DP30: ULTRAVIOLET LIGHT-ACTIVATED GABA DELIVERY USING ENGINEERED STEM CELLS
Natasha Doshi, BS; Andres Salvador, BS
Lake Erie College of Osteopathic Medicine
Introduction: Chronic pain is a complex condition with a significant impact on individuals and healthcare systems worldwide. Traditional treatments often fall short, leading to substantial physical, emotional, and economic burdens thereby highlighting the need for more effective therapies. Existing treatments often lead to limited efficacy, risk of tolerance, and a multitude of adverse side effects. Gamma-aminobutyric acid (GABA), the primary inhibitory neurotransmitter in the central nervous system, plays a crucial role in pain modulation. Dysregulated GABAergic signaling is implicated in chronic pain states, making it a promising therapeutic target. GABAergic dysfunction may be associated with impaired inhibitory signaling involving hyperalgesia and allodynia. This study explores an innovative approach using genetically engineered pluripotent stem cells (PSCs) differentiated into GABAergic neurons. By leveraging optogenetic technology, these neurons were designed to release GABA upon ultraviolet (UV) light stimulation, offering a precise and localized method for pain modulation.
Methods: Skin-derived induced pluripotent stem cells (iPSCs) were differentiated into GABAergic neurons in a neural induction medium. Genetic modification was performed using a modified adenoviral vector to introduce channelrhodopsin, enabling light-activated neurotransmitter release. Fluorescent staining techniques, including markers for GABAergic identity (GAD67) and neuronal lineage (βIII-tubulin, NeuN), confirmed successful differentiation. Blue light stimulation was applied using a UV LED device, and GABA levels were quantified using enzyme-linked immunosorbent assay (ELISA). Time-dependent responses were monitored to determine the duration and peak of GABA release.
Results: Experimental findings demonstrated a significant increase in GABA secretion upon light stimulation. The average GABA concentration in stimulated groups was 115 pg/µg, compared to 34 pg/µg in control groups (p < 0.05). Peak GABA release occurred approximately 210 seconds post-stimulation, with a drop-off observed after 7 minutes, suggesting an optimal therapeutic window for pain modulation. Fluorescent staining confirmed successful differentiation of iPSCs into functional GABAergic neurons.
Discussion/Conclusion: These results support the feasibility of using optogenetically engineered GABAergic neuronal stem cells for targeted pain management. The ability to achieve precise, on-demand GABA release through light stimulation offers a novel strategy to address chronic pain while minimizing systemic side effects associated with traditional analgesics. The observed drop in GABA secretion after 7 minutes highlights the need for further exploration of stimulation protocols to sustain therapeutic effects. Future research should focus on in vivo validation, long-term viability, and the integration of these cells within implantable delivery systems to enhance clinical applicability. This study provides a foundation for the development of patient-specific, stem cell-based therapies in chronic pain treatment.