Talks
Presenting Author Academic/Professional Position
Staff
Academic/Professional Position (Other)
Assistant Research Scientist
Academic Level (Author 1)
Staff
Discipline/Specialty (Author 1)
Immunology and Microbiology
Discipline/Specialty (Author 2)
Immunology and Microbiology
Presentation Type
Oral Presentation
Discipline Track
Biomedical Science
Abstract Type
Research/Clinical
Abstract
Background: MUC13, a recently identified oncoprotein, is aberrantly expressed in pancreatic ductal adenocarcinoma (PDAC) tumors and cells but absent in normal pancreatic tissues. Our laboratory has extensively documented its critical role in pancreatic cancer progression. Despite numerous studies, the molecular interactions mediated by MUC13 that contribute to cancer progression and metastasis remain poorly understood. A significant knowledge gap exists regarding the specific proteins that interact with MUC13. To address this, we conducted a comprehensive molecular interaction study using immunoprecipitation combined with liquid chromatography-mass spectrometry (IP-LCMS) to identify functional interactors of MUC13. Understanding these interactions is essential to elucidate the MUC13-driven biological mechanisms underlying both normal and pathological states. This study aimed to define the MUC13 interactome and uncover novel proteins that may contribute to MUC13-mediated oncogenic functions in pancreatic cancer.
Methodology & Results: Five PDAC cell lines (CaPan2, HPAF-II, Panc1, MiaPaCa, and HPNE) with differential grading were analyzed. Initial LCMS analyses revealed high MUC13 expression exclusively in HPAF-II cells, and undetectable levels in Panc1, MiaPaCa, CaPan2, and HPNE (normal cells). These findings were validated through ELISA, confocal microscopy, and immunoblotting. Immunoprecipitation was performed using HPAF-II cells (MUC13-positive) and Panc1 cells (MUC13-negative) with unique in-house generated MUC13 monoclonal antibodies (C-18 clones) as the test set, and IgG antibodies as controls. Proteins immunoprecipitated from these samples were subjected to LCMS analysis.
IP-LCMS analysis demonstrated a robust signal for MUC13 in HPAF-II cells, while minimal or non-significant signals were observed in the controls. Western blot validation further confirmed these results. After normalization, 54 unique proteins were identified as direct interactors of MUC13. Functional enrichment analysis revealed that the MUC13-associated interactome is involved in various oncogenic mechanisms, including central carbon metabolism, PD-1/PD-L1 checkpoint pathways, adherens junctions, cellular senescence, cell cycle regulation, apoptosis, viral carcinogenesis, and diabetic cardiomyopathy. Additionally, 304 proteins were identified in the second layer of interactions, collectively contributing to pancreatic cancer-associated pathways such as pancreatic neuroendocrine tumor signaling and growth factor pathways.
Conclusion: Proteins are key mediators of cellular processes, often functioning within multi-protein complexes. This study utilized an integrated IP-LCMS approach to identify the MUC13 interactome, uncovering a unique set of molecular partners that facilitate MUC13's oncogenic functions in pancreatic cancer cells. By identifying 54 direct interactors and 304 secondary interactors, we provide new insights into the MUC13-mediated pathways driving pancreatic cancer progression. These findings could aid in developing targeted therapies that disrupt specific MUC13-mediated molecular interactions, offering new avenues for treating pancreatic cancer.
Recommended Citation
Dhasmana, Anupam; Dhasmana, Swati; Baru, Rajasekhar; Singh, Shweta; Gomez, Abigail; Khan, Sheema; Yallapu, Murali M.; and Chauhan, Subhash, "MUC13-Driven Molecular Interactions in Pancreatic Cancer Cell lines" (2025). Research Symposium. 5.
https://scholarworks.utrgv.edu/somrs/2025/talks/5
Included in
MUC13-Driven Molecular Interactions in Pancreatic Cancer Cell lines
Background: MUC13, a recently identified oncoprotein, is aberrantly expressed in pancreatic ductal adenocarcinoma (PDAC) tumors and cells but absent in normal pancreatic tissues. Our laboratory has extensively documented its critical role in pancreatic cancer progression. Despite numerous studies, the molecular interactions mediated by MUC13 that contribute to cancer progression and metastasis remain poorly understood. A significant knowledge gap exists regarding the specific proteins that interact with MUC13. To address this, we conducted a comprehensive molecular interaction study using immunoprecipitation combined with liquid chromatography-mass spectrometry (IP-LCMS) to identify functional interactors of MUC13. Understanding these interactions is essential to elucidate the MUC13-driven biological mechanisms underlying both normal and pathological states. This study aimed to define the MUC13 interactome and uncover novel proteins that may contribute to MUC13-mediated oncogenic functions in pancreatic cancer.
Methodology & Results: Five PDAC cell lines (CaPan2, HPAF-II, Panc1, MiaPaCa, and HPNE) with differential grading were analyzed. Initial LCMS analyses revealed high MUC13 expression exclusively in HPAF-II cells, and undetectable levels in Panc1, MiaPaCa, CaPan2, and HPNE (normal cells). These findings were validated through ELISA, confocal microscopy, and immunoblotting. Immunoprecipitation was performed using HPAF-II cells (MUC13-positive) and Panc1 cells (MUC13-negative) with unique in-house generated MUC13 monoclonal antibodies (C-18 clones) as the test set, and IgG antibodies as controls. Proteins immunoprecipitated from these samples were subjected to LCMS analysis.
IP-LCMS analysis demonstrated a robust signal for MUC13 in HPAF-II cells, while minimal or non-significant signals were observed in the controls. Western blot validation further confirmed these results. After normalization, 54 unique proteins were identified as direct interactors of MUC13. Functional enrichment analysis revealed that the MUC13-associated interactome is involved in various oncogenic mechanisms, including central carbon metabolism, PD-1/PD-L1 checkpoint pathways, adherens junctions, cellular senescence, cell cycle regulation, apoptosis, viral carcinogenesis, and diabetic cardiomyopathy. Additionally, 304 proteins were identified in the second layer of interactions, collectively contributing to pancreatic cancer-associated pathways such as pancreatic neuroendocrine tumor signaling and growth factor pathways.
Conclusion: Proteins are key mediators of cellular processes, often functioning within multi-protein complexes. This study utilized an integrated IP-LCMS approach to identify the MUC13 interactome, uncovering a unique set of molecular partners that facilitate MUC13's oncogenic functions in pancreatic cancer cells. By identifying 54 direct interactors and 304 secondary interactors, we provide new insights into the MUC13-mediated pathways driving pancreatic cancer progression. These findings could aid in developing targeted therapies that disrupt specific MUC13-mediated molecular interactions, offering new avenues for treating pancreatic cancer.
