Poster F3 - Measurement of the Contraction of Cancer-Associated Fibroblasts with Cancer Cell Conditioning

Friday, October 25, 2024

10:00 AM – 11:00 AM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

Jose Hernandez, Jr.

PhD Student
Rice University
Houston, Texas, United States

Co-Author(s)

Aja A. Phan (she/they)

Undergraduate Researcher
University of Illinois at Urbana-Champaign
Morton, Illinois, United States
AK

Ayesha Khan

Undergraduate Researcher
University of Illinois at Urbana-Champaign, United States
BE

Bashar Emon

Postdoctoral Research Associate
University of Illinois at Urbana-Champaign
Champaign, Illinois, United States

Last Author(s)

MS

M. Taher A. Saif

Professor
University of Illinois at Urbana-Champaign
Urbana, Illinois, United States

Introduction:

Introduction: Cancer-associated fibroblasts (CAFs) are stromal cells that have been shown to accelerate tumorigenesis and metastatic progression [1]. CAFs are capable of restructuring the tumor ECM by both depositing matrix and reorganizing it. However, the effects of this restructuring are still not well understood. In this study, we used a novel micromechanical device to measure the force of contraction of cells within collagen based on deflection within the device. The measurement allows for quantification of the collective forces of cells in contracting a matrix. We compared the force of pancreatic cancer cells to the force created by pancreatic stellate cells (a precursor to CAFs) with and without conditioned media.

Materials and

Methods: Materials and

Methods: A novel microfabricated PDMS mechanosensor was used to measure the matrix stiffness and cellular force (Figure 1) [2]. Cells are seeded in a collagen-based gel placed between the mechanosensors, and the deflection of the sensors is measured based on time-lapse imaging using microscopy. MiaPaCa cells (an epithelial cell line derived from a pancreatic cancer patient) and pancreatic stellate cells (PSCs, which are known to give rise to CAFs[3]) are used. Three conditions were tested: MiaPaCa cells alone (control), PSC cells alone, PSC with MiaPaCa Conditioned Media (CM).

Results, Conclusions, and Discussions: Results and

Discussion: The device consists of two aligned, microfabricated beams and connected by a bridge of collagen containing cells (Figure 1A). One beam sits atop rigid springs (high stiffness) and the other sits atop soft springs (low stiffness), which enables the soft beam to produce large deflection during cell contraction while the rigid beam remains stationary. As the cells contract, the sensor grips atop the soft beam are pulled toward the stationary rigid beam and the deflection is measured (Figure 1B). Using the spring constants from the soft beam, the force of deflection (and cell contraction) is calculated.

Our data indicates that the pancreatic stellate cells are more contractile than cancer cells (Figure 1C). Additionally, our data suggest that the rate of pancreatic stellate contractility is similar for both the PSC+CM and PSC-only conditions, as the plot of force versus time is similar in steepness for both configurations.

Conclusion: Our findings demonstrate a novel platform for quantifying cell contractility. CAF precursors are more contractile than cancer cells, suggesting they are the primary mediators of tumor ECM remodeling. Moreover, the presence of cancer cell-conditioned media increases their contractility rate, suggesting there is a crosstalk between the cells based on soluble factors. Future work should investigate the specific soluble factors released by cancer cells that are contributing to CAF contractility. Additionally, additional cancer and CAF cell lines can be used to determine if the same mechanical relationship persists. Given that many CAF sub-types have emerged in the literature, it is particularly important to determine the CAF phenotypes that elicit the most robust mechanical responses.

Acknowledgements (Optional):