Designer DNA Hydrogels Stimulate 3D Cell Invasion by Enhanced Receptor Expression and Membrane Endocytosis.

Shanka Walia,Supriyo Naskar,Vinod Morya,Chinmay Ghoroi,Ankit Gangrade,Dhiraj Bhatia,Prabal K Maiti,Sameer Dalvi,Aditya Guduru Teja

doi:10.1021/acsbiomaterials.1c01085

Abstract

DNA has emerged as one of the smartest biopolymers to bridge the gap between chemical science and biology to design scaffolds like hydrogels by physical entanglement or chemical bonding with remarkable properties. We present here a completely new application of DNA-based hydrogels in terms of their capacity to stimulate membrane endocytosis, leading to enhanced cell spreading and invasion for cells in ex vivo 3D spheroids models. Multiscale simulation studies along with DLS data showed that the hydrogel formation was enhanced at lower temperature and it converts to liquid with increase in temperature. DNA hydrogels induced cell spreading as observed by the increase in cellular area by almost two-fold followed by an increase in the receptor expression, the endocytosis, and the 3D invasion potential of migrating cells. Our first results lay the foundation for upcoming diverse applications of hydrogels to probe and program various cellular and physiological processes that can have lasting applications in stem cell programming and regenerative therapeutics.

Highlights

The extracellular matrix (ECM) is a 3D natural scaffold produced by different proteins such as collagen, proteoglycans, fibronectin, elastin, and cell-binding glycoproteins.[1]
Different types of DNA hydrogels were assembled for motifs modified from previously published results.[21]
We modified the earlier reported oligonucleotide sequences to ensure higher thermal stability by increasing the G−C content in the sticky ends. Both four-way junction and three-way junction DNA hydrogels were prepared by the selfannealing method (Figure 1d)

Summary

Introduction

The extracellular matrix (ECM) is a 3D natural scaffold produced by different proteins such as collagen, proteoglycans, fibronectin, elastin, and cell-binding glycoproteins.[1]. Different polymers used to synthesize hydrogels include poly(vinyl alcohol) (PVA), poly(ethylene oxide) (PEO), and poly(acrylic acid) (PAA).[13] These materials are biocompatible as well as biodegradable, but on hydrolytic biodegradation, they release toxic acids that can be harmful for the cells Their hydrophobic nature can lead to nonspecific adsorption of proteins, resulting in uncontrolled and unwanted cell interactions.[13,14] to overcome these drawbacks, researchers have used biologically derived materials such as hyaluronic acid, elastin, and collagen as the cell matrix. Because of their inherent bioactivity, reduced stiffness, tedious synthesis protocols, and variability when synthesized in batches, they were difficult to control and can affect immune responses when implanted.[15,16]

Methods

Results

Conclusion