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Modhukur, V.

Publications and source records attributed to Modhukur, V..

5 recordsLinked to original sources

Aberrant epithelialization: A plausible factor for the development of endometrial polyps

Endometrial polyps (EPs) are localized overgrowths of endometrial glands and stroma, common in reproductive-age and postmenopausal women, and can cause abnormal uterine bleeding and infertility. Here, we investigated the cellular heterogeneity and molecular mechanisms of EPs by integrating bulk and single-cell RNA sequencing (scRNA-seq) of EPs and adjacent endometrial tissues (adENs) from 12 women. Bulk RNA-seq revealed high transcriptional similarity, with few differentially expressed genes including upregulated KMT2B and DLEC1 and downregulated COL9A1 and RAB3C. ScRNA-seq identified eight major cell clusters, such as stromal, epithelial, endothelial, immune, perivascular, macrophage, B, and ciliated cells. Pseudotime analysis showed aberrant stromal-to-epithelial transitions in EPs, marked by MECOM and EYA2 intermediate clusters, indicating incomplete epithelial maturation. These altered differentiation trajectories may disrupt perivascular and endothelial cell development, contributing to abnormal vascular remodeling in EPs, despite minimal overall transcriptomic changes compared with adENs.

molecular biology↗

OncoProExp: An Interactive Shiny Web Application for Comprehensive Cancer Proteomics and Phosphoproteomics Analysis

Cancer research has been revolutionized by mass spectrometry (MS)-based proteomics, enabling large-scale profiling of proteins and post-translational modifications (PTMs) to identify critical alterations in cancer signaling pathways. However, the lack of comprehensive, userfriendly platforms for integrative analysis limits efficient data exploration, biomarker identification, and translational insights. To address this gap, we developed OncoProExp, a Shiny-based interactive web application designed for in-depth exploration of cancer proteomes and phosphoproteomes. OncoProExp offers robust workflows for data preprocessing, including missing value imputation and statistical filtering. The platform features interactive visualizations such as principal component analysis (PCA), hierarchical clustering heatmaps, and gene set enrichment analysis (GSEA), enabling detailed functional annotation. Differential expression analysis to identify differentially expressed proteins (DEPs) and phosphoproteins (DEPPs) facilitating the discovery of potential biomarkers and therapeutic targets. The application supports survival analysis and pan-cancer exploration using clinical and proteome/phosphoproteomic datasets. OncoProExp incorporates state-of-the-art predictive modeling using machine learning algorithms, including Support Vector Machines (SVMs), Random Forests, and Artificial Neural Networks (ANNs) for cancer risk stratification, achieving near-perfect accuracy in multi-cancer and single-cancer classification. These models are enhanced by SHapley Additive exPlanations (SHAP) for interpretability. To enhance its translational utility, the platform supports user-uploaded data and enables protein-protein interaction analysis, pathway enrichment analysis, cancer drug relevance evaluation, and clinical annotation using curated cancer-specific datasets. OncoProExp is deployable via Docker containers, ensuring flexible and scalable integration into individual servers. Its utility has been demonstrated using Clinical Proteomic Tumor Analysis Consortium (CPTAC) datasets, showcasing its potential to advance cancer biomarker discovery, risk stratification, therapeutic target identification, and personalized treatment strategies. OncoProExp is freely accessible at https://oncopro.cs.ut.ee/ without login requirements, offering a comprehensive resource for translational cancer research.

bioinformatics↗

Proteome changes associated with effect of high-dose single-fractionation radiation on lung adenocarcinoma cell lines

Lung cancer is a leading cause of cancer-related mortality globally, with non-small cell lung cancer (NSCLC) representing 85% of cases. Advances in treatment modalities, including the emergence of antibody-drug conjugates and stereotactic radiation therapy, have improved outcomes. However, the possible synergistic effects of these therapies remain underexplored at the molecular level. This study investigated high-dose radiation-induced proteomic changes in lung adenocarcinoma cell line HCC-44 grown adherently and cell line A549, grown as adherent cells and 3D spheroids. Our hypothesis was that proteins upregulated by 10 Gy irradiation serve as resistance drivers in cancerous cells and can thus represent potential therapeutic targets. The label-free mass spectrometry revealed distinct proteomic responses to 10 Gy irradiation, varying by cell line and culturing conditions. Differentially expressed proteins elevated in the irradiated samples included ephrin type-A receptor 2 (EPHA2) in adherent cells and insulin-like growth factor 2 receptor (IGF2R), tetraspanin 3 (TSPAN3) as well as cathepsin D (CTSD) in spheroids. The validation of these targets was carried out via Western blot, immunofluorescence, viability assay and spheroid formation assay. The functional assays demonstrated that irradiation sensitized A549 cells to EPHA2 and CTSD inhibitors. These findings underscore the potential of integrating radiation and targeted therapies in NSCLC treatment, and highlight EPHA2 as a promising candidate for future therapeutic strategies.

cell biology↗

Endometriotic lesions exhibit distinct metabolic signature compared to paired eutopic endometrium at the single-cell level

Current therapeutics of endometriosis are limited to hormonal action on endometriotic lesions to disrupt their growth. Based on the recent findings of the high utilization of glycolysis over oxidative metabolism (Warburg-like effect) in endometriotic lesions, a new strategy of nonhormonal management by addressing cellular metabolism has been proposed. However, it remains unclear which cell types are metabolically altered and contribute to endometriotic lesion growth for targeting them with metabolic drugs. Using single-cell RNA-sequencing, we investigated the activity of twelve metabolic pathways and genes involved in steroidogenesis in paired samples of eutopic endometrium (EuE) and peritoneal lesions (ectopic endometrium, EcE) from women with confirmed endometriosis. We detected nine major cell clusters in both EuE and EcE. The metabolic pathways were differentially regulated in perivascular, stromal and to a lesser extent in endothelial cell clusters, with the highest changes in AMP-activated protein kinase signaling, Hypoxia-Inducible Factor-1 signaling, glutathione metabolism, oxidative phosphorylation, and glycolysis/gluconeogenesis. We identified a transcriptomic co-activation of glycolysis and oxidative metabolism in perivascular and stromal cells of EcE compared with EuE, suggesting that metabolic reprogramming may play a critical role in maintaining cell growth and survival of endometriotic lesions. Additionally, progesterone receptor was significantly downregulated in perivascular and endothelial cells of EcE. The expression of estrogen receptor 1 was significantly reduced in perivascular, stromal and endothelial cells of EcE. In parallel, perivascular cells exhibited a high expression of estrogen receptor 2 and HSD17B8 gene that encodes for protein converting estrone (E1) to estradiol (E2), while in endothelial cells HSD17B2 gene coding for enzyme converting E2 to E1 was downregulated. Overall, our results identified different expression patterns of energy metabolic pathways and steroidogenesis-related genes in perivascular, stromal, and endothelial cells in EcE compared with EuE. Perivascular cells, known to contribute to the restoration of endometrial stroma and angiogenesis, can be a potential target for non-hormonal treatment of endometriosis.

molecular biology↗

Inhibition of epigenetic and cell cycle-related targets in glioblastoma cell lines: onametostat reduces proliferation and viability in both normoxic and hypoxic conditions

The choice of targeted therapies for treatment of glioblastoma patients is currently limited, and most glioblastoma patients die from the disease recurrence. Thus, systematic studies in simplified model systems are required to pinpoint the choice of targets for further exploration in clinical settings. Here, we report screening of 5 compounds targeting epigenetic writers or erasers and 6 compounds targeting cell cycle-regulating protein kinases against 3 glioblastoma cell lines following incubation under normoxic or hypoxic conditions. The viability assay indicated that PRMT5 inhibitor onametostat was endowed with high potency under both normoxic and hypoxic conditions in both MGMT-positive and MGMT-negative cell lines. In U-251 MG and U-87 MG cells, onametostat also affected the spheroid formation at concentrations lower than the currently used chemotherapeutic drug lomustine. Furthermore, in T98-G cell line, treatment with onametostat led to dramatic changes in the transcriptome profile by inducing the cell cycle arrest, suppressing RNA splicing, and down-regulating several major glioblastoma cell survival pathways. In this way, we confirmed that inhibition of epigenetic targets might represent a viable strategy for glioblastoma treatment even in the case of decreased chemo- and radiation sensitivity, although further studies in clinically more relevant models are required.

cancer biology↗