La glycogénose de type Ia (GSDIa) est une maladie métabolique rare causée par une déficience en glucose-6-phosphatase (G6Pase), due à des mutations de la sous-unité catalytique (G6PC). Cette enzyme confère au foie, aux reins et à l’intestin la capacité de produire du glucose. Les patients atteints de GSDIa sont donc incapables de produire du glucose et souffrent d’hypoglycémies sévères lors de jeûnes courts. De plus, la déficience en G6Pase provoque une accumulation de glucose-6 phosphate dans le foie et les reins, conduisant à l’accumulation de glycogène et de lipides. A long terme, la plupart des patients souffre d’une maladie chronique rénale (MCR), qui peut évoluer en insuffisance rénale, nécessitant une mise sous dialyse ou une transplantation rénale. Cette MCR se caractérise par une fibrose, ainsi que par le développement de kystes dans les stades tardifs. Au niveau du foie, les patients développent une hépatomégalie et une stéatose hépatique qui peut évoluer vers le développement d’adénomes ou carcinomes hépatocellulaires. Le but de mes travaux de thèse a été d’identifier les mécanismes moléculaires impliqués dans l’établissement de la pathologie rénale et la formation des kystes, à l’aide de modèles murins invalidés pour le gène G6pc spécifiquement dans les reins (souris K.G6pc-/-). Alors que la GSDIa est une maladie caractérisée par l’accumulation hépatique et rénale de glycogène, nous avons d’abord montré que le développement de la fibrose, à l’origine de la perte de la fonction rénale, était induit par l’accumulation de lipides, indépendamment du contenu en glycogène. De plus, l’utilisation d’un agoniste de PPARα, le fénofibrate, en diminuant le contenu lipidique rénal, a ralenti l’installation de la fibrose et l’évolution de la MCR. Le mécanisme moléculaire impliqué est l’activation du système rénine angiotensine par les dérivés lipidiques, qui induit l’expression du facteur profibrotique TGFβ1. De même, le fénofibrate en limitant l’accumulation de lipides hépatiques a prévenu le développement d’atteintes hépatiques caractéristiques de la GSDI. Ainsi, l’activation du catabolisme des lipides par des agonistes de PPARα semble une stratégie thérapeutique intéressante pour réduire la progression des maladies rénales et hépatique de la GSDI. La deuxième partie de mes résultats suggèrent que le développement de kystes rénaux chez les patients atteints de la GSDI pourrait être causé par une altération du cil primaire, organelle jouant un rôle clé dans le maintien d’une structure et fonction normale des reins. En effet, une augmentation de la longueur du cil primaire a pu être observée dans les reins des souris K.G6pc-/- associée à une dérégulation de différentes protéines impliquées dans sa structure et sa fonction, par rapport aux souris contrôles. Nous avons également mis en évidence une reprogrammation métabolique de type Warburg, caractérisée par une activation accrue de la glycolyse aérobie, une inhibition de l’oxydation mitochondriale du pyruvate et une production de lipides. Ainsi, l’ensemble de ces perturbations va favoriser la prolifération cellulaire et le développement de kystes, et pourrait mener au développement de tumeur rénale comme observée chez une souris K.G6pc-/-. En conclusion nous avons démontré que, dans le cadre de la GSDI, l’accumulation de lipides dans les reins et le foie, secondaire à la déficience en G6Pase, joue un rôle clé dans le développement des complications hépatiques et rénales à long terme. Également, la reprogrammation métabolique rénale de type Warburg, prenant place dans le cadre de la GSDI, associée à un défaut du cil primaire pourrait être à l’origine de la formation des kystes et de tumeurs rénales. Ces études, en permettant une meilleure compréhension de la physiopathologie des complications à long terme de la GSDIa, offrent de nouvelles perspectives concernant les stratégies thérapeutiques à développer pour une meilleure prise en charge des patients atteints de GSDIa / Glycogen storage disease type Ia (GSDIa) is a rare metabolic disease caused by glucose-6-phosphatase (G6Pase) deficiency, due to mutations on the gene encoding G6Pase catalytic subunit (G6PC). This enzyme confers to the liver, kidneys and intestine the ability to produce glucose. Thus, patients with GSDIa are unable to ensure endogenous glucose production and suffer from severe hypoglycemia during fasting in the absence of nutritional control. In addition, G6Pase deficiency causes intracellular accumulation of glucose-6 phosphate in the liver and kidneys, leading to metabolic defects and the accumulation of glycogen and lipids. Over time, most adult patients suffer from chronic kidney disease (CKD), which can progress to kidney failure, requiring dialysis or kidney transplantation. This nephropathy is characterized in particular by tubulo-interstitial fibrosis and glomerulosclerosis, as well as by the development of cysts in the late stages. Moreover, patients develop hepatomegaly and hepatic steatosis that may progress to the development of hepatocellular adenomas or carcinomas. The aim of my thesis was to identify the molecular mechanisms involved in the establishment of renal pathology and cyst formation in GSDIa, by using mouse models where G6pc gene is specifically deleted in the kidneys (K.G6pc-/- mice). While GSDIa is a disease characterized by glycogen accumulation in the liver and kidneys, we first showed that the development of fibrosis, which causes progressive loss of kidney function, was induced by intracellular accumulation of lipids, regardless of glycogen content. The molecular mechanism probably involved is the activation of the renin angiotensin system by lipid derivatives such as diacylglycerol, which induced the expression of the profibrotic factor TGFβ1 and an epithelial-mesenchymal transition. In addition, the use of a PPARα agonist, i.e. fenofibrate, by decreasing renal lipid content, reduced the development of fibrosis and CKD evolution. Similarly, fenofibrate treatment prevented the accumulation of lipids in the liver and the development of liver damages that cause tumor development. Thus, the activation of lipid catabolism by PPARα agonists such as fenofibrate seems to be an interesting therapeutic strategy to reduce the progression of renal and hepatic diseases of GSDIa. The second part of my results suggest that the development of renal cysts in GSDI patients may be caused by an alteration of the primary cilia, a non-motile organelle that plays a key role in maintaining normal kidney structure and function. Indeed, defects in the primary cilia are involved in many polycystic kidney diseases. In summary, an increase in the length of the primary cilia was observed in the kidneys of K.G6pc-/- mice, which could be explained by a deregulation of the expression of different proteins involved in cilia structure and function, compared to control mice. We also demonstrated a metabolic reprogramming leading to a Warburg metabolism, characterized by the increased activation of aerobic glycolysis and the inhibition of mitochondrial pyruvate oxidation and lipid production in K.G6pc-/- mice. Thus, all these disorders would promote cell proliferation and cyst development, and could lead to the development of renal tumor, as recently observed in one K.G6pc-/- mouse (out of 36 studied mice). In conclusion, we have shown that, in GSDI, the accumulation of lipids in the kidneys and liver that occurs secondary to G6Pase deficiency plays a key role in the development of hepatic and renal long-term complications. In addition, the Warburg like metabolic reprogramming taking place in the GSDIa kidneys, associated with a defect in the primary cilia, could be at the origin of cysts formation and renal tumors. These new studies, by providing a better understanding of the pathophysiology of long-term complications of GSDIa, offer new perspectives on therapeutic strategies to be developed for better management of patients

Les omégas trois ont un bénéfice prouvé dans la prévention de maladie cardiovasculaire et l’inflammation. Un apport optimal peut être réalisé avec des modifications diététiques simples permettant d’avoir un enrichissement des membranes cellulaires et un effet métabolique. Le tissu adipeux de part son rôle important dans la genèse du syndrome métabolique semble être une cible importante du traitement par oméga trois. Les patients avec une maladie rénale chronique (MRC) ont un risque cardiovasculaire accru et cumulent les perturbations métaboliques comme le syndrome métabolique et un état micro inflammatoire. Des doses supra physiologiques d’oméga trois ont été utilisés dans le passé dans des études de prévention rénale ou traitement de dyslipidémie. Or l’effet métabolique en fonction de la dose d’oméga 3 n’est pas connu. En accord, avec les études chez le sujet sain, en fonction de la dose administrée, les omégas 3 ont un impact différent métabolique et sur l’expression génique. Des études complémentaires sont nécessaires pour vérifier la faisabilité et l’impact métabolique d’une modification de régime afin de diminuer le rapport n-6/n-3, ainsi que l’effet à long terme des omégas trois chez ces patients. Par ailleurs, les mécanismes impliqués dans les différences de dose réponse devront être caractérisés sur un modèle animal / Omega 3 fatty acids play an important modulatory role in metabolic and inflammatory responses, the progression of atherosclerosis and gene expression. Recent studies suggest their beneficial impact on adipocyte morphology and function. Chronic kidney disease (CKD) patients have an increased cardiovascular morbi-mortality and suffer from a cluster of metabolic disorders. On the basis of previous studies there are reasons to suggest that omega 3 supplementation may offer a host of benefits to CKD patients. Unfortunatly, published studies on the effect of such supplementation are characterized by supra physiological omega 3 doses, that may be difficult to implement for extended periods in one hand and in the other hand the metabolic effect of different doses of omega 3 hasn’t been studied in detail. Simple dietary modifications can help achieve the recommended n-6/n-3 ratio in healthy subjects. In CKD patients supplementation with n-3 shows a differential dose response effect. Further studies are required to test the faisability and metabolic impact of dietary modifications in order to decrease n-6/n-3 ratio and to assess the long term effect of omega supplementation in CKD patients. Finally the molecular pathways implicated in this differential dose response should be assessed in animal models

and key words Enhanced activation of renin-angiotensin system (RAS) and endothelin system (ES) plays the key pathophysiological role in the progression of hypertension and the chronic kidney disease (CKD). The aim of this study was to verify wheter the combined inhibition RAS and selective inhibition ETA receptor for endothelin 1 (ET-1) will show additive renoprotective effects in experimental model CKD. This therapeutic aproach was tested on the transgenic rats with mouse renin gen (TGR), to whome ablation nephrectomy (5/6 NX) was done in the age of 6 weeks. After next 6 weeks the relevant treatment was given in drinking-water: dual RAS blockade (trandolapril 6mg/L + losartan 100mg/L) or the combination of dual RAS blockade + inhibitor of ETA receptor (atrasentan 25mg/L). Results of the first series show 100 % mortality in untreated rats with 5/6 NX to the 30th week. Both type of treatments increased the survival rate up to 30 % in 5/6 NX TGR after the 50th week. In the second series influence of treatments on the blood pressure (BP) was monitored in 5/6 NX TGR, which had systolic BP over 210 mmHg. Both treatments decreased BP to the level normotensive rats and reduced heart hypertrophy. In the third series the results showed that treatment significantly decreased renal level of angiotensin II...

An increase in cellular inflammatory biomarkers directly increases the risk of cardiovascular disease (CVD). Using the social ecological and biomedical theories, the study examined quantitatively how specific inflammatory biomarkers are associated with cardiorenal syndrome (CRS), a potential complication of hypertension and diabetes, and how sociodemographic factors modify this association in the U.S. adult population. A retrospective secondary data analysis of the data collected from National Health and Nutrition Examination Survey (NHANES) 1999-2010 was utilized to evaluate these hypotheses. High sensitivity C-reactive protein, homocysteine (hcy), and fibrinogen had a modifying effect on Type 4 (chronic reno-cardiac etiology), Type 2 CRS (chronic cardio-renal etiology), and a significant additive effect on CRS even after controlling for known CVD and Chronic Kidney Disease (CKD) risk factors. For Type 4 CRS, the adjusted Odds Ratio of CVD in individuals with CKD was elevated, 2.29 (Confidence Interval [CI] 1.17-3.64, p < 0.05), among individuals with elevated hcy levels but close to 1.0 (0.65 CI 0.28-1.53, p > 0.05) among patients with normal hcy after the results were controlled for medical and demographic risk factors. Finally, race modified the effect of inflammatory markers on CRS. Out of all the biomarkers, income only modified the effect of hcy on CRS. Education level modified the effect of every inflammatory marker on CRS. While Ferritin-to-Transferrin ratio (F/T ratio) had a non-significant additive effect, due to the lack of adequate subjects, the modifying effect of F/T ratio could not be tested. This study can help initiate social change by urging healthcare professionals to monitor these biomarkers as a part of preventing hypertension, diabetes, and CRS.

Health literacy is widespread, but its potential is not recognized. By not recognizing health literacy, patients have the burden of coping with diabetes with renal complications without full knowledge of their responsibility to their health. The focus of the project was to assess participants with diabetes with low health literacy and low mean glomerular filtration rate (eGFR). The project goal was achieved by the assessment of the participants' health literacy and eGFR before and after education for their diabetes, then assessed to determine if teaching the participants would improve their health literacy, lab values, and overall health. Participants were recruited by being patients of the designated clinic and screened for diabetes and low eGFR, for a total of 30 participants. The Brief Health Literacy Screen was used to measure health literacy. The health of the participants was appraised by the laboratory values of eGFR and fasting glucose. The project methodology was an observational design using correlation and 2-sample t analysis with the variables eGFR, fasting glucose, and health literacy. The variables were compared before and after the participants' education. Results showed health literacy with patient education was associated with greater patient self-efficacy and improved fasting glucose numbers, eGFR flows, and health literacy scores. The current health climate shows value in different types of health providers. Social change was defined by the project launching a nurse practitioner as the leader for advancing the treatment plans of chronic kidney disease. This project impacts social change by showing patients in the process of improved health and empowering the patients to be advocates of their own health.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19599号 / 医博第4106号 / 新制||医||1014(附属図書館) / 32635 / 京都大学大学院医学研究科医学専攻 / (主査)教授 柳田 素子, 教授 妹尾 浩, 教授 浅野 雅秀 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Chronic kidney disease is a progressive kidney disease that affects approximately one tenth of the world population. In almost all cases of progressive, end-stage kidney disease, fibrosis is seen. Renal fibrosis is a condition of the kidneys in which collagens and other proteins accumulate in the extracellular matrix of the kidneys. The aim of the project was to explore the use of Fast Green to visualize collagen in renal tissue using light-sheet microscopy. A healthy sample and a diseased sample of renal tissue was imaged in aqueous and anhydrous conditions and a comparison of the difference in staining pattern between the conditions was performed. There was a clear difference in staining pattern between the two conditions, where the samples prepared in anhydrous conditions showed a higher staining specificity to collagen as described previously for other types of tissue. An evaluation of the differences in collagen expression between the healthy and the diseased sample was performed as well. There was a visible difference between them where a higher expression of collagen was observed in numerous tubuli, indicative of pathological scarring. / Kronisk njursjukdom är en progressiv njursjukdom som påverkar approximativt en tiondel av världens befolkning. I alla progressiva njursjukdomar ses så kallad njurfibros. Njurfibros är ett tillstånd då kollagen och andra proteiner ansamlas i njurarnas extracellulära matris. Målet med projektet var att utforska användningen av Fast Green för att visualisera kollagen i njurvävnad med ”Light sheet”-mikroskopi. Vävnadsproverfrån frisk vävnad och sjuk vävnad avbildades i vattenhaltiga och vattenfria förhållanden och en jämförelse av inmärkningen mellan förhållandena utfördes. Det var en tydlig skillnad i inmärkningen mellan de två förhållandena, där det i de vattenfria proven observerades att Fast Green märkte in kollagen med högre specificitet. En jämförelse av skillnaderna i kollagenuttryck mellan det friska och det sjuka provet utfördes. Det var tydliga skillnader mellan proven, där ett stort antalkollagen-påverkade tubuli kunde observeras i det sjuka provet.

Chronic Kidney Disease (CKD) is a common and dangerous health condition that requires early detection and treatment to be effective. Current diagnostic methods are time-consuming and expensive. In this research, we hope to construct a predictive model for CKD utilizing a combination of time series and static variables for early detection of CKD. In this study, we investigate the influence of multimodal approach by combining the predictions from multiple models that utilize different modalities. The ROCKET method is utilized for classification using time series features, whilst the Random Forest approach is employed for static data. XGBoost has been utilized to gain information about feature importance among labs and demographics-comorbidities data. In this study, we use the MIMIC-III database, adopting various strategies to handle data and class imbalance, such as stratification, balancing techniques, and backwards and forward fill for missing value imputation. The evaluation metrics for CKD and non-CKD class labels include precision, recall, F1, and accuracy. Our findings show that aggregating time series data produce contrasting results for labs compared to vitals data. We also addressed the significance of the different demographic, comorbidities and lab events features. The findings indicate that a multimodal approach did not show significant advantages over individual models when the individual models performed suboptimal. The study also found that Ethnicity is more significant than age and gender in predicting CKD. Furthermore, the study revealed some significant features from lab events and comorbidities. The study also provides some recommendations for future work to explore the potential of a multimodal approach further.

Diabetes mellitus is a metabolic disease largely due to lifestyle and nutritional imbalance, resulting in insulin resistance, hyperglycemia and vascular complications. Diabetic kidney disease (DKD) is a major cause of end-stage renal failure contributing to morbidity and mortality worldwide. Therapeutic options to prevent or reverse DKD progression are limited. Endothelial and glomerular filtration barrier (GFB) dysfunction and sterile inflammation are associated with DKD. Neutrophil extracellular traps (NETs), originally identified as an innate immune mechanism to combat infection, have been implicated in sterile inflammatory responses in non-communicable diseases. However, the contribution of NETs in DKD remains unknown. Here, we show that biomarkers of NETs are increased in diabetic mice and diabetic patients and that these changes correlate with DKD severity. Mechanistically, NETs promote NLRP3 inflammasome activation and glomerular endothelial dysfunction under high glucose stress in vitro and in vivo. Inhibition of NETs (PAD4 inhibitor) ameliorate endothelial dysfunction and renal injury in DKD. Taken together, NET-induced sterile inflammation promotes diabetes-associated endothelial dysfunction, identifying a new pathomechanism contributing to DKD. Inhibition of NETs may be a promising therapeutic strategy in DKD.

<p>  </p> <p>This research evaluates Kidney Health Initiative's (KHI)   four kidney replacement therapy (KRT) technology priority groups in the roadmap:</p> <p>1) Literature review of technology citations for each of the four KHI priority groups for safety and efficacy data with a patient centered focus. Additionally, the incorporation and integration of KHI minimum technical design criteria in six areas into the development process.</p> <p>2) Clinical PICO analysis of the critical clinical outcomes that the discovery and innovation from the specific technology priority group addresses</p> <p>3) Critical evaluation of KRT technologies on patients with ESKD by expert clinicians and scientists in KRT through Delphi method with targeted questionnaires</p>