Using DNA hydrogels to mimic, exploit, and fundamentally investigate extracellular matrices - Project Summary Synthetic hydrogel biomaterials hold great potential as cell culture scaffolds, but their complexity falls short of mimicking the intricacies of biological systems, limiting their clinical efficacy. To overcome this, the goal of this MIRA application is to develop a versatile, DNA-based synthetic extracellular matrix (ECM) mimic that can be precisely controlled, manipulated, and degraded. This research seeks to not only enhance regenerative medicine applications but also provide deeper insights into cell biology and ECM interactions. Our hypothesis is that by harnessing DNA's unique tertiary and quaternary structural properties, we can emulate complex mechanical features of the ECM in a controllable and orthogonal manner. DNA's distinct role in biology and its absence in native ECM make it biocompatible and well-suited as a synthetic cell matrix. However, cost and scalability have limited the use of DNA-based ECM mimics. Fortunately, recent advancements in large-scale production of cGMP-grade dsDNA for vaccines have made it dramatically more accessible and provide a starting material ripe for rapid clinical translation. We have already made significant progress in producing high-purity dsDNA and developing DNA-based hydrogels with ECM-like properties, including non-linear elasticity and tunable viscoelasticity. These hydrogels exhibit promise in encapsulating cells without cytotoxicity and offer the potential for controlling tissue-mimetic properties. Our future research program is divided into three thrusts. In Thrust 1, we aim to develop DNA hydrogels with tunable ECM-mimetic properties and study their interactions with embedded cells. We will independently explore tuning non-linear elasticity and viscoelasticity, providing valuable insights into cell behavior under hard to emulate mechanical conditions. Thrust 2 focuses on dynamic modulation of DNA hydrogel properties, allowing for the controllable actuation of changes in elasticity and viscoelasticity over time. This approach mimics the dynamic manipulation of ECM during biological processes such as disease onset or healing and aims to provide a deeper understanding of cell responses to changing mechanical properties. In Thrust 3, we investigate the expression and release of small proteins and growth factors directly from the DNA matrix, enhancing nutrient transport and enabling final release from the synthetic matrix. Overall, this research program aims to demonstrate the broad utility of DNA-based hydrogels as ECM mimics applications in regenerative medicine and fundamental studies of biophysics. These hydrogels have the potential to revolutionize other fields, from tissue engineering to drug delivery, biosensing, and beyond.
