Bayesian Two-Way Analysis of High-Dimensional Collinear Metabolomics Data

Kaksisuuntainen tehtävänasettelu on yleinen bioinformatiikan alalla. Tässä diplomityössä esitellään uusi bayesilaisen mallinnuksen menetelmä kaksisuuntaisen havaintoaineiston analysointiin. Menetelmä toimii myös vähän näytteitä sisältävillä korkeaulotteisilla havaintoaineistoilla. Havaintoaineiston oletetaan jakautuvan populaatioihin kovariaattien mukaan, jotka tyypillisessä biologisessa kokeessa ovat yksilön terveydentila, sukupuoli, lääketieteellinen hoito sekä yksilön ikä. Esiteltävä menetelmä on suunniteltu arvioimaan näiden kovariaattien vaikutus havaintoaineiston kontrolliryhmän perustasoon verrattuna. Menetelmä perustuu olettamukseen siitä, että havaintoaineiston piirteet muodostavat ryhmiä, joiden sisällä piirteet ovat voimakkaasti kollineaarisia. Tämä olettamus mahdollistaa piilomuuttajamalliin perustuvan dimensionaalisuuden pudotuksen, jonka ansiosta menetelmä on toimiva myös pienen näytemäärän havaintoaineistoille. Menetelmä käsittelee havaintoaineistoa täysin bayesilaisittain, Gibbsin otannan avulla. Bayesilainen lähestymistapa tuottaa arvion sekä mallin ja havaintoaineiston yhteisjakaumalle että mallin jokaisen parametrin marginaalijakaumalle. Tämä mahdollistaa tulosten epävarmuuden arvioinnin sekä vertailun toisiin malleihin. Uuden menetelmän toimivuutta esitellään metabolomiikan alalta olevan havaintoaineiston avulla. Aineisto sisältää lipidiprofiileja, jotka on mitattu terveistä lapsista ja lapsista, jotka myöhemmin sairastuvat tyypin 1 diabetekseen. Kahdessa erillisessä analyysissä tutkitaan sairauden ja sukupuolen sekä sairauden ja iän vaikutusta lipidiprofiileihin.Two-way experimental designs are common in bioinformatics. In this thesis, a new Bayesian model is proposed for the analysis of two-way data. The method also works for small sample-size data with a high number of features. The data set is assumed to be divided into populations according to covariates, which in the case of a typical biological experiment are the health status, the gender, the medical treatment and the age of the individual. The proposed method is designed to estimate the effect of these covariates compared to the ground level of a control group of the data. The method is based on the assumption that features of the data form groups that are highly collinear. This allows the use of a latent variable-based dimensionality reduction, which makes inference possible also for small sample-size data sets. The method treats the data in a completely Bayesian way, which produces an estimate for the joint distribution of the model and the data, and marginal posterior distributions of all model parameters. This allows one to evaluate the signicance and uncertainty of the results and to compare it to other models. Inference is carried out with Gibbs sampling. The performance of the new method is demonstrated with a metabolomic data set by comparing lipidomic profiles from children who remain healthy to those who will later develop type 1 diabetes. In two separate studies, the effect of the disease and gender, and the effect of the disease and time, are estimated

Stronger findings from mass spectral data through multi-peak modeling

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Lipidome as a predictive tool in progression to type 2 diabetes in Finnish men

Background. There is a need for early markers to track and predict the development of type 2 diabetes mellitus (T2DM) from the state of normal glucose tolerance through prediabetes. In this study we tested whether the plasma molecular lipidome has biomarker potential to predicting the onset of T2DM. Methods. We applied global lipidomic profiling on plasma samples from well-phenotyped men (107 cases, 216 controls) participating in the longitudinal METSIM study at baseline and at five-year follow-up. To validate the lipid markers, an additional study with a representative sample of adult male population (n = 631) was also conducted. A total of 277 plasma lipids were analyzed using the lipidomics platform based on ultra performance liquid chromatography coupled to time-of-flight mass spectrometry. Lipids with the highest predictive power for the development of T2DM were computationally selected, validated and compared to standard risk models without lipids. Results. A persistent lipid signature with higher levels of triacylglycerols and diacyl-phospholipids as well as lowerlevels of alkylacyl phosphatidylcholines was observed in progressors to T2DM. Lysophosphatidylcholine acyl C18:2 (LysoPC(18:2)), phosphatidylcholines PC(32:1), PC(34:2e) and PC(36:1), and triacylglycerol TG(17:1/18:1/18:2) were selected to the full model that included metabolic risk factors and FINDRISC variables. When further adjusting for BM and age, these lipids had respective odds ratios of 0.32, 2.4, 0.50, 2.2 and 0.31 (all p 0; p <0.05). Notably, the lipid models remained predictive of the development of T2DM in the fasting plasma glucose-matched subset of the validation study. Conclusion. This study indicates that a lipid signature characteristic of T2DM is present years before the diagnosis and improves prediction of progression to T2DM. Molecular lipid biomarkers were shown to have predictive power also in a high-risk group, where standard risk factors are not helpful at distinguishing progressors from non-progressors. (C) 2017 The Authors. Published by Elsevier Inc.Peer reviewe

Targeted Clinical Metabolite Profiling Platform for the Stratification of Diabetic Patients

Several small molecule biomarkers have been reported in the literature for prediction and diagnosis of (pre)diabetes, its co-morbidities, and complications. Here, we report the development and validation of a novel, quantitative method for the determination of a selected panel of 34 metabolite biomarkers from human plasma. We selected a panel of metabolites indicative of various clinically-relevant pathogenic stages of diabetes. We combined these candidate biomarkers into a single ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method and optimized it, prioritizing simplicity of sample preparation and time needed for analysis, enabling high-throughput analysis in clinical laboratory settings. We validated the method in terms of limits of detection (LOD) and quantitation (LOQ), linearity (R2), and intra- and inter-day repeatability of each metabolite. The method’s performance was demonstrated in the analysis of selected samples from a diabetes cohort study. Metabolite levels were associated with clinical measurements and kidney complications in type 1 diabetes (T1D) patients. Specifically, both amino acids and amino acid-related analytes, as well as specific bile acids, were associated with macro-albuminuria. Additionally, specific bile acids were associated with glycemic control, anti-hypertensive medication, statin medication, and clinical lipid measurements. The developed analytical method is suitable for robust determination of selected plasma metabolites in the diabetes clinic

Lipidome as a predictive tool in progression to type 2 diabetes in Finnish men

Background. There is a need for early markers to track and predict the development of type 2 diabetes mellitus (T2DM) from the state of normal glucose tolerance through prediabetes. In this study we tested whether the plasma molecular lipidome has biomarker potential to predicting the onset of T2DM.Methods. We applied global lipidomic profiling on plasma samples from well-phenotyped men (107 cases, 216 controls) participating in the longitudinal METSIM study at baseline and at five-year follow-up. To validate the lipid markers, an additional study with a representative sample of adult male population (n = 631) was also conducted. A total of 277 plasma lipids were analyzed using the lipidomics platform based on ultra performance liquid chromatography coupled to time-of-flight mass spectrometry. Lipids with the highest predictive power for the development of T2DM were computationally selected, validated and compared to standard risk models without lipids.Results. A persistent lipid signature with higher levels of triacylglycerols and diacyl-phospholipids as well as lowerlevels of alkylacyl phosphatidylcholines was observed in progressors to T2DM. Lysophosphatidylcholine acyl C18:2 (LysoPC(18:2)), phosphatidylcholines PC(32:1), PC(34:2e) and PC(36:1), and triacylglycerol TG(17:1/18:1/18:2) were selected to the full model that included metabolic risk factors and FINDRISC variables. When further adjusting for BM and age, these lipids had respective odds ratios of 0.32, 2.4, 0.50, 2.2 and 0.31 (all p 0; p < 0.05). Notably, the lipid models remained predictive of the development of T2DM in the fasting plasma glucose-matched subset of the validation study.Conclusion. This study indicates that a lipid signature characteristic of T2DM is present years before the diagnosis and improves prediction of progression to T2DM. Molecular lipid biomarkers were shown to have predictive power also in a high-risk group, where standard risk factors are not helpful at distinguishing progressors from non-progressors

Molecular Atlas of Postnatal Mouse Heart Development

Background The molecular mechanisms mediating postnatal loss of cardiac regeneration in mammals are not fully understood. We aimed to provide an integrated resource of mRNA, protein, and metabolite changes in the neonatal heart for identification of metabolism‐related mechanisms associated with cardiac regeneration. Methods and Results Methods and results Mouse ventricular tissue samples taken on postnatal day 1 (P01), P04, P09, and P23 were analyzed with RNA sequencing and global proteomics and metabolomics. Gene ontology analysis, KEGG pathway analysis, and fuzzy c‐means clustering were used to identify up‐ or downregulated biological processes and metabolic pathways on all 3 levels, and Ingenuity pathway analysis (Qiagen) was used to identify upstream regulators. Differential expression was observed for 8547 mRNAs and for 1199 of 2285 quantified proteins. Furthermore, 151 metabolites with significant changes were identified. Differentially regulated metabolic pathways include branched chain amino acid degradation (upregulated at P23), fatty acid metabolism (upregulated at P04 and P09; downregulated at P23) as well as the HMGCS (HMG‐CoA [hydroxymethylglutaryl‐coenzyme A] synthase)–mediated mevalonate pathway and ketogenesis (transiently activated). Pharmacological inhibition of HMGCS in primary neonatal cardiomyocytes reduced the percentage of BrdU‐positive cardiomyocytes, providing evidence that the mevalonate and ketogenesis routes may participate in regulating the cardiomyocyte cell cycle. Conclusions This study is the first systems‐level resource combining data from genomewide transcriptomics with global quantitative proteomics and untargeted metabolomics analyses in the mouse heart throughout the early postnatal period. These integrated data of molecular changes associated with the loss of cardiac regeneration may open up new possibilities for the development of regenerative therapiesPeer reviewe

Integrated lipidomics and proteomics point to early blood-based changes in childhood preceding later development of psychotic experiences: evidence from the Avon Longitudinal Study of Parents and Children

Background The identification of early biomarkers of psychotic experiences (PEs) is of interest as early diagnosis and treatment of those at risk of future disorder is associated with improved outcomes. The current study investigated early lipidomic and coagulation pathway protein signatures of later PEs in subjects from the Avon Longitudinal Study of Parents and Children cohort. Methods Plasma of 115 children (age 12) who were first identified as experiencing PEs at age 18 (48 cases and 67 controls) were assessed through integrated and targeted lipidomics and semi-targeted proteomics approaches. We assessed the lipids, LPCs (n=11) and PCs (n=61), and the protein members of the coagulation pathway (n=22) and integrated this data with complement pathway protein data already available on these subjects. Results Twelve PCs, four LPCs and the coagulation protein plasminogen were altered between the control and PE group after correction for multiple comparisons. Lipidomic and proteomic datasets were integrated into a multivariate network displaying a strong relationship between most lipids that were significantly associated to PEs and plasminogen. Finally, an unsupervised clustering approach identified four different clusters with one of the clusters presenting the highest ratio cases:controls (P < 0.01) and associated with a higher concentration of smaller LDL cholesterol particles. Conclusions Our findings indicate that the lipidome and proteome of subjects who report PEs at age 18 is already altered at age 12 indicating that metabolic dysregulation may contribute to an early vulnerability to PEs and suggesting cross-talk between these LPCs, PCs and coagulation and complement proteins