Regulatory Networks and Biomarkers of Beta-cell Dysfunction and Apoptosis
EMSL Project ID
48705
Abstract
In type 1 diabetes (T1D) research there remains a critical need for having a systems level understanding of the early stage molecular mechanisms that trigger the specific destruction of pancreatic β-cells by the autoimmune system. Thus, the overall objectives of this application are to discover novel signaling pathways and regulatory networks that contribute to early stage β-cell stress and death in vivo and to identify potential therapeutic targets for interventions and biomarkers for early diagnosis. These objectives will be pursued by applying enabling proteomics technologies available at EMSL and PNNL focusing on posttranslational protein modifications (PTMs) to unique sets of human islet samples. Specifically, we hypothesize that posttranslational regulation, involving phosphorylation, S-nitrosylation, and S-glutathionylation, represents a fundamental mechanism of triggering β-cell dysfunction preceding overt T1D. To address the limitation associated with clinical samples for studying dynamic signaling networks, our plan utilizes several complementary model systems: a) human islets isolated by laser-capture microdissection from clinical human pancreatic tissues, and b) human islets treated with cytokines in vitro. To pursue the objectives, studies in Aim 1 will identify signaling pathways and networks that are involved in early stage β-cell dysfunction and apoptosis using a global proteomics discovery approach. In Aim 2, we will apply a targeted quantification approach to verify the interesting regulatory networks and PTMs using samples from individual patients as well as time-course islet samples from "humanized" mice. In Aim 3, we will determine whether protein targets discovered in the first two aims can serve as potential beta-cell specific marker for early T1D diagnosis by performing serum proteomics as well as whether the selected targets have function roles in beta-cell apoptosis. Together, we anticipate that this project will demonstrate a new paradigm of systems level study of posttranslational regulation of β-cell dysfunction and apoptosis, and will provide a novel integrative view of the early stage regulatory networks that potentially trigger β-cell apoptosis and T1D. This work is enabled by the unique high sensitivity quantitative mass spectrometry-based proteomics capability at EMSL for analyzing very small sizes of human islet samples. It will demonstrate the application of advanced technologies to address national health burdens such as type 1 diabetes.
Project Details
Start Date
2014-12-01
End Date
2017-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Co-Investigator(s)
Team Members
Related Publications
Nakayasu E.S., F. Syed, S. Tersey, M.A. Gritsenko, H.D. Mitchell, C. Chan, and J. Turatsinze, et al. 2020. "Comprehensive proteomics analysis of stressed human islets identifies GDF15 as a target for type 1 diabetes intervention." Cell Metabolism 31, no. 2:363-374.e6. PNNL-SA-150166. doi:10.1016/j.cmet.2019.12.005
Quaamari A, E Dirice, N Gedeon, J Hu, JY Zhou, J Shirakawa, L Hou, J Goodman, C Karampelias, G Qiang, J Boucher, R Martinez, MA Gritsenko, DF De Jesus, S Kahraman, S Bhatt, RD Smith, HD Beer, P Jungtrakoon, Y Gong, AB Goldfine, CW Liew, A Doria, O Andersson, W Qian, E Remold-O'Donnell, and RN Kulkami. 2016. "SerpinB1 Promotes Pancreatic ? Cell Proliferation." Cell Metabolism 23(1):194-205. doi:10.1016/j.cmet.2015.12.001
Zhu Y., M. Dou, P.D. Piehowski, Y. Liang, F. Wang, R.K. Chu, and W.B. Chrisler, et al. 2018. "Spatially Resolved Proteome Mapping of Laser Capture Microdissected Tissue with Automated Sample Transfer to Nanodroplets." Molecular and Cellular Proteomics 17, no. 9:1864-1874. PNNL-SA-138815. doi:10.1074/mcp.TIR118.000686