821: Metabolically Glycoengineered Stem Cell Therapy Improved Brain Recovery After Cardiac Arrest

Introduction: Cardiac arrest (CA) is a leading cause of global morbidity and mortality, often resulting in severe hypoxic-ischemic brain injury. Neural stem cell (NSC) therapy is a promising strategy to restore neural function after brain injury, however, major challenges remain, including functional integration of NSCs into injured brain tissue post-transplantation. Using the emerging metabolic glycoengineering (MGE) technique to chemically modify surface glycans, the non-natural, thiol-modified ManNAc analog TProp enhances cell-cell interactions, survival signals, and immune evasion for effective brain repair. The Wnt/β-catenin signaling pathway plays a pivotal role in regulating cell survival and differentiation. This study aims to evaluate TProp’s effects on brain recovery and Wnt/β-catenin pathway modulation on MGE-mediated neuroregeneration. Methods: Wnt signaling proteins in NSCs treated with PBS, TProp, and TProp+IWR-1 (Wnt signaling inhibitor) were assessed in vitro via immunofluorescence staining. Wistar rats were randomized to receive TProp-NSCs or TProp-NSC+IWR-1 (n = 6, 4 × 105 cells in 10 µL PBS) delivered via intracerebroventricular (ICV) administration 3 hours post-CA. Neurological recovery was evaluated via neurological deficit scores (NDS). The brain sections were stained using Fluoro-Jade C (FJC) to assess degenerating neuron levels. Results: β-catenin and GSK-3β expression were upregulated in TProp-NSCs, indicating Wnt pathway activation in vitro, whereas Wnt inhibition with IWR-1 reverses the upregulation. ICV transplantation of TProp-NSCs following CA significantly improved neurological scores, whereas Wnt inhibition reduced these benefits. Hippocampal FJC positive area percentage of the TProp-NSC group exhibited significantly reduced neuronal degeneration compared to the TProp-NSC + IWR group. Conclusions: This study signifies Wnt pathway activation in neuroprotection enabled by MGE, suggesting that TProp enhances neuroprotective effects via Wnt/β-catenin signaling, revealing a promising strategy to improve stem cell-based therapies for ischemic brain injury after CA. Supported in part by the R01NS125232 and R01NS110387 from the National Institute of Neurological Disorders and Stroke and 2024-MSCRFD-6401 from the Maryland Stem Cell Research Fund (all to X Jia).