Advanced Proteomics and Metabolomics Studies of Type 1 Diabetes
EMSL Project ID
12500
Abstract
In this four year two phase (R21/R33) project we will apply advanced proteomic and metabolomic nanoflow liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry technologies in the study of both plasma and serum from Type 1 diabetes patients and isolated human pancreatic islets. The overall approach endeavors to advance the study of Type 1 diabetes and human islet transplantation by identifying biomarkers at the level of the proteome and metabolome that are predictive of both Type 1 diabetes and islet performance in vivo. Our approach will utilize proteome-wide stable isotope labeling of peptides, as well as quantitative cysteine-peptide enrichment technology (QCET) and N-linked glycopeptide enrichment strategies to obtain broad proteome coverage and enhance quantitation. We will also utilize very low nanoflow LC separations to minimize ionization suppression and eliminate background ions originating from the solvent, thereby improving normalization of metabolite peak intensities and improve quantitation. This approach will be capable of rapidly identifying and measuring expression levels for thousands of peptides or concentrations of metabolites in a single analysis.Phase 1 of this project will (a) define the sample processing and LC separation conditions necessary for broad metabolome and proteome coverage in human plasma/serum and pancreatic islets, (b) establish accurate mass and time tag databases for both peptides and metabolites detected in human plasma/serum and pancreatic islets, and (c) demonstrate the ability of the technology to distinguish plasma/serum from healthy control or recently diagnosed Type 1 diabetic patients. The refinement of this technological approach will provide the basis for high throughput studies of large numbers of samples. The application of this technology in Phase 2 of the project will involve (a) the high throughput studies of complete sample sets from the Diabetes Autoantibody Standardization Program (DASP), (b) validation of potential biomarkers via analysis of a blind DASP sample set, and (c) comparative studies of multiple human pancreatic islet preparations to identify potential biomarkers predictive of islet performance in vivo.
Project Details
Project type
Exploratory Research
Start Date
2005-02-25
End Date
2006-03-03
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Application of proteomics in the discovery of candidate protein biomarkers in a Diabetes Autoantibody Standardization Program sample subset. Metz TO, Qian W, Jacobs JM, Gritsenko MA, Moore RJ, Polpitiya AD, Monroe ME, Camp DG, Mueller PW, Smith RD. Journal of Proteome Research 2008, 7(2):698-707.
"Characterization of the Human Pancreatic Islet Proteome by Two- Dimensional LC/MS/MS" Metz, T.O., Jacobs, J.M., Gritsenko, M.A., Fontès, G., Qian, W.J., Camp II, D.G., Poitout, V., Smith, R.D. (2006) J. Proteome Res. 5, 3345-3354.
Deletion of GPR40 impairs glucose-induced insulin secretion in vivo in mice without affecting intracellular fuel metabolism in islets. Alquier T, Peyot M-L, Latour MG, Kebede M, Sorensen CM, Gesta S, Kahn R, Smith RD, Jetton TL, Metz TO, Prentki M, Poitout V. Diabetes 2009, 58:2607-2615.
Ding J, CM Sorensen, N Jaitly, H Jiang, D Orton, ME Monroe, RJ Moore, RD Smith, and T Metz. 2008. "Application of the accurate mass and time tag approach in studies of the human blood lipidome." Journal of Chromatography B 871(2):243-252. doi:10.1016/j.jchromb.2008.04.040
Metz TO. "Application of capillary LC-MS in metabolic fingerprinting studies of type 1 diabetes mellitus." Presented at the 7th Annual "Identifying and Validating Metabolic Markers for Drug Discovery and Clinical Studies" December 4-5 2006, Orlando, FL.
Metz TO, JM Jacobs, MA Gritsenko, G Fontes, W Qian, DG Camp, VJ Poitout, and RD Smith. 2006. "Characterization of the Human Pancreatic Islet Proteome by Two-Dimensional LC/MS/MS." Journal of Proteome Research 5(12):3345-3354.
Metz TO, Q Zhang, JS Page, Y Shen, SJ Callister, JM Jacobs, and RD Smith. 2007. "Future of liquid chromatography-mass spectrometry in metabolic profiling and metabolomic studies for biomarker discovery." Biomarkers in Medicine 1(1):159-185.
Metz, TO, WJ Qian, JM Jacobs, MA Gritsenko, RJ Moore, DG Camp, PW Mueller, and RD Smith. "Identification of biomarkers of type 1 diabetes mellitus utilizing capillary LC-FTICR MS and the accurate mass and time tag (AMT) approach. Presented at the 3rd Annual U.S. HUPO Conference March 4-9 2007, Seattle, WA.
Sorensen, CM, DS Daly, TO Metz, N Jaitly, SJ Callister, ME Monroe, JS Zimmer, RJ Moore, and RD Smith. "Screening of LC-MS-based metabolic profiling data utilizing statistical modeling to identify candidate biomarkers." Presented at the 55th Annual ASMS Conference on Mass Spectrometry, June 3-7 2007, Indianapolis, IN
Sorensen CM, J Ding, Q Zhang, T Alquier, R Zhao, PW Mueller, RD Smith, and TO Metz. 2010. "Perturbations in the Lipid Profile of Individuals with Newly Diagnosed Type 1 Diabetes Mellitus: Lipidomics Analysis of a Diabetes Antibody Standardization Program Sample Subset." Clinical Biochemistry 43(12):948-956.