This study presents an in-depth analysis of mitochondrial enzyme activities in Friedreich's ataxia (FA) patients, focusing on the Electron Transport Chain complexes I, II, and IV, the Krebs Cycle enzyme Citrate Synthase, and Coenzyme Q10 levels. It examines a cohort of 34 FA patients, comparing their mitochondrial enzyme activities and clinical parameters, including disease duration and cardiac markers, with those of 17 healthy controls. The findings reveal marked reductions in complexes II and, specifically, IV, highlighting mitochondrial impairment in FA. Additionally, elevated Neurofilament Light Chain levels and cardiomarkers were observed in FA patients. This research enhances our understanding of FA pathophysiology and suggests potential biomarkers for monitoring disease progression. The study underscores the need for further clinical trials to validate these findings, emphasizing the critical role of mitochondrial dysfunction in FA assessment and treatment.

• Klíčová slova
• Biomarkers, Complex I (NQR), Complex II (SQR), Complex IV (COX), Disease monitoring, Friedreich's ataxia, Mitochondrial dysfunction, Neurofilament light chain (NFL), Therapeutic monitoring, Ubiquinone (Coenzyme Q10),
• MeSH
• biologické markery * metabolismus   MeSH
• citrátsynthasa metabolismus   MeSH
• dospělí MeSH
• Friedreichova ataxie * diagnóza   MeSH
• kohortové studie MeSH
• lidé středního věku MeSH
• lidé MeSH
• mitochondrie metabolismus   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• ubichinon * analogy a deriváty   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery * MeSH
• citrátsynthasa MeSH
• coenzyme Q10 MeSH Prohlížeč
• ubichinon * MeSH

BACKGROUND: Mitochondrial diseases belong to the most severe inherited metabolic disorders affecting pediatric population. Despite detailed knowledge of mtDNA mutations and progress in identification of affected nuclear genes, diagnostics of a substantial part of mitochondrial diseases relies on clinical symptoms and biochemical data from muscle biopsies and cultured fibroblasts. METHODS: To investigate manifestation of oxidative phosphorylation defects in isolated lymphocytes, digitonin-permeabilized cells from 48 children were analyzed by high resolution respirometry, cytofluorometric detection of mitochondrial membrane potential and immunodetection of respiratory chain proteins with SDS and Blue Native electrophoreses. RESULTS: Evaluation of individual respiratory complex activities, ATP synthesis, kinetic parameters of mitochondrial respiratory chain and the content and subunit composition of respiratory chain complexes enabled detection of inborn defects of respiratory complexes I, IV and V within 2 days. Low respiration with NADH-dependent substrates and increased respiration with glycerol-3-phosphate revealed complex I defects; changes in p 50 for oxygen and elevated uncoupling control ratio pointed to complex IV deficiency due to SURF1 or SCO2 mutation; high oligomycin sensitivity of state 3-ADP respiration, upregulated mitochondrial membrane potential and low content of complex V were found in lymphocytes with ATP synthase deficiency due to TMEM70 mutations. CONCLUSION: Based on our results, we propose the best biochemical parameters predictive for defects of respiratory complexes I, IV and V manifesting in peripheral blood lymphocytes. GENERAL SIGNIFICANCE: The noninvasiveness, reliability and speed of an approach utilizing novel biochemical criteria demonstrate the high potential of isolated lymphocytes for diagnostics of oxidative phosphorylation disorders in pediatric patients.

• Klíčová slova
• AA, antimycin A, BNE, Blue Native PAGE, COX, cytochrome c oxidase, Diagnostics, FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, GP, glycerol-3-phosphate, GPDH, mitochondrial FAD-dependent glycerophosphate dehydrogenase, Lymphocytes, Mitochondrial diseases, OXPHOS, oxidative phosphorylation, Oxidative phosphorylation, PAGE, polyacrylamide gel electrophoresis, Respirometry, TMPD, tetramethylphenylenediamine, TMRM, tetramethylrhodamine methyl ester, cI–cV, respiratory chain complexes I–V, s3, state 3-ADP, s3u, state 3-uncoupled, s4o, state 4-oligomycin, ΔΨm, mitochondrial membrane potential,
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: Mitochondrial disorders (MD) may manifest in neonates, but early diagnosis is difficult. In this study, clinical and laboratory data were analyzed in 129 patients with neonatal onset of MD to identify any association between specific mitochondrial diseases and their symptoms with the aim of optimizing diagnosis. MATERIALS AND METHODS: Retrospective clinical and laboratory data were evaluated in 461 patients (331 families) with confirmed MD. RESULTS: The neonatal onset of MD was reported in 28% of the patients. Prematurity, intrauterine growth retardation and hypotonia necessitating ventilatory support were present in one-third, cardiomyopathy in 40%, neonatal seizures in 16%, Leigh syndrome in 15%, and elevated lactate level in 87%. Hyperammonemia was observed in 22 out of 52 neonates. Complex I deficiency was identified in 15, complex III in one, complex IV in 23, complex V in 31, combined deficiency of several complexes in 53, and PDH complex deficiency was identified in six patients. Molecular diagnosis was confirmed in 49 cases, including a newborn with a 9134A>G mutation in the MTATP6 gene, which has not been described previously. CONCLUSION: The most significant finding is the high incidence of neonatal cardiomyopathy and hyperammonemia. Based on our experience, we propose a diagnostic flowchart applicable to critically ill neonates suspicious for MD. This tool will allow for the use of direct molecular genetic analyses without the need for muscle biopsies in neonates with Alpers, Barth, MILS and Pearson syndromes, SCO1, SCO2, TMEM70, ATP5E, SUCLG1 gene mutations and PDH complex deficiency.

• MeSH
• hyperamonemie diagnóza   genetika   MeSH
• kardiomyopatie diagnóza   genetika   MeSH
• Leighova nemoc krev   diagnóza   genetika   MeSH
• lidé MeSH
• mitochondriální nemoci krev   diagnóza   genetika   MeSH
• mutace MeSH
• novorozenec nedonošený MeSH
• novorozenec MeSH
• retrospektivní studie MeSH
• růstová retardace plodu krev   diagnóza   genetika   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH