Microglial voltage-gated proton channel 1 ablation in diabetic mice mitigates diabetes-driven demyelination and cognitive decline

Table 1 Key strengths and innovations

Aspect	Summary	Supporting data
Multimodal approach	In vivo (HFD + STZ, Hv1 KO) and in vitro (high-glucose + siHv1) models converge on Hv1’s role	Behavioral (Figure 1 in the study of Li et al[1]), IF (Figures 2 and 6 in the study of Li et al[1]), TEM (Figure 5 in the study of Li et al[1]), ferroptosis markers (Figure 7 in the study of Li et al[1])
Functional rescue	Hv1 KO restores working/spatial memory deficits	40% fewer errors; 30% more entries (Figure 1C and D in the study of Li et al[1])
White matter protection	TEM shows normalization of g-ratio from 0.81 (diabetic) to 0.78 (KO)	Figure 5A-F in the study of Li et al[1]

HFD: High-fat diet; STZ: Streptozotocin; Hv1: Voltage-gated proton channel 1; KO: Knockout; siHv1: Small interfering RNA-mediated voltage-gated proton channel 1; IF: Immunofluorescence; TEM: Transmission electron microscope.

Table 2 Points for further consideration

Focus area	Action	Outcome measure
Longitudinal efficacy	Behavioral tests + diffusion MRI at 2, 8, 16 weeks	Memory scores; fractional anisotropy
Cell-type specificity	Generate CX3CR1-CreERT2 Hv1flox/flox for microglia-only KO; compare GFAP-Cre astrocyte KO	Cytokine levels; OPC survival; g-ratio
Pharmacological proof-of-concept	Screen Hv1 inhibitors for IC50, BBB penetration, and toxicity; test alone and with remyelinating agents	Patch-clamp IC50; brain/plasma ratio; maze performance
Mechanistic clarification	pH imaging in microglia under hyperglycemia; Co-IP of Hv1 and NOX2	Intracellular pH; ROS assays; protein interaction

MRI: Magnetic resonance imaging; Hv1: Voltage-gated proton channel 1; CX3CR1: CX3C chemokine receptor 1; KO: Knockout; OPC: Oligodendrocyte progenitor cell; BBB: Blood-brain barrier; Co-IP: Co-immunoprecipitation; NOX2: NADPH oxidase 2; ROS: Reactive oxygen species.

Table 3 Clinical implications and future directions

Goal	Strategy	Readouts
Early intervention window	Hv1 inhibitors initiated at MRI-detected white matter changes or mild cognitive impairment	Diffusion MRI metrics; cognitive test scores
Enhanced remyelination	Combine Hv1 blockade with OPC-differentiation drugs (e.g., clemastine)	Myelin thickness; OPC maturation markers
Biomarker development	PET tracer or CFS assay for Hv1/IL-1β/GRP78	Imaging signal intensity; CFS levels
Broader disease scope	Evaluate Hv1 targeting in MS, Alzheimer’s, and stroke models	Disease-specific lesion load; behavioral assays

Hv1: Voltage-gated proton channel 1; MRI: Magnetic resonance imaging; OPC: Oligodendrocyte progenitor cell; PET: Positron emission tomography; CFS: Chronic fatigue syndrome; IL-1β: Interleukin-1β; GRP78: Glucose-regulated protein 78; MS: Mass spectrometry.