Role of AMP-activated protein kinase in pancreatic islet beta-cell death during type 1 and type 2 diabetes

Lead Research Organisation: Imperial College London
Department Name: Dept of Chemistry

Abstract

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Technical Summary

Diabetes mellitus currently affects ~ 5 % of the population in westernised societies, an incidence expected to double by 2020. Destruction of beta-cells, achieved respectively through immune-mediated attack or other mechanisms, is involved in the aetiology of type 1 and type 2 diabetes respectively.
Our laboratories and others have demonstrated that AMP-activated protein kinase (AMPK) plays a key role in pancreatic islet beta-cells as a sensor of glucose. We propose here to explore the potential role of forced changes in AMPK activity, achieved using recombinant adenoviruses or transgenesis, in modulating beta-cell survival and function during a variety of pathophysiological pro-apoptotic assaults. Assessment of beta-cell function, necrosis and apoptosis will be achieved using a wide range of assays. Transplantation into mice of islets in which AMPK activity has been blocked or enhanced will also serve to assess the potential therapeutic value of these changes for human islet transplantation. Our principal aims are as follows:
Aim 1: The role of AMPK will be studied in three models relevant to beta cell death in type 1 diabetes:
(a) Non-immune attack in vitro: isolated mouse islets will be treated with the inflammatory cytokines, IL-1beta and TNFalpha
(b) Immune-mediated destruction of islet beta-cells in vitro: islets isolated from non-obese diabetic (NOD) mice and incubated with insulin-reactive CD8+ T cells.
(c) Recurrent immune-mediated destruction of islet beta-cells in vivo. Syngeneic islets that have been infected with viral constructs (or null virus control) will be transplanted under the kidney capsule of diabetic NOD mice, and changes in glycaemia monitored as a read-out of beta-cell survival and function.
Aim 2: We propose to examine the role of AMPK in ex vivo models of beta cell failure, as follows:
(a)?Glucolipotoxicity? in vitro: achieved either over-expression of the lipogenic transcription factor, sterol regulatory element binding protein 1c (SREBP1c) or by treatment of MIN6 beta cells with high glucose and saturated fatty acid concentrations
(b) Insulin or IGF-1 receptor depletion from cells. MIN6 cells will be deployed and receptor silencing achieved using specific, validated siRNAs
Aim 3: What mechanisms are involved in the execution of apoptosis during (a) pathophysiological stimulation (Aims 1 and 2) and (b) after activating AMPK? The role in both cases of downstream signalling pathways will be explored using classical biochemical approaches.
Aim 4: Generation and characterisation of transgenic mice expressing constitutively-active or dominant-negative AMPK selectively in pancreatic beta cells.

Publications

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publication icon
Da Silva Xavier G (2010) ChREBP regulates Pdx-1 and other glucose-sensitive genes in pancreatic ß-cells. in Biochemical and biophysical research communications

 
Description BBSRC
Amount £622,745 (GBP)
Funding ID BB/J015873/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2012 
End 10/2015
 
Description Defining the molecular and physiological mechanisms of pancreatic islet dysfunction which lead to type 2 diabetes
Amount £482,479 (GBP)
Funding ID MR/L020149/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 09/2013 
End 08/2018
 
Description Diabetes UK
Amount £232,543 (GBP)
Funding ID BDA 12/0004535 
Organisation Diabetes UK 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2013 
End 02/2016
 
Description Diabetes UK
Amount £223,220 (GBP)
Funding ID BDA 11/004210 
Organisation Diabetes UK 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2011 
End 09/2014
 
Description FP7
Amount £470,000 (GBP)
Organisation European Commission 
Department Seventh Framework Programme (FP7)
Sector Public
Country European Union (EU)
Start 10/2010 
End 09/2015
 
Description ICL ISSF award
Amount £50,000 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2012 
End 09/2014
 
Description MRC Experimental Challenge
Amount £3,085,359 (GBP)
Funding ID MR/L020149/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2014 
End 09/2018
 
Description MRC PRogramme
Amount £2,200,000 (GBP)
Funding ID MR/L02036X/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 06/2014 
End 05/2019
 
Description Programme Grant
Amount £1,500,000 (GBP)
Funding ID MR/J0003042/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2012 
End 09/2017
 
Description Programme Grant
Amount £500,000 (GBP)
Funding ID G0901521 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 10/2011 
End 09/2015
 
Description Targeting etiological molecular mechanisms to treat human diabetes
Amount £2,200,000 (GBP)
Funding ID MR/L02036X/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 11/2014 
End 10/2014
 
Description Wellcome Trust Senior Investigator Award
Amount £2,200,000 (GBP)
Funding ID WT098424AIA 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2012 
End 09/2018
 
Title Adenoviruses 
Description Adenoviruses encoding active or dominant-negative AMPK 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Has allowed collaboration and studies of the role of AMPK in other tissues and systems (outside the pancreas). 
 
Description AMPK in central control of hepatic glucose production 
Organisation University of Toronto
Country Canada 
Sector Academic/University 
PI Contribution Generation and use of adenoviral constructs
Impact Publication in Diabetes 2010 (x2)
Start Year 2008
 
Description Conditional knockout mice 
Organisation Cochin Institute
Country France 
Sector Academic/University 
PI Contribution Generation of tissue specifi KO mice
Impact Publication in Diabetologia, 2010 (Sun et al)
 
Description D Bosco (Université de Genève) 
Organisation University of Geneva
Department Faculty of Diabetes
Country Switzerland 
Sector Academic/University 
PI Contribution Experiments for publications
Collaborator Contribution Experiments for publications
Impact Sorcin links pancreatic ß cell lipotoxicity to ER Ca2+ stores - Marmugi A et al - PMID: 26822088 Hypoxia lowers SLC30A8/ZnT8 expression and free cytosolic Zn2+ in pancreatic beta cells. Gerber PA et al PMID: 24865615 Incretin-modulated beta cell energetics in intact islets of Langerhans. Hodson DJ et al - PMID: 24766140 ADCY5 couples glucose to insulin secretion in human islets. Hodson DJ et al PMID: 24740569 Lipotoxicity disrupts incretin-regulated human ß cell connectivity. Hodson DJ et al - PMID: 24018562
Start Year 2013
 
Description Denton 
Organisation University of Bristol
Country United Kingdom 
Sector Academic/University 
PI Contribution Experiments for publications
Collaborator Contribution Experiments for publications
Impact manuscript (in press) Denton RM et al Biochem J 2016
Start Year 2014
 
Description Denton 
Organisation University of Pisa
Country Italy 
Sector Academic/University 
PI Contribution Experiments for publications
Collaborator Contribution Experiments for publications
Impact Sorcin links pancreatic ß cell lipotoxicity to ER Ca2+ stores - Marmugi A et al - PMID: 26822088 Hypoxia lowers SLC30A8/ZnT8 expression and free cytosolic Zn2+ in pancreatic beta cells. Gerber PA et al PMID: 24865615 Incretin-modulated beta cell energetics in intact islets of Langerhans. Hodson DJ et al - PMID: 24766140 ADCY5 couples glucose to insulin secretion in human islets. Hodson DJ et al PMID: 24740569 Lipotoxicity disrupts incretin-regulated human ß cell connectivity. Hodson DJ et al - PMID: 24018562
Start Year 2014
 
Description Generator of b-cell specfic knock-out mice for PASK 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution Design of PASK flox'd mice
Impact None as yet
Start Year 2009
 
Description Gerald Gradwohl (IGBMC) 
Organisation Institute of Genetics and Molecular and Cellular Biology (IGBMC)
Country France 
Sector Academic/University 
PI Contribution Experiments for publications
Collaborator Contribution Experiments for publications
Impact Piccard et al - http://www.ncbi.nlm.nih.gov/pubmed/25497096
Start Year 2015
 
Description James Shapiro (Alberta) 
Organisation University of Alberta
Country Canada 
Sector Academic/University 
PI Contribution Experiments for publications
Collaborator Contribution Experiments for publications
Impact ADCY5 couples glucose to insulin secretion in human islets. Hodson DJ et al PMID: 24740569 Sorcin links pancreatic ß cell lipotoxicity to ER Ca2+ stores - Marmugi A et al - PMID: 26822088
Start Year 2014
 
Description Lorenzo Piemouti (Milan) 
Organisation University of Milan
Country Italy 
Sector Academic/University 
PI Contribution Experiments for publications
Collaborator Contribution Experiments for publications
Impact ADCY5 couples glucose to insulin secretion in human islets. Hodson DJ et al - PMID: 24740569 Sorcin links pancreatic ß cell lipotoxicity to ER Ca2+ stores. Marmugi A et al - PMID: 26822088
Start Year 2014
 
Description Piero Marchetti (Pisa) 
Organisation University of Pisa
Country Italy 
Sector Academic/University 
PI Contribution experiments for publications
Collaborator Contribution experiments for publications
Impact Sorcin links pancreatic ß cell lipotoxicity to ER Ca2+ stores - Marmugi A et al - PMID: 26822088 Hypoxia lowers SLC30A8/ZnT8 expression and free cytosolic Zn2+ in pancreatic beta cells. Gerber PA et al PMID: 24865615 Incretin-modulated beta cell energetics in intact islets of Langerhans. Hodson DJ et al - PMID: 24766140 ADCY5 couples glucose to insulin secretion in human islets. Hodson DJ et al PMID: 24740569 Lipotoxicity disrupts incretin-regulated human ß cell connectivity. Hodson DJ et al - PMID: 24018562
Start Year 2014
 
Description Role of AMPK in counter regulation 
Organisation Yale University
Country United States 
Sector Academic/University 
PI Contribution Provision of adenoviral constructs
Impact None as yet
Start Year 2008
 
Description Sekler 
Organisation Ben-Gurion University of the Negev
Country Israel 
Sector Academic/University 
PI Contribution Experiments for research paper
Collaborator Contribution Experiments for research paper
Impact Publication Pancreatic beta-cell Na+ channels control global Ca2+ signaling and oxidative metabolism by inducing Na+ and Ca2+ responses that are propagated into mitochondria. DOI - 10.1096/fj.13-248161
Start Year 2013
 
Description Yuval Dor (Israel) 
Organisation Open University of Israel
Country Israel 
Sector Academic/University 
PI Contribution Experiments for publications
Collaborator Contribution Experiments for publications
Impact LKB1 and AMPK differentially regulate pancreatic ß-cell identity - Kone M1 et al - FASEB J. 2014 Nov;28(11):4972-85. doi: 10.1096/fj.14-257667. Epub 2014 Jul 28. Loss of Liver Kinase B1 (LKB1) in Beta Cells Enhances Glucose-stimulated Insulin Secretion Despite Profound Mitochondrial Defects. Swisa A1et al - J Biol Chem. 2015 Aug 21;290(34):20934-46. doi: 10.1074/jbc.M115.639237. Epub 2015 Jul 2.
Start Year 2014
 
Description Inaugural lecture - Imperial College 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Participants in your research and patient groups
Results and Impact Inaugural lecture: 'Regulating insulin secretion: can be correct the deficiency in type 2 diabetes

None measured.
Year(s) Of Engagement Activity 2009
 
Description Talk to patient group (Brentwood, 2014) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact 50 patients attended a research talk, which sparked questions and discussion afterwards

informed research
Year(s) Of Engagement Activity 2014
 
Description Talk to patient group (Twickenham, 2013) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact 50 patients attended a research talk, which sparked questions and discussion afterwards

none
Year(s) Of Engagement Activity 2013