Insulin receptor based lymphocyte trafficking in the progression of type 1 diabetes
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Keywords
chemotaxis, Cre-recombinase, diabetes, insulin receptor, insulitis
Abstract
The insulin receptor (IR) is a transmembrane receptor which recognizes and binds the hormone insulin. We describe two models that were devised to explore the role of IR over-expression on T-lymphocytes and their chemotactic motility in the progression of type 1 diabetes. FVB/NJ-CD3-3×FLAG-mIR/MFM mice were generated to selectively over-express 3×FLAG tagged murine IR in T-lymphocytes via an engineered CD3 enhancer and promoter construct. Insertion of the 3×FLAG-mIR transgene into FVB/NJ mice, a known non-autoimmune prone strain, lead to a minor population of detectable 3×FLAG-mIR tagged T-lymphocytes in peripheral blood and the presence of a few lymphocytes in the pancreas of the Tg+/- compared to age matched Tg-/- control mice. In order to induce stronger murine IR over-expression then what was observed with the CD3 enhancer promoter construct, a second system utilizing the strong CAG viral promoter was generated. This system induces cell specific IR over-expression upon Cre-Lox recombination to afford functional 3×FLAG tagged murine IR with an internal eGFP reporter. The pPNTlox2-3×FLAG-mIR plasmid was constructed and validated in HEK-Cre-RFP cells to ensure selective Cre recombinase based 3×FLAG-mIR expression, receptor ligand affinity towards insulin, and functional initiation of signal transduction upon insulin stimulation.
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2. Kido Y (2001) The Insulin Receptor and Its Cellular Targets. Journal of Clinical Endocrinology & Metabolism 86(3):972-979.
3. Freychet P, Roth J, Neville D (1971) Insulin Receptors in the Liver: Specific Binding of 125I Insulin to the Plasma Membrane and Its Relation to Insulin Bioactivity. Proceedings of the National Academy of Sciences 68(8):1833-1837.
4. Olivares-Reyes J, Arellano-Plancarte A, Castillo-Hernandez J (2009) Angiotensin II and the development of insulin resistance: Implications for diabetes. Molecular and Cellular Endocrinology 302(2):128-139.
5. Blazquez E, Velazquez E, Hurtado-Carneiro V, Ruiz-Albusac J (2014) Insulin in the Brain: Its Pathophysiological Implications for States Related with Central Insulin Resistance, Type 2 Diabetes and Alzheimer’s Disease. Frontiers in Endocrinology 5(161): 1-21.
6. Boucher J, Kleinridders A, Kahn C (2014) Insulin Receptor Signaling in Normal and Insulin-Resistant States. Cold Spring Harbor Perspectives in Biology 6(1):a009191-a009191.
7. McInerney M, Flynn J, Goldblatt P, Najjar S, Sherwin R, et al (1996) High density insulin receptor-positive T lymphocytes from nonobese diabetic mice transfer insulitis and diabetes. The Journal of Immunology 157(8):3716-26.
8. Malaguarnera R, Sacco A, Voci C, Pandini G, Vigneri R, et al (2012) Proinsulin Binds with High Affinity the Insulin Receptor Isoform A and Predominantly Activates the Mitogenic Pathway. Endocrinology 153(5):2152-2163.
9. Yenush L, Kundra V, White MF, Zetter BR (1994) Functional domains of the insulin receptor responsible for chemotactic signaling. J. Biol. Chem. 269: 100.
10. Leiter EH (1987) Murine macrophages and pancreatic beta-cells: chemotactic properties of insulin and beta-cytostatic action of interleukin–1. J. Exp. Med. 166:1174.
11. Hendershot T, Liu H, Sarkar A, Giovannucci D, Clouthier D, et al (2006) Expression of Hand2 is sufficient for neurogenesis and cell type–specific gene expression in the enteric nervous system. Developmental Dynamics 236:1 93-105.
12. Belfiore A, Frasca F, Pandini G, Sciacca L, Vigneri R (2009) Insulin Receptor Isoforms and Insulin Receptor/Insulin-Like Growth Factor Receptor Hybrids in Physiology and Disease. Endocrine Reviews 30(6):586-623.
13. Lee J, Pilch P (1994) The insulin receptor: structure, function, and signaling. American Journal of Physiology 266: C319–334.
14. Kasuga M, Hedo J, Yamadall K, Kahn C (1982) The Structure of Insulin Receptor and its Subunits. Journal of Biological Chemistry 257(17):10392-10399.
15. Bravo D, Gleason J, Sanchez R, Roth R, Fuller R (1994) Accurate and efficient cleavage of the human insulin proreceptor by the human proprotein-processing protease furin. Characterization and kinetic parameters using the purified, secreted soluble protease expressed by a recombinant baculovirus. The Journal of Biological Chemistry. 269(41):25830-25837.
16. Kasuga M, Karlsson F, Kahn C (1982) Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor. Science 215(4529):185-187.
17. Leto D, Saltiel A (2012) Regulation of glucose transport by insulin: traffic control of GLUT4. Nature Reviews Molecular Cell Biology 13:6 383-396.
18. Morrison B, Swanson M, Sweet L, Pessin J (1988) Insulin-dependent covalent reassociation of isolated alpha beta heterodimericinsulin receptors into an alpha 2 beta 2 heterotetrameric disulfide-linked complex. The Journal of Biological Chemistry 263:16 7806-7813.
19. Ebina Y, Ellis L, Jarnagin K, Edery M, Graf L, et al (1985) The human insulin receptor cDNA: The structural basis for hormone-activated transmembrane signalling. Cell 40:4 747-758.
20. McInerney M, Najjar S, Brickley D, Lutzke M, Abou-Rjaily G, et al (2004) Anti-Insulin Receptor Autoantibodies Are Not Required for Type 2 Diabetes Pathogenesis in NZL/Lt Mice, a New Zealand Obese (NZO)-Derived Mouse Strain. Experimental Diab. Res 5 177–185.
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Marcia F McInerney, University of Toledo
Distinguished University Professor
