Evaluation of Polymer Induced Biostasis via MALDI Imaging Mass Spectrometry

Special circumstances surrounding any change from the Grantee's proposal: Receipt of instrument was delayed due to multiple COVID-associated shutdowns of the production facility in the UK. An instrument with some similar capability was provided on loan (MALDI 8020)allowing work to proceed until the MALDI 7090 instrument was received. A concise discussion of the use of the equipment including (a) any research work described in the proposal, and (b) any other research of interest to DoD: The labs of Drs. Christopher Bowman, Kristi Anseth and Sabrina Spencer are recipients of a grant from DARPA (W911NF-19-2-0024) to induce and reverse biostasis in living cells and tissues via intracellular crosslinking and degradation of hydrophilic polymers. Following cellular uptake of these polymers, an exogenous trigger causes them to form a network architecture that diminishes the diffusion of biomacromolecules (e.g. proteins, nucleic acids) and thereby mitigate the performance of cellular processes including those that might be deleterious (e.g. apoptosis, necrosis, differentiation) to biomedical interventions. An orthogonal trigger is subsequently employed to degrade the network, returning the cells to their prior state, effectively reversing biostasis. It is envisioned that this approach may be used to stabilize biological-based therapeutics at room temperature, mitigating the dependence of such materials on the cold chain for storage and transport. Moreover, this technology could be used to stabilize injury at the site of occurrence for subsequent movement to and treatment of patients at appropriate medical facilities. The characterization of these polymers both outside of and within biological contexts is key to accomplishing and progressing the objectives of this project and MALDI is a powerful tool in that respect.