Interfacial Curvature in Confined Coculture Directs Stromal Cell Activity with Spatial Corralling of Pancreatic Cancer Cells

Abstract

Interfacial cues in the tumor microenvironment direct the activity and assembly of multiple cell types. Pancreatic cancer, along with breast and prostate cancers, is enriched with cancer‐associated fibroblasts (CAFs) that activate to coordinate the deposition of the extracellular matrix, which can comprise over 90% of the tumor mass. While it is clear that matrix underlies the severity of the disease, the relationship between stromal‐tumor cell assembly and cell‐matrix dynamics remains elusive. Micropatterned hydrogels deconstruct the interplay between matrix stiffness and geometric confinement, guiding heterotypic cell populations and matrix assembly in pancreatic cancer. Interfacial cues at the perimeter of microislands guide CAF migration and direct cancer cell assembly. Computational modeling shows curvature‐stress dependent cellular localization for cancer and CAFs in coculture. Regions of convex curvature enhance edge stress that activates a myofibroblast phenotype in the CAFs with migration and increased collagen I deposition, ultimately leading to a central “corralling” of cancer cells. Inhibiting mechanotransduction pathways decreases CAF activation and the associated corralling phenotype. Together, this work reveals how interfacial biophysical cues underpin aspects of stromal desmoplasia, a hallmark of disease severity and chemoresistance in the pancreatic, breast, and prostate cancers, thereby providing a tool to expand stroma‐targeting therapeutic strategies.

Defined Microenvironments Trigger In Vitro Gastrulation in Human Pluripotent Stem Cells

“…Previously we demonstrated how changes in perimeter curvature would influence the behavior of microconfined cells, where geometry and stiffness both exerted an influence over cell phenotype in the context of cancer stemness. [ 30 , 31 ] To evaluate whether geometry would play a role in the observed differentiation with our microconfined hiPSC colonies, we cultured cells in shapes of the same area approximating a star, flower, square, and capital “I”—where positive and negative curvature and aspect ratio are varied—followed by immunostaining for germ layer markers. After 48 h there were no significant differences between mes‐endodermal marker expression across the shaped colonies, suggesting changes in geometry at the interface does not play a major role in the observed gastrulation‐like morphogenesis (Figure S8 , Supporting Information).…”

3D Printable Gelatin Methacryloyl (GelMA)‐Dextran Aqueous Two‐Phase System with Tunable Pores Structure and Size Enables Physiological Behavior of Embedded Cells In Vitro

“…[ 42 ] To summarize, we determined that pore size can shape the distribution of cells and their spread in 3D hydrogels confirming previous findings. [ 66 ]…”

Defined microenvironments trigger in vitro gastrulation in human pluripotent stem cells

“…Previously we demonstrated how changes in perimeter curvature would influence the behaviour of microconfined cells, where geometry and stiffness both exerted an influence over cell phenotype in the context of cancer stemness. 21, 22 . To evaluate whether geometry would play a role in the observed differentiation with our microconfined hiPSC colonies, we cultured cells in shapes of the same area approximating a star, flower, square, and capital ‘I’—where positive and negative curvature and aspect ratio are varied—followed by immunostaining for germ layer markers.…”

“…Mechanotransduction propagates through integrin-mediated adhesion, actomyosin contractility and mitogen activated protein kinase signalling, where numerous downstream effectors are employed by both matrix engagement and soluble morphogens 22 . Here we showed how adhesion to deformable hydrogels leads to enhanced cytoskeletal tension and YAP activity at the boundary which triggers EMT across the colony.…”