Role of immediate early genes in the development of salivary gland organoids in polyisocyanopeptide hydrogels

Abstract

Human salivary gland organoids have opened tremendous possibilities for regenerative medicine in patients undergoing radiotherapy for the treatment of head and neck cancer. However, their clinical translation is greatly limited by the current use of Matrigel for organoid derivation and expansion. Here, we envisage that the use of a fully, synthetic hydrogel based on the oligo (-ethylene glycol) functionalized polymer polyisocyanopeptides (PICs) can provide an environment suitable for the generation and expansion of salivary gland organoids (SGOs) after optimization of PIC polymer properties. We demonstrate that PIC hydrogels decorated with the cell-binding peptide RGD allow SGO formation from salivary gland (SG)-derived stem cells. This self-renewal potential is preserved for only 4 passages. It was found that SGOs differentiated prematurely in PIC hydrogels affecting their self-renewal capacity. Similarly, SGOs show decreased expression of immediate early genes (IEGs) after culture in PIC hydrogels. Activation of multiple signalling pathways involved in IEG expression by β-adrenergic agonist isoproterenol, led to increased stem cell self-renewal capacity as measured by organoid forming efficiency (OFE). These results indicate that PIC hydrogels are promising 3D matrices for SGOs, with the option to be used clinically, after further optimization of the hydrogel and culture conditions.

Conductive Polyisocyanide Hydrogels Inhibit Fibrosis and Promote Myogenesis

“…We use hydrogels based on polyisocyanides (PICs), which form a relatively new class of tunable soft materials . By tailoring the molecular weight and polymer concentration, the mechanical properties and network architecture can be controlled for native tissue. , As a result, the material has been successfully applied as a matrix material for 3D in vitro studies to study single cell , and organoid behavior. , Additional in vivo experiments indicate that PIC hydrogels are nontoxic and biocompatible. , In these applications, PIC is biofunctionalized with the commonly used cell-adhesive peptide Gly-Arg-Gly-Asp-Ser (RGD) that allows cell–matrix interactions via integrin binding, but other biological cues can be inserted similarly . These and many other studies demonstrate the capacity of PIC gels to combine a high degree of tunability and similar mechanical properties as native tissue, which makes them particularly suitable for in vitro studies and different tissue engineering purposes.…”

“… 14 , 16 As a result, the material has been successfully applied as a matrix material for 3D in vitro studies to study single cell 17 , 18 and organoid behavior. 19 , 20 Additional in vivo experiments indicate that PIC hydrogels are nontoxic and biocompatible. 21 , 22 In these applications, PIC is biofunctionalized with the commonly used cell-adhesive peptide Gly-Arg-Gly-Asp-Ser (RGD) that allows cell–matrix interactions via integrin binding, but other biological cues can be inserted similarly.…”

TempEasy 3D Hydrogel Coculture System Provides Mechanistic Insights into Prostate Cancer Bone Metastasis

“…Earlier, we developed TempEasy, a user-friendly indirect 3D coculture system. This system employs the synthetic oligo­(ethylene glycol)-substituted polyisocyanide (PIC) gels. , Mounting experimental evidence from a wide range of in vitro cell culture and organoid − studies as well as in vivo animal experiments − support the outstanding biocompatibility of PIC hydrogel. TempEasy capitalizes on the thermoreversible gelation of the PIC gels.…”

Organoid-based personalized medicine: from tumor outcome prediction to autologous transplantation

“… 18 ). However, synthetic hydrogels do not always possess the same growth promoting properties as Matrigel, such as not supporting extensive self-renewal and promoting stem cell differentiation ( 19 , 20 ). Recent efforts have yielded the first fully synthetic matrices supporting serial passaging of epithelial organoids, 21 , 22 but their adoption by more research groups and their usage in broader organoid types remains to be determined.…”