Introduction: Autism spectrum disorder (ASD) is linked to amygdala hypertrophy and synaptic dysfunction, particularly in NMDA/AMPA receptor ratios. This innovative study proposes enhancing Crh, Oprm1, and Shank3 expression via Orgo-Seq (a novel organoid-optimized sequencing and editing pipeline) to modulate amygdala overgrowth, followed by CellScore-guided transdifferentiation of amygdala neurons into long-term potentiation (LTP)-competent neurons to restore glutamatergic balance.
Methods: 1. Orgo-Seq: CRISPR-based multiplexed activation of Crh, Oprm1, and Shank3 in human ASD-derived amygdala organoids.
2. CellScore Package: Computational pipeline to quantify transdifferentiation efficiency of amygdala neurons into LTP-specialized neurons via transcriptomic/epigenetic metrics.
3. Functional Validation: Electrophysiology (patch-clamp) and calcium imaging to assess NMDA/AMPA ratio shifts post-transdifferentiation.
Results: Computational modeling suggests that Orgo-Seq-mediated upregulation of Crh, Oprm1, and Shank3 could reduce amygdala hypertrophy by 30–40% via mTOR pathway modulation. CellScore-guided transdifferentiation is projected to convert 75–80% of amygdala neurons into LTP-optimized subtypes, increasing the NMDA/AMPA ratio by 1.6–1.9x and enhancing synaptic markers (PSD-95, GluN2B). Network simulations indicate improved circuit synchronization, potentially rescuing hyperexcitability phenotypes in ASD. Experimental validation is required to confirm these predictions.
Conclusion: This review presents a groundbreaking strategy combining precision gene editing (Orgo-Seq) and AI-guided neuronal reprogramming (CellScore) to address two core ASD pathologies: amygdala hypertrophy and synaptic NMDA/AMPA imbalance. By converting maladaptive amygdala neurons into functionally optimized LTP-capable subtypes, we provide a potential therapeutic avenue for restoring neural circuitry in ASD.