Pyruvate kinase (PK) deficiency is the most common cause of chronic congenital non-spherocytic haemolytic anaemia worldwide, with an estimated prevalence of one in 100 000 to one in 300 000 people. PK deficiency results in chronic haemolytic anaemia, with wide ranging and serious consequences affecting health, quality of life, and mortality. The goal of the International Guidelines for the Diagnosis and Management of Pyruvate Kinase Deficiency was to develop evidence-based guidelines for the clinical care of patients with PK deficiency. These clinical guidelines were developed by use of GRADE methodology and the AGREE II framework. Experts were invited after consideration of area of expertise, scholarly contributions in PK deficiency, and country of practice for global representation. The expert panel included 29 expert physicians (including adult and paediatric haematologists and other subspecialists), geneticists, laboratory specialists, nurses, a guidelines methodologist, patients with PK deficiency, and caregivers from ten countries. Five key topic areas were identified, the panel prioritised key questions, and a systematic literature search was done to generate evidence summaries that were used in the development of draft recommendations. The expert panel then met in person to finalise and vote on recommendations according to a structured consensus procedure. Agreement of greater than or equal to 67% among the expert panel was required for inclusion of a recommendation in the final guideline. The expert panel agreed on 31 total recommendations across five key topics: diagnosis and genetics, monitoring and management of chronic complications, standard management of anaemia, targeted and advanced therapies, and special populations. These new guidelines should facilitate best practices and evidence-based PK deficiency care into clinical practice.
- MeSH
- hemolytická nesférocytická kongenitální anemie * diagnóza terapie MeSH
- kvalita života MeSH
- lidé MeSH
- pyruvátkinasa * nedostatek MeSH
- vrozené poruchy metabolismu pyruvátu * diagnóza terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- směrnice pro lékařskou praxi MeSH
- MeSH
- enzymy metabolismus MeSH
- erytrocyty metabolismus MeSH
- genetické nemoci vrozené MeSH
- glykolýza fyziologie MeSH
- hemolytická nesférocytická kongenitální anemie genetika MeSH
- hemolytické anemie * diagnóza terapie MeSH
- hemolýza genetika MeSH
- lidé MeSH
- pyruvátkinasa genetika nedostatek MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
INTRODUCTION: Pyruvate kinase (PK) deficiency is a rare, under-recognised, hereditary condition that leads to chronic haemolytic anaemia and potentially serious secondary complications, such as iron overload, cholecystitis, pulmonary hypertension and extramedullary haematopoiesis. It is an autosomal recessive disease caused by homozygous or compound heterozygous mutations in the PKLR gene. Due to its rarity and clinical heterogeneity, information on the natural history and long-term clinical course of PK deficiency is limited, presenting major challenges to patient management, the development of new therapies and establishing disease-specific treatment recommendations. The Pyruvate Kinase Deficiency Global Longitudinal (Peak) Registry is an initiative to address the gaps in the knowledge of PK deficiency. This manuscript describes the objectives, study design and methodology for the Peak Registry. METHODS AND ANALYSIS: The Peak Registry is an observational, longitudinal, global registry of adult and paediatric patients with a genetically confirmed diagnosis of PK deficiency. The Peak Steering Committee is composed of 11 clinicians and researchers with experience in the diagnosis and management of PK deficiency from 10 countries, a patient representative and representatives from the sponsor (Agios Pharmaceuticals). The registry objective is to foster an understanding of the longitudinal clinical implications of PK deficiency, including its natural history, treatments and outcomes, and variability in clinical care. The aim is to enrol up to 500 participants from approximately 60 study centres across 20 countries over 7 years, with between 2 and 9 years of follow-up. Data will include demographics, diagnosis history, genotyping, transfusion history, relevant clinical events, medications, emergency room visits and hospitalisations. ETHICS AND DISSEMINATION: Registry protocol and informed consent forms are approved by institutional review boards/independent ethics committees at each study site. The study is being conducted in accordance with the Declaration of Helsinki. Registry data will be published in peer-reviewed journal articles and conference publications. TRIAL REGISTRATION NUMBER: NCT03481738.
- MeSH
- dítě MeSH
- dospělí MeSH
- hemolytická nesférocytická kongenitální anemie * diagnóza genetika MeSH
- homozygot MeSH
- lidé MeSH
- pyruvátkinasa genetika MeSH
- vrozené poruchy metabolismu pyruvátu * genetika MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Mechanisms of right ventricular (RV) dysfunction in heart failure (HF) are poorly understood. RV response to volume overload (VO), a common contributing factor to HF, is rarely studied. The goal was to identify interventricular differences in response to chronic VO. Rats underwent aorto-caval fistula (ACF)/sham operation to induce VO. After 24 weeks, RV and left ventricular (LV) functions, gene expression and proteomics were studied. ACF led to biventricular dilatation, systolic dysfunction and hypertrophy affecting relatively more RV. Increased RV afterload contributed to larger RV stroke work increment compared to LV. Both ACF ventricles displayed upregulation of genes of myocardial stress and metabolism. Most proteins reacted to VO in a similar direction in both ventricles, yet the expression changes were more pronounced in RV (pslope: < 0.001). The most upregulated were extracellular matrix (POSTN, NRAP, TGM2, CKAP4), cell adhesion (NCAM, NRAP, XIRP2) and cytoskeletal proteins (FHL1, CSRP3) and enzymes of carbohydrate (PKM) or norepinephrine (MAOA) metabolism. Downregulated were MYH6 and FAO enzymes. Therefore, when exposed to identical VO, both ventricles display similar upregulation of stress and metabolic markers. Relatively larger response of ACF RV compared to the LV may be caused by concomitant pulmonary hypertension. No evidence supports RV chamber-specific regulation of protein expression in response to VO.
- MeSH
- extracelulární matrix - proteiny genetika metabolismus MeSH
- krysa rodu rattus MeSH
- molekuly buněčné adheze genetika metabolismus MeSH
- myokard metabolismus MeSH
- potkani Sprague-Dawley MeSH
- protein-glutamin:amin-gama-glutamyltransferasa 2 MeSH
- proteom genetika metabolismus MeSH
- pyruvátkinasa genetika metabolismus MeSH
- remodelace komor * MeSH
- srdeční komory metabolismus patologie patofyziologie MeSH
- srdeční selhání metabolismus patologie patofyziologie MeSH
- tepový objem MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tuberculosis (TB) remains one of the major health concerns worldwide. Mycobacterium tuberculosis (Mtb), the causative agent of TB, can flexibly change its metabolic processes during different life stages. Regulation of key metabolic enzyme activities by intracellular conditions, allosteric inhibition or feedback control can effectively contribute to Mtb survival under different conditions. Phosphofructokinase (Pfk) is one of the key enzymes regulating glycolysis. Mtb encodes two Pfk isoenzymes, Pfk A/Rv3010c and Pfk B/Rv2029c, which are differently expressed upon transition to the hypoxia-induced non-replicating state of the bacteria. While pfkB gene and protein expression are upregulated under hypoxic conditions, Pfk A levels decrease. Here, we present biochemical characterization of both Pfk isoenzymes, revealing that Pfk A and Pfk B display different kinetic properties. Although the glycolytic activity of Pfk A is higher than that of Pfk B, it is markedly inhibited by an excess of both substrates (fructose-6-phosphate and ATP), reaction products (fructose-1,6-bisphosphate and ADP) and common metabolic allosteric regulators. In contrast, synthesis of fructose-1,6-bisphosphatase catalyzed by Pfk B is not regulated by higher levels of substrates, and metabolites. Importantly, we found that only Pfk B can catalyze the reverse gluconeogenic reaction. Pfk B thus can support glycolysis under conditions inhibiting Pfk A function.
- MeSH
- adenosindifosfát metabolismus farmakologie MeSH
- adenosintrifosfát metabolismus farmakologie MeSH
- alosterická regulace MeSH
- bakteriální proteiny antagonisté a inhibitory metabolismus MeSH
- enzymová indukce MeSH
- fosfofruktokinasy antagonisté a inhibitory metabolismus MeSH
- fruktosadifosfáty biosyntéza farmakologie MeSH
- fruktosafosfáty metabolismus farmakologie MeSH
- glukoneogeneze MeSH
- glykolýza MeSH
- hexosafosfáty metabolismus MeSH
- izoenzymy antagonisté a inhibitory metabolismus MeSH
- katalýza MeSH
- kinetika MeSH
- kyslík farmakologie MeSH
- L-laktátdehydrogenasa metabolismus MeSH
- Mycobacterium tuberculosis účinky léků enzymologie MeSH
- pyruvátkinasa metabolismus MeSH
- rekombinantní proteiny metabolismus MeSH
- substrátová specifita MeSH
- zpětná vazba fyziologická MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- MeSH
- dědičná eliptocytóza genetika krev MeSH
- dědičná sférocytóza genetika krev MeSH
- diferenciální diagnóza MeSH
- dítě MeSH
- kongenitální hemolytická anemie * genetika krev MeSH
- lidé MeSH
- nedostatek glukóza-6-fosfátdehydrogenázy genetika krev MeSH
- novorozenec MeSH
- novorozenecká žloutenka diagnóza MeSH
- pyruvátkinasa nedostatek MeSH
- srpkovitá anemie genetika krev MeSH
- talasemie genetika krev MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Mycobacterium tuberculosis (Mtb) causes both acute tuberculosis and latent, symptom-free infection that affects roughly one-third of the world's population. It is a globally important pathogen that poses multiple dangers. Mtb reprograms its metabolism in response to the host niche, and this adaptation contributes to its pathogenicity. Knowledge of the metabolic regulation mechanisms in Mtb is still limited. Pyruvate kinase, involved in the late stage of glycolysis, helps link various metabolic routes together. Here, we demonstrate that Mtb pyruvate kinase (Mtb PYK) predominantly catalyzes the reaction leading to the production of pyruvate, but its activity is influenced by multiple metabolites from closely interlinked pathways that act as allosteric regulators (activators and inhibitors). We identified allosteric activators and inhibitors of Mtb PYK originating from glycolysis, citrate cycle, nucleotide/nucleoside inter-conversion related pathways that had not been described so far. Enzyme was found to be activated by fructose-1,6-bisphosphate, ribose-5-phosphate, adenine, adenosine, hypoxanthine, inosine, L-2-phosphoglycerate, l-aspartate, glycerol-2-phosphate, glycerol-3-phosphate. On the other hand thiamine pyrophosphate, glyceraldehyde-3-phosphate and L-malate were identified as inhibitors of Mtb PYK. The detailed kinetic analysis indicated a morpheein model of Mtb PYK allosteric control which is strictly dependent on Mg2+ and substantially increased by the co-presence of Mg2+ and K+.
- MeSH
- biologické markery MeSH
- krevní transfuze MeSH
- lidé MeSH
- magnetická rezonanční tomografie MeSH
- management nemoci MeSH
- mutace MeSH
- mutační analýza DNA MeSH
- přetížení železem diagnóza epidemiologie etiologie metabolismus MeSH
- prevalence MeSH
- pyruvátkinasa nedostatek MeSH
- železo metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- dopisy MeSH
Erytrocytární enzymopatie je genetické onemocnění vyznačující se vrozenými kvantitativními či kvalitativními abnormalitami erytrocytárních enzymů, které jsou jednou z příčin vzniku vrozené nesférocytární hemolytické anémie. V případě zralých erytrocytů, které neobsahují jádro, mitochondrie ani endoplazmatické retikulum, se enzymopatie týkají glykolýzy a pentózafosfátového cyklu, popř. metabolismu glutathionu a nukleotidů. Mezi nejrozšířenější abnormality patří deficit pyruvátkinázy a glukóza-6-fosfátdehydrogenázy, které jsou vzácnou příčinou vzniku hemolytické anémie v české i slovenské populaci. Enzymopatie jsou charakterizovány normo-/lehce makrocytární normochromní anémií s obecnými znaky hemolýzy. Při negativní rodinné anamnéze, je diagnostika enzymopatií vzhledem k minimálním charakteristickým abnormalitám v morfologii erytrocytů komplikovaná (proto byly dříve označovány jako vrozené nesférocytární hemolytické anémie). Základní diferenciální diagnostika nedokáže rozlišit jednotlivé enzymové defekty. Přesná diagnóza může být určena až na základě snížené specifické aktivity daného enzymu a potvrzena dále na molekulárně-genetické úrovni. Deficit glukóza-6-fosfátdehydrogenázy (G6PD) a pyruvátkinázy (PK) byl v české a slovenské populaci popsán již v 80. letech. Ucelený přehled o výskytu tohoto onemocnění v české a slovenské populaci, zvláště pak o jeho vzácnějších formách, doposud chybí. Od roku 2013 se naše laboratoř zabývá metodou přímého stanovení aktivity enzymů s následným genetickým vyšetřením u pacientů se suspektní enzymopatií. V současné době jsme jedinou specializovanou laboratoří zabývající se touto diagnostikou v České republice, resp. ve střední Evropě. Doposud byl enzymový deficit diagnostikován u dvaceti čtyř pacientů. Vedle nejčastějších deficitů G6PD a PK (9 a 12 případů), byly identifikovány 2 rodiny s deficitem glukózafosfátizomerázy (GPI) a 1 rodina s velmi vzácným deficitem hexokinázy (HK). Deficit GPI a HK jsme v české a slovenské populaci diagnostikovali poprvé. Celkově bylo dosud identifikováno dvacet dva kauzálních mutací, z nichž pět bylo v literatuře popsáno poprvé – G6PD p.(Phe216Tyr), PK p.(Arg518Leufs*12), p.(Asp293Val), GPI p.(Ser160Pro) a p.(Arg472Cys).
Hereditary red blood cell (RBC) enzymopathies are genetic disorders affecting genes encoding red blood cell enzymes. They cause a specific type of anaemia designated hereditary nonspherocytic haemolytic anaemia. Enzymopathies affect cellular metabolism, which in the RBC mainly consists of anaerobic glycolysis, the hexose monophosphate shunt, glutathione metabolism and nucleotide metabolism. Enzymopathies are commonly associated with normocytic/slightly macrocytic normochromic haemolytic anaemia. In general, RBC enzymopathies are not characterized by specific abnormalities in RBC morphology. Routinely used haematology methods are unable to pinpoint a specific enzyme defect. Definitive diagnosis is based on the detection of reduced specific enzyme activity and molecular characterization of the defect at DNA level. This work represents advanced and up-to-date information regarding RBC enzyme deficiencies in the Czech and Slovak populations dating from the 1980s, when mutations in Czech and Slovak subjects resulting in glucose-6-phosphate dehydrogenase (G6PD) and pyruvate kinase (PK) deficiencies were first described. Both deficiencies represent the most common erythroenzymopathies worldwide. Since 2013, our laboratory has focused on the introduction of direct enzyme assays supplemented by genetic testing in patients with haemolytic anaemia and suspected erythroenzymopathy. To date, several different enzyme defects have been diagnosed in twenty-four patients. Apart from G6PD and PK deficiencies (9 and 12 cases), 2 families with glucose phosphate isomerase (GPI) defect and 1 family with the very rare hexokinase (HK) deficiency have been identified. The two latter deficiencies were diagnosed in the Czech and Slovak populations for the first time. Among the 22 identified mutations, were novel and have not been previously reported in literature: 5, namely G6PD p.(Phe216Tyr), PK p.(Arg518Leufs*12), p.(Asp293Val) and GPI p.(Ser160Pro), p.(Arg472Cys).
- Klíčová slova
- erytrocytární enzymopatie,
- MeSH
- enzymy metabolismus MeSH
- erytrocyty metabolismus MeSH
- glykolýza fyziologie MeSH
- hemolytická nesférocytická kongenitální anemie genetika MeSH
- hemolytické anemie * diagnóza terapie MeSH
- lidé MeSH
- pyruvátkinasa genetika nedostatek MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- souhrny MeSH
