Project description:This set has around 6000 samples of human plasma from The Environmental Determinants of Diabetes in the Young (TEDDY) Study, which comprises of time points from 3 months to case endpoint visit from children who developed type 1 diabetes or islet autoimmunity and their matched controls. In total, 3506 transitions were monitored to measure the abundance of 167 proteins.
Project description:<p>The Environmental Determinants of Diabetes in the Young (TEDDY) Study investigates genetic and genetic-environmental interactions, including gestational events, childhood infections, dietary exposures, and other environmental factors after birth, in relation to the development of islet autoimmunity and type 1 diabetes (T1D). A consortium of six clinical centers assembled to participate in the development and implementation of the study to identify environmental triggers for the development of islet autoimmunity and T1D in genetically susceptible individuals. Beginning in 2004, the TEDDY study screened over 400,000 newborns for high-risk HLA-DR, DQ genotypes from both the general population and families already affected by T1D. The TEDDY study enrolled around 8,676 participants across six clinical centers worldwide (Finland, Germany, Sweden and three in the United States) in the 15-year prospective follow-up.</p> <p>Participants are followed every three months for islet autoantibody (IA) measurements with blood sampling until four years of age and then at least every six months until the age of 15. After the age of four, autoantibody positive participants continue to be followed at three month intervals and autoantibody negative participants are followed at six-month intervals. In addition to the analysis of autoantibodies, additional data and sample collection are performed at each visit. Parents collect monthly stool samples in early childhood. The parents also fill out questionnaires at regular intervals in connection with study visits and record information about diet and health status in the child's TEDDY Book between visits. Continued long-term follow-up of the currently active TEDDY participants will provide important scientific information on early childhood diet, reported and measured infections, vaccinations, and psychosocial stressors that may contribute to the development of type 1 diabetes and islet autoimmunity.</p> <p>Additional information on the TEDDY study is available in the following articles: The Environmental Determinants of Diabetes in the Young (TEDDY) Study. TEDDY Study Group. Annals of the New York Academy of Science, 2008 and TEDDY - The Environmental Determinants of Diabetes in the Young - An Observational Clinical Trial. Annals of the New York Academy of Science, 2006 <a href="http://www.ncbi.nlm.nih.gov/pubmed/?term=19120261">Annals of the New York Academy of Science, 2008</a> and TEDDY - The Environmental Determinants of Diabetes in the Young - An Observational Clinical Trial. <a href="http://www.ncbi.nlm.nih.gov/pubmed/?term=17130573">Annals of the New York Academy of Science, 2006</a></p> <p> Details of the TEDDY protocol can be found in The Environmental Determinants of Diabetes in the Young (TEDDY): Genetic Criteria and International Diabetes Risk Screening of 421,000 infants. <a href="http://www.ncbi.nlm.nih.gov/pubmed/?term=21564455">Pediatric Diabetes, 2011</a></p> <p> There are three substudies associated with the current TEDDY project release in dbGaP: <ul> <li>TEDDY SNP Array - <a href="study.cgi?study_id=phs001037">phs001037</a> with targeted SNP array data.</li> <li>TEDDY Microbiome - <a href="study.cgi?study_id=phs001443">phs001443</a> with metagenomic and 16S rRNA sequencing data.</li> <li>TEDDY Gene Expression - <a href="study.cgi?study_id=phs001562">phs001562</a> with gene expression data.</li> </ul> </p>
Project description:In this set, we analyzed human plasma samples from The Environmental Determinants of Diabetes in the Young (TEDDY) Study. In total, 2,252 samples from 184 individuals were concatenated into 364 pooled samples from individuals that developed type 1 diabetes or islet autoimmunity and their matched controls. Samples were analyzed from the same individuals pre and post-autoimmunity onset paired against controls. Each pooled sample was depleted with MARS abundant protein depletion column (Agilent), digested with trypsin and labeled with 8-plex iTRAQ reagent. Samples were then multiplexed and fractionated into 24 fractions by high-pH reverse-phase chromatography (DOI: 10.1038/s41596-018-0006-9). Each fraction was analyzed by liquid chromatography tandem mass spectrometry.
Project description:Analysis of ex vivo isolated lymphatic endothelial cells from the dermis of patients to define type 2 diabetes-induced changes. Results preveal aberrant dermal lymphangiogenesis and provide insight into its role in the pathogenesis of persistent skin inflammation in type 2 diabetes. The ex vivo dLEC transcriptome reveals a dramatic influence of the T2D environment on multiple molecular and cellular processes, mirroring the phenotypic changes seen in T2D affected skin. The positively and negatively correlated dLEC transcripts directly cohere to prolonged inflammatory periods and reduced infectious resistance of patients´ skin. Further, lymphatic vessels might be involved in tissue remodeling processes during T2D induced skin alterations associated with impaired wound healing and altered dermal architecture. Hence, dermal lymphatic vessels might be directly associated with T2D disease promotion.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.