Interrogating Matrix Stiffness and Metabolomics in Pancreatic Ductal Carcinoma Using an Openable Microfluidic...

Area of Science:

• Oncology
• Biomedical Engineering
• Microfluidics

Background:

• Pancreatic ductal adenocarcinoma (PDAC) features a dense fibrotic stroma, promoting tumor aggressiveness and treatment resistance.
• Existing in vitro models inadequately support visualization of fibrous networks, mechanical stiffness measurement, and metabolomic profiling of the stroma.
• Understanding tumor-stromal interactions is crucial for developing effective PDAC therapies.

Purpose of the Study:

• To develop and validate an advanced microfluidic platform for studying pancreatic cancer stroma.
• To enable detailed analysis of fibrous network structure, mechanical properties, and cellular metabolism within a PDAC model.
• To investigate the impact of pancreatic stellate cells (PSCs) on pancreatic tumor cell (PTC) behavior and the tumor microenvironment.

Main Methods:

• An openable multilayer microfluidic chip was engineered with pancreatic tumor cells (PTCs) and pancreatic stellate cells (PSCs) in a 3D collagen matrix.
• Reflected light confocal (RLC) microscopy was used for fibrous network visualization.
• Atomic force microscopy (AFM) assessed in situ mechanical stiffness.
• Compartmentalized hydrogel extraction coupled with mass spectrometry (MS) enabled PSC metabolomic profiling.

Main Results:

• RLC microscopy revealed decreased pore size and increased fiber density in cocultures compared to monocultures.
• AFM demonstrated significant increases in matrix stiffness in the presence of cocultured PSCs and PTCs.
• Fluorescence microscopy confirmed hallmark PSC activation, localized fiber alignment, densification, and increased collagen production.
• Mass spectrometry analysis identified metabolic profiles consistent with in vivo PDAC studies.

Conclusions:

• The developed microfluidic PDAC-on-a-chip platform effectively mimics the pancreatic tumor stroma.
• This platform facilitates advanced analysis of fibrous network dynamics, mechanical properties, and cellular metabolism.
• It offers a powerful tool for investigating tumor-stromal interactions and discovering novel therapeutic strategies for pancreatic ductal adenocarcinoma.