Browsing by Author "El-Shinawi, Mohamed"

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Abstract 3302: High incidence of MAC387 positive cells in the carcinoma tissues of inflammatory breast cancer patients correlate with the detection of multiple human Cytomegalovirus genotypes and invasive properties of the disease
(American Association for Cancer Research, 2016) Taha Mohamed, Hossam; Gadalla, Ramy; Abdel Aziz Ibrahim, Sherif; Akram Nouh, M.; El-Shinawi, Mohamed; J. Schneider, Robert; Mostafa Mohamed, Mona
Introduction: Previously we showed that the incidence of multiple human cytomegalovirus (HCMV) genotypes in the carcinoma tissues of inflammatory breast cancer (IBC) patients plays essential role in the disease progression. Primary HCMV infection to monocytes induces differentiation and biological turnover of monocytes to macrophages. In addition infected macrophages serves as “mobile vectors” for virus spreading and dissemination to different organs mainly by transendothelial migration. In addition we screened for the infiltration of CD14+ and CD68+ monocytes/macrophages markers in the carcinoma tissues of IBC versus non-IBC patients we showed that of CD14+ cells highly infiltrate tumor microenvironment (TME) of IBC patients compared to non-IBC. Aims: In the present study we aim to 1) Assess the level of expression of MAC387 protein by monocytes/macrophages infiltrating TME of IBC versus non-IBC patients; 2) Test the correlation between the density of infiltrated MAC387+ cells and the incidence of different HCMV genotypes in carcinoma tissues of IBC versus non-IBC tissues. Since MAC387 found to be more common in cancers characterized by high metastatic properties we will also 3) determine whether the expression of MCA387 correlate with lymph-node metastasis and lymphovascular invasion in IBC versus non-IBC breast cancer patients. Materials and Methods: A total of 135 breast cancer patients (91 non-IBC and 44 IBC) were enrolled to the present study during the period of January 2012 to September 2015 from Ain Shams university Hospitals. Detection of MAC387 marker was assessed by immunohistochemistry and HCMV genotypes were detected using multiplex PCR methodology. Results: MAC387+ positive cells were more prevalent in IBC tissues than non-IBC tissues (p = 0.4). Incidence of higher number of MAC387+ cells were positively correlate with higher number of metastatic lymph nodes in both IBC and non-IBC patients r = 0.807 and 0.779 respectively. Moreover, Incidence of higher number of MAC387+ cells found to be positively correlate with lymphovascular invasion in IBC patients r = 0622. Detection of multiple HCMV genotypes was statistically higher (p = 0.04) in IBC tissues in comparison with non-IBC tissues. Moreover, triple negative non-IBC and IBC tissues showed higher incidence of multiple HCMV genotypes in comparison with hormonal positive non-IBC and IBC tissues. of the monocytes/macrophages MAC387+ positive cells were more prevalent in IBC tissues showed multiple HCMV genotypes in comparison with IBC tissues showed single HCMV genotype (p = 0.46). Conclusion: MAC387+ positive cells were more prevalent in IBC tissues and correlate with presence of multiple HCMV genotypes and high invasive properties of the disease.
Abstract 4788: Detection of different genotypes of Human Cytomegalovirus in breast cancer patients.
(American Association for Cancer Research, 2013) Taha Mohamed, Hossam; El-Shinawi, Mohamed; Bashtar, Abdel-Rahman; Tarek abdel Salam, El-Said; J. Schneider, Robert; Mostafa Mohamed, Mona
Background: Human Cytomegalovirus (HCMV) is an endemic herpes virus that re-emerges in cancer patients enhancing oncogenic potential. Recent studies have shown that HCMV infection is associated with certain types of cancer morbidity such as glioblastoma; Although HCMV has been detected in breast cancer tissues. The ability of HCMV to infect diverse organs and cell types in vivo has been attributed to strain variations in certain genes of the virus. The relationship between genetic variants of the envelope glycoprotein genes of HCMV and disease outcome has been studied recently, among these variable genes HCMV ORF UL73 (envelope glycoprotein N). Aims: The aims of the present study were to 1) Screen for the infection of Human cytomegalovirus infection in inflammatory versus non-inflammatory breast cancer patients and 2) test the frequency of occurrence of multiple genotypes of HCMV glycoprotein N in IBC versus Non-IBC patients, to determine if there is any mixed infections by multiple genotypes in paired clinical specimens obtained from patients. Material and Methods: A total 93 (62 Non-IBC and 31 IBC) women diagnosed with breast cancer by clinical examination, ultrasound, mammography, and confirmed by biopsy (tru-cut) were enrolled into this study from Ain Shams university Hospitals. During modified radical mastectomy or conservative breast surgery carcinoma and non carcinoma tissues with peripheral blood were collected to detect the presence of Human cytomegalovirus DNA using nested PCR. HCMV glycoprotein N polymorphism (gN) was detected using multiplex PCR by using specific primers to each gN genotype which enable us to detect any mixed genotypes infection. Results: Analysis of the HCMV nested PCR to the carcinoma tissues showed that 63% of non-IBC carcinoma tissues were HCMV-DNA positive and 37% were HCMV-DNA negative. In IBC 80% of carcinoma tissues were HCMV-DNA positive and 20% were HCMV-DNA negative. Application of multiplex PCR for gN gene on HCMV positive carcinoma tissues (25 IBC and 32 Non-IBC) revealed that gN-1 genotype was detected in 25% of the non-IBC patients and 32% of the IBC patients, While gN-3b was detected in 6.25% of non-IBC patients and 4% of the IBC patients. Genotype gN-4a was detected in 9.38% of Non-IBC patients and in 4% of the IBC patients, while gN-4b\c was detected in 59.37% of Non-IBC patients and in 60% of the IBC patients. Also revealed mixed infection of gN-1+ gN-4b\c, gN-3b + gN-4b\c and gN-4a + gN-4b\c in Non-IBC patients and gN-1+ gN-4b\c, gN-3b + gN-4b\c in IBC patients. Conclusion: The frequency of HCMV infection in IBC was higher than Non-IBC patients; however the most dominant HCMV gN genotypes in IBC and Non-IBC are similar.
Adipocyte of Obese Breast Cancer Patients Is Characterized by The Overexpression of Caveolin-1 Protein/Mediator the Main Constituent of the Plasma Membrane Vesicles Caveolae That Contain Proteins Contribute to Breast Cancer Progression
(Egyptian Society of Biological Sciences, 2019) Saber, Aya; Abdelaziz Ibrahim, Sherif; Hosney, Mohamed; Taha Mohamed, Hossam; Fares, Mohamed; Sabet, Salwa; El-Shinawi, Mohamed; Mostafa Mohamed, Mona
Breast cancer (BC) is the second leading mortality cause due to poor survival rates compared to lung cancer all over the world. Recently, lifestyle increased obesity among the population globally. Since, the adipose tissues (AT) are the major contributor to the volume of the breast and adipocytes cells, which constitute AT are one of the major prominent cells play an effective role in cancer progression via releasing different mediators and adipokines. Thus, AT may display a crucial role in BC progression, especially in obese patients compared to non-obese patients, which characterized by increased AT. Interestingly, adipocytes are characterized by expressing caveolin-1 (Cav-1) protein. Cav-1 constitutes the lipid raft of caveola which contains different proteolytic enzymes inducing cancer metastasis. In this regard, the aim of the present study was to explore the level of expression of Cav-1 protein in the tissue specimen of 5 non-obese vs. 15 obese patients using immunohistochemistry (IHC) and immunoblotting techniques. Our finding demonstrates that the level of Cav-1expression was statistically significantly low in non-obese compared to obese BC patients (p < 0.05). Herein, our results revealed that the highest expression of Cav-1 in obese patients compared to non-obese (control) patients can be considered as a biomarker for BC patients.
Characterization of inflammatory breast cancer: a vibrational microspectroscopy and imaging approach at the cellular and tissue level
(Royal Society of Chemistry, 2018) Taha Mohamed, Hossam; Untereiner, Valérie; Proult, Isabelle; Abdelaziz Ibrahim, Sherif; Götte, Martin; El-Shinawi, Mohamed; Mostafa Mohamed, Mona; D. Sockalingum, Ganesh; Brézillon, Stéphane
Inflammatory breast cancer (IBC) has a poor prognosis because of the lack of specific biomarkers and its late diagnosis. An accurate and rapid diagnosis implemented early enough can significantly improve the disease outcome. Vibrational spectroscopy has proven to be useful for cell and tissue characterization based on the intrinsic molecular information. Here, we have applied infrared and Raman microspectroscopy and imaging to differentiate between non-IBC and IBC at both cell and tissue levels. Two human breast cancer cell lines (MDA-MB-231 and SUM-149), 20 breast cancer patients (10 non-IBC and 10 IBC), and 4 healthy volunteer biopsies were investigated. Fixed cells and tissues were analyzed by FTIR microspectroscopy and imaging, while live cells were studied by Raman microspectroscopy. Spectra were analyzed by hierarchical cluster analysis (HCA) and images by common k-means clustering algorithms. For both cell suspensions and single cells, FTIR spectroscopy showed sufficient high inter-group variability to delineate MDA-MB-231 and SUM-149 cell lines. Most significant differences were observed in the spectral regions of 1096–1108 and 1672–1692 cm−1. Analysis of live cells by Raman microspectroscopy gave also a good discrimination of these cell types. The most discriminant regions were 688–992, 1019–1114, 1217–1375 and 1516–1625 cm−1. Finally, k-means cluster analysis of FTIR images allowed delineating non-IBC from IBC tissues. This study demonstrates the potential of vibrational spectroscopy and imaging to discriminate between non-IBC and IBC at both cell and tissue levels.
Differential gene expression of Fresh Tissue and Patient-Derived Explants' Matricellular Proteins Augment Inflammatory Breast Cancer Metastasis: The Possible Role of IL-6 and MCP-1.
(Dove Medical Press Ltd., 2023-01) Tarek, Alshaimaa; Mohamed, Hossam Taha; El-Sharkawy, Aya Ali; El-Sayed, Shrouk Khalaf; Hirshon, Jon Mark; Woodward, Wendy A; El-Shinawi, Mohamed; Mohamed, Mona Mostafa
Background Matricellular proteins comprising matrisome and adhesome are responsible for structure integrity and interactions between cells in the tumour microenvironment of breast cancer. Changes in the gene expression of matrisome and adhesome augment metastasis. Since inflammatory breast cancer (IBC) is characterized by high metastatic behavior. Herein we compared the gene expression profile of matrisome and adhesome in non-IBC and IBC in fresh tissue and ex-vivo patients derived explants (PDEs), we also compared the secretory inflammatory mediators of PDEs in non-IBC and IBC to identify secretory cytokines participate in cross-talk between cells via interactions with matrisome and adhisome. Methods Fifty patients (31 non-IBC; 19 IBC) were enrolled in the present study. To test their validation in clinical studies, PDEs were cultured as an ex-vivo model. Gene expression and cytokine array were used to identify candidate genes and cytokines contributing to metastasis in the examined fresh tissues and PDEs. Bioinformatics analysis was applied on identified differentially expressed genes (DEGs) using GeneMANIA and Metascape gene annotation and analysis resource to identify pathways involved in IBC metastasis. Results Normal and cancer fresh tissues and PDEs of IBC were characterized by overexpression of CDH1 and MMP14 and downregulation of CTNNA1 and TIMP1 compared to non-IBC. The secretome of IBC cancer PDEs is characterized by significantly high expression of interleukin 6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1/CCL2) compared to non-IBC. Conclusion Genes expressed by adhisome and matrisome play a significant role in IBC metastasis and should be considered novel target therapy.
Editorial: Inflammatory tumor microenvironment: role of cytokines and virokines in breast cancer progression and metastasis
(Frontiers Media SA, 2024-06) Mohamed, Hossam Taha; El-Shinawi, Mohamed; Mohamed, Mona Mostafa
Various factors contributing to breast cancer progression and metastasis (Feng et al., 2018; Park et al., 2022). One of these factors is the presence of inflammatory tumor microenvironment (TME), which composed of cellular components (e.g., cancer cells, immune cells, endothelial cells, fibroblasts, mast cells) and non-cellular components (e.g., extracellular matrix proteins, cytokines, chemokines, signal molecules), and it differs significantly from the normal tissue microenvironment in terms of low vascular density, hypoxia, weak acidity, and reducibility (Zarrilli et al., 2020). Breast cancer cells control the function of TME components via the expression of cytokines that can increase selfproliferation, growth, and treatment resistance in an autocrine form, and encourage recruitment, activation, and differentiation of other cells in the TME in a paracrine approach as IL-6, IL-8, and even VEGF (Malla and Kiran, 2022; Nengroo et al., 2022; Habanjar et al., 2023).
Human cytomegalovirus infection enhances NF-κB/p65 signaling in inflammatory breast cancer patients
(Public Library of Science, 2013) El-Shinawi, Mohamed; Taha Mohamed, Hossam; A. El-Ghonaimy, Eslam; Tantawy, Marwa; Younis, Amal; J. Schneider, Robert; Mostafa Mohamed, Mona
Human Cytomegalovirus (HCMV) is an endemic herpes virus that re-emerges in cancer patients enhancing oncogenic potential. Recent studies have shown that HCMV infection is associated with certain types of cancer morbidity such as glioblastoma. Although HCMV has been detected in breast cancer tissues, its role, if any, in the etiology of specific forms of breast cancer has not been investigated. In the present study we investigated the presence of HCMV infection in inflammatory breast cancer (IBC), a rapidly progressing form of breast cancer characterized by specific molecular signature. We screened for anti-CMV IgG antibodies in peripheral blood of 49 non-IBC invasive ductal carcinoma (IDC) and 28 IBC patients. In addition, we screened for HCMV-DNA in postsurgical cancer and non-cancer breast tissues of non-IBC and IBC patients. We also tested whether HCMV infection can modulate the expression and activation of transcriptional factor NF-κB/p65, a hallmark of IBC. Our results reveal that IBC patients are characterized by a statistically significant increase in HCMV IgG antibody titers compared to non-IBC patients. HCMV-DNA was significantly detected in cancer tissues than in the adjacent non-carcinoma tissues of IBC and IDC, and IBC cancer tissues were significantly more infected with HCMV-DNA compared to IDC. Further, HCMV sequence analysis detected different HCMV strains in IBC patients tissues, but not in the IDC specimens. Moreover, HCMV-infected IBC cancer tissues were found to be enhanced in NF-κB/p65 signaling compared to non-IBC patients. The present results demonstrated a correlation between HCMV infection and IBC. Etiology and causality of HCMV infection with IBC now needs to be rigorously examined.
IL-10 correlates with the expression of carboxypeptidase B2 and lymphovascular invasion in inflammatory breast cancer: the potential role of tumor infiltrated macrophages
(Mosby, 2018) Taha Mohamed, Hossam; El-Husseiny, Noura; A. El-Ghonaimy, Eslam; Abdelaziz Ibrahim, Sherif; A. Bazzi, Zainab; Cavallo-Medved, Dora; B. Boffa, Michael; El-Shinawi, Mohamed; Mostafa Mohamed, Mona
Background: Pro-carboxypeptidase B2 (pro-CPB2) or thrombin-activatable fibrinolysis inhibitor (TAFI) is a glycoprotein encoded by the CPB2 gene and deregulated in several cancer types, including breast cancer. Thrombin binding to thrombomodulin (TM), encoded by THBD, is important for TAFI activation. CPB2 gene expression is influenced by genetic polymorphism and cytokines such as interleukin 10 (IL-10). Our previous results showed that tumor infiltrating monocytes/macrophages (CD14+ /CD16+ ) isolated from inflammatory breast cancer (IBC) patients’ secrete high levels of IL-10. The aim of the present study is to test genetic polymorphism and expression of CPB2 in healthy breast tissues and carcinoma tissues of 2 non-IBC and IBC patients. Furthermore, to investigate whether IL-10 modulates the expression of CPB2 and THBD in vivo and in-vitro. Materials and methods We tested CPB2 Thr325Ile polymorphism using restriction fragment length polymorphism, (RFLP) technique in healthy and carcinoma breast tissues. The mRNA expression of CPB2, THBD and IL10 were assessed by RT-qPCR. Infiltration of CD14+ cells was assessed by immunohistochemistry. We investigated the correlation between infiltration of CD14+ cells and expression of IL10 and CPB2. Furthermore, we correlated IL10 expression with the expression of both CPB2 and THBD in breast carcinoma tissues. Finally, we validated the role of recombinant IL-10 in regulating the expression of CPB2 and THBD using different breast cancer cell lines. Results: Our data showed that CPB2 genotypes carrying the high-risk allele [Thr/Ile (CT) and Ile/Ile (TT)] were more frequent in both IBC and non-IBC patients compared to control group. CPB2 genotypes did not show any statistical correlation with CPB2 mRNA expression levels or patients’ clinical pathological properties. Interestingly, CPB2 and IL10 expression were significantly higher and positively correlated with the incidence of CD14+ cells in carcinoma tissues of IBC as compared to non-IBC. On the other hand, THBD expression was significantly lower in IBC carcinoma versus non-IBC tissues. Based on molecular subtypes, CPB2 and IL10 expression were significantly higher in triple negative (TN) as compared to hormonal positive (HP) carcinoma tissues of IBC. Moreover, CPB2 expression was positively correlated with presence of lymphovascular invasion and the expression of IL10 in carcinoma tissues of IBC patients. Furthermore, recombinant human IL-10 stimulated CPB2 expression in SUM-149 (IBC cell line) but not in MDA-MB-231 (non-IBC cell line), while there was no significant effect THBD expression. 3 Conclusion: Carcinoma tissues of IBC patients are characterized by higher expression of CPB2 and lower expression of THBD. Moreover, CPB2 positively correlates with IL10 mRNA expression, incidence of CD14+ cells and lymphovascular invasion in IBC patients. IL-10 stimulated CPB2 expression in TN-IBC cell line suggests a relevant role of CPB2 in the aggressive phenotype of IBC.
IL-8 and MCP-1/CCL2 regulate proteolytic activity in triple negative inflammatory breast cancer a mechanism that might be modulated by Src and Erk1/2
(Academic Press Inc., 8/15/2020) Taha Mohamed, Hossam; El-Ghonaimy, Eslam A; El-Shinawi, Mohamed; Hosney, Mohamed; Götte, Martin; Woodward, Wendy A; El-Mamlouk, Tahani; Mostafa Mohamed, Mona
Inflammatory breast cancer (IBC) is a highly metastatic and lethal breast cancer. As many as 25–30% of IBCs are triple negative (TN) and associated with low survival rates and poor prognosis. We found that the microenvironment of IBC is characterized by high infiltration of tumor associated macrophages (TAMs) and by over-expression of the cysteine protease cathepsin B (CTSB). TAMs in IBC secrete high levels of the cytokines interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1/CCL2) compared to non-IBC patients. Herein, we tested the roles of IL-8 and MCP-1/CCL2 in modulating proteolytic activity and invasiveness of TN-non-IBC as compared to TN-IBC and addressed the underlying molecular mechanism(s) for both cytokines. Quantitative real time PCR results showed that IL-8 and MCP-1/CCL2 were significantly overexpressed in tissues of TN-IBCs. IL-8 and MCP-1/CCL2 induced CTSB expression and activity of the p-Src and p-Erk1/2 signaling pathways relevant for invasion and metastasis in TN-non-IBC, HCC70 cells and TN-IBC, SUM149 cells. Dasatinib, an inhibitor of p-Src, and U0126, an inhibitor of p-Erk1/2, down-regulated invasion and expression of CTSB by HCC70 and SUM149 cells, a mechanism that is reversed by IL-8 and MCP-1/CCL2. Our study shows that targeting the cytokines IL-8 and MCP-1/CCL2 and associated signaling molecules may represent a promising therapeutic strategy in TN-IBC patients
IL-8 secreted by tumor associated macrophages contribute to lapatinib resistance in HER2-positive locally advanced breast cancer via activation of Src/STAT3/ERK1/2-mediated EGFR signaling
(Elsevier, 03/02/2021) Ahmed, Shaza; Taha Mohamed, Hossam; El-Husseiny, Noura; El Mahdy, Manal M; Safwat, Gehan; Diab, Ayman A; El-Sherif, Ahmed A; El-Shinawi, Mohamed; Mohamed, Mona Mostafa
Locally advanced breast cancer (LABC) is an aggressive disease characterized by late clinical presentation, large tumor size, treatment resistance and low survival rate. Expression of EGFR/HER2 and activation of intracellular tyrosine kinase domains in LABC are associated with poor prognosis. Thus, target therapies such as the anti-receptor tyrosine kinases lapatinib drug have been more developed in the past decade. The response to lapatinib involves the inhibition of RTKs and subsequently signaling molecules such as Src/STAT3/Erk1/2 known also to be activated by the cytokines in the tumor microenvironment (TME). The aim of the present study is to identify the major cytokine that might contribute to lapatinib resistance in EGFR+/HER2+ LABC patients. Indeed, tumor associated macrophages (TAMs) are the main source of cytokines in the TME. Herein, we isolated TAMs from LABC during modified radical mastectomy (MRM). Cytokine profile of TAMs revealed that IL-8 is the most prominent highly secreted cytokine by TAMs of LABC patients. Using in-vitro cell culture model we showed that recombinant IL-8 (50 and 100 ng/mL) at different time intervals interfere with lapatinib action via activation of Src/EGFR and signaling molecules known to be inhibited during treatment. We proposed that to improve LABC patients' response to lapatinib treatment it is preferred to use combined therapy that neutralize or block the action of IL-8.
Incidence of Human Cytomegalovirus in Breast Carcinoma Tissues is Associated with A Higher Expression of Growth Factor Receptor-Bound Protein 2
(The Egyptian Journal of Hospital Medicine, 2019) Fares, Mohamed; Taha Mohamed, Hossam; Abdelaziz Ibrahim, Sherif; Hosney, Mohamed; I. Rady, Mohamed; El-Shinawi, Mohamed; Mostafa Mohamed, Mona
Background: female mammary carcinoma is the second most common cancer incidence among women and the fifth most common leading cause of cancer death worldwide. Premenopausal young women are more frequently targeted by inflammatory breast cancer (IBC), which is the most lethal form of breast cancer. The human cytomegalovirus (HCMV) has been identified as one of the viral infection with a higher frequency in carcinoma tissues of IBC than in non-IBC. The adaptor protein growth factor receptor-bound protein 2 (Grb2), was found to be upregulated in HCMV-infected cells and play as crucial role in cancer progression. Objective: this study aimed to assess the expression level of Grb2 in carcinoma tissues of IBC and non-IBC with HCMV infection. Patients and Methods: overall, 135 female diagnosed with breast carcinoma were enrolled in this study. Using conventional and real time polymerase chain reaction (PCR), we determined the incidence of HCMV and assessed the expression level of Grb2 mRNA in the breast cancer tissue samples. Results: Grb2 mRNA was significantly upregulated in HCMV+ IBC higher than in HCMV+ non-IBC. According to the molecular subtype, Grb2 mRNA was significantly higher upregulated in breast carcinoma tissues of HCMV+ hormonal positive (HP) than in triple negative (TN) counterparts. Conclusion: HCMV infection is associated with a high expression of Grb2 mRNA in IBC and that HP HCMV+ mammary carcinoma tissues confer upregulated Grb2 mRNA, suggesting a potential role of HCMV infection in enhancing of Grb2 mRNA expression in breast cancer with HP.
Inflammatory and Non-inflammatory Breast Cancer: A Potential Role for Detection of Multiple Viral DNAs in Disease Progression
(Springer US, 2016) El-Shinawi, Mohamed; Taha Mohamed, Hossam; Hesham Abdel-Fattah, Hadeer; Abdel Aziz Ibrahim, Sherif; S. El-Halawany, Medhat; Akram Nouh, M.; J. Schneider, Robert; Mostafa Mohamed, Mona
Background Inflammatory breast cancer (IBC) is the most lethal form of breast cancer. Multiple viral infections in IBC tissues were found to be associated with disease pathogenesis. Objective The aim of the present study was to correlate the incidence of viral DNA with breast cancer progression. Materials and Methods Overall, 135 women diagnosed with breast cancer were enrolled in this study. Using polymerase chain reaction and sequencing assays, we determined the incidence of human papillomavirus types 16 and 18 (HPV-16 and -18), human cytomegalovirus (HCMV), Epstein–Barr virus, human herpes simplex virus type 1 and 2, and human herpes virus type 8 (HHV-8) in breast carcinoma tissue biopsies. We also assessed the expression of the cell proliferation marker Ki-67 by immunohistochemistry in association with the incidence of viral DNA. Results HCMV and HPV-16 were the most detected viral DNAs in breast carcinoma tissues; however, the frequency of HCMV and HHV-8 DNA were significantly higher in IBC than non-IBC tissues. Moreover, the prevalence of multiple viral DNAs was higher in IBC than non-IBC tissues. The incidence of multiple viral DNAs positively correlates with tumor size and number of metastatic lymph nodes in both non-IBC and IBC patients. The expression of Ki-67 was found to be significantly higher in both non-IBC and IBC tissues in which multiple viral DNAs were detected. Conclusions The incidence of multiple viral DNAs in IBC tissues was higher compared with non-IBC tissues. The present results suggest the possibility of a functional relationship between the presence of multiple viral DNAs and disease pathogenesis.
Inflammatory breast cancer: high incidence of detection of mixed human cytomegalovirus genotypes associated with disease pathogenesis
(Frontiers, 2014) Taha Mohamed, Hossam; El-Shinawi, Mohamed; Akram Nouh, M.; Bashtar, Abdel-Rahman; Tarek Elsayed, Elsayed; J. Schneider, Robert; Mostafa Mohamed, Mona
Inflammatory breast cancer (IBC) is a highly metastatic, aggressive, and fatal form of breast cancer. Patients presenting with IBC are characterized by a high number of axillary lymph node metastases. Recently, we found that IBC carcinoma tissues contain significantly higher levels of human cytomegalovirus (HCMV) DNA compared to other breast cancer tissues that may regulate cell signaling pathways. In fact, HCMV pathogenesis and clinical outcome can be statistically associated with multiple HCMV genotypes within IBC. Thus, in the present study, we established the incidence and types of HCMV genotypes present in carcinoma tissues of infected non-IBC versus IBC patients. We also assessed the correlation between detection of mixed genotypes of HCMV and disease progression. Genotyping of HCMV in carcinoma tissues revealed that glycoprotein B (gB)-1 and glycoprotein N (gN)-1 were the most prevalent HCMV genotypes in both non-IBC and IBC patients with no significant difference between patients groups. IBC carcinoma tissues, however, showed statistically significant higher incidence of detection of the gN-3b genotype compared to non-IBC patients. The incidence of detection of mixed genotypes of gB showed that gB-1 + gB-3 was statistically significantly higher in IBC than non-IBC patients. Similarly, the incidence of detection of mixed genotypes of gN showed that gN-1 + gN-3b and gN-3 + gN-4b/c were statistically significant higher in the carcinoma tissues of IBC than non-IBC. Mixed presence of different HCMV genotypes was found to be significantly correlated with the number of metastatic lymph nodes in non-IBC but not in IBC patients. In IBC, detection of mixed HCMV different genotypes significantly correlates with lymphovascular invasion and formation of dermal lymphatic emboli, which was not found in non-IBC patients.
Inflammatory breast cancer: High incidence of GCC haplotypes (−1082A/G, −819T/C, and −592A/C) in the interleukin-10 gene promoter correlates with over-expression of interleukin-10 in patients’ carcinoma tissues
(SAGE Publications, 2017) Sabet, Salwa; Khalaf El-Sayed, Shrouk; Taha Mohamed, Hossam; El-Shinawi, Mohamed; M. Mohamed, Mona
Interleukin-10 is involved in carcinogenesis by supporting tumor escape from the immune response. The aim of this study was to assess the single nucleotide polymorphisms, −1082A/G, −819T/C and −592A/C, in interleukin-10 gene promoter in inflammatory breast cancer compared to non–inflammatory breast cancer and association of these polymorphisms with interleukin-10 gene expression. We enrolled 105 breast cancer tissue (72 non–inflammatory breast cancer and 33 inflammatory breast cancer) patients and we determined the three studied single nucleotide polymorphisms in all samples by polymerase chain reaction restriction fragment length polymorphism and investigated their association with the disease and with various prognostic factors. In addition, we assessed the expression of interleukin-10 gene by realtime quantitative reverse transcription polymerase chain reaction and the correlation between studied single nucleotide polymorphisms and interleukin-10 messenger RNA expression. We found co-dominant effect as the best inheritance model (in the three studied single nucleotide polymorphisms in non–inflammatory breast cancer and inflammatory breast cancer samples), and we didn’t identify any association between single nucleotide polymorphisms genotypes and breast cancer prognostic factors. However, GCC haplotype was found highly associated with inflammatory breast cancer risk (p<0.001, odds ratio=43.05). Moreover, the expression of interleukin-10 messenger RNA was significantly higher (p<0.001) by 5.28-fold and 8.95-fold than non–inflammatory breast cancer and healthy control, respectively, where GCC haplotype significantly increased interleukin-10 gene expression (r=0.9, p<0.001).
Inflammatory breast cancer: Mixed viral infections within carcinoma tissues and the expression of Ki-67 proliferation marker.
(American Society of Clinical Oncology, 2017) taha Mohamed, Hossam; Hesham Abdel Fattah, Hadeer; El-Shinawi, Mohamed; Abdelaziz Ibrahim, Sherif; S. El-Halawany, Medhat; El Ghazaly, Hesham; Schneider, Robert; Mohamed, Mona
Background: Inflammatory breast cancer (IBC) is the most lethal form of breast cancer. Our previous results showed that IBC carcinoma tissues possess mixed human cytomegalovirus genotypes than non-IBC carcinoma tissues. However, the role of viral infection in breast cancer is poorly understood. Methods: We enrolled 135 women diagnosed breast cancer (91 Non-IBC and 44 IBC). The incidence of different viral DNA (Herpes viruses and HPV) was performed using nested and multiplex PCR and DNA sequencing. The expression of Ki-67 proliferation index was assessed by immunohistochemistry. Results: DNA of HCMV and HPV-16 were the most detected in breast tissues of both IBC and non-IBC patients. However, as a single infection the incidence of HCMV-DNA and HHV-8 DNA were significantly higher in carcinoma tissues of IBC in comparison with non-IBC (p = 0.035, p= 0.039, respectively). Moreover, the prevalence of mixed infection of different viral DNA was higher in IBC than non-IBC carcinoma tissues (P= 0.003). HCMV and HPV-16 were the dominant mixed infection in both non-IBC and IBC tissues. Interestingly, although no significant difference in expression of Ki67 has been detected in tissues of IBC and non-IBC, we found that Ki-67 was significantly higher in mixed than single viral infected tissues of both non-IBC and IBC (p = 0.04 and p = 0.03 respectively). Conclusions: The incidence of mixed viral DNA detected in carcinoma tissues of IBC is higher than non-IBC. Moreover, mixed viral DNA is positively correlated with upregulation of Ki67 expression in breast carcinoma tissues.
Inflammatory Breast Cancer: The Secretome of HCMV+ Tumor-Associated Macrophages Enhances Proliferation, Invasion, Colony Formation, and Expression of Cancer Stem Cell Markers
(Frontiers Media S.A., 2022-06) Mohamed, Hossam Taha; El-Sharkawy, Aya Ali; El-Shinawi, Mohamed; Schneider, Robert J; Mohamed, Mona Mostafa
Inflammatory breast cancer (IBC) is a highly aggressive phenotype of breast cancer that is characterized by a high incidence early metastasis. We previously reported a significant association of human cytomegalovirus (HCMV) DNA in the carcinoma tissues of IBC patients but not in the adjacent normal tissues. HCMV-infected macrophages serve as “mobile vectors” for spreading and disseminating virus to different organs, and IBC cancer tissues are highly infiltrated by tumor-associated macrophages (TAMs) that enhance IBC progression and promote breast cancer stem cell (BCSC)-like properties. Therefore, there is a need to understand the role of HCMV-infected TAMs in IBC progression. The present study aimed to test the effect of the secretome (cytokines and secreted factors) of TAMs derived from HCMV+ monocytes isolated from IBC specimens on the proliferation, invasion, and BCSC abundance when tested on the IBC cell line SUM149. HCMV+ monocytes were isolated from IBC patients during modified radical mastectomy surgery and tested in vitro for polarization into TAMs using the secretome of SUM149 cells. MTT, clonogenic, invasion, real-time PCR arrays, PathScan Intracellular Signaling array, and cytokine arrays were used to characterize the secretome of HCMV+ TAMs for their effect on the progression of SUM149 cells. The results showed that the secretome of HCMV+ TAMs expressed high levels of IL-6, IL-8, and MCP-1 cytokines compared to HCMV- TAMs. In addition, the secretome of HCMV+ TAMs induced the proliferation, invasion, colony formation, and expression of BCSC-related genes in SUM149 cells compared to mock untreated cells. In addition, the secretome of HCMV+ TAMs activated the phosphorylation of intracellular signaling molecules p-STAT3, p-AMPKa, p-PRAS40, and p-SAPK/JNK in SUM149 cells. In conclusion, this study shows that the secretome of HCMV+ TAMs enhances the proliferation, invasion, colony formation, and BCSC properties by activating the phosphorylation of p-STAT3, p-AMPKa, p-PRAS40, and p- SAPK/JNK intracellular signaling molecules in IBC cells. UV
Synchrotron Fourier-Transform Infrared Microspectroscopy: Characterization of In Vitro Polarized Tumor-associated Macrophages Stimulated by The Secretome of Inflammatory and Non-Inflammatory Breast Cancer cells
(Elsevier, 2022-09) Mohamed, Hossam Taha; Kamel, Gihan; El-Husseiny, Noura; El-Sharkawy, Aliya; El-Sherif, Ahmed A.; El-Shinawi, Mohamed; Mohamed, Mona Mostafa
ct: Studies suggested that the pathogenesis of inflammatory breast cancer (IBC) is related to inflammatory manifestations accompanied by specific cellular and molecular mechanisms in the IBC tumor microenvironment (TME). IBC is characterized by significantly higher infiltration of tumor-associated macrophages (TAMs) that contribute to its metastatic process via secreting many cytokines such as TNF, IL-6, IL-8, and IL-10 that enhance invasion and angiogenesis. Thus, there is a need to first understand how IBC-TME modulates the polarization of TAMs to better understand the role of TAMs in IBC. Herein, we used gene expression signature and Synchrotron Fourier-Transform Infrared Microspectroscopy (SR- µFTIR) to study the molecular and biochemical changes, respectively of in vitro polarized TAMs stimulated by the secretome of IBC and non-IBC cells. The gene expression signature showed significant differences in the macrophage‟s polarization-related genes between stimulated TAMs. FTIR spectra showed absorption bands in the region of 1700- 1500 cm-1 attributed to the amide I (ν(C=O), & νAS (C-N), δ (N-H), and amide II ν(C-N), δ (N-H) proteins bands. Moreover, three peaks of different intensities and areas were detected in the lipid region of the vCH2 and vCH3 stretching modes positioned within the 3000 - 2800 cm-1 range. The PCA analysis for the second derivative spectra of the amide regions discriminates between stimulated IBC and non-IBC TAMs. This study showed that IBC and non-IBC TMEs differentially modulate the polarization of TAMs and SR-µFTIR can determine these biochemical changes which will help to better understand the potential role of TAMs in IBC.
Syndecan-1 is a novel molecular marker for triple negative inflammatory breast cancer and modulates the cancer stem cell phenotype via the IL-6/STAT3, Notch and EGFR signaling pathways
(BioMed Central, 2017) Abdelaziz Ibrahim, Sherif; Gadalla, Ramy; A. El-Ghonaimy, Eslam; Samir, Omnia; Taha Mohamed, Hossam; Hassan, Hebatallah; Greve, Burkhard; El-Shinawi, Mohamed; Mostafa Mohamed, Mona; Götte, Martin
Background Inflammatory breast cancer (IBC), a particularly aggressive form of breast cancer, is characterized by cancer stem cell (CSC) phenotype. Due to a lack of targeted therapies, the identification of molecular markers of IBC is of major importance. The heparan sulfate proteoglycan Syndecan-1 acts as a coreceptor for growth factors and chemokines, modulating inflammation, tumor progression, and cancer stemness, thus it may emerge as a molecular marker for IBC. Methods We characterized expression of Syndecan-1 and the CSC marker CD44, Notch-1 & -3 and EGFR in carcinoma tissues of triple negative IBC (n = 13) and non-IBC (n = 17) patients using qPCR and immunohistochemistry. Impact of siRNA-mediated Syndecan-1 knockdown on the CSC phenotype of the human triple negative IBC cell line SUM-149 and HER-2-overexpressing non-IBC SKBR3 cells employing qPCR, flow cytometry, Western blotting, secretome profiling and Notch pharmacological inhibition experiments. Data were statistically analyzed using Student’s t-test/Mann-Whitney U-test or one-way ANOVA followed by Tukey’s multiple comparison tests. Results Our data indicate upregulation and a significant positive correlation of Syndecan-1 with CD44 protein, and Notch-1 & -3 and EGFR mRNA in IBC vs non-IBC. ALDH1 activity and the CD44(+)CD24(-/low) subset as readout of a CSC phenotype were reduced upon Syndecan-1 knockdown. Functionally, Syndecan-1 silencing significantly reduced 3D spheroid and colony formation. Intriguingly, qPCR results indicate downregulation of the IL-6, IL-8, CCL20, gp130 and EGFR mRNA upon Syndecan-1 suppression in both cell lines. Moreover, Syndecan-1 silencing significantly downregulated Notch-1, -3, -4 and Hey-1 in SUM-149 cells, and downregulated only Notch-3 and Gli-1 mRNA in SKBR3 cells. Secretome profiling unveiled reduced IL-6, IL-8, GRO-alpha and GRO a/b/g cytokines in conditioned media of Syndecan-1 knockdown SUM-149 cells compared to controls. The constitutively activated STAT3 and NFκB, and expression of gp130, Notch-1 & -2, and EGFR proteins were suppressed upon Syndecan-1 ablation. Mechanistically, gamma-secretase inhibition experiments suggested that Syndecan-1 may regulate the expression of IL-6, IL-8, gp130, Hey-1, EGFR and p-Akt via Notch signaling. Conclusions Syndecan-1 acts as a novel tissue biomarker and a modulator of CSC phenotype of triple negative IBC via the IL-6/STAT3, Notch and EGFR signaling pathways, thus emerging as a promising therapeutic target for IBC.
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(2014) M Mohamed, Mona; Al-Raawi, Diaa; Sabet, Salwa F; El-Shinawi, Mohamed
Inflammatory breast cancer (IBC) is a highly metastatic and fatal form of breast cancer. In fact, IBC is characterized by specific morphological, phenotypic, and biological properties that distinguish it from non-IBC. The aggressive behavior of IBC being more common among young women and the low survival rate alarmed researchers to explore the disease biology. Despite the basic and translational studies needed to understand IBC disease biology and identify specific biomarkers, studies are limited by few available IBC cell lines, experimental models, and paucity of patient samples. Above all, in the last decade, researchers were able to identify new factors that may play a crucial role in IBC progression. Among identified factors are cytokines, chemokines, growth factors, and proteases. In addition, viral infection was also suggested to participate in the etiology of IBC disease. In this review, we present novel factors suggested by different studies to contribute to the etiology of IBC and the proposed new therapeutic insights.