X inactive specific transcript drives mitochondrial metabolic rewiring and neural stem cell fate after spinal cord injury

Table 2 Prioritized research agenda and experimental strategies for X-chromosome inactivation-centered metabolic interventions in spinal cord injury

Research priority	Current gap (critique of evidence)	Suggested experimental approach (specific parameters)	Expected outcome	Ref.
Precision histomorphology and lineage specificity	Insufficiency of specialized histological characterization; lack of specific parameters to confirm NSC terminal differentiation	Confocal co-localization analysis (e.g., BrdU/NeuN for neurogenesis; synaptophysin for synaptic integration); integration of snRNA-seq and spatial transcriptomics	High-resolution mapping of XIST-mediated lineage commitment and functional integration across neural/glial subpopulations	Li et al[7], 2024; Zhang et al[32], 2024
Long-term structural and functional stability	Absence of 12-24 week chronic-phase data; limited correlation between short-term metabolic gains and longitudinal functional trajectories	Standardized chronic injury paradigms (compression/contusion); 12-24 weeks follow-up; multiparametric MRI/DTI coupled with composite behavioral metrics (gait analysis, grid walking)	Verification of sustained regenerative benefits, scar-stabilizing effects, and long-term structure-function correlations in chronic SCI	Rosenzweig et al[35], 2018; Chen et al[33], 2023
Metabolic safety and targeted delivery systems	Systemic metabolic risks of non-specific CPT1A intervention; demand for scalable, CNS-oriented and bio-responsive delivery platforms	Cell-type-specific Cpt1a cKO or knock-in models; engineered exosomes (EVs) or LNPs integrated into hydrogel scaffolds	Establishment of an optimized therapeutic window and clinically feasible “gene-cell-vector” platforms for spinal cord repair	Morant-Ferrando et al[12], 2023; Williams et al[34], 2025

NSC: Neural stem cell; BrdU: 5-bromo-2’-deoxyuridine; NeuN: Neuronal nuclei; snRNA-seq: Single-nucleus RNA sequencing; XIST: X-inactive specific transcript; MRI: Magnetic resonance imaging; DTI: Diffusion tensor imaging; SCI: Spinal cord injury; CPT1A: Carnitine palmitoyl transferase 1A; CNS: Central nervous system; cKO: Conditional knockout; EVs: Extracellular vesicles; LNPs: Lipid nanoparticles.