Abstract DP196: Sex-specific Efficacy of Intestinal Organoid Secretomes in Stroke Recovery: Linking the Gut to Neuroprotection

Background: Stroke induces acute intestinal epithelial dysfunction and increased gut permeability, contributing to systemic inflammation and neurotoxicity. We previously demonstrated that transplantation of intestinal epithelial stem cells (IESCs) from young donors mitigates both acute and chronic stroke deficits in aging animals. However, IESCs rarely engraft or differentiate in vivo, suggesting that their therapeutic efficacy is mediated largely by paracrine secretions. Here, we tested the hypothesis that IESC organoid–derived conditioned media (IESC-CM) are sufficient to confer neuroprotection by restoring gut barrier integrity and attenuating systemic inflammation, thereby improving acute and chronic stroke outcomes. Methods: IESCs isolated from young male or female rats were expanded as intestinal oganoids and conditioned media (IESC-CM) was collected. Middle-aged male and female rats were subjected to endothelin-1–induced middle cerebral artery occlusion (MCAo) and randomized to vehicle or IESC-CM treatment groups. IESC-CM was administered intraperitoneally at 4, 24, and 48 h post-stroke. Infarct volume and sensorimotor function were assessed at 4 d post-stroke. At 30 d, animals were tested for Social Interaction, Novel Object Recognition test, and Barnes maze . Circulating gut permeability markers and cytokines were quantified at acute and chronic phases. Results: IESC-CM significantly reduced infarct volume, improved sensorimotor recovery, and attenuated gut permeability in both sexes in the acute phase. In the chronic phase, vehicle-treated MCAo animals exhibited depressive-like behaviors and impaired cognition, whereas IESC-CM treatment normalized affective behaviors in both sexes. Strikingly, cognitive benefits were sex-dependent: IESC-CM improved episodic memory, spatial learning, and retrieval in males, but not in females. This sex-specific effect was paralleled by differential cytokine modulation, with IL-17A levels significantly reduced by IESC-CM in males but unchanged in females. Conclusion: These findings demonstrate that IESC-derived secretomes confer robust neuroprotection acutely in both sexes, but promote long-term cognitive recovery preferentially in males. The divergence may reflect sex differences in the composition of the IESC secretome, resulting in greater suppression of proinflammatory cytokines and and enhanced gut restoration in males as compared to females.