Cells expressing the stem cell factor receptor,c-kit,contribute to neoangiogenesis in diabetes

Abstract

While neoangiogenesis in diabetes is of greatest clinical significance in the retina, the pathological formation of new blood vessels also develops in other vascular beds in diabetes, including the mesentery of the streptozotocin-induced diabetic rat. However, the contribution of bone marrow-derived cells to this process of vasculogenesis is unknown.

In this study, male Sprague-Dawley rats were randomised to receive either streptozotocin or vehicle, and their mesenteric vasculature was examined after three weeks. Origins from bone marrow and endothelial cell differentiation were identified by immunolabelling for the stem cell factor receptor, c-kit and von Willebrand factor (vWF), respectively. Expression for the angiogenic chemokine, stromal derived factor-1 (SDF-1) was assessed by quantitative real-time polymerase chain reaction (PCR).

At three weeks, rats with diabetes had a dramatic (190-fold) increase in lectin-labelled blood vessel profiles in the mesenteric bed (p<0.0001) in association with a five-fold increase in SDF-1 mRNA (p<0.01). Immunohistochemical studies identified abundant large, ovoid, lumen-forming, c-kit⁺ cells in the mesentery of diabetic, but not control, rats. Many of these c-kit⁺ cells also showed positive immunolabelling for vWF, consistent with endothelial differentiation.

In conclusion, cells of bone marrow origin contribute to new vessel formation in the diabetic mesentery. This phenomenon may also apply to the neovascularisation that develops at clinically important sites such as in the retina.

Keywords

diabetes stem cell factor stromal derived factor neoangiogenesis mesentery

References

1. Williamson JR , Kilo C. Basement-membrane thickening and diabetic microangiopathy. Diabetes 1976;25:925–7.

2. Bollinger A , Frey J Jager K , Furrer J , Seglias J , Siegenthaler W. Patterns of diffusion through skin capillaries in patients with long-term diabetes. N Engl J Med 1982;307:1305–10.

3. Stephan CC , Chang KC , LeJeune W et al. Role for heparin-binding growth factors in glucose-induced vascular dysfunction. Diabetes 1998;47:1771–8.

4. Rumble JR , Cooper ME , Soulis T et al. Vascular hypertrophy in experimental diabetes: role of advanced glycation end products. J Clin Invest 1997;99:1016–27.

5. Arshi K , Bendayan M , Ghitescu LD. Alterations of the rat mesentery vasculature in experimental diabetes. Lab Invest 2000;80:1171–84.

6. Norrby K , Jakobsson A , Sorbo J. Quantitative angiogenesis in spreads of intact rat mesenteric windows. Microvasc Res 1990;39:341–8.

7. Conway EM , Collen D , Carmeliet P. Molecular mechanisms of blood vessel growth. Cardiovasc Res 2001;49:507–21.

8. Kocher AA , Schuster MD , Szabolcs MJ et al. Neovascularization of ischemic myocardium by human bone-marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function. Nat Med 2001;7:430–6.

9. Otani A , Kinder K , Ewalt K , Otero FJ , Schimmel P , Friedlander M. Bone marrow-derived stem cells target retinal astrocytes and can promote or inhibit retinal angiogenesis. Nat Med 2002;8:1004–10.

10.

10. Ogawa M , Matsuzaki Y , Nishikawa S et al. Expression and function of c-kit in hemopoietic progenitor cells. J Exp Med 1991;174(1):63–71.

11.

11. Strieter RM , Polverini PJ , Kunkel SL et al. The functional role of the ELR motif in CXC chemokine-mediated angiogenesis. J Biol Chem 1995;270:27348–57.

12.

12. Heidemann J , Ogawa H , Rafiee P et al. Mucosal angiogenesis regulation by CXCR4 and its ligand CXCL12 expressed by human intestinal microvascular endothelial cells. Am J Physiol Gastrointest Liver Physiol 2004;286:G1059–68.

13.

13. Bunag RD. Validation in awake rats of a tail-cuff method for measuring systolic pressure. J Applied Physiol 1973;34:279–82.

14.

14. Moravski CJ , Skinner SL , Stubbs AJ et al. The Renin-Angiotensin system influences ocular endothelial cell proliferation in diabetes: transgenic and interventional studies. Am J Pathol 2003;162:151–60.

15.

15. Shintani S , Murohara T , Ikeda H et al. Augmentation of postnatal neovascularization with autologous bone marrow transplantation. Circulation 2001;103:897–903.

16.

16. Gilbert RE , Rumble JR , Cao ZM et al. Endothelin receptor antagonism ameliorates mast cell infiltration, vascular hypertrophy, and epidermal growth factor expression in experimental diabetes. Circulation Res 2000;86:158–65.

17.

17. Ceponis A , Konttinen YT , Takagi M et al. Expression of stem cell factor (SCF) and SCF receptor (c-kit) in synovial membrane in arthritis: correlation with synovial mast cell hyperplasia and inflammation. J Rheumatol 1998;25:2304–14.

18.

18. Kelly DJ , Aaltonen P , Cox AJ et al. Expression of the slit-diaphragm protein, nephrin, in experimental diabetic nephropathy: differing effects of anti-proteinuric therapies. Nephrol Dial Transplant 2002;17:1327–32.

19.

19. Asahara T , Masuda H , Takahashi T et al. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res 1999;85:221–8.

20.

20. Hu Y , Zhang Z , Torsney E et al. Abundant progenitor cells in the adventitia contribute to atherosclerosis of vein grafts in ApoE-deficient mice. J Clin Invest 2004;113:1258–65.

21.

21. Segal MS , Bihorac A , Koc M. Circulating endothelial cells: tea leaves for renal disease. Am J Physiol Renal Physiol 2002;283(1):F11–19.

22.

22. Lammie A , Drobnjak M , Gerald W , Saad A , Cote R , Cordon-Cardo C. Expression of c-kit and kit ligand proteins in normal human tissues. J Histochem Cytochem 1994;42:1417–25.

23.

23. Murayama T , Tepper OM , Silver M et al. Determination of bone marrow-derived endothelial progenitor cell significance in angiogenic growth factor-induced neovascularization in vivo. Exp Hematol 2002;30:967–72.

24.

24. Welsh M , Anneren C , Lindholm C , Kriz V , Oberg-Welsh C. Role of tyrosine kinase signaling for beta-cell replication and survival. Ups J Med Sci 2000;105(2):7–15.

25.

25. Tachibana K , Hirota S , Iizasa H et al. The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract. Nature 1998;393:591–4.

26.

26. Salcedo R , Wasserman K , Young HA et al. Vascular endothelial growth factor and basic fibroblast growth factor induce expression of CXCR4 on human endothelial cells: In vivo neovascularization induced by stromal-derived factor-1alpha. Am J Pathol 1999;154:1125–35.

27.

27. Hiasa K , Ishibashi M , Ohtani K et al. Gene transfer of stromal cell-derived factor-1alpha enhances ischemic vasculogenesis and angiogenesis via vascular endothelial growth factor/endothelial nitric oxide synthase-related pathway: next-generation chemokine therapy for therapeutic neovascularization. Circulation 2004;109:2454–61.