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Ubiquitin specific peptidase (USP37) mediated effects in microscaffold-encapsulated cells: a comprehensive study on
Shreemoyee De1, Ravi Chauhan2, Mayank Singh2
1Centre for Biomedical Engineering, Indian Institute of Technology Delhi Hauz Khas New Delhi 110016 India shreemoyee.de@iitd.ac.in sneetu@iitd.ac.in.
Abstract:
Though significant advances have been made in developing therapeutic strategies for cancer, suitable in vitro models for mechanistically identifying relevant drug targets and understanding disease progression are still lacking. Most studies are generally performed using two-dimensional (2D) models, since these models can be readily established and allow high throughput assays. However, these models have also been reported as the reason for unreliable pre-clinical information. To avoid this discrepancy, three-dimensional (3D) cell culture models have been established and have demonstrated the potential to provide alternative ways to study tissue behavior. However, most of these models first require optimization and cell cultures with a certain density, thus adding a prepping step in the platform before it can be used for any studies. This limits their use in studies where the fundamental understanding of biological processes must be carried out in a short time frame. In this study, we developed a 3D cell culture system that tests a less explored cancer therapeutic target-the deubiquitinating enzyme ubiquitin specific peptidase 37 (USP37)-in different cancer cell lines using sensitive carbon dot pH nanosensors, which provides a rapid model for studies compared to the parallel model available commercially. This enzyme is found to be elevated in different cancers and has been reported to play a role in cell cycle regulation, oncogenesis and metastasis. However, the confirmation of the role of USP37 downregulation in cellular proliferation via appropriate in vitro 3D models has not been demonstrated. To establish the applicability of the developed 3D platform in studying such oncogenes, classical 2D models have been used in this study for identifying the role of USP37 in tumor progression and metastasis. The data clearly suggests that this ingeniously developed 3D cell culture system is a better alternative to 2D models to study the growth and migration of different cancer cell lines on depletion of oncogenic proteins like USP37 and its effect on epithelial-mesenchymal transition (EMT) markers, and it can further be targeted as a viable therapeutic option.
Insights
A novel 3D cell culture system rapidly models cancer progression and tests ubiquitin specific peptidase 37 (USP37) as a therapeutic target. This system offers a superior alternative to 2D models for studying oncogene depletion and its effects on cancer cell growth and metastasis.
Area of Science:
- Oncology
- Biotechnology
- Biomedical Engineering
Background:
- Current two-dimensional (2D) in vitro cancer models offer limited predictive value for clinical outcomes due to their inability to replicate the complex tumor microenvironment.
- Three-dimensional (3D) cell culture models show promise but often require extensive optimization and cell density adjustments, hindering rapid experimental application.
- The deubiquitinating enzyme ubiquitin specific peptidase 37 (USP37) is implicated in cancer progression, yet its therapeutic targeting requires robust in vitro validation models.
Purpose of the Study:
- To develop and validate a rapid 3D cell culture system for studying cancer progression and therapeutic targets.
- To investigate the role of ubiquitin specific peptidase 37 (USP37) in cancer cell proliferation, migration, and epithelial-mesenchymal transition (EMT) using the developed 3D model.
- To assess the utility of carbon dot pH nanosensors within the 3D system for real-time monitoring of cellular processes.
Main Methods:
- Development of a novel, user-friendly 3D cell culture platform enabling rapid experimental setup.
- Utilized sensitive carbon dot pH nanosensors for real-time monitoring of cellular behavior within the 3D model.
- Investigated the effects of ubiquitin specific peptidase 37 (USP37) depletion on cancer cell lines using both the novel 3D system and traditional 2D models.
- Assessed changes in cancer cell growth, migration, and epithelial-mesenchymal transition (EMT) markers.
Main Results:
- The developed 3D cell culture system demonstrated significantly faster experimental throughput compared to existing commercial models.
- Depletion of ubiquitin specific peptidase 37 (USP37) in cancer cell lines led to reduced proliferation and migration, as observed in the 3D model.
- The 3D model effectively captured the impact of USP37 downregulation on epithelial-mesenchymal transition (EMT) markers, correlating with reduced metastatic potential.
- Carbon dot pH nanosensors provided sensitive and real-time data on cellular responses within the 3D microenvironment.
Conclusions:
- The novel 3D cell culture system provides a rapid, reliable, and superior alternative to 2D models for studying cancer biology and evaluating therapeutic targets like USP37.
- Targeting ubiquitin specific peptidase 37 (USP37) represents a promising therapeutic strategy for various cancers, supported by evidence from this advanced in vitro model.
- This 3D platform facilitates mechanistic studies of oncogene function and drug response, accelerating the development of new cancer therapies.
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