Adipocyte-derived microvesicles are associated with multiple angiogenic factors and induce angiogenesis in vivo and in vitro.

We previously reported that 3T3-L1 and rat primary adipocytes secreted microvesicles, known as adipocyte-derived microvesicles (ADMs). In the present study, we further characterized the 3T3-L1 ADMs and found that they exhibited angiogenic activity in vivo. Antibody arrays and gelatin zymography analyses revealed that several angiogenic and antiangiogenic proteins, including leptin, TNFalpha, acidic fibroblast growth factor (FGFa), interferon-gamma, and matrix metalloprotease (MMP)-2 and MMP-9, were present in the ADMs. Gene expression of most of these angiogenic factors was induced in the adipose tissue of diet-induced obese mice. Furthermore, leptin, TNFalpha, and MMP-2 were up-regulated at the protein level in the adipocyte fractions prepared from epididymal adipose tissues of high-fat-diet-induced obese mice. ADMs induced cell migration and tube formation of human umbilical vein endothelial cells, which were partially suppressed by neutralizing antibodies to leptin, TNFalpha, or FGFa but not to interferon-gamma. Supporting these data, a mixture of leptin, TNFalpha, and FGFa induced tube formation. ADMs also promoted cell invasion of human umbilical vein endothelial cells through Matrigel, which was suppressed by the addition of the MMP inhibitor 1,10'-phenanthroline and a neutralizing antibody to MMP-2 but not to MMP-9. These results suggest that ADMs are associated with multiple angiogenic factors and play a role in angiogenesis in adipose tissue.

[1]  P. Kopelman.  Obesity as a medical problem , 2000, Nature.

[2]  R. Pifarré,et al.  MYOCARDIAL REVASCULARIZATION BY OMENTAL GRAFT WITHOUT PEDICLE: EXPERIMENTAL BACKGROUND AND REPORT ON 25 CASES FOLLOWED 6 TO 16 MONTHS. , 1965, The Journal of thoracic and cardiovascular surgery.

[3]  P. Lin,et al.  Adipokine resistin promotes in vitro angiogenesis of human endothelial cells. , 2006, Cardiovascular research.

[4]  Jinping Li,et al.  Gene expression profile of rat adipose tissue at the onset of high-fat-diet obesity. , 2002, American journal of physiology. Endocrinology and metabolism.

[5]  D. Loskutoff,et al.  Angiogenesis in an in vivo model of adipose tissue development , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[6]  N. Aoki,et al.  Identification and characterization of microvesicles secreted by 3T3-L1 adipocytes: redox- and hormone-dependent induction of milk fat globule-epidermal growth factor 8-associated microvesicles. , 2007, Endocrinology.

[7]  D. Hicklin,et al.  Granulocyte colony‐stimulating factor promotes neovascularization by releasing vascular endothelial growth factor from neutrophils , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  G. Garcı́a-Cardeña,et al.  Biological action of leptin as an angiogenic factor. , 1998, Science.

[9]  J. Slot,et al.  Proteomic and Biochemical Analyses of Human B Cell-derived Exosomes , 2003, The Journal of Biological Chemistry.

[10]  A. Saiki,et al.  Hepatocyte growth factor secreted by cultured adipocytes promotes tube formation of vascular endothelial cells in vitro , 2006, International Journal of Obesity.

[11]  B. Lowell,et al.  Adipose tissue mass can be regulated through the vasculature , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[12]  H. Shepard,et al.  Macrophage-induced angiogenesis is mediated by tumour necrosis factor-α , 1987, Nature.

[13]  D. L. Crandall,et al.  A Review of the Microcirculation of Adipose Tissue: Anatomic, Metabolic, and Angiogenic Perspectives , 1997, Microcirculation.

[14]  P. Carmeliet,et al.  Impaired Adipose Tissue Development in Mice With Inactivation of Placental Growth Factor Function , 2006, Diabetes.

[15]  N. Aoki,et al.  Stage specific expression of milk fat globule membrane glycoproteins in mouse mammary gland: comparison of MFG-E8, butyrophilin, and CD36 with a major milk protein, β-casein , 1997 .

[16]  T. Rosengart,et al.  Vascular endothelial growth factor is the major angiogenic factor in omentum: mechanism of the omentum-mediated angiogenesis. , 1997, The Journal of surgical research.

[17]  C. Beck,et al.  REVASCULARIZATION OF THE HEART BY VEIN GRAFT FROM AORTA TO CORONARY SINUS , 1951, Annals of surgery.

[18]  Yihai Cao,et al.  Angiogenesis Inhibitor, TNP-470, Prevents Diet-Induced and Genetic Obesity in Mice , 2004, Circulation research.

[19]  W. Risau,et al.  Tumor necrosis factor type alpha, a potent inhibitor of endothelial cell growth in vitro, is angiogenic in vivo. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[20]  R. Lijnen,et al.  Modulation of angiogenesis during adipose tissue development in murine models of obesity. , 2005, Endocrinology.

[21]  E. Van Obberghen,et al.  Matrix Metalloproteinases Are Differentially Expressed in Adipose Tissue during Obesity and Modulate Adipocyte Differentiation* , 2003, The Journal of Biological Chemistry.

[22]  P. Grimaldi,et al.  Cellular and molecular aspects of adipose tissue development. , 1992, Annual review of nutrition.

[23]  D. Loskutoff,et al.  Tissue factor gene expression in the adipose tissues of obese mice. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[24]  W. Kiess,et al.  Adipocytokines: leptin--the classical, resistin--the controversical, adiponectin--the promising, and more to come. , 2005, Best practice & research. Clinical endocrinology & metabolism.

[25]  T. Maciag,et al.  Models of in vitro angiogenesis: endothelial cell differentiation on fibrin but not matrigel is transcriptionally dependent. , 1995, Biochemical and biophysical research communications.

[26]  Y. Matsuzawa.  White adipose tissue and cardiovascular disease. , 2005, Best practice & research. Clinical endocrinology & metabolism.

[27]  M. Lafontan,et al.  Angiogenesis in adipose tissue. , 2002, Annales d'endocrinologie.

[28]  Johan Skog,et al.  Glioblastoma microvesicles transport RNA and protein that promote tumor growth and provide diagnostic biomarkers , 2008, Nature Cell Biology.

[29]  Bruce M. Spiegelman,et al.  Obesity and the Regulation of Energy Balance , 2001, Cell.

[30]  Yihai Cao.  Angiogenesis modulates adipogenesis and obesity. , 2007, The Journal of clinical investigation.

[31]  R. Busse,et al.  Leptin, the product of Ob gene, promotes angiogenesis. , 1998, Circulation research.

[32]  Sebastian Amigorena,et al.  Lactadherin promotes VEGF-dependent neovascularization , 2005, Nature Medicine.

[33]  S. Rafii,et al.  Impaired recruitment of bone-marrow–derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth , 2001, Nature Medicine.

[34]  Anna Janowska-Wieczorek,et al.  Microvesicles derived from activated platelets induce metastasis and angiogenesis in lung cancer , 2005, International journal of cancer.