Browsing by Author "Mohamed, Hossam Taha"

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Antiviral activity of chitosan nanoparticles encapsulating curcumin against hepatitis C virus genotype 4a in human hepatoma cell lines
(Dove Medical Press Ltd., 05/09/2020) Loutfy, Samah A; Elberry, Mostafa H; Farroh, Khaled Yehia; Mohamed, Hossam Taha; Mohamed, Aya A; Mohamed, ElChaimaa B; Faraag, Ahmed Hassan Ibrahim; Mousa, Shaker A
View references (46) Purpose: Current direct-acting antiviral agents for treatment of hepatitis C virus genotype 4a (HCV-4a) have been reported to cause adverse effects, and therefore less toxic antivirals are needed. This study investigated the role of curcumin chitosan (CuCs) nanocomposite as a potential anti-HCV-4a agent in human hepatoma cells Huh7. Methods: Docking of curcumin and CuCs nanocomposite and binding energy calculations were carried out. Chitosan nanoparticles (CsNPs) and CuCs nanocomposite were prepared with an ionic gelation method and characterized with TEM, zeta size and potential, and HPLC to calculate encapsulation efficiency. Cytotoxicity studies were performed on Huh7 cells using MTT assay and confirmed with cellular and molecular assays. Anti-HCV-4a activity was determined using real-time PCR and Western blot. Results: The strength of binding interactions between protein ligand complexes gave scores with NS3 protease, NS5A polymerase, and NS5B polymerase of-124.91,-159.02, and-129.16, for curcumin respectively, and-68.51,-54.52, and-157.63 for CuCs nanocomposite, respec-tively. CuCs nanocomposite was prepared at sizes 29–39.5 nm and charges of 33 mV. HPLC detected 4% of curcumin encapsulated into CsNPs. IC50 was 8 µg/mL for curcumin and 25 µg/ mL for the nanocomposite on Huh7 but was 25.8 µg/mL and 34 µg/mL on WISH cells. CsNPs had no cytotoxic effect on tested cell lines. Apoptotic genes’ expression revealed the caspase-dependent pathway mechanism. CsNPs and CuCs nanocomposite demonstrated 100% inhibition of viral entry and replication, which was confirmed with HCV core protein expression. Conclusion: CuCs nanocomposite inhibited HCV-4a entry and replication compared to curcumin alone, suggesting its potential role as an effective therapeutic agent. © 2020 Loutfy et al
Aspergillus Niger thermostable Cytosine deaminase‑dextran conjugates with enhanced structure stability, proteolytic resistance, and Antiproliferative activity
(BioMed Central Ltd, 2023-01) El‑Sayed, Ashraf S. A; Rady, Amgad M; Mohamed, Hossam Taha; Zein, Nabila; Yassin, Marwa A; Mohamed, Nabil Z; Hassan, Abdallah; Amer, MahmoudReyad El‑Sharakawy5 M; El‑Sharkawy, Aya Ali; El‑Sayed, Nesma; Ali, Mostafa G
Cytosine deaminase (CDA) is a prodrug mediating enzyme converting 5-furocytosine into 5-furouracil with profound broad-range anticancer activity towards various cell lines. Availability, molecular stability, and catalytic efciency are the main limiting factors halting the clinical applications of this enzyme on prodrug and gene therapies, thus, screen‑ ing for CDA with unique biochemical and catalytic properties was the objective. Thermotolerant/ thermophilic fungi could be a distinctive repertoire for enzymes with afordable stability and catalytic efciency. Among the recovered thermotolerant isolates, Aspergillus niger with optimal growth at 45 °C had the highest CDA productivity. The enzyme was purifed, with purifcation 15.4 folds, molecular mass 48 kDa and 98 kDa, under denaturing and native PAGE, respectively. The purifed CDA was covalently conjugated with dextran with the highest immobilization yield of 75%. The free and CDA-dextran conjugates have the same optimum pH7.4, reaction temperature 37 °C, and pI 4.5, and similar response to the inhibitors and amino acids suicide analogues, ensuring the lack of efect of dextran conjuga‑ tion on the CDA conformational structure. CDA-Dextran conjugates had more resistance to proteolysis in response to proteinase K and trypsin by 2.9 and 1.5 folds, respectively. CDA-Dextran conjugates displayed a dramatic structural and thermal stability than the free enzyme, authenticating the acquired structural and catalytic stability upon dextran conjugation. The thermal stability of CDA was increased by about 1.5 folds, upon dextran conjugation, as revealed from the half-life time (T1/2). The afnity of CDA-conjugates (Km 0.15mM) and free CDA (Km 0.22mM) to deaminate 5-fuorocytosine was increased by 1.5 folds. Upon dextran conjugation, the antiproliferative activity of the CDA towards the diferent cell lines “MDA-MB, HepG-2, and PC-3” was signifcantly increased by mediating the prodrug 5-FC. The CDA-dextran conjugates strongly reduce the tumor size and weight of the Ehrlich cells (EAC), dramatically increase the titers of Caspase-independent apoptotic markers PARP-1 and AIF, with no cellular cytotoxic activity, as revealed from the hematological and biochemical parameters.
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).
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
Infrared microspectroscopy and imaging analysis of inflammatory and non-inflammatory breast cancer cells and their GAG secretome
(MDPI AG, 9/19/2020) Mohamed, Hossam Taha; Untereiner, Valérie; Cinque, Gianfelice; Ibrahim, Sherif Abdelaziz; Götte, Martin; Nguyen, Nguyet Que; Rivet, Romain; Sockalingum, Ganesh D.; Brézillon, Stéphane
Glycosaminoglycans (GAGs)/proteoglycans (PGs) play a pivotal role in the metastasis of inflammatory breast cancer (IBC). They represent biomarkers and targets in diagnosis and treatment of different cancers including breast cancer. Thus, GAGs/PGs could represent potential prognostic/diagnostic biomarkers for IBC. In the present study, non-IBC MDA-MB-231, MCF7, SKBR3 cells and IBC SUM149 cells, as well as their GAG secretome were analyzed. The latter was measured in toto as dried drops with high-throughput (HT) Fourier Transform InfraRed (FTIR) spectroscopy and imaging. FTIR imaging was also employed to investigate single whole breast cancer cells while synchrotron-FTIR microspectroscopy was used to specifically target their cytoplasms. Data were analyzed by hierarchical cluster analysis and principal components analysis. Results obtained from HT-FTIR analysis of GAG drops showed that the inter-group variability enabled us to delineate between cell types in the GAG absorption range 1350–800 cm−1. Similar results were obtained for FTIR imaging of GAG extracts and fixed single whole cells. Synchrotron-FTIR data from cytoplasms allowed discrimination between non-IBC and IBC. Thus, by using GAG specific region, not only different breast cancer cell lines could be differentiated, but also non-IBC from IBC cells. This could be a potential diagnostic spectral marker for IBC detection useful for patient management. © 2020 by the authors. Licensee MDPI, Basel, Switzerland
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.