Md Ashiq Mahmud

PhD

PhD Student

Animal cell culture animal experimentation Animal Handling biotech Biotechnology Cancer Bioinformatics Cancer Biology Cancer cell line maintenance Cancer Cell Models 2D Cancer genomics CAR-T cell development cell and molecular biology techniques Cell Biology Epigenetics Immunoassays immunofluroscence assay Immunohistochemistry Immunology immunostaining In Vitro Diagnostics in vitro molecular and cellular assays in vitro transcription In vivo experiment H & E In Vivo Imaging In Vivo mice handling and experiementation In Vivo pharmacology microRNA MicroRNA Analysis Mouse Xenograft Models mRNA in vitro transcription oncology research RNA extraction RNA extraction and quantification RNA Interference RNA Isolation RNA microarrays RNA Sequencing RNA Sequencing and Data Analysis RNA-Seq RNA-seq analysis RNA-Seq Data Analysis Screening of chemical libraries against cancer. scRNA-Seq scRNA-Seq (Single-cell RNA Sequencing) Translational in vitro studies Translational oncology

1st author peer-reviewed publication(s)

Melanoma is among the most abundant malignancies in the US and worldwide. Ligstroside aglycone (LA) is a rare extra-virgin olive oil-derived monophenolic secoiridoid with diverse bioactivities. LA dose-response screening at the NCI 60 cancer cells panel identified the high sensitivity of the Malme-3M cell line, which harbors a BRAF V600E mutation. Daily oral 10 mg/kg LA exhibited potent in vivo antitumor effects against Malme-3M cells xenograft in a nude mouse model by targeting the BRAF signaling pathway. A human Clariom S microarray analysis of the collected Malme- 3M tumors identified 571 dysregulated genes, with the downregulation of pathways critical for melanoma cells growth and survival. A Western blot analysis of the collected animal tumors further validated the downregulation of the mutated BRAF-MAPK axis, as well as the GPD1 and ELOVL6 expression levels. A histopathological analysis of Malme-3M tumor sections showed extensive focal tumor necrosis in treated mice. An immunofluorescence study of tumor sections showed notable reductions in proliferation marker ki67 and the vasculogenesis marker CD31 in treated tumors. These findings promote LA as a potential nutraceutical lead for the control of the BRAF V600E mutant melanoma.

Psoriasis is a chronic immune-mediated skin disease characterized by keratinocyte hyperproliferation and persistent inflammation driven by cytokines signaling. This study aimed to identify novel anti-psoriasis natural products and evaluate their potential molecular attributes. Transcriptomics analysis of psoriatic lesions (GSE54456) and in vitro IL-17A-stimulated HaCaT keratinocytes demonstrated significant upregulation of the lysine methyltransferase SMYD2, suggesting its possible involvement in psoriasis pathology. Treatments of HaCaT cells in vitro with the olive tree (Olea europaea) fruit oil phenolics S-(–)-oleocanthal (OC), S-(–)-hydroxy-oleocanthal (HOC, oleacein), and S-(–)-ligstroside aglycone (LA) at a single 10 µM treatment concentration significantly suppressed the SMYD2 expression and reduced markers of psoriatic inflammation, validated by quantitative PCR and Western blot analysis. The RAW 264.7 macrophages stimulated with LPS and IFN-γ exhibited diminished expression of the pro-inflammatory cytokines, including iNOS, IL-6, and TNF-α, following OC, HOC, and LA treatments. Molecular and protein-protein interaction modeling identified the IL-17A dimer selective binding pocket where OC displayed near-benchmark complementarity binding to the co-crystallized ligand (PubChem CID 153616520), engaging key residues Gln94, Trp67, and Leu97. OC induced the largest pocket expansion (1.52-fold) among all ligands, suggesting strong allosteric interference with IL-17RA binding. Consistently, AlphaLISA assays confirmed the OC ability to effectively inhibit the IL-17A–IL-17RA interaction, unlike LA, which showed modest inhibition. Further, surface plasmon resonance demonstrated dose-dependent blockade of IL-17A–IL-17F binding by OC, validating its IL-17A-IL-17F protein-protein heterodimerization interaction inhibitory capacity. Collectively, findings identify OC as a potent natural modulator for the IL-17 signaling that downregulates SMYD2 and disrupts IL-17A protein-protein interactions, evidencing its anti-psoriatic therapeutic potential.