Biology subjects

Shafiq, A.

Publications and source records attributed to Shafiq, A..

3 recordsLinked to original sources

bioRxiv2026

Small Extracellular Vesicle Cargoes Associated with Lymph Node Metastasis in Head and Neck Cancer

Head and neck squamous cell carcinoma (HNSCC) is a highly aggressive malignancy in which cervical lymph node metastasis critically determines patient prognosis. Despite advances in clinical staging based on tumor size, depth of invasion, and nodal status, these parameters fail to capture the biological heterogeneity of HNSCC, leading to overtreatment or undertreatment, and significant morbidity. Emerging evidence implicates small extracellular vesicles (SEVs) as key mediators of tumor progression and promising biomarkers for metastatic potential. In this study, we performed iTRAQ-based proteomic profiling of SEVs from metastatic (MOC2) and nonmetastatic (MOC1) HNSCC mouse cell lines and identified multiple cargoes associated with metastatic processes, including angiogenesis, extracellular matrix remodeling, immune modulation, and perineural invasion. Further, we employed an immune-competent orthotopic mouse oral carcinoma (MOC) model to investigate how exosome biogenesis affects HNSCC metastasis. Loss of the exosome/SEV biogenesis protein HRS in MOC2 HNSCC cells impaired SEV formation in vitro and tumor growth and metastasis in vivo. Consistent with the SEV cargoes enriched in MOC2 EVs, immunohistochemical analyses of MOC2 tumors revealed reduced blood vessel and nerve density in HRS-deficient tumors. Analysis of candidate biomarker SEV cargoes in the circulating EVs from HPV-negative HNSCC patients revealed significant correlations of these proteins with metastatic status. Collectively, these findings identify SEV cargoes as potential functional mediators of metastasis and liquid biopsy biomarkers in HNSCC.

cancer biology↗

bioRxiv2025

Oncogenic H-Ras Reprograms Madin-Darby Canine Kidney (MDCK) Cell-derived Midbody Remnant Proteins Following Epithelial-Mesenchymal Transition

Epithelial-mesenchymal transition (EMT) is a highly conserved morphogenic process that allows highly polarized, immotile epithelial cells to transform into motile mesenchymal cells: it is a fundamental cellular process involved in embryonic development, tumour cell metastasis, organ fibrosis and tissue regeneration. To assess the contribution of secreted midbody remnants (MBRs) - a new class of membranous extracellular vesicle (EV) molecularly distinct from exosomes/small EVs - to the EMT process, we conducted a proteomic analysis of MBRs released from Madin-Darby canine kidney (MDCK) cells, and MDCK cells transformed with oncogenic H-Ras (21D1 cells). MBRs were harvested from cell culture media in milligram quantities using a continuous culture bioreactor device and purified using sequential centrifugation and buoyant density gradient centrifugation (OptiPrep). Gel-MS/MS protein profiling showed MDCK cell-MBRs reflect their epithelial origin (e.g., enriched CDH1, DSP, THBS1, OLCN, EPCAM proteins) and 21D1 cell-MBRs their mesenchymal phenotype (e.g., HRAS, VIM, MMP14, CDH2, WNT5A and enriched invasive and cell motility proteins). Prominent findings were the unique expression of the immune checkpoint protein NT5E/CD73 (ecto-5'-nucleotidase), and ser/thr kinases LIMK1/K2 in 21D1-MBRs (not present in MDCK cell-MBRs), and enrichment in Wnt signalling network proteins. Collectively, our findings suggest MBRs might play a previously unrecognized role in the EMT process. SignificanceO_LIEpithelial-to-mesenchymal transition (EMT) is a critical cell biological process that occurs during normal embryonic development and cancer progression. Our study describes, for the first time, the large-scale sequential purification of secreted midbody remnants (MBRs) and exosomes/sEVs from the in vitro cell line EMT model Madin-Darby canine kidney (MDCK) cells and MDCK cells transformed with oncogenic H-Ras (21D1 cells): GeLC-MS/MS protein profiling identified the repertoire of enriched MDCK-MBR proteins following EMT. C_LIO_LIMBRs display a proteome profile distinct from sEVs that is enriched with factors of the centralspindlin complex (KIF23.1, KIF4A, INCENP, CEP55, PLK1) and further include components of the mitochondrial network, cytokinesis, microtubule movement, and intercellular connection. C_LIO_LIIn the context of EMT, our data reveal simultaneous activation of EMT signalling pathways in MBRs including signalling receptor binding, regulation of cell differentiation, and Wnt, VEGF and PDGF signalling. C_LIO_LIWe identify several mesenchymal enriched networks in MBRs associated with focal adhesion, cell matrix, kinase activity, and cell shape/organisation, while epithelial derived MBRs are show enriched networks predominately associated with mitochondrial (processing/transport), midbody, and plasma membrane annotation. C_LIO_LIOur study sheds light on the signalling architecture of MBRs following oncogenic H- Ras-induced EMT: collectively, our data informs ongoing efforts to delineate oncogenic drivers of cancer initiation, progression, and metastasis. C_LI

cell biology↗

bioRxiv2020

Elevated soluble Galectin-3 as a marker of chemotherapy efficacy in Breast cancer patients: a prospective study

PurposeGalectin-3 (Gal-3) is a glycan-binding lectin with a debated role in cancer progression due to its various functions and patterns of expression. The current study investigates the relationship between breast cancer prognosis and secreted Gal-3. MethodsBreast cancer patients with first time cancer diagnosis and no prior treatment (n=88) were placed in either adjuvant or neoadjuvant setting based on their treatment modality. Stromal and plasma Gal-3 levels were measured in each patient at the time of diagnosis and then throughout treatment using immunohistochemistry (IHC) and ELISA respectively. Healthy women (>18 years of age, n=63) were used to establish baseline levels of plasma Gal-3. Patients were followed for 84 months for disease free survival analysis. ResultsEnhanced levels of plasma (adjuvant) and stromal (neo-adjuvant) Gal-3 were found to be markers of chemotherapy efficacy. The patients with chemotherapy induced increase in extracellular Gal-3 had longer disease-free interval and significantly lower rate of recurrence during 84-month follow-up compared to patients with unchanged or decreased secretion. ConclusionThe findings support the use of plasma Gal-3 as a marker for chemotherapy efficacy when no residual tumor is visible through imaging. Furthermore, stromal levels in any remaining tumors post chemotherapy can also be used to predict long term prognosis in patients. Key pointsIncreased Gal-3 secretion due to chemotherapy leads to better prognosis and longer disease-free survival. The analysis of soluble Gal-3 expression could be useful as a support tool in predicting treatment efficacy in patients with no visible tumor remaining for follow up through imaging.

cancer biology↗