Assembling and decorating hyaluronan hydrogels with twin protein superglues to mimic cell-cell interactions

Abstract

Simple polymeric scaffolds have yielded dramatic effects on cell behavior. For more sophisticated phenotypes, precise and efficient chemistries are desired to incorporate proteins into these scaffolds. Here we derivatize hyaluronan with an elastin-like polypeptide containing telechelic SpyTags (HA-SpyTag). Our second network component, the TriCatcher protein, had two SpyCatchers and a terminal SnoopCatcher. Mixing HA-SpyTag with TriCatcher led to rapid hydrogel formation, via spontaneous amidation. SnoopCatcher allowed modular network decoration with SnoopTagJr-linked adhesion molecules, through orthogonal transamidation. This programmed scaffold enables the testing of how individual matrix-anchored protein interactions affect cell behavior. Epithelial cell adhesion molecule (EpCAM) regulates cell behavior and migration, with important effects in cancer. EpCAM-anchoring to the hydrogel induced disassembly of non-malignant mammary spheres in 3D culture. Integrating signaling proteins into biomaterials via efficient biocompatible chemistry should reveal key cues to control cell behavior.

Glycosaminoglycan-Inspired Biomaterials for the Development of Bioactive Hydrogel Networks

“…Later, the authors developed a similar system based on a different bacterial protein (RrgA), yielding the modules SnoopTag and SnoopCatcher ( Figure 4D(i), right), which do not cross-react with SpyTag/SpyCatcher [55]. More recently, the group combined these systems with peptide sequences for cell adhesion (RGD) and MMP-sensitive motifs cleavage to produce a HA hydrogel [56]. SpyTag/SpyCatcher containing these peptide sequences were used for covalent crosslinking between HA chains, and SnoopTag/SnoopCatcher were used to introduce any ligand of interest, such as epithelial cell adhesion molecule (EpCAM) or E-cadherin ( Figure 4D(ii)) [56].…”

“…More recently, the group combined these systems with peptide sequences for cell adhesion (RGD) and MMP-sensitive motifs cleavage to produce a HA hydrogel [56]. SpyTag/SpyCatcher containing these peptide sequences were used for covalent crosslinking between HA chains, and SnoopTag/SnoopCatcher were used to introduce any ligand of interest, such as epithelial cell adhesion molecule (EpCAM) or E-cadherin ( Figure 4D(ii)) [56]. Embedded fibroblasts were viable within these hydrogels and able to spread when RGD was present [56].…”

“…SpyTag/SpyCatcher containing these peptide sequences were used for covalent crosslinking between HA chains, and SnoopTag/SnoopCatcher were used to introduce any ligand of interest, such as epithelial cell adhesion molecule (EpCAM) or E-cadherin ( Figure 4D(ii)) [56]. Embedded fibroblasts were viable within these hydrogels and able to spread when RGD was present [56]. Non-malignant mammary cell spheres were also embedded in the hydrogels as the authors aimed at mimicking the interface between cancer cells and healthy cells by mediating the display of EpCAM [56].…”

“…Embedded fibroblasts were viable within these hydrogels and able to spread when RGD was present [56]. Non-malignant mammary cell spheres were also embedded in the hydrogels as the authors aimed at mimicking the interface between cancer cells and healthy cells by mediating the display of EpCAM [56]. EpCAM is a marker for cancer stem cells (CSC), and EpCAM positive breast cells are associated with CSC-like phenotype with increased metastatic ability [57].…”

“…EpCAM is a marker for cancer stem cells (CSC), and EpCAM positive breast cells are associated with CSC-like phenotype with increased metastatic ability [57]. Even though cells were viable, presentation of bound EpCAM led to the dissociation of the embedded mammary cell spheres, as opposed to hydrogels with no EpCAM [56]. Thus, this type of strategy can be a way to produce GAG covalent hydrogels and effectively present biomolecules of interest.…”