The composition and mechanical compliance of the extracellular matrix (ECM) have been shown to serve as regulators of tumor growth and invasive behavior. These effects may be particularly relevant in tumors of the pancreas, noted for a profound desmoplastic reaction and an abundance of stroma rich in ECM. In view of recent progress in the clinical implementation of photodynamic therapy (PDT) for pancreatic tumors, in this report we examine how ECM composition and rheological properties impact upon invasive behavior and response to PDT in 3D multicellular pancreatic tumor spheroids in ECM environments with characterized rheological properties. Tumor spheroids were cultured initially in attachment-free conditions to form millimeter-sized spheroids that were transplanted into reconstituted ECM microenvironments (Matrigel and Type I Collagen) that were characterized using bulk oscillatory shear rheology. Analysis of growth behavior shows that the soft collagen ECM promoted growth and extensive invasion and this microenvironment was used in subsequent assessment of PDT and chemotherapy response. Evaluation of treatment response revealed that primary tumor nodule growth is inhibited more effectively with PDT, while verteporfin PDT response is significantly enhanced in the ECM-infiltrating populations that are non-responsive to oxaliplatin chemotherapy. This finding is potentially significant, suggesting the potential for PDT to target these clinically problematic invasive populations that are associated with aggressive metastatic progression and chemoresistance. Experiments to further validate and identify the mechanistic basis of this observation are ongoing.
Particle tracking microrheology (PTM) has recently been employed as a non-destructive way to longitudinally track physical changes in 3D pancreatic tumor co-culture models concomitant with tumor growth and invasion into the extracellular matrix (ECM). While the primary goal of PTM is to quantify local viscoelasticity via the Generalized Stokes-Einstein Relation (GSER), a more simplified way of describing local tissue mechanics lies in the tabulation and subsequent visualization of the spread of probe displacements in a given field of view. Proper analysis of this largely untapped byproduct of standard PTM has the potential to yield valuable insight into the structure and integrity of the ECM. Here, we use clustering algorithms in R to analyze the trajectories of probes in 3D pancreatic tumor/fibroblast co-culture models in an attempt to differentiate between probes that are effectively constrained by the ECM and/or contractile traction forces, and those that exhibit uninhibited mobility in local water-filled pores. We also discuss the potential pitfalls of this method. Accurately and reproducibly quantifying the boundary between these two categories of probe behavior could result in an effective method for measuring the average pore size in a given region of ECM. Such a tool could prove useful for studying stromal depletion, physical impedance to drug delivery, and degradation due to cellular invasion.