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Keane FM, Yao TW, Seelk S, et al. Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs. FEBS Open Bio. 2013;4:43-54doi: 10.1016/j.fob.2013.12.001.
Keane, F. M., Yao, T. W., Seelk, S., Gall, M. G., Chowdhury, S., Poplawski, S. E., Lai, J. H., Li, Y., Wu, W., Farrell, P., Vieira de Ribeiro, A. J., Osborne, B., Yu, D. M., Seth, D., Rahman, K., Haber, P., Topaloglu, A. K., Wang, C., Thomson, S., Hennessy, A., Prins, J., Twigg, S. M., McLennan, S. V., McCaughan, G. W., Bachovchin, W. W., & Gorrell, M. D. (2013). Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs. FEBS open bio, 443-54. https://doi.org/10.1016/j.fob.2013.12.001
Keane, Fiona M, et al. "Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs." FEBS open bio vol. 4 (2013): 43-54. doi: https://doi.org/10.1016/j.fob.2013.12.001
Keane FM, Yao TW, Seelk S, Gall MG, Chowdhury S, Poplawski SE, Lai JH, Li Y, Wu W, Farrell P, Vieira de Ribeiro AJ, Osborne B, Yu DM, Seth D, Rahman K, Haber P, Topaloglu AK, Wang C, Thomson S, Hennessy A, Prins J, Twigg SM, McLennan SV, McCaughan GW, Bachovchin WW, Gorrell MD. Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs. FEBS Open Bio. 2013 Dec 08;4:43-54. doi: 10.1016/j.fob.2013.12.001. eCollection 2013. PMID: 24371721; PMCID: PMC3871272.
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FEBS Open Bio. 2013 Dec 08;4:43-54. doi: 10.1016/j.fob.2013.12.001. eCollection 2013.

Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs.

FEBS open bio

Fiona M Keane, Tsun-Wen Yao, Stefanie Seelk, Margaret G Gall, Sumaiya Chowdhury, Sarah E Poplawski, Jack H Lai, Youhua Li, Wengen Wu, Penny Farrell, Ana Julia Vieira de Ribeiro, Brenna Osborne, Denise M T Yu, Devanshi Seth, Khairunnessa Rahman, Paul Haber, A Kemal Topaloglu, Chuanmin Wang, Sally Thomson, Annemarie Hennessy, John Prins, Stephen M Twigg, Susan V McLennan, Geoffrey W McCaughan, William W Bachovchin, Mark D Gorrell

Affiliations

  1. Centenary Institute, Camperdown, NSW, Australia ; Sydney Medical School, University of Sydney, NSW, Australia.
  2. Centenary Institute, Camperdown, NSW, Australia.
  3. Sackler School of Biomedical Sciences, Tufts University School of Medicine, Boston, MA, USA.
  4. Department of Renal Medicine, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
  5. Centenary Institute, Camperdown, NSW, Australia ; Drug Health Services, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
  6. Drug Health Services, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
  7. Sydney Medical School, University of Sydney, NSW, Australia ; Drug Health Services, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
  8. Pediatric Endocrinology, Faculty of Medicine, Cukurova University, Adana, Turkey.
  9. Sydney Medical School, University of Sydney, NSW, Australia ; Collaborative Transplantation Research Group, Bosch Institute, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
  10. Sydney Medical School, University of Sydney, NSW, Australia ; Department of Renal Medicine, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
  11. Department of Renal Medicine, Royal Prince Alfred Hospital, Camperdown, NSW, Australia ; School of Medicine, University of Western Sydney, NSW, Australia.
  12. Mater Medical Research Institute, University of Queensland, and Department of Endocrinology, Princess Alexandra Hospital, Brisbane, QLD, Australia.
  13. Sydney Medical School, University of Sydney, NSW, Australia ; Department of Endocrinology, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.

PMID: 24371721 PMCID: PMC3871272 DOI: 10.1016/j.fob.2013.12.001

Abstract

The protease fibroblast activation protein (FAP) is a specific marker of activated mesenchymal cells in tumour stroma and fibrotic liver. A specific, reliable FAP enzyme assay has been lacking. FAP's unique and restricted cleavage of the post proline bond was exploited to generate a new specific substrate to quantify FAP enzyme activity. This sensitive assay detected no FAP activity in any tissue or fluid of FAP gene knockout mice, thus confirming assay specificity. Circulating FAP activity was ∼20- and 1.3-fold less in baboon than in mouse and human plasma, respectively. Serum and plasma contained comparable FAP activity. In mice, the highest levels of FAP activity were in uterus, pancreas, submaxillary gland and skin, whereas the lowest levels were in brain, prostate, leukocytes and testis. Baboon organs high in FAP activity included skin, epididymis, bladder, colon, adipose tissue, nerve and tongue. FAP activity was greatly elevated in tumours and associated lymph nodes and in fungal-infected skin of unhealthy baboons. FAP activity was 14- to 18-fold greater in cirrhotic than in non-diseased human liver, and circulating FAP activity was almost doubled in alcoholic cirrhosis. Parallel DPP4 measurements concorded with the literature, except for the novel finding of high DPP4 activity in bile. The new FAP enzyme assay is the first to be thoroughly characterised and shows that FAP activity is measurable in most organs and at high levels in some. This new assay is a robust tool for specific quantitation of FAP enzyme activity in both preclinical and clinical samples, particularly liver fibrosis.

Keywords: ALD, alcoholic liver disease; AMC, amino-4-methylcoumarin; Biomarker; DMSO, dimethyl sulfoxide; DPP4, dipeptidyl peptidase 4; Dipeptidyl peptidase; EDTA, ethylene diamine tetra acetic acid; FAP, fibroblast activation protein-α; Fibroblast; Fibrosis; HCV, hepatitis C virus; LDS, lithium dodecyl sulphate; LN, lymph node; Liver disease; ND, non-diseased; PBC, primary biliary cirrhosis; PBMC, peripheral blood mononuclear cells; PBS, phosphate-buffered saline; PEP, prolyl endopeptidase; PVDF, polyvinylidene fluoride; Protease activity; Protease substrates; STLV, simian T-cell lymphotrophic virus; gko, gene knock out; het, heterozygous; mAb, monoclonal antibody; wt, wild type; yrs, years

References

  1. Mol Cancer Ther. 2012 Feb;11(2):257-66 - PubMed
  2. Scand J Immunol. 2001 Sep;54(3):249-64 - PubMed
  3. Cancer Biol Ther. 2012 Feb 1;13(3):123-9 - PubMed
  4. Cancer Immunol Immunother. 2009 Nov;58(11):1723-47 - PubMed
  5. Arthritis Res Ther. 2006;8(6):R171 - PubMed
  6. Eur J Biochem. 1993 Jun 15;214(3):829-35 - PubMed
  7. PLoS One. 2010 May 25;5(5):e10817 - PubMed
  8. Diabetologia. 2010 Mar;53(3):572-83 - PubMed
  9. Biol Chem. 2011 Mar;392(3):199-207 - PubMed
  10. Hepatology. 2005 Oct;42(4):935-45 - PubMed
  11. Dis Markers. 2003-2004;19(6):267-72 - PubMed
  12. Clin Sci (Lond). 2005 Apr;108(4):277-92 - PubMed
  13. Diabetologia. 2005 Jun;48(6):1168-72 - PubMed
  14. J Clin Invest. 2011 Jan;121(1):308-17 - PubMed
  15. Diabetes Res Clin Pract. 2008 Jan;79(1):79-85 - PubMed
  16. J Exp Med. 2013 Jun 3;210(6):1125-35 - PubMed
  17. Diabetes. 2011 Jul;60(7):1917-25 - PubMed
  18. Front Biosci. 2008 Jan 01;13:3168-80 - PubMed
  19. J Exp Med. 2013 Jun 3;210(6):1137-51 - PubMed
  20. FEBS Lett. 1978 Jul 15;91(2):360-4 - PubMed
  21. Biochim Biophys Acta. 2014 Jul;1844(7):1248-59 - PubMed
  22. Hepatology. 1992 Jun;15(6):1048-53 - PubMed
  23. Am J Obstet Gynecol. 2003 Mar;188(3):670-6 - PubMed
  24. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7235-9 - PubMed
  25. Immunol Cell Biol. 2002 Aug;80(4):391-7 - PubMed
  26. Regul Pept. 1993 Dec 10;49(2):133-44 - PubMed
  27. Am J Pathol. 2011 Mar;178(3):1134-44 - PubMed
  28. Mol Ther. 2013 Aug;21(8):1611-20 - PubMed
  29. J Histochem Cytochem. 2009 Nov;57(11):1025-40 - PubMed
  30. Am J Physiol Gastrointest Liver Physiol. 2012 Apr 15;302(8):G759-61 - PubMed
  31. Diagn Pathol. 2011 Nov 08;6:111 - PubMed
  32. World J Gastroenterol. 2013 May 21;19(19):2883-93 - PubMed
  33. Regul Pept. 2013 Sep 10;186:26-35 - PubMed
  34. Oncogene. 2012 Feb 23;31(8):992-1000 - PubMed
  35. Liver. 2002 Apr;22(2):93-101 - PubMed
  36. FEBS J. 2011 Apr;278(8):1316-32 - PubMed
  37. J Thromb Haemost. 2011 May;9(5):987-96 - PubMed
  38. J Gastroenterol Hepatol. 2008 Feb;23(2):166-7 - PubMed
  39. Neoplasia. 2013 Apr;15(4):348-58 - PubMed
  40. Blood. 2006 Feb 15;107(4):1397-404 - PubMed
  41. Bioconjug Chem. 2012 Aug 15;23(8):1704-11 - PubMed
  42. Hepatology. 1990 Apr;11(4):534-44 - PubMed
  43. Int J Dev Biol. 2001 Apr;45(2):445-7 - PubMed
  44. Cancer Sci. 2010 Nov;101(11):2325-32 - PubMed
  45. Eur Heart J. 2011 Nov;32(21):2713-22 - PubMed
  46. J Exp Med. 1997 Dec 1;186(11):1865-72 - PubMed
  47. World J Gastroenterol. 2012 Feb 28;18(8):840-6 - PubMed
  48. J Biol Chem. 1999 Dec 17;274(51):36505-12 - PubMed
  49. Mol Cell Biol. 2000 Feb;20(3):1089-94 - PubMed
  50. J Clin Invest. 2009 Dec;119(12):3613-25 - PubMed
  51. Blood. 2004 May 15;103(10):3783-8 - PubMed
  52. Diabet Med. 2000 May;17(5):346-50 - PubMed
  53. Mol Cancer Ther. 2009 May;8(5):1378-86 - PubMed
  54. FEBS Lett. 2012 Jul 30;586(16):2507-12 - PubMed
  55. Clin Cancer Res. 2010 Dec 15;16(24):6111-21 - PubMed
  56. Dis Markers. 2012;32(5):309-20 - PubMed
  57. Curr Mol Med. 2012 Dec;12(10):1220-43 - PubMed
  58. Clin Cancer Res. 2012 Nov 15;18(22):6208-18 - PubMed
  59. J Surg Oncol. 2013 Sep;108(3):157-62 - PubMed
  60. J Gastroenterol Hepatol. 2008 Feb;23(2):244-51 - PubMed
  61. Int J Dermatol. 2011 Aug;50(8):948-55 - PubMed
  62. EMBO J. 2012 Apr 4;31(7):1630-43 - PubMed
  63. Hepatology. 1999 Jun;29(6):1768-78 - PubMed
  64. Oncol Rep. 2011 Dec;26(6):1369-75 - PubMed
  65. Int J Cancer. 1997 May 2;71(3):383-9 - PubMed
  66. Blood. 2000 Sep 1;96(5):1674-80 - PubMed
  67. Eur J Endocrinol. 2006 Sep;155(3):485-93 - PubMed
  68. BMB Rep. 2013 May;46(5):252-7 - PubMed
  69. Diagn Microbiol Infect Dis. 2009 Dec;65(4):365-71 - PubMed

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