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The study by Chen et al. is the first to apply the revolutionary genetic engineering tool, base editing, in a rat model for the treatment of primary hyperoxaluria type 1, a disease that originates in the liver but in which the kidney is the main organ affected. This commentary contextualizes and describes the gene-editing technology applied by the authors, provides an interpretation and opinion of their results, and indicates possible future applications.
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Edição de Genes , Nefropatias , Ratos , Animais , Edição de Genes/métodos , Sistemas CRISPR-Cas , Engenharia Genética , Nefropatias/genética , Nefropatias/terapia , RimRESUMO
BACKGROUND: In autosomal dominant polycystic kidney disease (ADPKD), cyst development and enlargement lead to ESKD. Macrophage recruitment and interstitial inflammation promote cyst growth. TWEAK is a TNF superfamily (TNFSF) cytokine that regulates inflammatory responses, cell proliferation, and cell death, and its receptor Fn14 (TNFRSF12a) is expressed in macrophage and nephron epithelia. METHODS: To evaluate the role of the TWEAK signaling pathway in cystic disease, we evaluated Fn14 expression in human and in an orthologous murine model of ADPKD. We also explored the cystic response to TWEAK signaling pathway activation and inhibition by peritoneal injection. RESULTS: Meta-analysis of published animal-model data of cystic disease reveals mRNA upregulation of several components of the TWEAK signaling pathway. We also observed that TWEAK and Fn14 were overexpressed in mouse ADPKD kidney cysts, and TWEAK was significantly high in urine and cystic fluid from patients with ADPKD. TWEAK administration induced cystogenesis and increased cystic growth, worsening the phenotype in a murine ADPKD model. Anti-TWEAK antibodies significantly slowed the progression of ADPKD, preserved renal function, and improved survival. Furthermore, the anti-TWEAK cystogenesis reduction is related to decreased cell proliferation-related MAPK signaling, decreased NF-κB pathway activation, a slight reduction of fibrosis and apoptosis, and an indirect decrease in macrophage recruitment. CONCLUSIONS: This study identifies the TWEAK signaling pathway as a new disease mechanism involved in cystogenesis and cystic growth and may lead to a new therapeutic approach in ADPKD.
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Citocina TWEAK/metabolismo , Rim Policístico Autossômico Dominante/metabolismo , Rim Policístico Autossômico Dominante/patologia , Receptor de TWEAK/metabolismo , Adulto , Animais , Anticorpos Neutralizantes/farmacologia , Apoptose , Proliferação de Células/efeitos dos fármacos , Cistos/metabolismo , Cistos/patologia , Citocina TWEAK/antagonistas & inibidores , Citocina TWEAK/genética , Citocina TWEAK/farmacologia , Modelos Animais de Doenças , Progressão da Doença , Feminino , Fibrose , Expressão Gênica , Humanos , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos , Masculino , Camundongos , Pessoa de Meia-Idade , NF-kappa B/metabolismo , Rim Policístico Autossômico Dominante/fisiopatologia , Transdução de Sinais , Receptor de TWEAK/genéticaRESUMO
The Polycystic Kidney Disease (PKD) is characterized by progressive renal cyst development and other extrarenal manifestation including Polycystic Liver Disease (PLD). Phenotypical characterization of animal models mimicking human diseases are commonly used, in order to, study new molecular mechanisms and identify new therapeutic approaches. The main biomarker of disease progression is total volume of kidney and liver in both human and mouse, which correlates with organ function. For this reason, the estimation of the number and area of the tissue occupied by cysts, is critical for the understanding of physiological mechanisms underlying the disease. In this regard, cystic index is a robust parameter commonly used to quantify the severity of the disease. To date, the vast majority of biomedical researchers use ImageJ as a software tool to estimate the cystic index by quantifying the cystic areas of histological images after thresholding. This tool has imitations of being inaccurate, largely due to incorrectly identifying non-cystic regions. We have developed a new software, named CystAnalyser (register by Universidade de Santiago de Compostela-USC, and Fundación Investigación Sanitaria de Santiago-FIDIS), that combines automatic image processing with a graphical user friendly interface that allows investigators to oversee and easily correct the image processing before quantification. CystAnalyser was able to generate a cystic profile including cystic index, number of cysts and cyst size. In order to test the CystAnalyser software, 795 cystic kidney, and liver histological images were analyzed. Using CystAnalyser there were no differences calculating cystic index automatically versus user input, except in specific circumstances where it was necessary for the user to distinguish between mildly cystic from non-cystic regions. The sensitivity and specificity of the number of cysts detected by the automatic quantification depends on the type of organ and cystic severity, with values 76.84-78.59% and 76.96-89.66% for the kidney and 87.29-93.80% and 63.42-86.07% for the liver. CystAnalyser, in addition, provides a new tool for estimating the number of cysts and a more specific measure of the cystic index than ImageJ. This study proposes CystAnalyser is a new robust and freely downloadable software tool for analyzing the severity of disease by quantifying histological images of cystic organs for routine biomedical research. CystAnalyser can be downloaded from https://citius.usc.es/transferencia/software/cystanalyser (for Windows and Linux) for research purposes.
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Cistos , Interpretação de Imagem Assistida por Computador/métodos , Hepatopatias , Doenças Renais Policísticas , Software , Algoritmos , Animais , Cistos/classificação , Cistos/diagnóstico por imagem , Cistos/patologia , Histocitoquímica , Humanos , Rim/diagnóstico por imagem , Rim/patologia , Fígado/diagnóstico por imagem , Fígado/patologia , Hepatopatias/classificação , Hepatopatias/diagnóstico por imagem , Hepatopatias/patologia , Camundongos , Doenças Renais Policísticas/classificação , Doenças Renais Policísticas/diagnóstico por imagem , Doenças Renais Policísticas/patologiaRESUMO
Autosomal recessive polycystic kidney disease (ARPKD) is a rare disorder and one of the most severe forms of polycystic kidney disease, leading to end-stage renal disease (ESRD) in childhood. PKHD1 is the gene that is responsible for the vast majority of ARPKD. However, some cases have been related to a new gene that was recently identified (DZIP1L gene), as well as several ciliary genes that can mimic a ARPKD-like phenotypic spectrum. In addition, a number of molecular pathways involved in the ARPKD pathogenesis and progression were elucidated using cellular and animal models. However, the function of the ARPKD proteins and the molecular mechanism of the disease currently remain incompletely understood. Here, we review the clinics, treatment, genetics, and molecular basis of ARPKD, highlighting the most recent findings in the field.
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Suscetibilidade a Doenças , Rim Policístico Autossômico Recessivo/etiologia , Rim Policístico Autossômico Recessivo/metabolismo , Animais , Biomarcadores , Ensaios Clínicos como Assunto , Terapia Combinada , Diagnóstico Diferencial , Gerenciamento Clínico , Estudos de Associação Genética/métodos , Predisposição Genética para Doença , Humanos , Mutação , Fenótipo , Rim Policístico Autossômico Recessivo/patologia , Locos de Características Quantitativas , Receptores de Superfície Celular/química , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Transdução de Sinais , Relação Estrutura-Atividade , Resultado do TratamentoRESUMO
Gitelman and Bartter syndromes are rare inherited diseases that belong to the category of renal tubulopathies. The genes associated with these pathologies encode electrolyte transport proteins located in the nephron, particularly in the Distal Convoluted Tubule and Ascending Loop of Henle. Therefore, both syndromes are characterized by alterations in the secretion and reabsorption processes that occur in these regions. Patients suffer from deficiencies in the concentration of electrolytes in the blood and urine, which leads to different systemic consequences related to these salt-wasting processes. The main clinical features of both syndromes are hypokalemia, hypochloremia, metabolic alkalosis, hyperreninemia and hyperaldosteronism. Despite having a different molecular etiology, Gitelman and Bartter syndromes share a relevant number of clinical symptoms, and they have similar therapeutic approaches. The main basis of their treatment consists of electrolytes supplements accompanied by dietary changes. Specifically for Bartter syndrome, the use of non-steroidal anti-inflammatory drugs is also strongly supported. This review aims to address the latest diagnostic challenges and therapeutic approaches, as well as relevant recent research on the biology of the proteins involved in disease. Finally, we highlight several objectives to continue advancing in the characterization of both etiologies.
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Síndrome de Bartter/patologia , Síndrome de Gitelman/patologia , Túbulos Renais Distais/patologia , Alça do Néfron/patologia , Equilíbrio Hidroeletrolítico/fisiologia , Síndrome de Bartter/diagnóstico , Síndrome de Bartter/genética , Síndrome de Bartter/terapia , Eletrólitos/análise , Eletrólitos/uso terapêutico , Síndrome de Gitelman/diagnóstico , Síndrome de Gitelman/genética , Síndrome de Gitelman/terapia , Humanos , Hiperaldosteronismo/patologia , Hipercalciúria/patologia , Hipopotassemia/patologia , Hiponatremia/patologia , Nefrocalcinose/patologia , Erros Inatos do Transporte Tubular Renal/patologiaRESUMO
PURPOSE: This study aimed to assess the effect of vibration frequency (f out) on the electromyographic (EMG) activity of the biceps brachii (BB) and triceps brachii (TB) muscles when acting as agonist and antagonist during static exercises with different loads. METHODS: Fourteen healthy men were asked to hold a vibratory bar as steadily as possible for 10 s during lying row (pulling) and bench press (pushing) exercise at f out of 0 (non-vibration condition), 18, 31 and 42 Hz with loads of 20, 50, and 80 % of the maximum sustainable load (MSL). The root mean square of the EMG activity (EMGRMS) of the BB and TB muscles was expressed as a function of the maximal EMGRMS for respective muscles to characterize agonist activation and antagonist coactivation. RESULTS: We found that (1) agonist activation was greater during vibration (42 Hz) compared to non-vibration exercise for the TB but not for the BB muscle (p < 0.05); (2) antagonist activation was greater during vibration compared to non-vibration exercise for both BB (p < 0.01) and TB (p < 0.05) muscles; (3) the vibration-induced increase in antagonist coactivation was proportional to vibration f out in the range 18-42 Hz and (4) the vibration-induced increase in TB agonist activation and antagonist coactivation occurred at all loading conditions in the range 20-80 % MSL. CONCLUSION: The use of high vibration frequencies within the range of 18-42 Hz can maximize TB agonist activation and antagonist activation of both BB and TB muscles during upper limb vibration exercise.
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Braço/fisiologia , Contração Muscular , Músculo Esquelético/fisiologia , Vibração , Adulto , Humanos , MasculinoRESUMO
This study aimed to analyze the influence of different vibration frequencies, loading conditions, and exercise types on the mechanical behavior of a novel vibratory bar (VB). Fourteen healthy men were asked to hold the VB during lying row (pulling) and bench press (pushing) static exercise as steadily as possible for 10 seconds with loads of 20, 50, and 80% of the maximum sustained load (MSL) and at preset vibration frequencies (f(in)) of 20, 35, and 50 Hz. Root mean square vibration acceleration (a(RMS)), peak-to-peak displacement (D), and frequency (f(out)) were gained from a 3-dimensional accelerometer fixed to the VB. Increasing vibration frequency (from 20 to 50 Hz) resulted in a progressive and sizeable increase in VB a(RMS) and f(out) (both p ≤ 0.001) with smaller variations of D (≤5.9%, p ≤ 0.001). Adding weight to the VB (progressive overload from 20 to 80% MSL) did not affect D and minimally altered a(RMS) (<4.2%, p = 0.014) and f(out) (<1.7%, p = 0.002). Altering the type of exercise (pulling vs. pushing) did not affect VB a(RMS), D, and f(out). In conclusion, this study establishes the validity of a novel VB and legitimates its use for effective and safe upper-body static exercise with a wide range of vibration frequencies and loading conditions in the context of physical training or rehabilitation.
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Tolerância ao Exercício/fisiologia , Exercício Físico/fisiologia , Músculo Esquelético/fisiologia , Vibração , Levantamento de Peso/fisiologia , Aceleração , Adulto , Antropometria , Humanos , Contração Isométrica/fisiologia , Masculino , Reprodutibilidade dos Testes , Estudos de Amostragem , Suporte de Carga , Adulto JovemRESUMO
Focal and segmental glomerulosclerosis (FSGS) is a histological pattern that has several etiologies, including genetics. The autosomal dominant form of FSGS is a heterogenic disease caused by mutations within three known genes: α-actinin 4 (ACTN4), canonical transient receptor potential 6 (TRPC6), and the inverted formin 2 (INF2) gene. More recently, INF2 mutations have also been attributed to Charcot-Marie-Tooth neuropathy associated with FSGS. Here we performed direct sequencing, histological characterization, and functional studies in a cohort of families with autosomal dominant FSGS. We detected a novel mutation in exon 6 of the INF2 gene outside of the exon 2-4 candidate region used for rapid diagnosis of autosomal dominant FSGS. This new mutation is predicted to alter a highly conserved amino-acid residue within the 17th α-helix of the diaphanous inhibitory domain of the protein. A long-term follow-up of this family indicated that all patients were diagnosed in adulthood, as opposed to early childhood, and progression to end-stage renal disease was at different times without clinical or electrodiagnostic evidence of neuropathy. Thus, this novel mutation in INF2 linked to nonsyndromic FSGS indicates the necessity for full gene sequencing if no mutation is found in the current rapid-screen region of the gene.
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Testes Genéticos , Glomerulosclerose Segmentar e Focal/diagnóstico , Glomerulosclerose Segmentar e Focal/genética , Mutação/genética , Proteínas do Tecido Nervoso/genética , Adulto , Sequência de Aminoácidos , Sequência de Bases , Doença de Charcot-Marie-Tooth/genética , Estudos de Coortes , Progressão da Doença , Éxons/genética , Feminino , Seguimentos , Glomerulosclerose Segmentar e Focal/epidemiologia , Humanos , Masculino , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/química , Linhagem , Espanha/epidemiologiaRESUMO
The intestinal epithelium is a complex system characterized by massive and continuous cell renewal and differentiation. In this context, cell-type-specific transcription factors are thought to play a crucial role by modulating specific transcription networks and signalling pathways. Hnf1alpha and beta are closely related atypical homeoprotein transcription factors expressed in several epithelia, including the gut. With the use of a conditional inactivation system, we generated mice in which Hnf1b is specifically inactivated in the intestinal epithelium on a wild-type or Hnf1a(-/-) genetic background. Whereas the inactivation of Hnf1a or Hnf1b alone did not lead to any major intestinal dysfunction, the concomitant inactivation of both genes resulted in a lethal phenotype. Double-mutant animals had defective differentiation and cell fate commitment. The expression levels of markers of all the differentiated cell types, both enterocytes and secretory cells, were affected. In addition, the number of goblet cells was increased, whereas mature Paneth cells were missing. At the molecular level, we show that Hnf1alpha and beta act upstream of the Notch pathway controlling directly the expression of two crucial components: Jag1 and Atoh1. We demonstrate that the double-mutant mice present with a defect in intestinal water absorption and that Hnf1alpha and beta directly control the expression of Slc26a3, a gene whose mutations are associated with chloride diarrhoea in human patients. Our study identifies new direct target genes of the Hnf1 transcription factors and shows that they play crucial roles in both defining cell fate and controlling terminal functions in the gut epithelium.
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Diferenciação Celular/fisiologia , Fator 1-alfa Nuclear de Hepatócito/metabolismo , Fator 1-beta Nuclear de Hepatócito/metabolismo , Mucosa Intestinal/citologia , Mucosa Intestinal/metabolismo , Animais , Antiporters/genética , Antiporters/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Diferenciação Celular/genética , Imunoprecipitação da Cromatina , Fator 1-alfa Nuclear de Hepatócito/genética , Fator 1-beta Nuclear de Hepatócito/genética , Imuno-Histoquímica , Hibridização In Situ , Técnicas In Vitro , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteína Jagged-1 , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Knockout , Reação em Cadeia da Polimerase , Proteínas Serrate-Jagged , Transportadores de SulfatoRESUMO
Wearables are being increasingly used to monitor heart rate (HR). However, their usefulness for analyzing continuous HR in research or at clinical level is questionable. The aim of this study is to analyze the level of agreement between different wearables in the measurement of HR based on photoplethysmography, according to different body positions and physical activity levels, and compared to a gold-standard ECG. The proposed method measures agreement among several time scales since different wearables obtain HR at different sampling rates. Eighteen university students (10 men, 8 women; 22 ± 2.45 years old) participated in a laboratory study. Participants simultaneously wore an Apple Watch and a Polar Vantage watch. ECG was measured using a BIOPAC system. HR was recorded continuously and simultaneously by the three devices, for consecutive 5-min periods in 4 different situations: lying supine, sitting, standing and walking at 4 km/h on a treadmill. HR estimations were obtained with the maximum precision offered by the software of each device and compared by averaging in several time scales, since the wearables obtained HR at different sampling rates, although results are more detailed for 5 s and 30 s epochs. Bland-Altman (B-A) plots show that there is no noticeable difference between data from the ECG and any of the smartwatches while participants were lying down. In this position, the bias is low when averaging in both 5 s and 30 s. Differently, B-A plots show that there are differences when the situation involves some level of physical activity, especially for shorter epochs. That is, the discrepancy between devices and the ECG was greater when walking on the treadmill and during short time scales. The device showing the biggest discrepancy was the Polar Watch, and the one with the best results was the Apple Watch. We conclude that photoplethysmography-based wearable devices are suitable for monitoring HR averages at regular intervals, especially at rest, but their feasibility is debatable for a continuous analysis of HR for research or clinical purposes, especially when involving some level of physical activity. An important contribution of this work is a new methodology to synchronize and measure the agreement against a gold standard of two or more devices measuring HR at different and not necessarily even paces.
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Fotopletismografia , Dispositivos Eletrônicos Vestíveis , Adulto , Exercício Físico/fisiologia , Feminino , Frequência Cardíaca/fisiologia , Humanos , Masculino , Monitorização Fisiológica/métodos , Fotopletismografia/métodos , Adulto JovemRESUMO
Genetic kidney diseases (GKDs) are a group of rare diseases, affecting approximately about 60 to 80 per 100,000 individuals, for which there is currently no treatment that can cure them (in many cases). GKDs usually leads to early-onset chronic kidney disease, which results in patients having to undergo dialysis or kidney transplant. Here, we briefly describe genetic causes and phenotypic effects of six GKDs representative of different ranges of prevalence and renal involvement (ciliopathy, glomerulopathy, and tubulopathy). One of the shared characteristics of GKDs is that most of them are monogenic. This characteristic makes it possible to use site-specific nuclease systems to edit the genes that cause GKDs and generate in vitro and in vivo models that reflect the genetic abnormalities of GKDs. We describe and compare these site-specific nuclease systems (zinc finger nucleases (ZFNs), transcription activator-like effect nucleases (TALENs) and regularly clustered short palindromic repeat-associated protein (CRISPR-Cas9)) and review how these systems have allowed the generation of cellular and animal GKDs models and how they have contributed to shed light on many still unknown fields in GKDs. We also indicate the main obstacles limiting the application of these systems in a more efficient way. The information provided here will be useful to gain an accurate understanding of the technological advances in the field of genome editing for GKDs, as well as to serve as a guide for the selection of both the genome editing tool and the gene delivery method most suitable for the successful development of GKDs models.
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Edição de Genes , Nefropatias , Animais , Sistemas CRISPR-Cas/genética , Endonucleases/genética , Feminino , Edição de Genes/métodos , Humanos , Nefropatias/genética , Nefropatias/terapia , Masculino , Nucleases de Dedos de ZincoRESUMO
(1) Background: Polycystic liver disease (PLD) is a heterogeneous group of congenital disorders characterized by bile duct dilatation and cyst development derived from cholangiocytes. Nevertheless, the cystogenesis mechanism is currently unknown and the PLD treatment is limited to liver transplantation. Novel and efficient therapeutic approaches are th6us needed. In this context, the present work has a principal aim to find novel molecular pathways, as well as new therapeutic targets, involved in the hepatic cystogenesis process. (2) Methods: Quantitative proteomics based on SWATH-MS technology were performed comparing hepatic proteomes of Wild Type and mutant/polycystic livers in a polycystic kidney disease (PKD) murine model (Pkd1cond/cond;Tam-Cre-/+). (3) Results: We identified several proteins altered in abundance, with two-fold cut-off up-regulation or down-regulation and an adjusted p-value significantly related to hepatic cystogenesis. Then, we performed enrichment and a protein-protein analysis identifying a cluster focused on hepatic fibrinogens. Finally, we validated a selection of targets by RT-qPCR, Western blotting and immunohistochemistry, finding a high correlation with quantitative proteomics data and validating the fibrinogen complex. (4) Conclusions: This work identified a novel molecular pathway in cystic liver disease, highlighting the fibrinogen complex as a possible new therapeutic target for PLD.
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Polycystin-1 (PC1) has an essential function in renal tubular morphogenesis and disruption of its function causes cystogenesis in human autosomal dominant polycystic kidney disease. We have previously shown that recombinant human PC1 is cis-autoproteolytically cleaved at the G protein-coupled receptor proteolytic site domain. To investigate the role of cleavage in vivo, we generated by gene targeting a Pkd1 knockin mouse (Pkd1(V/V)) that expresses noncleavable PC1. The Pkd1(V/V) mice show a hypomorphic phenotype, characterized by a delayed onset and distal nephron segment involvement of cystogenesis at postnatal maturation stage. We show that PC1 is ubiquitously and incompletely cleaved in wild-type mice, so that uncleaved and cleaved PC1 molecules coexist. Our study establishes a critical but restricted role of cleavage for PC1 function and suggests a differential function of the two types of PC1 molecules in vivo.
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Túbulos Renais/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Canais de Cátion TRPP/metabolismo , Envelhecimento/fisiologia , Animais , Animais Recém-Nascidos , Sequência de Bases , Células Cultivadas , Deleção de Genes , Regulação da Expressão Gênica no Desenvolvimento , Genótipo , Doenças Renais Císticas/genética , Doenças Renais Císticas/metabolismo , Doenças Renais Císticas/patologia , Camundongos , Camundongos Transgênicos , Receptores Acoplados a Proteínas G/genéticaRESUMO
Programmed cell senescence during embryo development is a recently described process that opens a new perspective to understand the senescence response and that adds a new player whose contribution to development needs to be addressed. Identifying developmental syndromes with a root in deregulated programmed cell senescence will undoubtedly reinforce our view of senescence and could provide a new angle to confront disease. One of the structures that was initially reported to undergo cellular senescence is the mesonephros. During E12.5-E14.5, before regression, mesonephric tubules are positive for the most widely used marker of cell senescence, SAßG, and negative for proliferation marker, Ki67, in a p21Cip1-dependent manner. PKD2 is one of the genes defective in autosomal dominant polycystic kidney disease (ADPKD). Inherited mutations in this gene result in cyst formation in adults after a secondary hit. Polycystin-2 (PC2) protein, the product of PKD2 gene expression, inhibits cell cycle progression by inducing p21Cip1, whereas mutated PKD2 results in increased proliferation and defective differentiation of kidney epithelial cells. Here, we addressed the possibility of defective programmed cell senescence as a consequence of Pkd2 deletion in mice. We analyzed embryos for the expression of the senescence marker SAßG, for the proliferative status of mesonephric tubule cells, and for the expression of p21Cip1, without identifying any noticeable deregulation of cell senescence. Our results exclude defective programmed cell senescence upon Pkd2 ablation as an initial event in ADPKD.
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Senescência Celular , Desenvolvimento Embrionário , Canais de Cátion TRPP/fisiologia , Ductos Mesonéfricos/citologia , Animais , Camundongos , Camundongos Knockout , Ductos Mesonéfricos/metabolismoRESUMO
A method for estimating the uncertainty in time-domain indices of RR time series is described. The method relies on the central limit theorem that states that the distribution of a sample average of independent samples has an uncertainty that asymptotically approaches to the sample standard deviation divided by the square root of the number of samples. Because RR time series cannot be characterized by a set of independent samples, we propose to estimate the uncertainty of indices by computing them in blocks that satisfy that the obtained partial indices are independent. We propose a methodology to search sets of independent partial indices and apply this methodology to the estimation of the uncertainty in the mean RR, SDRR, and r-msDD indices. The results show that the uncertainty can be higher than the 10% of the index for the SDRR and even higher for the r-msDD. Moreover, a statistical test for the difference of two indices is proposed.
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Algoritmos , Diagnóstico por Computador/métodos , Sistema de Condução Cardíaco/fisiologia , Frequência Cardíaca/fisiologia , Modelos Cardiovasculares , Simulação por Computador , Interpretação Estatística de Dados , Humanos , Modelos Estatísticos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common genetic disorders worldwide. In recent decades, the field has undergone a revolution, starting with the identification of causal ADPKD genes, including PKD1, PKD2, and the recently identified GANAB. In addition, advances defining the genetic mechanisms, protein localization and function, and the identification of numerous pathways involved in the disease process, have contributed to a better understanding of this illness. Together, this has led to a better prognosis, diagnosis, and treatment in clinical practice. In this mini review, we summarize and discuss new insights about the molecular mechanisms underlying ADPKD, including its genetics, protein function, and cellular pathways.
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Autosomal recessive polycystic kidney disease (ARPKD) is an important childhood nephropathy, occurring 1 in 20,000 live births. The major clinical phenotypes are expressed in the kidney with dilatation of the collecting ducts, systemic hypertension, and progressive renal insufficiency, and in the liver with biliary dysgenesis, portal tract fibrosis, and portal hypertension. The systemic hypertension has been attributed to enhanced distal sodium reabsorption in the kidney, the structural defects have been ascribed to altered cellular morphology, and fibrosis to increased TGF-ß signaling in the kidney and biliary tract, respectively. The pathogenic mechanisms underlying these abnormalities have not been determined. In the current report, we find that disrupting PKHD1 results in altered sub-cellular localization and function of the C2-WWW-HECT domain E3 family of ligases regulating these processes. We also demonstrate altered activity of RhoA and increased TGF-ß signaling and ENaC activity. Linking these phenomena, we found that vesicles containing the PKHD1/Pkhd1 gene product, FPC, also contain the NEDD4 ubiquitin ligase interacting protein, NDFIP2, which interacts with multiple members of the C2-WWW-HECT domain E3 family of ligases. Our results provide a mechanistic explanation for both the cellular effects and in vivo phenotypic abnormalities in mice and humans that result from Pkhd1/PKHD1 mutation.
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Ubiquitina-Proteína Ligases Nedd4/metabolismo , Rim Policístico Autossômico Recessivo/genética , Rim Policístico Autossômico Recessivo/metabolismo , Receptores de Superfície Celular/deficiência , Animais , Biomarcadores , Linhagem Celular , Modelos Animais de Doenças , Ativação Enzimática , Expressão Gênica , Humanos , Espaço Intracelular/metabolismo , Masculino , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Modelos Biológicos , Mutação , Rim Policístico Autossômico Recessivo/patologia , Transporte Proteico , Ratos , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Transdução de Sinais , Proteínas rho de Ligação ao GTP/metabolismoRESUMO
Maturity Onset Diabetes of the Young type 3 (MODY3), linked to mutations in the transcription factor HNF1A, is the most prevalent form of monogenic diabetes mellitus. HNF1alpha-deficiency leads to defective insulin secretion via a molecular mechanism that is still not completely understood. Moreover, in MODY3 patients the severity of insulin secretion can be extremely variable even in the same kindred, indicating that modifier genes may control the onset of the disease. With the use of a mouse model for HNF1alpha-deficiency, we show here that specific genetic backgrounds (C3H and CBA) carry a powerful genetic suppressor of diabetes. A genome scan analysis led to the identification of a major suppressor locus on chromosome 3 (Moda1). Moda1 locus contains 11 genes with non-synonymous SNPs that significantly interacts with other loci on chromosomes 4, 11 and 18. Mechanistically, the absence of HNF1alpha in diabetic-prone (sensitive) strains leads to postnatal defective islets growth that is remarkably restored in resistant strains. Our findings are relevant to human genetics since Moda1 is syntenic with a human locus identified by genome wide association studies of fasting glycemia in patients. Most importantly, our results show that a single genetic locus can completely suppress diabetes in Hnf1a-deficiency.
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We describe the case of a young woman who was diagnosed with advanced kidney disease, with an incidental finding of nephrocalcinosis of unknown aetiology, having been found asymptomatic throughout her life. The genetic study by panels of known genes associated with tubulointerstitial disease allowed us to discover autosomal dominant distal renal tubular acidosis associated with a de novo mutation in exon 14 of the SLC4A1 gene, which would have been impossible to diagnose clinically due to the advanced nature of the kidney disease when it was discovered.
Assuntos
Acidose Tubular Renal/diagnóstico , Acidose Tubular Renal/genética , Proteína 1 de Troca de Ânion do Eritrócito/genética , Adulto , Éxons , Feminino , Humanos , Rim/fisiopatologia , Mutação , Nefrocalcinose/etiologiaRESUMO
Acute kidney injury (AKI) and chronic kidney disease (CKD) are associated with decreased renal function and increased mortality risk, while the therapeutic armamentarium is unsatisfactory. The availability of adequate animal models may speed up the discovery of biomarkers for disease staging and therapy individualization as well as design and testing of novel therapeutic strategies. Some longstanding animal models have failed to result in therapeutic advances in the clinical setting, such as kidney ischemia-reperfusion injury and diabetic nephropathy models. In this regard, most models for diabetic nephropathy are unsatisfactory in that they do not evolve to renal failure. Satisfactory models for additional nephropathies are needed. These include anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, IgA nephropathy, anti-phospholipase-A2-receptor (PLA2R) membranous nephropathy and Fabry nephropathy. However, recent novel models hold promise for clinical translation. Thus, the AKI to CKD translation has been modeled, in some cases with toxins of interest for human CKD such as aristolochic acid. Genetically modified mice provide models for Alport syndrome evolving to renal failure that have resulted in clinical recommendations, polycystic kidney disease models that have provided clues for the development of tolvaptan, that was recently approved for the human disease in Japan; and animal models also contributed to target C5 with eculizumab in hemolytic uremic syndrome. Some ongoing trials explore novel concepts derived from models, such TWEAK targeting as tissue protection for lupus nephritis. We now review animal models reproducing diverse, genetic and acquired, causes of AKI and CKD evolving to kidney failure and discuss the contribution to clinical translation and prospects for the future.