to visualize the strength of reproducibility. Methods: Here, we present an intra‐ and inter‐observer study on evaluation of OSCC using some of the most common histopathological parameters. The observers were from different Norwegian university hospitals, and calibration to ensure accuracy was first performed. Percentage of the agreement was calculated for the score made by the individual observer at different times, as well as between pairs of observers. Results: The evaluation made by the same observer at two different time points (intrarater) correlated better than observations made by different participants (interrater). In an attempt to increase the rate of agreement, many of the parameters were either dichotomized into simply low‐ and high grade, or to a three‐tier system when more than three options in the original design. This increased the concurrence with 15.4% for the intrarater and with 23% for the interrater comparisons. Conclusion: High agreement for histopathological parameters can be difficult to obtain on hematoxylin and eosin staining in scoring systems with many options. A simpler system might be more advantageous to achieve higher degree of reproducibility.

• Nginamau, Elisabeth Sivy; Mæhle, Bjørn Ove & Jonsson, Roland (2011). An experimental protocol for the fractionation and 2DE separation of HeLa and A-253 cell lysates suitable for the identification of the individual antigenic proteome in Sjögren's syndrome. Autoimmunity. ISSN 0891-6934. 44(8), p. 652–663. doi: 10.3109/08916934.2011.593598. Full text in Research Archive
• Jonsson, Roland; Nginamau, Elisabeth Sivy; Szyszko, Ewa Alicja & Brokstad, Karl Albert (2007). Role of B cells in Sjögren's syndrome - from benign lymphoproliferation to overt malignancy. Frontiers in Bioscience. ISSN 1093-9946. 12, p. 2159–2170. doi: 10.2741/2219. Full text in Research Archive

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• Nginamau, Elisabeth Sivy & Jonsson, Roland (2008). Autoantigens Ro52 and Ro60 but not La48 are associated to membrane structures in HeLa cells. Scandinavian Journal of Immunology. ISSN 0300-9475. 68(2), p. 202–202. Full text in Research Archive
• Nginamau, Elisabeth Sivy & Jonsson, Roland (2007). Identification of cell surface autoantigens in Sjögren's syndrome. Inflammation Research. ISSN 1023-3830. 56, p. S457–S457. Full text in Research Archive
• Costea, Daniela Elena; Johannessen, Anne Christine; Suleiman, Ahmed & Nginamau, Elisabeth Sivy (2020). Biomarker identification in oral squamous cell carcinoma : study on cohorts of patients from Sudan. Universitetet i Bergen. Full text in Research Archive
• Nginamau, Elisabeth Sivy (2015). Expression and possible mechanisms of autoantigenic selection of Ro52, Ro60 and La48 in Sjögren’s syndrome. Universitetet i Bergen. ISSN 9788230826980. Full text in Research Archive

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