Cargando…

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

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 specifi...

Descripción completa

Detalles Bibliográficos
Autores principales: Keane, Fiona M., Yao, Tsun-Wen, Seelk, Stefanie, Gall, Margaret G., Chowdhury, Sumaiya, Poplawski, Sarah E., Lai, Jack H., Li, Youhua, Wu, Wengen, Farrell, Penny, Vieira de Ribeiro, Ana Julia, Osborne, Brenna, Yu, Denise M.T., Seth, Devanshi, Rahman, Khairunnessa, Haber, Paul, Topaloglu, A. Kemal, Wang, Chuanmin, Thomson, Sally, Hennessy, Annemarie, Prins, John, Twigg, Stephen M., McLennan, Susan V., McCaughan, Geoffrey W., Bachovchin, William W., Gorrell, Mark D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3871272/
https://www.ncbi.nlm.nih.gov/pubmed/24371721
http://dx.doi.org/10.1016/j.fob.2013.12.001
_version_ 1782296792766873600
author Keane, Fiona M.
Yao, Tsun-Wen
Seelk, Stefanie
Gall, Margaret G.
Chowdhury, Sumaiya
Poplawski, Sarah E.
Lai, Jack H.
Li, Youhua
Wu, Wengen
Farrell, Penny
Vieira de Ribeiro, Ana Julia
Osborne, Brenna
Yu, Denise M.T.
Seth, Devanshi
Rahman, Khairunnessa
Haber, Paul
Topaloglu, A. Kemal
Wang, Chuanmin
Thomson, Sally
Hennessy, Annemarie
Prins, John
Twigg, Stephen M.
McLennan, Susan V.
McCaughan, Geoffrey W.
Bachovchin, William W.
Gorrell, Mark D.
author_facet Keane, Fiona M.
Yao, Tsun-Wen
Seelk, Stefanie
Gall, Margaret G.
Chowdhury, Sumaiya
Poplawski, Sarah E.
Lai, Jack H.
Li, Youhua
Wu, Wengen
Farrell, Penny
Vieira de Ribeiro, Ana Julia
Osborne, Brenna
Yu, Denise M.T.
Seth, Devanshi
Rahman, Khairunnessa
Haber, Paul
Topaloglu, A. Kemal
Wang, Chuanmin
Thomson, Sally
Hennessy, Annemarie
Prins, John
Twigg, Stephen M.
McLennan, Susan V.
McCaughan, Geoffrey W.
Bachovchin, William W.
Gorrell, Mark D.
author_sort Keane, Fiona M.
collection PubMed
description 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.
format Online
Article
Text
id pubmed-3871272
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-38712722013-12-26 Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs() Keane, Fiona M. Yao, Tsun-Wen Seelk, Stefanie Gall, Margaret G. Chowdhury, Sumaiya Poplawski, Sarah E. Lai, Jack H. Li, Youhua Wu, Wengen Farrell, Penny Vieira de Ribeiro, Ana Julia Osborne, Brenna Yu, Denise M.T. Seth, Devanshi Rahman, Khairunnessa Haber, Paul Topaloglu, A. Kemal Wang, Chuanmin Thomson, Sally Hennessy, Annemarie Prins, John Twigg, Stephen M. McLennan, Susan V. McCaughan, Geoffrey W. Bachovchin, William W. Gorrell, Mark D. FEBS Open Bio Article 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. Elsevier 2013-12-08 /pmc/articles/PMC3871272/ /pubmed/24371721 http://dx.doi.org/10.1016/j.fob.2013.12.001 Text en © 2014 The Authors http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Article
Keane, Fiona M.
Yao, Tsun-Wen
Seelk, Stefanie
Gall, Margaret G.
Chowdhury, Sumaiya
Poplawski, Sarah E.
Lai, Jack H.
Li, Youhua
Wu, Wengen
Farrell, Penny
Vieira de Ribeiro, Ana Julia
Osborne, Brenna
Yu, Denise M.T.
Seth, Devanshi
Rahman, Khairunnessa
Haber, Paul
Topaloglu, A. Kemal
Wang, Chuanmin
Thomson, Sally
Hennessy, Annemarie
Prins, John
Twigg, Stephen M.
McLennan, Susan V.
McCaughan, Geoffrey W.
Bachovchin, William W.
Gorrell, Mark D.
Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs()
title Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs()
title_full Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs()
title_fullStr Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs()
title_full_unstemmed Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs()
title_short Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs()
title_sort quantitation of fibroblast activation protein (fap)-specific protease activity in mouse, baboon and human fluids and organs()
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3871272/
https://www.ncbi.nlm.nih.gov/pubmed/24371721
http://dx.doi.org/10.1016/j.fob.2013.12.001
work_keys_str_mv AT keanefionam quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT yaotsunwen quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT seelkstefanie quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT gallmargaretg quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT chowdhurysumaiya quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT poplawskisarahe quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT laijackh quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT liyouhua quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT wuwengen quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT farrellpenny quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT vieiraderibeiroanajulia quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT osbornebrenna quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT yudenisemt quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT sethdevanshi quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT rahmankhairunnessa quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT haberpaul quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT topalogluakemal quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT wangchuanmin quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT thomsonsally quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT hennessyannemarie quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT prinsjohn quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT twiggstephenm quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT mclennansusanv quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT mccaughangeoffreyw quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT bachovchinwilliamw quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans
AT gorrellmarkd quantitationoffibroblastactivationproteinfapspecificproteaseactivityinmousebaboonandhumanfluidsandorgans