Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 Mar 1;101(5):982–992. doi: 10.1172/JCI1684

Thrombospondin-1 is required for normal murine pulmonary homeostasis and its absence causes pneumonia.

J Lawler 1, M Sunday 1, V Thibert 1, M Duquette 1, E L George 1, H Rayburn 1, R O Hynes 1
PMCID: PMC508649  PMID: 9486968

Abstract

The thrombospondins are a family of extracellular calcium-binding proteins that modulate cellular phenotype. Thrombospondin-1 (TSP-1) reportedly regulates cellular attachment, proliferation, migration, and differentiation in vitro. To explore its function in vivo, we have disrupted the TSP-1 gene by homologous recombination in the mouse genome. Platelets from these mice are completely deficient in TSP-1 protein; however, thrombin-induced platelet aggregation is not diminished. TSP-1-deficient mice display a mild and variable lordotic curvature of the spine that is apparent from birth. These mice also display an increase in the number of circulating white blood cells, with monocytes and eosinophils having the largest percent increases. The brain, heart, kidney, spleen, stomach, intestines, aorta, and liver of TSP-1-deficient mice showed no major abnormalities. However, consistent with high levels of expression of TSP-1 in lung, we observe abnormalities in the lungs of mice that lack the protein. Although normal at birth, histopathological analysis of lungs from 4-wk-old TSP-1-deficient mice reveals extensive acute and organizing pneumonia, with neutrophils and macrophages. The macrophages stain for hemosiderin, indicating that diffuse alveolar hemorrhage is occurring. At later times, the number of neutrophils decreases and a striking increase in the number of hemosiderin-containing macrophages is observed associated with multiple-lineage epithelial hyperplasia and the deposition of collagen and elastin. A thickening and ruffling of the epithelium of the airways results from increasing cell proliferation in TSP-1-deficient mice. These results indicate that TSP-1 is involved in normal lung homeostasis.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams J. C., Lawler J. Cell-type specific adhesive interactions of skeletal myoblasts with thrombospondin-1. Mol Biol Cell. 1994 Apr;5(4):423–437. doi: 10.1091/mbc.5.4.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adams J. C., Lawler J. Diverse mechanisms for cell attachment to platelet thrombospondin. J Cell Sci. 1993 Apr;104(Pt 4):1061–1071. doi: 10.1242/jcs.104.4.1061. [DOI] [PubMed] [Google Scholar]
  3. Asch A. S., Leung L. L., Polley M. J., Nachman R. L. Platelet membrane topography: colocalization of thrombospondin and fibrinogen with the glycoprotein IIb-IIIa complex. Blood. 1985 Oct;66(4):926–934. [PubMed] [Google Scholar]
  4. Asch A. S., Liu I., Briccetti F. M., Barnwell J. W., Kwakye-Berko F., Dokun A., Goldberger J., Pernambuco M. Analysis of CD36 binding domains: ligand specificity controlled by dephosphorylation of an ectodomain. Science. 1993 Nov 26;262(5138):1436–1440. doi: 10.1126/science.7504322. [DOI] [PubMed] [Google Scholar]
  5. Bagavandoss P., Wilks J. W. Specific inhibition of endothelial cell proliferation by thrombospondin. Biochem Biophys Res Commun. 1990 Jul 31;170(2):867–872. doi: 10.1016/0006-291x(90)92171-u. [DOI] [PubMed] [Google Scholar]
  6. Boivin G. P., O'Toole B. A., Orsmby I. E., Diebold R. J., Eis M. J., Doetschman T., Kier A. B. Onset and progression of pathological lesions in transforming growth factor-beta 1-deficient mice. Am J Pathol. 1995 Jan;146(1):276–288. [PMC free article] [PubMed] [Google Scholar]
  7. Bornstein P. Diversity of function is inherent in matricellular proteins: an appraisal of thrombospondin 1. J Cell Biol. 1995 Aug;130(3):503–506. doi: 10.1083/jcb.130.3.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brookfield J. Can genes be truly redundant? Curr Biol. 1992 Oct;2(10):553–554. doi: 10.1016/0960-9822(92)90036-a. [DOI] [PubMed] [Google Scholar]
  9. Corless C. L., Mendoza A., Collins T., Lawler J. Colocalization of thrombospondin and syndecan during murine development. Dev Dyn. 1992 Apr;193(4):346–358. doi: 10.1002/aja.1001930408. [DOI] [PubMed] [Google Scholar]
  10. DeFreitas M. F., Yoshida C. K., Frazier W. A., Mendrick D. L., Kypta R. M., Reichardt L. F. Identification of integrin alpha 3 beta 1 as a neuronal thrombospondin receptor mediating neurite outgrowth. Neuron. 1995 Aug;15(2):333–343. doi: 10.1016/0896-6273(95)90038-1. [DOI] [PubMed] [Google Scholar]
  11. Dejana E., Callioni A., Quintana A., de Gaetano G. Bleeding time in laboratory animals. II - A comparison of different assay conditions in rats. Thromb Res. 1979;15(1-2):191–197. doi: 10.1016/0049-3848(79)90064-1. [DOI] [PubMed] [Google Scholar]
  12. DiCesare P. E., Mörgelin M., Mann K., Paulsson M. Cartilage oligomeric matrix protein and thrombospondin 1. Purification from articular cartilage, electron microscopic structure, and chondrocyte binding. Eur J Biochem. 1994 Aug 1;223(3):927–937. doi: 10.1111/j.1432-1033.1994.tb19070.x. [DOI] [PubMed] [Google Scholar]
  13. DiPietro L. A., Nissen N. N., Gamelli R. L., Koch A. E., Pyle J. M., Polverini P. J. Thrombospondin 1 synthesis and function in wound repair. Am J Pathol. 1996 Jun;148(6):1851–1860. [PMC free article] [PubMed] [Google Scholar]
  14. Dixit V. M., Haverstick D. M., O'Rourke K. M., Hennessy S. W., Grant G. A., Santoro S. A., Frazier W. A. A monoclonal antibody against human thrombospondin inhibits platelet aggregation. Proc Natl Acad Sci U S A. 1985 May;82(10):3472–3476. doi: 10.1073/pnas.82.10.3472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Doetschman T. C., Eistetter H., Katz M., Schmidt W., Kemler R. The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol. 1985 Jun;87:27–45. [PubMed] [Google Scholar]
  16. Gao A. G., Lindberg F. P., Finn M. B., Blystone S. D., Brown E. J., Frazier W. A. Integrin-associated protein is a receptor for the C-terminal domain of thrombospondin. J Biol Chem. 1996 Jan 5;271(1):21–24. doi: 10.1074/jbc.271.1.21. [DOI] [PubMed] [Google Scholar]
  17. George E. L., Georges-Labouesse E. N., Patel-King R. S., Rayburn H., Hynes R. O. Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin. Development. 1993 Dec;119(4):1079–1091. doi: 10.1242/dev.119.4.1079. [DOI] [PubMed] [Google Scholar]
  18. Huber A. R., Ellis S., Johnson K. J., Dixit V. M., Varani J. Monocyte diapedesis through an in vitro vessel wall construct: inhibition with monoclonal antibodies to thrombospondin. J Leukoc Biol. 1992 Nov;52(5):524–528. doi: 10.1002/jlb.52.5.524. [DOI] [PubMed] [Google Scholar]
  19. Hughes J., Liu Y., Van Damme J., Savill J. Human glomerular mesangial cell phagocytosis of apoptotic neutrophils: mediation by a novel CD36-independent vitronectin receptor/thrombospondin recognition mechanism that is uncoupled from chemokine secretion. J Immunol. 1997 May 1;158(9):4389–4397. [PubMed] [Google Scholar]
  20. Hynes R. O. Targeted mutations in cell adhesion genes: what have we learned from them? Dev Biol. 1996 Dec 15;180(2):402–412. doi: 10.1006/dbio.1996.0314. [DOI] [PubMed] [Google Scholar]
  21. Iruela-Arispe M. L., Liska D. J., Sage E. H., Bornstein P. Differential expression of thrombospondin 1, 2, and 3 during murine development. Dev Dyn. 1993 May;197(1):40–56. doi: 10.1002/aja.1001970105. [DOI] [PubMed] [Google Scholar]
  22. Jetten A. M., Shirley J. E., Stoner G. Regulation of proliferation and differentiation of respiratory tract epithelial cells by TGF beta. Exp Cell Res. 1986 Dec;167(2):539–549. doi: 10.1016/0014-4827(86)90193-x. [DOI] [PubMed] [Google Scholar]
  23. Kao K. J., Shaut D. M., Klein P. A. Functional involvement of thrombospondin in platelet aggregation induced by low versus high concentrations of thrombin. Thromb Haemost. 1986 Feb 28;55(1):136–142. [PubMed] [Google Scholar]
  24. Kuhn C., Mason R. J. Immunolocalization of SPARC, tenascin, and thrombospondin in pulmonary fibrosis. Am J Pathol. 1995 Dec;147(6):1759–1769. [PMC free article] [PubMed] [Google Scholar]
  25. Kulkarni A. B., Huh C. G., Becker D., Geiser A., Lyght M., Flanders K. C., Roberts A. B., Sporn M. B., Ward J. M., Karlsson S. Transforming growth factor beta 1 null mutation in mice causes excessive inflammatory response and early death. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):770–774. doi: 10.1073/pnas.90.2.770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. Laird P. W., Zijderveld A., Linders K., Rudnicki M. A., Jaenisch R., Berns A. Simplified mammalian DNA isolation procedure. Nucleic Acids Res. 1991 Aug 11;19(15):4293–4293. doi: 10.1093/nar/19.15.4293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lawler J., Derick L. H., Connolly J. E., Chen J. H., Chao F. C. The structure of human platelet thrombospondin. J Biol Chem. 1985 Mar 25;260(6):3762–3772. [PubMed] [Google Scholar]
  29. Lawler J., Duquette M., Ferro P., Copeland N. G., Gilbert D. J., Jenkins N. A. Characterization of the murine thrombospondin gene. Genomics. 1991 Nov;11(3):587–600. doi: 10.1016/0888-7543(91)90066-n. [DOI] [PubMed] [Google Scholar]
  30. Lawler J., Duquette M., Whittaker C. A., Adams J. C., McHenry K., DeSimone D. W. Identification and characterization of thrombospondin-4, a new member of the thrombospondin gene family. J Cell Biol. 1993 Feb;120(4):1059–1067. doi: 10.1083/jcb.120.4.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lawler J., Weinstein R., Hynes R. O. Cell attachment to thrombospondin: the role of ARG-GLY-ASP, calcium, and integrin receptors. J Cell Biol. 1988 Dec;107(6 Pt 1):2351–2361. doi: 10.1083/jcb.107.6.2351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Legrand C., Thibert V., Dubernard V., Bégault B., Lawler J. Molecular requirements for the interaction of thrombospondin with thrombin-activated human platelets: modulation of platelet aggregation. Blood. 1992 Apr 15;79(8):1995–2003. [PubMed] [Google Scholar]
  33. Leung L. L. Role of thrombospondin in platelet aggregation. J Clin Invest. 1984 Nov;74(5):1764–1772. doi: 10.1172/JCI111595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Magdaleno S. M., Wang G., Jackson K. J., Ray M. K., Welty S., Costa R. H., DeMayo F. J. Interferon-gamma regulation of Clara cell gene expression: in vivo and in vitro. Am J Physiol. 1997 Jun;272(6 Pt 1):L1142–L1151. doi: 10.1152/ajplung.1997.272.6.L1142. [DOI] [PubMed] [Google Scholar]
  35. Majack R. A., Cook S. C., Bornstein P. Control of smooth muscle cell growth by components of the extracellular matrix: autocrine role for thrombospondin. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9050–9054. doi: 10.1073/pnas.83.23.9050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Majack R. A., Goodman L. V., Dixit V. M. Cell surface thrombospondin is functionally essential for vascular smooth muscle cell proliferation. J Cell Biol. 1988 Feb;106(2):415–422. doi: 10.1083/jcb.106.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mansfield P. J., Boxer L. A., Suchard S. J. Thrombospondin stimulates motility of human neutrophils. J Cell Biol. 1990 Dec;111(6 Pt 2):3077–3086. doi: 10.1083/jcb.111.6.3077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Mansfield P. J., Suchard S. J. Thrombospondin promotes chemotaxis and haptotaxis of human peripheral blood monocytes. J Immunol. 1994 Nov 1;153(9):4219–4229. [PubMed] [Google Scholar]
  39. Mansour S. L., Thomas K. R., Capecchi M. R. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature. 1988 Nov 24;336(6197):348–352. doi: 10.1038/336348a0. [DOI] [PubMed] [Google Scholar]
  40. McGregor B., Colon S., Mutin M., Chignier E., Zech P., McGregor J. Thrombospondin in human glomerulopathies. A marker of inflammation and early fibrosis. Am J Pathol. 1994 Jun;144(6):1281–1287. [PMC free article] [PubMed] [Google Scholar]
  41. Mosher D. F., Misenheimer T. M., Stenflo J., Hogg P. J. Modulation of fibrinolysis by thrombospondin. Ann N Y Acad Sci. 1992 Dec 4;667:64–69. doi: 10.1111/j.1749-6632.1992.tb51598.x. [DOI] [PubMed] [Google Scholar]
  42. Murphy-Ullrich J. E., Gurusiddappa S., Frazier W. A., Hök M. Heparin-binding peptides from thrombospondins 1 and 2 contain focal adhesion-labilizing activity. J Biol Chem. 1993 Dec 15;268(35):26784–26789. [PubMed] [Google Scholar]
  43. O'Shea K. S., Dixit V. M. Unique distribution of the extracellular matrix component thrombospondin in the developing mouse embryo. J Cell Biol. 1988 Dec;107(6 Pt 2):2737–2748. doi: 10.1083/jcb.107.6.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. O'Shea K. S., Rheinheimer J. S., Dixit V. M. Deposition and role of thrombospondin in the histogenesis of the cerebellar cortex. J Cell Biol. 1990 Apr;110(4):1275–1283. doi: 10.1083/jcb.110.4.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Raugi G. J., Mullen J. S., Bark D. H., Okada T., Mayberg M. R. Thrombospondin deposition in rat carotid artery injury. Am J Pathol. 1990 Jul;137(1):179–185. [PMC free article] [PubMed] [Google Scholar]
  46. Raugi G. J., Olerud J. E., Gown A. M. Thrombospondin in early human wound tissue. J Invest Dermatol. 1987 Dec;89(6):551–554. doi: 10.1111/1523-1747.ep12461198. [DOI] [PubMed] [Google Scholar]
  47. Reed M. J., Iruela-Arispe L., O'Brien E. R., Truong T., LaBell T., Bornstein P., Sage E. H. Expression of thrombospondins by endothelial cells. Injury is correlated with TSP-1. Am J Pathol. 1995 Oct;147(4):1068–1080. [PMC free article] [PubMed] [Google Scholar]
  48. Rudnicki M. A., Braun T., Hinuma S., Jaenisch R. Inactivation of MyoD in mice leads to up-regulation of the myogenic HLH gene Myf-5 and results in apparently normal muscle development. Cell. 1992 Oct 30;71(3):383–390. doi: 10.1016/0092-8674(92)90508-a. [DOI] [PubMed] [Google Scholar]
  49. Sage H., Farin F. M., Striker G. E., Fisher A. B. Granular pneumocytes in primary culture secrete several major components of the extracellular matrix. Biochemistry. 1983 Apr 26;22(9):2148–2155. doi: 10.1021/bi00278a015. [DOI] [PubMed] [Google Scholar]
  50. Savill J., Hogg N., Ren Y., Haslett C. Thrombospondin cooperates with CD36 and the vitronectin receptor in macrophage recognition of neutrophils undergoing apoptosis. J Clin Invest. 1992 Oct;90(4):1513–1522. doi: 10.1172/JCI116019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Schultz-Cherry S., Ribeiro S., Gentry L., Murphy-Ullrich J. E. Thrombospondin binds and activates the small and large forms of latent transforming growth factor-beta in a chemically defined system. J Biol Chem. 1994 Oct 28;269(43):26775–26782. [PubMed] [Google Scholar]
  52. Shull M. M., Ormsby I., Kier A. B., Pawlowski S., Diebold R. J., Yin M., Allen R., Sidman C., Proetzel G., Calvin D. Targeted disruption of the mouse transforming growth factor-beta 1 gene results in multifocal inflammatory disease. Nature. 1992 Oct 22;359(6397):693–699. doi: 10.1038/359693a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Silverstein R. L., Leung L. L., Harpel P. C., Nachman R. L. Platelet thrombospondin forms a trimolecular complex with plasminogen and histidine-rich glycoprotein. J Clin Invest. 1985 Jun;75(6):2065–2073. doi: 10.1172/JCI111926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Silverstein R. L., Nachman R. L., Pannell R., Gurewich V., Harpel P. C. Thrombospondin forms complexes with single-chain and two-chain forms of urokinase. J Biol Chem. 1990 Jul 5;265(19):11289–11294. [PubMed] [Google Scholar]
  55. Stern M., Savill J., Haslett C. Human monocyte-derived macrophage phagocytosis of senescent eosinophils undergoing apoptosis. Mediation by alpha v beta 3/CD36/thrombospondin recognition mechanism and lack of phlogistic response. Am J Pathol. 1996 Sep;149(3):911–921. [PMC free article] [PubMed] [Google Scholar]
  56. Sunday M. E., Willett C. G., Graham S. A., Oreffo V. I., Linnoila R. I., Witschi H. Histochemical characterization of non-neuroendocrine tumors and neuroendocrine cell hyperplasia induced in hamster lung by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone with or without hyperoxia. Am J Pathol. 1995 Sep;147(3):740–752. [PMC free article] [PubMed] [Google Scholar]
  57. Sunday M. E., Willett C. G., Patidar K., Graham S. A. Modulation of oncogene and tumor suppressor gene expression in a hamster model of chronic lung injury with varying degrees of pulmonary neuroendocrine cell hyperplasia. Lab Invest. 1994 Jun;70(6):875–888. [PubMed] [Google Scholar]
  58. Taraboletti G., Roberts D., Liotta L. A., Giavazzi R. Platelet thrombospondin modulates endothelial cell adhesion, motility, and growth: a potential angiogenesis regulatory factor. J Cell Biol. 1990 Aug;111(2):765–772. doi: 10.1083/jcb.111.2.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Thompson E. M., Hughes J., Van Noorden S., Sharpe J., Savill J. Expression of the multifunctional extracellular matrix protein thrombospondin in crescentic glomerulonephritis. J Pathol. 1996 Jan;178(1):89–94. doi: 10.1002/(SICI)1096-9896(199601)178:1<89::AID-PATH457>3.0.CO;2-4. [DOI] [PubMed] [Google Scholar]
  60. Tolsma S. S., Volpert O. V., Good D. J., Frazier W. A., Polverini P. J., Bouck N. Peptides derived from two separate domains of the matrix protein thrombospondin-1 have anti-angiogenic activity. J Cell Biol. 1993 Jul;122(2):497–511. doi: 10.1083/jcb.122.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Tucker R. P., Hagios C., Chiquet-Ehrismann R., Lawler J. In situ localization of thrombospondin-1 and thrombospondin-3 transcripts in the avian embryo. Dev Dyn. 1997 Mar;208(3):326–337. doi: 10.1002/(SICI)1097-0177(199703)208:3<326::AID-AJA4>3.0.CO;2-K. [DOI] [PubMed] [Google Scholar]
  62. Vorbroker D. K., Profitt S. A., Nogee L. M., Whitsett J. A. Aberrant processing of surfactant protein C in hereditary SP-B deficiency. Am J Physiol. 1995 Apr;268(4 Pt 1):L647–L656. doi: 10.1152/ajplung.1995.268.4.L647. [DOI] [PubMed] [Google Scholar]
  63. Watkins S. C., Lynch G. W., Kane L. P., Slayter H. S. Thrombospondin expression in traumatized skeletal muscle. Correlation of appearance with post-trauma regeneration. Cell Tissue Res. 1990 Jul;261(1):73–84. doi: 10.1007/BF00329440. [DOI] [PubMed] [Google Scholar]
  64. Wight T. N., Raugi G. J., Mumby S. M., Bornstein P. Light microscopic immunolocation of thrombospondin in human tissues. J Histochem Cytochem. 1985 Apr;33(4):295–302. doi: 10.1177/33.4.3884704. [DOI] [PubMed] [Google Scholar]
  65. Yabkowitz R., Dixit V. M., Guo N., Roberts D. D., Shimizu Y. Activated T-cell adhesion to thrombospondin is mediated by the alpha 4 beta 1 (VLA-4) and alpha 5 beta 1 (VLA-5) integrins. J Immunol. 1993 Jul 1;151(1):149–158. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES