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Updated: Jun 25, 2025
A Three-dimensional Model of Spheroids to Study Colon Cancer Stem Cells
Published on: January 22, 2021
Fluorescence-Guided Spatial Drug Screening in 3D Colorectal Cancer Spheroids
Jia Ning Nicolette Yau1,2, Thirumal Yempala1, Ram Pravin Kumar Muthuramalingam1
1Department of Pharmacy and Pharmaceutical Sciences, Faculty of Science, National University of Singapore, Singapore, 117544, Singapore.
Abstract:
The limited recapitulation of critical cancer features in 2D cultures causes poor translatability of preclinical results from in vitro assays to in vivo tumor models. This contributes to slow drug development with a low success rate. 3D cultures better recapitulate the tumor microenvironment, enabling more accurate predictions when screening drug candidates and improving the development of chemotherapeutics. Platinum (Pt) (IV) compounds are promising prodrugs designed to reduce the severe systemic toxicity of widely used Food and Drug Administration (FDA)-approved Pt(II) drugs such as cisplatin. Here, this work presents spatiotemporal evaluations in 3D colorectal cancer (CRC) spheroids of mitochondria-targeting Pt(IV) complexes. CRC spheroids provide a greater pathophysiological recapitulation of in vivo tumors than 2D cultures by a marked upregulation of the ABCG2 chemoresistance marker expression. Furthermore, new 3D-staining protocols are introduced to evaluate the real-time decrease in mitochondria membrane potential (ΔΨ) in CRC spheroids, and a Pt-sensing dye to quantify the Pt mitochondrial accumulation. Finally, this work demonstrates a correlation between in vitro results and the efficacy of the compounds in vivo. Overall, the CRC spheroids represent a fast and cost-effective model to assess the behavior of Pt compounds in vitro and predict their translational potential in CRC treatment.
Insights
This study uses 3D colorectal cancer (CRC) spheroids to test novel platinum (Pt) (IV) compounds, improving drug development. These 3D models accurately predict how Pt compounds perform in vivo, enhancing chemotherapy development.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Pharmacology
Background:
- 2D cell cultures poorly mimic cancer, hindering drug development success.
- 3D cultures better represent the tumor microenvironment and drug responses.
- Platinum (IV) compounds offer a less toxic alternative to platinum (II) drugs.
Purpose of the Study:
- To evaluate mitochondria-targeting platinum (IV) complexes in 3D colorectal cancer (CRC) spheroids.
- To develop new 3D staining methods for assessing drug effects on mitochondria.
- To correlate in vitro findings with in vivo efficacy for predicting drug translation.
Main Methods:
- Utilized 3D CRC spheroids to model tumor pathophysiology, including ABCG2 marker expression.
- Developed novel 3D staining protocols to measure real-time mitochondrial membrane potential (ΔΨ) changes.
- Employed a platinum-sensing dye to quantify platinum accumulation in mitochondria.
Main Results:
- 3D CRC spheroids exhibit enhanced chemoresistance marker expression compared to 2D cultures.
- New protocols successfully monitored mitochondrial dysfunction and platinum uptake in 3D models.
- Demonstrated a strong correlation between in vitro results and in vivo compound efficacy.
Conclusions:
- 3D CRC spheroids are a valuable model for assessing platinum compound behavior and predicting clinical success.
- This approach accelerates drug development and improves the translational potential of new chemotherapeutics.
- Mitochondria-targeting Pt(IV) complexes show promise for colorectal cancer treatment.
