Project description:BackgroundGestational diabetes mellitus (GDM) is a metabolic disease that can have long-term adverse effects on the cognitive function of mothers. In our study, we explored the changes in metabolic health and cognitive function in mice of middle- and old- age after exposure to GDM, and whether metformin therapy during pregnancy provided long-term benefits.ResultsMice with GDM demonstrated significant cognitive impairment in old age, which was associated with insulin resistance. Gestational metformin therapy was shown to increase insulin sensitivity and improve cognition. The ovarian aging rate was also accelerated in mice exposed to GDM during pregnancy, which may be related to fatty acid metabolism in the ovaries.ConclusionTreatment with metformin during pregnancy was shown to improve fatty acid metabolism in ovarian tissues.MethodDuring pregnancy, mice were fed with a high-fat diet (GDM group) or a low-fat diet (Control group), and a third group received metformin while receiving a high-fat diet (Treatment group). At 12 months old, the mice completed an oral glucose tolerance test, insulin tolerance test, Morris water maze test, female sex hormones were measured, and metabolite profiles of tissue from the ovaries, hypothalamus, and pituitary glands were analysed using gas chromatography-mass spectrometry.
Project description:We report a global adeno-associated virus (AAV)9-based gene therapy protocol to deliver therapeutic galactosylceramidase (GALC), a lysosomal enzyme that is deficient in Krabbe's disease. When globally administered via intrathecal, intracranial, and intravenous injections to newborn mice affected with GALC deficiency (twitcher mice), this approach largely surpassed prior published benchmarks of survival and metabolic correction, showing long-term protection of demyelination, neuroinflammation, and motor function. Bone marrow transplantation, performed in this protocol without immunosuppressive preconditioning, added minimal benefits to the AAV9 gene therapy. Contrasting with other proposed pre-clinical therapies, these results demonstrate that achieving nearly complete correction of GALC's metabolic deficiencies across the entire nervous system via gene therapy can have a significant improvement to behavioral deficits, pathophysiological changes, and survival. These results are an important consideration for determining the safest and most effective manner for adapting gene therapy to treat this leukodystrophy in the clinic.
Project description:Adverse long-term cardiovascular (CV) consequences of PE are well established in women. However, the mechanism responsible for that risk remains unknown. Here, we mated wild-type female mice of the FVB/N strain to STOX1A-overexpressing mice to mimic severe PE and investigated the long-term consequences on the maternal cardiovascular system. Ultrasonography parameters were analyzed in mice before pregnancy and at 3 and 6 months post-pregnancy. At 6 months post-pregnancy, cardiac stress test induced by dobutamine injection revealed an abnormal ultrasonography Doppler profile in mice with previous PE. Eight months post-pregnancy, the heart, endothelial cells (ECs) and plasma of females were analyzed and compared to controls. The heart of mice with PE showed left-ventricular hypertrophy associated with altered histology (fibrosis). Transcriptomic analysis revealed the deregulation of 1149 genes in purified ECs and of 165 genes in the hearts, many being involved in heart hypertrophy. In ECs, the upregulated genes were associated with inflammation and cellular stress. Systems biology analysis identified interleukin 6 (IL-6) as a hub gene connecting these pathways. Plasma profiling of 33 cytokines showed that, 8 of them (Cxcl13, Cxcl16, Cxcl11, IL-16, IL-10, IL-2, IL-4 and Ccl1) allowed to discriminate mice with previous PE from controls. Thus, PE triggers female long-term CV consequences on the STOX1 mouse model.
Project description:Long-term consequences of preeclampsia were studied by high-throughput approaches (Transcriptomics, Luminex cytokine profile) in mice 8 months after the disease in the heart, in the endothelial cells and in the plasma. In the heart we found a persisting ventricular hypertrophy with an altered histology and an abnormal ultrasound Doppler profile under effort simulation by dobutamine injection. The transcriptomic profile of the endothelium revealed a deregulation for 1149 genes (327 down-regulated and 822 up-regulated, with a threshold of 1.5, p<0.05). The up-regulated genes could be grouped consistently in gene pathways and gene ontology terms mainly focused on Inflammation and Stress sensu lato. A cytokine profile of the mouse serum was carried out. Using a combination of 8 cytokines (Cxcl13, Cxcl16, Cxcl11, Il-16, Il-10, Il-2, Il-4 and Ccl1) in a Discriminant Analysis, we were able to separate unambiguously the mice having had a preeclamptic pregnancy from the controls. Seven out of eight of these cytokines (with the exception of Il-16) varied in the same direction in the endothelium and in the plasma. In sum, our study shows that preeclampsia alone is able to trigger considerable long-term consequences, and suggests that specific cytokines could help to improve the follow-up of patients long after a preeclamptic pregnancy.
Project description:ObjectivesThe purpose of this study was to identify predictors of healthy arterial aging (long-term coronary artery calcification [CAC] of 0) among individuals with metabolic syndrome (MetS) or type 2 diabetes (T2D), which may improve primary prevention strategies.BackgroundIndividuals with MetS or T2D have a heterogeneously increased risk of atherosclerotic cardiovascular disease and not all have a high-intermediate risk.MethodsWe included 574 participants from the MESA (Multi-Ethnic Study of Atherosclerosis) with MetS or T2D who had CAC=0 at baseline and a repeat CAC scan 10 years later. Multivariable logistic regression assessed the association of traditional and novel atherosclerotic cardiovascular disease risk factors and the MetS severity score (based on the 5 MetS criteria) with healthy arterial aging.ResultsThe mean age of participants was 58.9 years, 67% were women, 422 participants had MetS, and 152 had T2D. The proportion with long-term CAC=0 was similar for MetS (42%) and T2D (44%). A younger age was the only individual low/normal traditional risk factor associated with an increased likelihood of long-term CAC=0 (odds ratio [OR]: 1.50; 95% confidence interval [CI]: 1.22 to 1.85 per 10-years younger). The strongest associations of nontraditional risk factors were observed for an absence of thoracic calcification (OR: 2.42; 95% CI: 1.24 to 4.72), absence of carotid plaque (OR: 1.81; 95% CI: 1.25 to 2.61), and among persons with a high sensitivity troponin <3 ng/ml (OR: 1.55; 95% CI: 1.01 to 2.38). In addition, persons with the lowest quartile MetS severity score had a substantially higher odds of healthy long-term CAC=0 (OR: 2.71; 95% CI: 1.27 to 5.76).ConclusionsMore than 40% of adults with MetS or T2D and baseline CAC=0 had long-term absence of CAC, which was most strongly associated with an absence of extracoronary atherosclerosis and a low MetS score. An optimal overall cardiovascular profile appears to be more important than an ideal value of any individual risk factor to maintain healthy arterial aging.
Project description:Asthma bronchiale is an inflammatory disease of the respiratory airways and a major factor of increasing health care costs worldwide. The molecular actors leading to asthma are not fully understood and require further investigation. The aim of this study was to monitor the proteome during asthma development from early inflammatory to late fibrotic stages. A time-course-based ovalbumin (OVA) mouse model was applied to establish an asthma phenotype and the lung proteome was analysed at four time points during asthma development (0 weeks = control, 5 weeks, 8 weeks and 12 weeks of OVA treatment).
Project description:The aim of the project was the analysis of the bulk proteome and phosphoproteome in 3D organoids (tubuloids) from the adult mouse kidney. Proteome (nearly 9000 proteins) and phosphoproteome (nearly 16000 phospho sites) were compared in three biological (independent) replicates of epithelial organoids (early passage and long-term, 3-month cultures) and freshly isolated, MAC-sorted, Epcam-positive kidney epithelial cells. After collection of the organoids, Matrigel was removed using a non-enzymatic solution.
Project description:RNA interference (RNAi) gene silencing is a potential therapeutic strategy for dominant retinal degeneration disorders. We used self-complementary (sc) AAV2/8 vector to develop an RNAi-based gene therapy in a dominant retinal degeneration mouse model expressing bovine GCAP1(Y99C). We established an in vitro shRNA screening assay based on EGFP-tagged bovine GCAP1, and identified a shRNA that effectively silenced the bovine GCAP1 transgene with ?80% efficiency. Subretinal injection of scAAV2/8 carrying shRNA expression cassette showed robust expression as early as 1 wk after injection. The gene silencing significantly improved photoreceptor survival, delayed disease onset, and increased visual function. Our results provide a promising strategy toward effective RNAi-based gene therapy by scAAV2/8 delivery for dominant retinal diseases.
Project description:BackgroundElderly patients undergoing cardiac operation often suffer various metabolic comorbidities, such as diabetes mellitus (DM) and obesity. The metabolic disorders in these individuals are widely considered to be possible predisposing factors for unfavourable prognosis. This retrospective study was aimed to determine the association of metabolic diseases with the mortality of elderly patients after coronary artery bypass grafting (CABG) and to identify the protective or risk factors related to their short- and long-term survival.MethodsTotally 684 patients aged 75 years or above undergoing isolated CABG were evaluated retrospectively. There were two groups depending on the body mass index (BMI): an overweight and obesity group (n = 354) and a normal weight and lean group (n = 330). Propensity score matching (PSM) was performed to adjust baseline clinical characteristics, which reduced confounding bias. The short-term postoperative mortality was tested via logistic regression. Kaplan-Meier and Cox regression analyses were done to compute the overall survival in each group and to identify relevant variables associated with all-cause mortality, respectively.ResultsThe prevalence rates of metabolic comorbidities in the total cohort were: diabetes mellitus (32.5%), overweight or obesity (51.8%) and hypertension (72.8%). The 30-day postoperative mortality was 5.1% and the long-term mortality was 15.25% at a median 46.2-month follow-up (1.0-178.6 months). The 30-day postoperative mortality was relevant to DM, diseased coronary arteries, New York Heart Association class, intra-aortic balloon pump and emergency surgery. The long-term mortality was negatively associated with overweight and obesity. Univariate and multivariate logistic regression recognized DM as an adverse factor related with 30-day postoperative mortality whether before or after PSM. The long-term mortality was not significantly relevant with DM (HR = 0.753, 95% CI 0.402-1.411). Overweight or obesity was not the risk factor of 30-day postoperative mortality (OR = 1.284, 95% CI 0.426-3.868), but was the protective factor of long-term survival (HR = 0.512, 95% CI 0.279-0.939).ConclusionsThe "obesity paradox" exists regarding the prognosis of individuals aged ≥ 75, which was presented as lower long-term mortality no matter from all cause or cardio-cerebrovascular cause in patients with BMI ≥ 24. Trial registration ChiCTR2200061869 (05/07/2022).