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1.
PLoS One ; 15(7): e0236788, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32735629

RESUMO

This study aimed to establish and reproduce transgenic pigs expressing human growth hormone (hGH) in their milk. We also aimed to purify hGH from the milk, to characterize the purified protein, and to assess the potential of our model for mass production of therapeutic proteins using transgenic techniques. Using ~15.5 L transgenic pig milk, we obtained proteins with ≥ 99% purity after three pre-treatments and five column chromatography steps. To confirm the biosimilarity of our milk-derived purified recombinant hGH (CGH942) with commercially available somatropin (Genotropin), we performed spectroscopy, structural, and biological analyses. We observed no difference between the purified protein and Genotropin samples. Furthermore, rat models were used to assess growth promotion potential. Our results indicate that CGH942 promotes growth, by increasing bone development and body weight. Toxicity assessments revealed no abnormal findings after 4 weeks of continuous administration and 2 weeks of recovery. The no-observed-adverse-effect level for both males and females was determined to be 0.6 mg/kg/day. Thus, no toxicological differences were observed between commercially available somatropin and CGH942 obtained from transgenic pig milk. In conclusion, we describe a transgenic technique using pigs, providing a new platform to produce human therapeutic proteins.


Assuntos
Animais Geneticamente Modificados/metabolismo , Hormônio do Crescimento Humano , Proteínas Recombinantes , Animais , Cromatografia de Afinidade , Feminino , Técnicas de Transferência de Genes , Hormônio do Crescimento Humano/química , Hormônio do Crescimento Humano/genética , Hormônio do Crescimento Humano/metabolismo , Humanos , Ratos , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/toxicidade , Suínos
2.
Genes (Basel) ; 10(11)2019 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-31698873

RESUMO

Prader-Willi syndrome (PWS) is an imprinting genetic disorder characterized by lack of expression of genes on the paternal chromosome 15q11-q13 region. Growth hormone (GH) replacement positively influences stature and body composition in PWS. Our hypothesis was that early diagnosis delays onset of obesity in PWS. We studied 352 subjects with PWS, recruited from the NIH Rare Disease Clinical Research Network, to determine if age at diagnosis, ethnicity, gender, and PWS molecular class influenced the age they first become heavy, as determined by their primary care providers, and the age they first developed an increased appetite and began seeking food. The median ages that children with PWS became heavy were 10 years, 6 years and 4 years for age at diagnosis < 1 year, between 1 and 3 years, and greater than 3 years of age, respectively. The age of diagnosis and ethnicity were significant factors influencing when PWS children first became heavy (p < 0.01), however gender and the PWS molecular class had no influence. Early diagnosis delayed the onset of becoming heavy in individuals with PWS, permitting early GH and other treatment, thus reducing the risk of obesity-associated co-morbidities. Non-white individuals had an earlier onset of becoming heavy.


Assuntos
Diagnóstico Precoce , Síndrome de Prader-Willi/complicações , Síndrome de Prader-Willi/diagnóstico , Fatores Etários , Criança , Pré-Escolar , Cromossomos Humanos Par 15/genética , Comorbidade , Feminino , Impressão Genômica/genética , Terapia de Reposição Hormonal , Hormônio do Crescimento Humano/genética , Humanos , Lactente , Masculino , Obesidade/genética , Obesidade/prevenção & controle , Doenças Raras/genética
3.
Arch. endocrinol. metab. (Online) ; 63(6): 608-617, Nov.-Dec. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1055027

RESUMO

ABSTRACT The somatotropic axis is the main hormonal regulator of growth. Growth hormone (GH), also known as somatotropin, and insulin-like growth factor 1 (IGF-1) are the key components of the somatotropic axis. This axis has been studied for a long time and the knowledge of how some molecules could promote or impair hormones production and action has been growing over the last decade. The enhancement of large-scale sequencing techniques has expanded the spectrum of known genes and several other candidate genes that could affect the GH-IGF1-bone pathway. To date, defects in more than forty genes were associated with an impairment of the somatotropic axis. These defects can affect from the secretion of GH to the bioavailability and action of IGF-1. Affected patients present a large heterogeneous group of conditions associated with growth retardation. In this review, we focus on the description of the GH-IGF axis genetic defects reported in the last decade. Arch Endocrinol Metab. 2019;63(6):608-17


Assuntos
Humanos , Fator de Crescimento Insulin-Like I/genética , Hormônio do Crescimento Humano/deficiência , Hormônio do Crescimento Humano/genética , Transtornos do Crescimento/genética , Mutação/genética , Fenótipo , Fator de Crescimento Insulin-Like I/metabolismo , Transdução de Sinais , Genótipo , Transtornos do Crescimento/metabolismo
4.
PLoS One ; 14(10): e0223024, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31647849

RESUMO

AIM: To examine global gene expression response to profound metabolic and hormonal stress induced by acute sprint exercise. METHODS: Healthy men and women (n = 14) performed three all-out cycle sprints interspersed by 20 min recovery. Muscle biopsies were obtained before the first, and 2h and 20 min after last sprint. Microarray analysis was performed to analyse acute gene expression response and repeated blood samples were obtained. RESULTS: In skeletal muscle, a set of immediate early genes, FOS, NR4A3, MAFF, EGR1, JUNB were markedly upregulated after sprint exercise. Gene ontology analysis from 879 differentially expressed genes revealed predicted activation of various upstream regulators and downstream biofunctions. Gene signatures predicted an enhanced turnover of skeletal muscle mass after sprint exercise and some novel induced genes such as WNT9A, FZD7 and KLHL40 were presented. A substantial increase in circulating free fatty acids (FFA) was noted after sprint exercise, in parallel with upregulation of PGC-1A and the downstream gene PERM1 and gene signatures predicting enhanced lipid turnover. Increase in growth hormone and insulin in blood were related to changes in gene expressions and both hormones were predicted as upstream regulators. CONCLUSION: This is the first study reporting global gene expression in skeletal muscle in response to acute sprint exercise and several novel findings are presented. First, in line with that muscle hypertrophy is not a typical finding after a period of sprint training, both hypertrophy and atrophy factors were regulated. Second, systemic FFA and hormonal and exposure might be involved in the sprint exercise-induced changes in gene expression.


Assuntos
Ácidos Graxos não Esterificados/genética , Proteínas Musculares/genética , Músculo Esquelético/fisiopatologia , Transcriptoma/genética , Adulto , Glicemia/metabolismo , Exercício Físico/fisiologia , Ácidos Graxos não Esterificados/metabolismo , Feminino , Hormônio do Crescimento Humano/genética , Hormônio do Crescimento Humano/metabolismo , Humanos , Insulina/metabolismo , Masculino , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Corrida/fisiologia
5.
Horm Res Paediatr ; 92(1): 15-27, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31509834

RESUMO

BACKGROUND: The reason for the insufficient catch-up growth seen in 10% of children born small for gestational age (SGA) is poorly understood. Disturbances in the growth hormone (GH) - insulin-like growth factor (IGF) axis might underlie this failure to show sufficient catch-up growth. CONCLUSION: This review summarizes insights gained in the molecular and (epi) genetic mechanisms of the GH-IGF axis in short children born SGA. The most notable anomalies of the IGF system are the lowered IGF-I levels in both cord blood and the placenta, and the increased expression of IGF-binding proteins (IGFBP)-1 and IGFBP-2, which inhibit IGF-I, in the placenta of SGA neonates. These observations suggest a decreased bioactivity of IGF-I in utero. IGF-I levels remain reduced in SGA children with short stature, as well as IGFBP-3 and acid-labile subunit levels. Proteolysis of IGFBP-3 appears to be increased.


Assuntos
Epigênese Genética , Hormônio do Crescimento Humano , Recém-Nascido Pequeno para a Idade Gestacional/metabolismo , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina , Fator de Crescimento Insulin-Like I , Proteólise , Criança , Feminino , Hormônio do Crescimento Humano/genética , Hormônio do Crescimento Humano/metabolismo , Humanos , Recém-Nascido , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/genética , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/metabolismo , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Masculino
6.
Hum Mutat ; 40(11): 2033-2043, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31231873

RESUMO

Isolated growth hormone deficiency (IGHD) is a rare condition mainly caused by mutations in GH1. The aim of this study was to assess the contribution of GHRHR mutations to IGHD in an unusually large group of patients. All GHRHR coding exons and flanking intronic regions were sequenced in 312 unrelated patients with nonsyndromic IGHD. Functional consequences of all newly identified missense variants were assessed in vitro (i.e., study of the expression of recombinant GHRHRs and their ability to activate the cyclic adenosine monophosphate (cAMP) signaling pathway). Genotype-phenotype correlation analyses were performed according to the nature of the identified mutation. We identified 20 different disease-causing GHRHR mutations (truncating and missense loss-of-function mutations), among which 15 are novel, in 24 unrelated patients. Of note, about half (13/24) of those patients represent sporadic cases. The clinical phenotype of patients with at least one missense GHRHR mutation was found to be indistinguishable from that of patients with bi-allelic truncating mutations. This study, which unveils disease-causing GHRHR mutations in 8% (24/312) of IGHD cases, identifies GHRHR as the second IGHD gene most frequently involved after GH1. The finding that 8% of IGHD cases without GH1 mutations are explained by GHRHR molecular defects (including missense mutations), together with the high proportion of sporadic cases among those patients, has important implications for genetic counseling.


Assuntos
Nanismo Hipofisário/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Mutação , Receptores de Neuropeptídeos/genética , Receptores de Hormônios Reguladores de Hormônio Hipofisário/genética , Alelos , Sequência de Aminoácidos , Substituição de Aminoácidos , AMP Cíclico , Análise Mutacional de DNA , Nanismo Hipofisário/diagnóstico , Feminino , Genótipo , Hormônio do Crescimento Humano/genética , Humanos , Masculino , Linhagem , Receptores de Neuropeptídeos/química , Receptores de Hormônios Reguladores de Hormônio Hipofisário/química
7.
Endocr J ; 66(9): 807-816, 2019 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-31189758

RESUMO

CDK5 regulatory subunit associated protein 1-like 1 (CDKAL1) is a tRNA-modifying enzyme that catalyzes 2-methylthiolation (ms2) and has been implicated in the development of type 2 diabetes (T2D). CDKAL1-mediated ms2 is important for efficient protein translation and regulates insulin biosynthesis in pancreatic cells. Interestingly, an association between T2D and release of growth hormone (GH) has been reported in humans. However, it is unknown whether CDKAL1 is important for hormone production in the pituitary gland. The present study investigated the role of CDKAL1 in GH-producing pituitary adenomas (GHPAs). CDKAL1 activity was suppressed in GHPAs, as evidenced by a decrease in ms2, compared with non-functioning pituitary adenomas (NFPAs), which do not produce specific hormones. Downregulation of Cdkal1 using small interfering and short hairpin RNAs increased the biosynthesis and secretion of GH in rat GH3 cells. Depletion of Cdkal1 increased the cytosolic calcium level via downregulation of DnaJ heat shock protein family (Hsp40) member C10 (Dnajc10), which is an endoplasmic reticulum protein related to calcium homeostasis. This stimulated transcription of GH via upregulation of Pit-1. Moreover, CDKAL1 activity was highly sensitive to proteostatic stress and was upregulated by suppression of this stress. Taken together, these results suggest that dysregulation of CDKAL1 is involved in the pathogenesis of GHPAs, and that modulation of the proteostatic stress response might control CDKAL1 activity and facilitate treatment of GHPAs.


Assuntos
Adenoma/genética , Hormônio do Crescimento/biossíntese , Neoplasias Hipofisárias/genética , tRNA Metiltransferases/fisiologia , Adenoma/metabolismo , Adenoma/patologia , Animais , Células Cultivadas , Estresse do Retículo Endoplasmático/fisiologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Hormônio do Crescimento/genética , Adenoma Hipofisário Secretor de Hormônio do Crescimento/genética , Adenoma Hipofisário Secretor de Hormônio do Crescimento/metabolismo , Adenoma Hipofisário Secretor de Hormônio do Crescimento/patologia , Hormônio do Crescimento Humano/biossíntese , Hormônio do Crescimento Humano/genética , Humanos , Neoplasias Hipofisárias/metabolismo , Neoplasias Hipofisárias/patologia , RNA Interferente Pequeno/farmacologia , Ratos , Resposta a Proteínas não Dobradas/fisiologia , tRNA Metiltransferases/genética
8.
Horm Metab Res ; 51(4): 248-255, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31022740

RESUMO

The objective of the study is the functional characterization of a novel POU1F1 c.605delC mutation in combined pituitary hormone deficiency (CPHD) and to report the clinical and genetic details of 160 growth hormone deficiency patients. Screening of GH1, GHRHR, POU1F1, PROP1, and HESX1 genes by Sanger sequencing was carried out in 160 trios and 100 controls followed by characterization of the POU1F1 c.605delC mutation by expression studies including site directed mutagenesis, co-transfection, protein degradation, and luciferase assays to compare the wild type and mutant POU1F1. In vitro studies showed that the POU1F1 c.605delC mutation codes for a truncated protein with reduced transactivation capacity on its downstream effectors, viz., growth hormone (GH) and prolactin (PRL) causing severe CPHD. Experiments using different protease inhibitors reveal rescue of the protein upon blockage of the lysosomal pathway that might be useful in novel drug designing using targeted approach thereby maintaining the milieu and preventing/delaying the disease. The study provides an insight into the disease causing mechanism of POU1F1 c.605delC mutation identified in a CPHD child with severe short stature and failure to thrive. It also shows mutation effect on the expression, function and turnover of protein and highlights mechanistic details by which these potent regulators may operate.


Assuntos
Nanismo Hipofisário/genética , Testes Genéticos , Mutação/genética , Fator de Transcrição Pit-1/genética , Criança , Feminino , Hormônio do Crescimento Humano/genética , Humanos , Hipopituitarismo/genética , Masculino , Proteínas Mutantes/metabolismo , Taxa de Mutação , Prolactina/genética , Domínios Proteicos , Proteólise , Fator de Transcrição Pit-1/química , Ativação Transcricional/genética
9.
J Pediatr Endocrinol Metab ; 32(4): 415-419, 2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-30893054

RESUMO

Background Monoallelic mutations of GHR have been described in idiopathic short stature (ISS), although the significance of these remain unclear. We report a case of ISS with novel monoallelic S219L mutation of GHR and discuss the possible significance of monoallelic GHR mutation in ISS. Case presentation The proband, a 13.9-year-old Japanese boy, had severe short stature (-3.8 standard deviation [SD]). Serum insulin-like growth factor (IGF)-I level and growth hormone (GH) secretion was normal. His parents were nonconsanguineous and had normal stature. Genetic analyses revealed a novel monoallelic missense variation in exon 7 of GHR (S219L). The proband's mother had the same variation. S219L might be the novel mutation judging from there being no registration of it as a single-nucleotide polymorphism (SNP) in any database, evolutional conservation of Ser219, in silico analyses, and computational molecular visualization analysis. Furthermore, a review of the literature showed that the median height of missense mutation carriers of GHR was relatively low. Conclusions We propose the possibility that monoallelic mutation of GHR increases the susceptibility to short stature.


Assuntos
Estatura/genética , Transtornos do Crescimento/genética , Transtornos do Crescimento/patologia , Hormônio do Crescimento Humano/genética , Mutação , Adolescente , Alelos , Humanos , Masculino , Prognóstico
10.
Arch Endocrinol Metab ; 63(1): 70-78, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30864634

RESUMO

Short stature is a common feature, and frequently remains without a specific diagnosis after conventional clinical and laboratorial evaluation. Longitudinal growth is mainly determined by genetic factors, and hundreds of common variants have been associated to height variability among healthy individuals. Although isolated short stature may be caused by the combination of variants, with a deleterious impact on the growth of individuals with polygenic inheritance, recent studies have pointed out some monogenic defects as the cause of the growth disorder observed in nonsyndromic children. The majority of these defects are in genes related to the growth plate cartilage and in the growth hormone (GH) - insulin-like growth factor 1 (IGF-1) axis. Affected patients usually present the mildest spectrum of some forms of skeletal dysplasia, or subtle abnormalities of laboratory tests, suggesting hormonal resistance or insensibility. The lack of specific characteristics, however, does not allow formulation of a definitive diagnosis without the use of broad genetic studies. Thus, molecular genetic studies including panels of genes or exome analysis will become essential in investigating and identifying the causes of isolated short stature in children, with a crucial impact on treatment and follow-up.


Assuntos
Estatura/genética , Variação Genética/genética , Transtornos do Crescimento/genética , Hormônio do Crescimento Humano/genética , Fator de Crescimento Insulin-Like I/genética , Humanos
11.
Radiat Res ; 191(4): 360-368, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30759046

RESUMO

Exposure to ionizing radiation combined with traumatic tissue injury is an important life-threatening condition found in the civilian populations after nuclear and radiological events. The significance feature of radiation combined injury (RCI) is the severe combined effect, which makes the injury more complicated. At present, there are limited measures available to treat RCI. Here we show that a chimeric protein dTMP-GH, fusing human growth hormone (hGH) with a tandem dimer of thrombopoietin mimetic peptide (dTMP), could be an effective therapy agent for RCI in a mice model. In this study, using a RCI mouse model exposed to 60Co γ-ray photons (6.0 Gy, 0.3 Gy/min) followed by a 20% total-body-surface-area burns (henceforth called: RB-CI) was established. Administration of dTMP-GH (200 ug/kg) for 10 consecutive days beginning at 24 h after injury improved survival rate during a 30-day observation period compared with the control vehicle group. dTMP-GH treatment also showed enhanced bone marrow hematopoiesis recovery determined by peripheral blood analysis and bone marrow histopathology. Meanwhile, dTMP-GH treatment accelerated skin wound closure and mitigated ileum injury in the RCI model. These results suggest that dTMP-GH may prove to be an effective therapeutic drug for RCI.


Assuntos
Queimaduras/complicações , Hormônio do Crescimento Humano/uso terapêutico , Peptídeos/genética , Lesões Experimentais por Radiação/complicações , Lesões Experimentais por Radiação/tratamento farmacológico , Proteínas Recombinantes de Fusão/uso terapêutico , Pele/patologia , Animais , Hormônio do Crescimento Humano/genética , Humanos , Íleo/efeitos dos fármacos , Íleo/efeitos da radiação , Masculino , Camundongos , Peptídeos/química , Multimerização Proteica , Lesões Experimentais por Radiação/patologia , Lesões Experimentais por Radiação/fisiopatologia , Proteínas Recombinantes de Fusão/genética , Análise de Sobrevida , Cicatrização/efeitos dos fármacos , Cicatrização/efeitos da radiação
12.
Microb Cell Fact ; 18(1): 10, 2019 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-30657054

RESUMO

BACKGROUND: Many valuable biopharmaceutical and biotechnological proteins have been produced in Escherichia coli, however these proteins are almost exclusively localised in the cytoplasm or periplasm. This presents challenges for purification, i.e. the removal of contaminating cellular constituents. One solution is secretion directly into the surrounding media, which we achieved via the 'hijack' of the flagellar type III secretion system (FT3SS). Ordinarily flagellar subunits are exported through the centre of the growing flagellum, before assembly at the tip. However, we exploit the fact that in the absence of certain flagellar components (e.g. cap proteins), monomeric flagellar proteins are secreted into the supernatant. RESULTS: We report the creation and iterative improvement of an E. coli strain, by means of a modified FT3SS and a modular plasmid system, for secretion of exemplar proteins. We show that removal of the flagellin and HAP proteins (FliC and FlgKL) resulted in an optimal prototype. We next developed a high-throughput enzymatic secretion assay based on cutinase. This indicated that removal of the flagellar motor proteins, motAB (to reduce metabolic burden) and protein degradation machinery, clpX (to boost FT3SS levels intracellularly), result in high capacity secretion. We also show that a secretion construct comprising the 5'UTR and first 47 amino acidsof FliC from E. coli (but no 3'UTR) achieved the highest levels of secretion. Upon combination, we show a 24-fold improvement in secretion of a heterologous (cutinase) enzyme over the original strain. This improved strain could export a range of pharmaceutically relevant heterologous proteins [hGH, TrxA, ScFv (CH2)], achieving secreted yields of up to 0.29 mg L-1, in low cell density culture. CONCLUSIONS: We have engineered an E. coli which secretes a range of recombinant proteins, through the FT3SS, to the extracellular media. With further developments, including cell culture process strategies, we envision further improvement to the secreted titre of recombinant protein, with the potential application for protein production for biotechnological purposes.


Assuntos
Escherichia coli/metabolismo , Engenharia Metabólica , Sistemas de Secreção Tipo III/metabolismo , Regiões 5' não Traduzidas , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , Escherichia coli/crescimento & desenvolvimento , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Flagelos/metabolismo , Flagelina/genética , Hormônio do Crescimento Humano/genética , Hormônio do Crescimento Humano/metabolismo , Humanos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Tiorredoxinas/genética , Tiorredoxinas/metabolismo
13.
J Clin Res Pediatr Endocrinol ; 11(4): 426-431, 2019 11 22.
Artigo em Inglês | MEDLINE | ID: mdl-30678423

RESUMO

Isolated growth hormone (GH) deficiency (IGHD) type 2 is a rare autosomal dominant disorder characterized by severe short stature with low GH level. Timely diagnosis is important for optimal results of recombinant human GH (rhGH) treatment and detection of additional pituitary deficiencies in affected relatives. A male child presented at the age of one year with severe, proportionate short stature [-4.9 standard deviation score (SDS)] and with a normal body mass index (-1.1 SDS). Physical examination revealed frontal bossing, midfacial hypoplasia, normal external genitalia and no dysmorphic features. Paternal and maternal heights were -6.1 and -1.9 SDS. Serum insulin-like growth factor-1 (IGF-1) and IGF-binding protein-3 were undetectable and the peak GH concentration by clonidine stimulation test was extremely low (0.18 ng/mL). Brain magnetic resonance imaging showed anterior pituitary hypoplasia. Genetic analysis identified a novel heterozygous mutation (c.291+2T>G) expected to lead to splicing out exon 3 of GH1. rhGH from age 2.4 years led to appropriate catch-up. In conclusion, we identified a novel GH1 gene mutation in an infant with classical IGHD type 2 presentation.


Assuntos
Estatura/genética , Nanismo Hipofisário/genética , Transtornos do Crescimento/genética , Hormônio do Crescimento Humano/deficiência , Hormônio do Crescimento Humano/genética , Mutação , Fatores Etários , Estatura/efeitos dos fármacos , Desenvolvimento Infantil , Pré-Escolar , Nanismo Hipofisário/sangue , Nanismo Hipofisário/diagnóstico , Nanismo Hipofisário/tratamento farmacológico , Predisposição Genética para Doença , Transtornos do Crescimento/sangue , Transtornos do Crescimento/diagnóstico , Transtornos do Crescimento/tratamento farmacológico , Hereditariedade , Terapia de Reposição Hormonal , Hormônio do Crescimento Humano/administração & dosagem , Humanos , Lactente , Masculino , Linhagem , Fenótipo , Resultado do Tratamento
14.
Arch. endocrinol. metab. (Online) ; 63(1): 70-78, Jan.-Feb. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-989290

RESUMO

ABSTRACT Short stature is a common feature, and frequently remains without a specific diagnosis after conventional clinical and laboratorial evaluation. Longitudinal growth is mainly determined by genetic factors, and hundreds of common variants have been associated to height variability among healthy individuals. Although isolated short stature may be caused by the combination of variants, with a deleterious impact on the growth of individuals with polygenic inheritance, recent studies have pointed out some monogenic defects as the cause of the growth disorder observed in nonsyndromic children. The majority of these defects are in genes related to the growth plate cartilage and in the growth hormone (GH) - insulin-like growth factor 1 (IGF-1) axis. Affected patients usually present the mildest spectrum of some forms of skeletal dysplasia, or subtle abnormalities of laboratory tests, suggesting hormonal resistance or insensibility. The lack of specific characteristics, however, does not allow formulation of a definitive diagnosis without the use of broad genetic studies. Thus, molecular genetic studies including panels of genes or exome analysis will become essential in investigating and identifying the causes of isolated short stature in children, with a crucial impact on treatment and follow-up.


Assuntos
Humanos , Variação Genética/genética , Estatura/genética , Fator de Crescimento Insulin-Like I/genética , Hormônio do Crescimento Humano/genética , Transtornos do Crescimento/genética
15.
Endocr Rev ; 40(2): 476-505, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30265312

RESUMO

GH insensitivity (GHI) presents in childhood with growth failure and in its severe form is associated with extreme short stature and dysmorphic and metabolic abnormalities. In recent years, the clinical, biochemical, and genetic characteristics of GHI and other overlapping short stature syndromes have rapidly expanded. This can be attributed to advancing genetic techniques and a greater awareness of this group of disorders. We review this important spectrum of defects, which present with phenotypes at the milder end of the GHI continuum. We discuss their clinical, biochemical, and genetic characteristics. The objective of this review is to clarify the definition, identification, and investigation of this clinically relevant group of growth defects. We also review the therapeutic challenges of mild GHI.


Assuntos
Transtornos do Crescimento , Hormônio do Crescimento Humano , Fator de Crescimento Insulin-Like I , Adolescente , Criança , Pré-Escolar , Feminino , Transtornos do Crescimento/genética , Transtornos do Crescimento/metabolismo , Transtornos do Crescimento/patologia , Transtornos do Crescimento/fisiopatologia , Hormônio do Crescimento Humano/genética , Hormônio do Crescimento Humano/metabolismo , Humanos , Lactente , Fator de Crescimento Insulin-Like I/administração & dosagem , Fator de Crescimento Insulin-Like I/deficiência , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Masculino
16.
Am J Physiol Endocrinol Metab ; 316(2): E196-E209, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30532991

RESUMO

Fluorescent protein reporter genes are widely used to identify and sort murine pancreatic ß-cells. In this study, we compared use of the MIP-GFP transgene, which exhibits aberrant expression of human growth hormone (hGH), with a newly derived Ins2Apple allele that lacks hGH expression on the expression of sex-specific genes. ß-Cells from MIP-GFP transgenic mice exhibit changes in the expression of 7,733 genes, or greater than half of their transcriptome, compared with ß-cells from Ins2Apple/+ mice. To determine how these differences might affect a typical differential gene expression study, we analyzed the effect of sex on gene expression using both reporter lines. Six hundred fifty-seven differentially expressed genes were identified between male and female ß-cells containing the Ins2Apple allele. Female ß-cells exhibit higher expression of Xist, Tmed9, Arpc3, Eml2, and several islet-enriched transcription factors, including Nkx2-2 and Hnf4a, whereas male ß-cells exhibited a generally higher expression of genes involved in cell cycle regulation. In marked contrast, the same male vs. female comparison of ß-cells containing the MIP-GFP transgene revealed only 115 differentially expressed genes, and comparison of the 2 lists of differentially expressed genes revealed only 17 that were common to both analyses. These results indicate that 1) male and female ß-cells differ in their expression of key transcription factors and cell cycle regulators and 2) the MIP-GFP transgene may attenuate sex-specific differences that distinguish male and female ß-cells, thereby impairing the identification of sex-specific variations.


Assuntos
Proteínas de Fluorescência Verde/genética , Hormônio do Crescimento Humano/genética , Células Secretoras de Insulina/metabolismo , Insulina/genética , Animais , Feminino , Expressão Gênica , Genes Reporter/genética , Proteínas de Fluorescência Verde/metabolismo , Hormônio do Crescimento Humano/metabolismo , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Regiões Promotoras Genéticas , RNA Mensageiro/metabolismo , Fatores Sexuais , Transgenes
17.
Arch Endocrinol Metab ; 63(6): 608-617, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31939486

RESUMO

The somatotropic axis is the main hormonal regulator of growth. Growth hormone (GH), also known as somatotropin, and insulin-like growth factor 1 (IGF-1) are the key components of the somatotropic axis. This axis has been studied for a long time and the knowledge of how some molecules could promote or impair hormones production and action has been growing over the last decade. The enhancement of large-scale sequencing techniques has expanded the spectrum of known genes and several other candidate genes that could affect the GH-IGF1-bone pathway. To date, defects in more than forty genes were associated with an impairment of the somatotropic axis. These defects can affect from the secretion of GH to the bioavailability and action of IGF-1. Affected patients present a large heterogeneous group of conditions associated with growth retardation. In this review, we focus on the description of the GH-IGF axis genetic defects reported in the last decade. Arch Endocrinol Metab. 2019;63(6):608-17.


Assuntos
Transtornos do Crescimento/genética , Hormônio do Crescimento Humano/deficiência , Hormônio do Crescimento Humano/genética , Fator de Crescimento Insulin-Like I/genética , Mutação/genética , Genótipo , Transtornos do Crescimento/metabolismo , Humanos , Fator de Crescimento Insulin-Like I/metabolismo , Fenótipo , Transdução de Sinais
18.
FEMS Microbiol Lett ; 365(23)2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30476048

RESUMO

Methodologies that exploit the durability of Bacillus subtilis spores by displaying heterologous proteins or antigenic molecules on the spore surface for mucosal vaccine delivery and other applications are well established. Here we extend the concept by engineering spores intended as oral delivery vehicles for therapeutic proteins. The method is exemplified by the expression and deposition of human growth hormone in the developing spore core, where the protein is shielded from physicochemical and biological degradation by the protective spore structure. Lysates from physically disrupted spores are shown to stimulate differentiation of a pre-adipocyte cell line to mature adipocyte cells, indicating that the spore-core located human growth hormone is folded correctly and functional. We also introduce a methodology for controlled release of heterologous proteins from the spore core, which utilises components of the PBSX prophage to lyse spores during germination and outgrowth. With further development, spore core expression, coupled with an engineered autolytic germination mechanism, may permit the use of spores as oral delivery carriers of therapeutic proteins.


Assuntos
Bacillus subtilis/genética , Produtos Biológicos/metabolismo , Expressão Gênica , Hormônio do Crescimento Humano/genética , Hormônio do Crescimento Humano/metabolismo , Esporos Bacterianos/genética , Células 3T3 , Adipócitos/efeitos dos fármacos , Animais , Bacillus subtilis/metabolismo , Bacteriólise , Diferenciação Celular/efeitos dos fármacos , Técnicas de Visualização da Superfície Celular , Sistemas de Liberação de Medicamentos , Humanos , Lisogenia , Camundongos , Prófagos/genética , Prófagos/crescimento & desenvolvimento , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Esporos Bacterianos/metabolismo
19.
Pediatr Endocrinol Rev ; 16(Suppl 1): 17-27, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30378779

RESUMO

Recombinant human growth hormones were the products of a revolution in biotechnology that took place in the San Francisco Bay area of California in the 1970's. A combination of Herb Boyer's restriction enzymes with Stanley Cohen's bacterial plasmids provided the power to select and amplify virtually any gene. The complementary personalities and talents of Herb Boyer and Robert Swanson led to formation of Genentech and with it the development of a product that overcame the limitations of scarcity and the risks of slow virus contamination inherent in extracted pituitary growth hormone. The extra amino acid in metGH was dropped and other manufacturers joined in the effort to explore indications for rhGH beyond the replacement of a missing hormone. After more than thirty years of availability and careful study, we still have much to learn about the safe and effective use of rhGH.


Assuntos
Hormônio do Crescimento Humano/genética , Biotecnologia , Humanos , Proteínas Recombinantes , Análise de Sequência
20.
Biofactors ; 44(5): 480-484, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30175865

RESUMO

Biological functions of immunoglobulin-free light chains (FLCs), other than in chronic inflammatory diseases, are still poorly defined; the field of insulin resistance (IR) has not been investigated, despite the strict relationships with oxidative stress (OS) and inflammation. Therefore, we evaluated FLCs levels and their relationships with metabolic parameters in adult growth hormone deficiency (GHD) and metabolic syndrome (MetS), both characterized by IR. One hundred subjects were enrolled: group A, patients with GHD [n =31, 24-69 years, mean ± SEM body mass index (BMI) 26.8 ± 1.5 kg/m2 ]; group B, patients with MetS (n = 29, 21-70 years, BMI 31.9 ± 1.3); group C, controls (N = 40, 21-62 years, BMI 21.6 ± 1.1). Groups were matched by age range and, for patients, by BMI. Morning blood sample was collected for metabolic parameters and FLCs, assessed by turbidimetric assay. GHD patients show levels of FLCs significantly higher than MetS and controls (mean ± SEM κ 37.21 ± 6.97, 15.27 ± 0.86, 12.34 ± 0.85 mg/l; λ 19.44 ± 2.61, 11.78 ± 0.72 and 11.67 ± 0.77 mg/l; κ/λ ratio 1.77 ± 0.13, 1.38 ± 0.09; and 1.10 ± 0.06, respectively); only κ were higher in MetS versus controls. Therefore, the ratio showed progressive declining values in GHD versus MetS versus controls. Our data show increased FLCs levels in GHD and MetS, with the highest values in the former. Both conditions show OS, but with different molecular patterns. FLCs may contribute to chronic inflammation, leading to OS, and cardiovascular complications of GHD. Prognostic and therapeutic implications require further investigation. © 2018 BioFactors, 44(5):480-484, 2018.


Assuntos
Nanismo Hipofisário/imunologia , Hormônio do Crescimento Humano/genética , Inflamação/imunologia , Síndrome Metabólica/imunologia , Plasmócitos/imunologia , Adulto , Idoso , Índice de Massa Corporal , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/complicações , Doenças Cardiovasculares/imunologia , Nanismo Hipofisário/sangue , Nanismo Hipofisário/complicações , Nanismo Hipofisário/patologia , Feminino , Hormônio do Crescimento Humano/sangue , Hormônio do Crescimento Humano/deficiência , Humanos , Cadeias Leves de Imunoglobulina/sangue , Cadeias Leves de Imunoglobulina/genética , Cadeias Leves de Imunoglobulina/imunologia , Inflamação/sangue , Inflamação/patologia , Resistência à Insulina/genética , Masculino , Síndrome Metabólica/sangue , Síndrome Metabólica/patologia , Pessoa de Meia-Idade , Estresse Oxidativo
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