RESUMEN
Duchenne muscular dystrophy (DMD) is the most prevalent herediatry disease in men, characterized by dystrophin deficiency, progressive muscle wasting, cardiac insufficiency, and premature mortality, with no effective therapeutic options. Here, we investigated whether adenine base editing can correct pathological nonsense point mutations leading to premature stop codons in the dystrophin gene. We identified 27 causative nonsense mutations in our DMD patient cohort. Treatment with adenine base editor (ABE) could restore dystrophin expression by direct A-to-G editing of pathological nonsense mutations in cardiomyocytes generated from DMD patient-derived induced pluripotent stem cells. We also generated two humanized mouse models of DMD expressing mutation-bearing exons 23 or 30 of human dystrophin gene. Intramuscular administration of ABE, driven by ubiquitous or muscle-specific promoters could correct these nonsense mutations in vivo, albeit with higher efficiency in exon 30, restoring dystrophin expression in skeletal fibers of humanized DMD mice. Moreover, a single systemic delivery of ABE with human single guide RNA (sgRNA) could induce body-wide dystrophin expression and improve muscle function in rotarod tests of humanized DMD mice. These findings demonstrate that ABE with human sgRNAs can confer therapeutic alleviation of DMD in mice, providing a basis for development of adenine base editing therapies in monogenic diseases.
RESUMEN
OBJECTIVE: To identify metabolites and metabolic pathways affected in dogs with struvite and calcium oxalate urolithiasis compared to healthy dogs. To explore the candidate urinary biomarkers to distinguish dogs with struvite and calcium oxalate urolithiasis. ANIMALS: 13 dogs with calcium oxalate urolithiasis, 7 dogs with struvite urolithiasis, and 13 healthy dogs were recruited between September 2021 and January 2023. METHODS: Metabolomic profiles were analyzed from urine samples using UPLC-MS MS. According to the variable importance in the projection (> 1) and correlation coefficient (P < .05) obtained by orthogonal partial least squares discriminant analysis, the differential metabolites were screened. The Kyoto Encyclopedia of Genes and Genomes database was used to identify the metabolic pathways involved. RESULTS: Compared to healthy dogs, those with calcium oxalate urolithiasis exhibited distinct metabolites primarily associated with phenylalanine metabolism, nicotinic acid, and nicotinamide metabolic pathways. Conversely, dogs with struvite urolithiasis demonstrated variations in metabolites mainly linked to tryptophan metabolism and glycerophospholipid metabolic pathways. Between calcium oxalate and struvite groups, pyocyanin, glycylprolylarginine, traumatin, cysteinyl-leucine, and 8-hydroxydodecylcarnitine are candidate urinary biomarkers. CLINICAL RELEVANCE: Our findings provide an in-depth analysis of metabolic perturbations associated with calcium oxalate and struvite urolithiasis in dogs. We also identified candidate urinary biomarkers distinguishing between dogs with calcium oxalate and struvite urolithiasis, which can be subsequently validated to assist in stone diagnosis and guide treatment choices.
RESUMEN
PURPOSE: Fibroblast activating protein (FAP) is highly expressed in the synovial tissues of rheumatoid arthritis (RA) patients. The aim of this study was to determine the feasibility of PET imaging with an Al[18F] F-NOTA-labeled FAP inhibitor 04(18F-FAPI-04) for the evaluation of arthritic progression and therapeutic response in experimental arthritis. METHODS: Fibroblast-like synoviocytes (FLSs) were obtained from patients with RA or osteoarthritis (OA), and the relationship between 18F-FAPI-04 uptake and the inflammatory activity of RA FLSs was investigated. Collagen-induce arthritis (CIA) mice models were established and treated with methotrexate (MTX) or etanercept (ETC). Then, PET imaging was performed 24 h following 18F-FAPI-04 injection. The imaging results were compared by assessing macroscopic arthritis scores and histological staining. RESULTS: 18F-FAPI-04 uptake was obvious in RA FLSs that characterizing FAP activation. The higher the uptake of 18F-FAPI-04, the more severity of the inflammatory phenotype in RA FLS. Furthermore, the uptake of 18F-FAPI-04 in inflamed joints could be found even before the deformity of the parental joints could be observed by histological examination. Both MTX and ETC were effective in inhibiting the progression of arthritis in CIA mice was confirmed by macroscopic, histological, and radiographic pathology scores. Importantly, 18F-FAPI-04 uptake declined accordingly in CIA models following MTX and ETC treatment. CONCLUSIONS: These findings suggest that PET imaging of 18F-FAPI-04 can be used to monitor treatment response in RA, and is more sensitive in disease speculation than macroscopic arthritis scoring.