Abstract
Pathogenic variants in ATP6V1B2, which encodes a critical subunit of vacuolar-type H+-ATPases (V-ATPases), disrupt lysosomal acidification via haploinsufficiency and clinically manifest as intellectual disability and seizure disorders. Despite significant morbidity, mechanism-based therapies remain an unmet need. Through integrated clinical analysis of a Chinese cohort and systematic literature review, we delineated genotype-phenotype correlations in ATP6V1B2-related syndromes. Isogenic HEK293T models (ATP6V1B2R506X/+ and ATP6V1B2R506X/R506X) were generated using CRISPR/Cas9 for dynamic lysosomal pH monitoring via ratiometric RpH-LAMP1-3×flag imaging to evaluate pathophysiological mechanisms. Parallel investigations in Atp6v1b2R506X/R506X mice incorporated continuous video-EEG monitoring, behavioral assessments, western blot analyses, and transmission electron microscopy to evaluate therapeutic responses. Drug concentrations in plasma and brain homogenates were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Clinical analysis revealed central nervous system manifestations (epilepsy, intellectual disability, developmental delay) as primary morbidity determinants. Cellular studies demonstrated significant increase of lysosomal pH in mutant cells compared to wild-type control. Remarkably, treatment with the cAMP analog CPT-cAMP restored lysosomal acidification in a concentration-dependent manner. In vivo studies confirmed spontaneous seizure activity in mutant mice and CPT-cAMP’s penetration of the BBB was confirmed by LC-MS/MS. Intraperitoneal CPT-cAMP administration (20 mg/kg) exerted triple therapeutic effects: (1) significant reduction in seizure frequency, (2) improved cognitive performance in behavioral paradigms, and (3) restoration of autophagic flux through resolution of autophagosome accumulation. These findings establish proof-of-concept for cAMP-mediated lysosomal pH modulation as a viable therapeutic strategy. Our results position CPT-cAMP as a promising candidate for addressing both neurological and cognitive manifestations in ATP6V1B2-related disorders.
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We appreciate the support from all patients who participated in this study.
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Our research is supported by the grants from National Natural Science Foundation of China (82271177, 82271185), and Beijing Natural Science Foundation of (7242137). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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WX, YY, and PD conceived of the study and were responsible for submission of the manuscript for publication. LZ, WZ, GY, and PD participated in its design and drafting. LZ performed the experiments. LZ, WZ, GY, SQ, YL, LG, GW, YM, JX, XG, LC, XL, and RL participated in the literature search, data collection, and data analysis.
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Zheng, L., Zhao, W., Yang, G. et al. Therapeutic potential of cAMP-mediated lysosomal pH modulation in ATP6V1B2-related neuropathology. Cell Death Discov. (2026). https://doi.org/10.1038/s41420-026-03056-4
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DOI: https://doi.org/10.1038/s41420-026-03056-4
