Loss of Neuropilin-2 in Murine Mesenchymal-like Colon Cancer Organoids Causes Mesenchymal-to-Epithelial Transition and an Acquired Dependency on Insulin-Receptor Signaling and Autophagy
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. In Vitro 3D Organoid Culture and Treatment
2.2. RNA Sequencing and Data Analysis
2.3. In Vivo Subcutaneous Tumorigenesis
2.4. Immunohistochemistry (IHC) and Immunofluorescence (IF)
2.5. Western Blot Analysis
2.6. Statistical Analyzes
3. Results
3.1. Nrp2 Knockout in Mouse CRC Organoids Induces Tight Junction Stabilization and EMT Reduction
3.2. Nrp2 Knockout in Mouse CRC Organoids Reduces Adult Intestinal Stem Cell Gene Expression without Influencing Fetal Intestinal Stem Cell Signature
3.3. Nrp2 Knockout in Mouse CRC Organoids Induces Insulin Receptor Signaling Upregulation
3.4. Nrp2 Depletion in Mouse CRC Organoids Activates Autophagy
3.5. Combined Inhibition of IR Signaling and Early Autophagy Reduces Nrp2−/− CRC Organoid Viability
3.6. Nrp2 Expression Is Associated with a Mesenchymal Molecular Subtype of Human CRC and Inversely Correlated with Autophagy Marker Expression
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Poghosyan, S.; Frenkel, N.; Lentzas, A.; Laoukili, J.; Rinkes, I.B.; Kranenburg, O.; Hagendoorn, J. Loss of Neuropilin-2 in Murine Mesenchymal-like Colon Cancer Organoids Causes Mesenchymal-to-Epithelial Transition and an Acquired Dependency on Insulin-Receptor Signaling and Autophagy. Cancers 2022, 14, 671. https://doi.org/10.3390/cancers14030671
Poghosyan S, Frenkel N, Lentzas A, Laoukili J, Rinkes IB, Kranenburg O, Hagendoorn J. Loss of Neuropilin-2 in Murine Mesenchymal-like Colon Cancer Organoids Causes Mesenchymal-to-Epithelial Transition and an Acquired Dependency on Insulin-Receptor Signaling and Autophagy. Cancers. 2022; 14(3):671. https://doi.org/10.3390/cancers14030671
Chicago/Turabian StylePoghosyan, Susanna, Nicola Frenkel, Aristeidis Lentzas, Jamila Laoukili, Inne Borel Rinkes, Onno Kranenburg, and Jeroen Hagendoorn. 2022. "Loss of Neuropilin-2 in Murine Mesenchymal-like Colon Cancer Organoids Causes Mesenchymal-to-Epithelial Transition and an Acquired Dependency on Insulin-Receptor Signaling and Autophagy" Cancers 14, no. 3: 671. https://doi.org/10.3390/cancers14030671