Cíl studie: Acidolabilní pojednotka (ALS) jako glykoproteinová součást ternárního komplexu IGF-1/IGFBP-3/ALS reguluje bioaktivitu a prodlužuje poločas IGF-1, čímž přispívá k řízení tělesného růstu. Proteinovou složku ALS kóduje gen IGFALS. Zkoumali jsme vliv růstového hormonu (GH) na hladiny ALS a dalších složek ternárního komplexu a možnost identifikovat podle nízkých hladin ALS pacienty s patogenními variantami genu IGFALS. Metody: U 511 dětí s deficitem GH (GHD) a/nebo u dětí s poruchou růstu navazující na malou velikost při narození (SGA-SS) jsme při léčbě GH analyzovali hladiny a vzájemnou korelaci všech tří složek ternárního komplexu. U tří dětí s nízkou hladinou ALS jsme následně sekvenovali gen IGFALS. Vliv podání GH na složky ternárního komplexu jsme studovali u 23 dětí s GHD nebo SGA-SS neurčené etiologie. Výsledky: U 511 dětí s GHD a/nebo SGA-SS na dlouhodobé léčbě GH byla hladina ALS 8750 μg/l (medián; Q1-Q3: 6353–10 963), hladina IGF-1 233 μg/l (Q1-Q3: 147–329), což odpovídalo +1,21 SD (Q1-Q3: -0,11 až +2,45), a hladina IGFBP-3 5660 μg/l (Q1-Q3: 4668–6800), což odpovídalo +3,65 SD (Q1-Q3: +1,81 až +5,97 SD). Hladiny ALS těsně korelovaly s IGF-1 (r = 0,70; p <0,0001) i s IGFBP-3 (r = 0,61; p <0,0001). Sangerova sekvenace IGFALSu dětí s nízkou hladinou ALS (369, 487 a 1490 μg/l) neprokázala patogenní ani jinou variantu genu. U 23 dětí s GHD nebo SGA-SS stoupla po 3–4 měsících podávání GH hladina ALS z 4859 (medián; Q1-Q3: 4176–6240) na 6681 (5413–8332) μg/l (p = 0,0004), hladina IGF-1 z 68 (46–114) na 146 (84–178) μg/l (p <0,0001) a hladina IGFBP-3 z 3390 (2820–4030) na 4700 (3940–5300) μg/l (p <0,0001). Vliv GH na IGF-1 (213 % výchozí hodnoty) je výraznější než na IGFBP-3 (132 %; p <0,0001) a ALS (139 %; p <0,0001). Závěry: Měření hladin ALS není efektivní pro detekci mutací IGFALS. GH zvyšuje hladiny ALS, ale biochemické stanovení ALS má jen nevýznamnou přidanou hodnotu pro diagnostiku a sledování dětí s malým vzrůstem.

Aims: Acid-labile subunit (ALS) as a glycoprotein component of the ternary complex IGF-1/IGFBP-3/ALS co-regulates bioactivity and prolongs half-life of IGF-1 and thus substantially contributes to regulation of statural growth. Protein component of ALS is encoded by IGFALS gene. We studied the impact of growth hormone (GH) on circulating levels of ALS and additional ternary complex components, and the option to identify carriers of pathogenic variants of IGFALS gene according to low ALS levels. Methods: We studied interrelations between ternary complex components in 511 children on GH therapy. Children with low ALS levels underwent IGFALS sequencing. We monitored the effect of GH therapy on the ternary complex in 23 children treated for GH deficiency (GHD) or for short stature after having been born small for gestational age (SGA-SS). Results: In 511 children with GHD and/or SGA-SS on long-term GH therapy ALS level was 8750 μg/l (median; Q1-Q3: 6353–10963), IGF-1 was 233 μg/l (Q1-Q3: 147–329), corresponding to +1.21 SD (Q1-Q3: from -0.11 to +2.45), and IGFBP-3 was 5660 μg/l (Q1-Q3: 4668–6800), corresponding to +3.65 SD (Q1-Q3: from +1.81 to +5.97 SD). ALS is strongly interrelated with IGF-1 (r=0.70; p<0.0001) and with IGFBP-3 (r=0.61; p<0.0001). IGFALS Sanger sequencing in three children with low ALS levels (369, 487 and 1490 μg/l) displayed normal results. In 23 children following 3–4 months of GH therapy, ALS increased from 4859 (median; Q1-Q3: 4176-6240) to 6681 (5413-8332) μg/l (p=0.0004), IGF-1 from 68 (46-114) to 146 (84-178) μg/l (p<0.0001) and IGFBP-3 from 3390 (2820–4030) to 4700 (3940–5300) μg/l (p<0.0001). The impact of GH is more pronounced in IGF-1 (213% increase) compared to IGFBP-3 (132%; p<0.0001) and ALS (139%; p<0.0001). Conclusions: Our results suggest that estimation of ALS levels is ineffective to detect IGFALS gene mutations. Biochemical measurements of ALS do not substantially contribute to diagnostic work-up and to follow-up in short stature children, according to our experience.

• Klíčová slova
• acidolabilní podjednotka,
• MeSH
• dítě MeSH
• faktory ternárních komplexů analýza   MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• lidský růstový hormon MeSH
• poruchy růstu * genetika   patofyziologie   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

BACKGROUND: Congenital skeletal abnormalities are a heterogeneous group of diseases most commonly associated with small or disproportionate growth, cranial and facial dysmorphisms, delayed bone maturation, etc. Nonetheless, no detailed genotype-phenotype correlation in patients with specific genetic variants is readily available. Ergo, this study focuses on the analysis of patient phenotypes with candidate variants in genes involved in bone growth as detected by molecular genetic analysis. METHODS: In this study we used molecular genetic methods to analyse the ACAN, COL2A1, FGFR3, IGFALS, IGF1, IGF1R, GHR, NPR2, STAT5B and SHOX genes in 128 Czech children with suspected congenital skeletal abnormalities. Pathogenic variants and variants of unclear clinical significance were identified and we compared their frequency in this study cohort to the European non-Finnish population. Furthermore, a prediction tool was utilised to determine their possible impact on the final protein. All clinical patient data was obtained during pre-test genetic counselling. RESULTS: Pathogenic variants were identified in the FGFR3, GHR, COL2A1 and SHOX genes in a total of six patients. Furthermore, we identified 23 variants with unclear clinical significance and high allelic frequency in this cohort of patients with skeletal abnormalities. Five of them have not yet been reported in the scientific literature. CONCLUSION: Congenital skeletal abnormalities may lead to a number of musculoskeletal, neurological, cardiovascular problems. Knowledge of specific pathogenic variants may help us in therapeutic procedures.

• MeSH
• dítě MeSH
• frekvence genu genetika   MeSH
• genetické asociační studie MeSH
• kostra * metabolismus   MeSH
• lidé MeSH
• poruchy růstu * epidemiologie   genetika   metabolismus   MeSH
• protein SHOX genetika   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Events occurring in the chicken caecum following Salmonella Enteritidis infection are relatively well-described. However, mechanisms of the immune response and defence beyond the intestinal tract are less well-described. In this study, we therefore determined changes in protein abundance in the liver and blood serum in response to S. Enteritidis infection using the unbiased approach of shotgun proteomics. Complement and coagulation cascades, TNF signalling, antigen processing and presentation was activated in the liver following infection with S. Enteritidis. Chicken proteins that decreased in the liver were involved in glycolysis, the citrate cycle, oxidative phosphorylation and fatty acid metabolism. No functional category was significantly activated or suppressed in the serum. Concerning individual proteins, VNN1, SAA, AVD, SERPINA3, SERPINB10, AGT, MRP126 or CP increased in abundance both in the liver and serum. MT4, MT3, PTGDS, GLRX and TGM4, though highly inducible in the liver, did not increase in the serum. PIGR, SERPINF2 and IGJ increased in the serum but not in the liver. SERPINA4, apoAIV, CLEC3B, SERPINF1, HRG, AHSG and ALB decreased both in the liver and serum. Avidin-like LOC431660, THRSP, GATM, GGACT, ACOX1, ALDOB or FABP7 decreased in the liver but not in the serum. Finally, CKM, CKB, PLTP, COMP, IGFALS, AMY1A or SERPIND1 decreased in the serum after S. Enteritidis infection but not in the liver. Differently abundant proteins characterise the chicken's response to infection and can be also used as markers of chicken health status.

• MeSH
• cékum imunologie   MeSH
• játra imunologie   metabolismus   mikrobiologie   MeSH
• kur domácí krev   imunologie   MeSH
• nemoci drůbeže imunologie   mikrobiologie   MeSH
• prezentace antigenu MeSH
• proteiny akutní fáze analýza   MeSH
• proteomika * MeSH
• Salmonella enteritidis MeSH
• salmonelová infekce u zvířat krev   imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Deoxynivalenol (DON) is an unavoidable contaminant in human food, animal feeds, and agricultural products. Growth retardation in children caused by extensive DON pollution has become a global problem that cannot be ignored. Previous studies have shown that DON causes stunting in children through intestinal dysfunction, insulin-like growth factor-1 (IGF-1) axis disorder and peptide YY (PYY). Galanin-like peptide (GALP) is an important growth regulator, but its role in DON-induced growth retardation is unclear. In this study, we report the important role of GALP during DON-induced growth inhibition in the rat pituitary tumour cell line GH3. DON was found to increase the expression of GALP through hypomethylationin the promoter region of the GALP gene and upregulate the expression of proinflammatory factors, while downregulate the expression of growth hormone (GH). Furthermore, GALP overexpression promoted proinflammatory cytokines, including TNF-α, IL-1β, IL-11 and IL-6, and further reduced cell viability and cell proliferation, while the inhibitory effect of GALP was the opposite. The expression of GALP and insulin like growth factor binding protein acid labile subunit (IGFALS) showed the opposite trend, which was the potential reason for the regulation of cell proliferation by GALP. In addition, GALP has anti-apoptotic effects, which could not eliminate the inflammatory damage of cells, thus aggravating cell growth inhibition. The present findings provide new mechanistic insights into the toxicity of DON-induced growth retardation and suggest a therapeutic potential of GALP in DON-related diseases.

• MeSH
• apoptóza MeSH
• epigeneze genetická účinky léků   MeSH
• galanin genetika   metabolismus   MeSH
• glykoproteiny genetika   metabolismus   MeSH
• hypofýza cytologie   MeSH
• krysa rodu rattus MeSH
• nádorové buněčné linie MeSH
• proliferace buněk MeSH
• transportní proteiny genetika   metabolismus   MeSH
• trichotheceny farmakologie   MeSH
• umlčování genů MeSH
• up regulace účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

CONTEXT: Familial short stature (FSS) is a term describing a growth disorder that is vertically transmitted. Milder forms may result from the combined effect of multiple genes; more severe short stature is suggestive of a monogenic condition. The etiology of most FSS cases has not been thoroughly elucidated to date. OBJECTIVES: To identify the genetic etiology of severe FSS in children treated with GH because of the diagnosis of small for gestational age or GH deficiency (SGA/GHD). DESIGN, SETTINGS, AND PATIENTS: Of 736 children treated with GH because of GHD/SGA, 33 with severe FSS (life-minimum height -2.5 SD or less in both the patient and shorter parent) were included in the study. The genetic etiology was known in 5 of 33 children prior to the study [ACAN (in 2], NF1, PTPN11, and SOS1). In the remaining 28 of 33, whole-exome sequencing was performed. The results were evaluated using American College of Medical Genetics and Genomics standards and guidelines. RESULTS: In 30 of 33 children (90%), we found at least one variant with potential clinical significance in genes known to affect growth. A genetic cause was elucidated in 17 of 33 (52%). Of these children, variants in growth plate-related genes were found in 9 of 17 [COL2A1, COL11A1, and ACAN (all in 2), FLNB, FGFR3, and IGF1R], and IGF-associated proteins were affected in 2 of 17 (IGFALS and HMGA2). In the remaining 6 of 17, the discovered genetic mechanisms were miscellaneous (TRHR, MBTPS2, GHSR, NF1, PTPN11, and SOS1). CONCLUSIONS: Single-gene variants are frequent among families with severe FSS, with variants affecting the growth plate being the most prevalent.

• MeSH
• dítě MeSH
• genetická predispozice k nemoci epidemiologie   MeSH
• genetická variace * MeSH
• kohortové studie MeSH
• kojenec MeSH
• lidé MeSH
• lidský růstový hormon terapeutické užití   MeSH
• metaloendopeptidasy genetika   MeSH
• mladiství MeSH
• poruchy růstu farmakoterapie   epidemiologie   genetika   MeSH
• předškolní dítě MeSH
• prognóza MeSH
• prospektivní studie MeSH
• receptor fibroblastových růstových faktorů, typ 3 genetika   MeSH
• receptor IGF typ 1 genetika   MeSH
• rodokmen MeSH
• růstová ploténka účinky léků   MeSH
• sekvenování exomu metody   MeSH
• stupeň závažnosti nemoci MeSH
• výsledek terapie MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

INTRODUCTION: Among children born small for gestational age, 10-15% fail to catch up and remain short (SGA-SS). The underlying mechanisms are mostly unknown. We aimed to decipher genetic aetiologies of SGA-SS within a large single-centre cohort. METHODS: Out of 820 patients treated with growth hormone (GH), 256 were classified as SGA-SS (birth length and/or birth weight <-2 SD for gestational age and life-minimum height <-2.5 SD). Those with the DNA triplet available (child and both parents) were included in the study (176/256). Targeted testing (karyotype/FISH/MLPA/specific Sanger sequencing) was performed if a specific genetic disorder was clinically suggestive. All remaining patients underwent MS-MLPA to identify Silver-Russell syndrome, and those with unknown genetic aetiology were subsequently examined using whole-exome sequencing or targeted panel of 398 growth-related genes. Genetic variants were classified using ACMG guidelines. RESULTS: The genetic aetiology was elucidated in 74/176 (42%) children. Of these, 12/74 (16%) had pathogenic or likely pathogenic (P/LP) gene variants affecting pituitary development (LHX4, OTX2, PROKR2, PTCH1, POU1F1), the GH-IGF-1 or IGF-2 axis (GHSR, IGFALS, IGF1R, STAT3, HMGA2), 2/74 (3%) the thyroid axis (TRHR, THRA), 17/74 (23%) the cartilaginous matrix (ACAN, various collagens, FLNB, MATN3), and 7/74 (9%) the paracrine chondrocyte regulation (FGFR3, FGFR2, NPR2). In 12/74 (16%), we revealed P/LP affecting fundamental intracellular/intranuclear processes (CDC42, KMT2D, LMNA, NSD1, PTPN11, SRCAP, SON, SOS1, SOX9, TLK2). SHOX deficiency was found in 7/74 (9%), Silver-Russell syndrome in 12/74 (16%) (11p15, UPD7), and miscellaneous chromosomal aberrations in 5/74 (7%) children. CONCLUSIONS: The high diagnostic yield sheds a new light on the genetic landscape of SGA-SS, with a central role for the growth plate with substantial contributions from the GH-IGF-1 and thyroid axes and intracellular regulation and signalling.

• MeSH
• dítě MeSH
• gestační stáří MeSH
• hypotrofický novorozenec MeSH
• insulinu podobný růstový faktor I MeSH
• lidé MeSH
• lidský růstový hormon * genetika   MeSH
• nanismus * MeSH
• novorozenec MeSH
• poruchy růstu genetika   diagnóza   MeSH
• protein SHOX MeSH
• Silverův-Russellův syndrom * genetika   MeSH
• tělesná výška genetika   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: The growth hormone deficiency (GHD) diagnosis is controversial especially due to low specificity of growth hormone (GH) stimulation tests. It is therefore believed that children diagnosed with GHD form a heterogeneous group with growth disorder frequently independent on GH function. No study evaluating the complex etiology of growth failure in children with diagnosed GHD has been performed thus far. AIMS: To discover genetic etiology of short stature in children with diagnosed GHD from families with short stature. METHODS: Fifty-two children diagnosed with primary GHD and vertically transmitted short stature (height SDS in the child and his/her shorter parent <-2 SD) were included to our study. The GHD diagnosis was based on growth data suggestive of GHD, absence of substantial disproportionality (sitting height to total height ratio <-2 SD or >+2 SD), IGF-1 levels <0 for age and sex specific SD and peak GH concentration <10 ug/L in two stimulation tests. All children were examined using next-generation sequencing methods, and the genetic variants were subsequently evaluated by American College of Medical Genetics standards and guidelines. RESULTS: The age of children at enrollment into the study was 11 years (median, IQR 9-14 years), their height prior to GH treatment was -3.0 SD (-3.6 to -2.8 SD), IGF-1 concentration -1.4 SD (-2.0 to -1.1 SD), and maximal stimulated GH 6.3 ug/L (4.8-7.6 ug/L). No child had multiple pituitary hormone deficiency or a midbrain region pathology. Causative variant in a gene that affects growth was discovered in 15/52 (29%) children. Of them, only 2 (13%) had a genetic variant affecting GH secretion or function (GHSR and OTX2). Interestingly, in 10 (67%) children we discovered a primary growth plate disorder (ACAN, COL1A2, COL11A1, COL2A1, EXT2, FGFR3, NF1, NPR2, PTPN11 [2x]), in one (7%) a genetic variant impairing IGF-1 action (IGFALS) and in two (12%) a variant in miscellaneous genes (SALL4, MBTPS2). CONCLUSIONS: In children with vertically transmitted short stature, genetic results frequently did not correspond with the clinical diagnosis of GH deficiency. These results underline the doubtful reliability of methods standardly used to diagnose GH deficiency.

• MeSH
• dítě MeSH
• hypofyzární nanismus * diagnóza   genetika   farmakoterapie   MeSH
• insulinu podobný růstový faktor I genetika   MeSH
• lidé MeSH
• lidský růstový hormon * MeSH
• mladiství MeSH
• reprodukovatelnost výsledků MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH