Divya Murthy, PhD

Tumor microenvironment-mediated metabolic reprogramming in pancreatic cancer. I work on understanding the role of metabolites on the survival and aggressiveness of pancreatic cancer. My work deals with: • Identification of metabolites by Liquid Chromatography based metabolomics (LC-MS/MS). • Identification of genes that potentially promote an aggressive cancer phenotype in pancreatic cancer using Chromatin Immunoprecipitation and RNA-Seq • Proteome analysis using mass spectrometry. • Developing CRISPR-Cas9-based knockout models. Technical Skill set: • Phospho-proteomic studies in pancreatic cancer cells using Reverse Phase Protein Arrays • Tumor cell culture and in-vitro drug testing using spectroscopic and fluorescence microplate assay • Biomarker assay using immunohistochemistry, western blot, and quantitative real-time PCR assays. • LC-MS/MS-based metabolomic studies • Development of tumor organoid-based model system. • GSEA of biological datasets to determine biologically and statistically significant set of genes. https://aacrjournals.org/cancerdiscovery/article/doi/10.1158/2159-8290.CD-23-0334/730024/Vitamin-B6-competition-in-the-tumor https://www.mdpi.com/1422-0067/24/8/7304 https://www.sciencedirect.com/science/article/pii/S2213231722000738?via%3Dihub https://www.nature.com/articles/s41388-021-02132-6 https://www.gastrojournal.org/article/S0016-5085(21)03159-0/fulltext?referrer=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F https://www.sciencedirect.com/science/article/abs/pii/S0304383520303815?via%3Dihub https://rupress.org/jem/article/217/7/e20190745/151806/SIRT1-NOX4-signaling-axis-regulates-cancer https://www.frontiersin.org/articles/10.3389/fphys.2018.00335/full https://link.springer.com/protocol/10.1007%2F978-1-4939-7665-2_8 https://www.sciencedirect.com/science/article/pii/S1535610817302544?via%3Dihub https://www.sciencedirect.com/science/article/pii/S0304383517302902?via%3Dihub https://www.bioscience.org/2017/v22/af/4543/fulltext.htm

The project focused on understanding the melanosome and mitochondria dynamics during pigmentation under the guidance of Dr. Rajesh S. Gokhale. • Delineated the role of autophagic machinery in melanosome biogenesis and degradation. • Characterized a non-canonical role of autophagic protein MAP1LC3B in mediating movement of melanosomes during melanogenesis. • Identified the motor protein (kinesin) that mediate translocation of melanosomes in melanocytes. • Explored the melanosome-mitochondria crosstalk during melanogenesis using a compendium of tools and techniques including mass spectrometry, mammalian cell culture, and extensive confocal based microscopy. • Utilized organotypic culture derived from human subjects for isolation of melanocytes to validate cell line-based findings. Technical Skill set: • Organelle isolation and its biochemical characterization • In vitro biochemical assays • Molecular biology techniques: Cloning of mammalian genes, site directed mutagenesis, recombinant protein expression in E.coli, protein purification and enzyme activity studies, protein-protein interaction studies by co-immunoprecipitation and protein pull down assays, DNA and RNA isolation and purification. • FACS-based studies • Extensive training in confocal based microscopic analyses of subcellular organelles and structures; and live cell imaging to understand organelle dynamics and kinetics. https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3001634 https://www.tandfonline.com/doi/full/10.1080/15548627.2017.1327509 https://www.sciencedirect.com/science/article/pii/S0006349515009923?via%3Dihub https://www.sciencedirect.com/science/article/pii/S0958166913000232?via%3Dihub

Elucidating the role of PLEKHM2 in melanosome trafficking on cytoskeletal tracks in melanocytes. • Collaborated on a research project to elucidate the role of the adaptor protein, PLEKHM2 (SKIP), in mediating movement of melanosomes within melanocyte. • Performed exogenous overexpression and confocal-based experiments to study subcellular localization of PLEKHM2 on melanosomes. • Extensively employed microscopy to analyze subcellular organelles and structures, and live cell imaging to understand organelle dynamics and kinetics. https://www.biorxiv.org/content/10.1101/2021.03.11.434917v1

Cytokine expression profiling in Caco-2 and RAW 264.7 cell lines exposed to Lactobacilli isolates to evaluate their probiotic effect. • The project aimed at understanding the modulation in the cytokine profiles of pro- and anti-inflammatory cytokines in Caco-2 colon carcinoma cell lines and RAW 264.7 macrophage cell lines stimulated with various Lactobacilli isolates. • Isolated various Lactobacilli strain from different sources, stimulation of Caco-2 and RAW 264.7 cell lines with the Lactobacilli isolates. • Performed bacterial adhesion assays and semi-quantitative PCR-based cytokine profiling. The project yielded interesting findings. I observed significant immunomodulation on stimulation with various Lactobacilli isolates implicating them as potential probiotic candidates. Technical expertise attained: • Mammalian cell culture, semi-quantitative PCR, bacterial adhesion assays.