Related Experiment Video
Updated: Aug 11, 2026
A Mouse Model to Investigate the Role of Cancer-Associated Fibroblasts in Tumor Growth
Published on: December 22, 2020
In vitro models of cancer-associated fibroblast heterogeneity uncover subtype-specific effects of CRISPR
Elysia Saputra1, Shamsudheen Karuthedath Vellarikkal1, Lixia Li1
1Department of Data, AI, and Genome Sciences, Merck & Co., Inc., Rahway, NJ, USA.
Abstract:
Cancer-associated fibroblasts (CAFs) are sought after as potential therapeutic targets due to their pro- and antitumorigenic functions, which are attributed to specializations in CAF subtypes. A precise targeting of specific subtypes would be required to design therapies that effectively modulate CAF phenotypes, necessitating translatable model systems to support target discovery efforts. However, not only is our knowledge of CAF heterogeneity in solid tumors lacking, particularly in pancreatic tumors, but the translatability of CAF models has also not been rigorously evaluated. Here, we develop a coculturing model with primary CAFs and immortalized tumor cell lines that can reliably represent CAF phenotypes observed in tumors, with correlations to immuno-resistant and immunomodulatory phenotypes. Using single-cell transcriptomics, we characterize the CAF subtype heterogeneity in the in vitro CAF cell lines isolated from pancreatic cancer patients and investigate the impact of perturbing potential stromal genes on different CAF subtypes. We also infer the continuum of state changes underlying the interconvertibility of CAF subtypes. Finally, we use immortalized CAF cell lines to perform single-cell CRISPR perturbations of stromal targets, revealing the subtype-specific effects of perturbations and the impact of model-type selection on the translatability of insights.
Insights
Cancer-associated fibroblasts (CAFs) exhibit diverse subtypes with varying roles in cancer. This study develops a novel model to accurately represent CAF heterogeneity and subtype-specific responses to therapeutic targets in pancreatic cancer.
Area of Science:
- Oncology
- Cancer Biology
- Fibroblast Research
Background:
- Cancer-associated fibroblasts (CAFs) are crucial in tumor progression, exhibiting diverse subtypes with pro- and anti-tumorigenic functions.
- Understanding CAF heterogeneity is vital for developing targeted cancer therapies, yet knowledge, especially in pancreatic cancer, and model translatability remain limited.
Purpose of the Study:
- To develop and validate a co-culturing model for CAFs that accurately reflects in vivo heterogeneity.
- To characterize CAF subtypes in pancreatic cancer and assess their phenotypes, including immunomodulatory roles.
- To investigate the impact of targeting stromal genes on CAF subtypes and evaluate model translatability.
Main Methods:
- Development of a co-culturing model using primary CAFs and immortalized tumor cell lines.
- Single-cell transcriptomics to analyze CAF subtype heterogeneity in pancreatic cancer.
- Single-cell CRISPR perturbations to assess the effects of stromal gene targets on CAF subtypes.
Main Results:
- The co-culturing model reliably represents CAF phenotypes, correlating with immuno-resistant and immunomodulatory characteristics.
- Single-cell transcriptomics revealed significant CAF subtype heterogeneity in pancreatic cancer.
- CRISPR perturbations demonstrated subtype-specific effects of stromal gene targeting, highlighting the importance of model selection.
Conclusions:
- A novel co-culturing model effectively recapitulates CAF heterogeneity and phenotypes relevant to pancreatic cancer.
- Targeting specific CAF subtypes offers a promising therapeutic strategy, but requires careful consideration of model systems for translatability.
- Further research into CAF interconvertibility and subtype-specific targeting is warranted for effective cancer therapy development.
Related Concept Videos
01:59CRISPR
02:31Homologous Recombination
01:28CRISPR/Cas9 Genome Editing