Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation: A feasibility study

Johan Malmström, Kristoffer Larsen, Lennart Hansson, Claes-Göran Löfdahl, Ole Nørregaard Jensen, György Marko-Varga, Gunilla Westergren-Thorsson

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Udgivelsesdato: 2002-Apr
OriginalsprogEngelsk
TidsskriftProteomics
Vol/bind2
Udgave nummer4
Sider (fra-til)394-404
ISSN1615-9853
DOI
StatusUdgivet - 2002

Fingeraftryk

Biopsy
Feasibility Studies
Proteoglycans
Proteome
Tissue
Lung
Fibroblasts
Proteins
Versicans
Biglycan
Decorin
Biomarkers
Electrophoresis
Cell culture
Ionization
Mass spectrometry
Muscle
Actins
Desorption
Adhesion

Citer dette

Malmström, Johan ; Larsen, Kristoffer ; Hansson, Lennart ; Löfdahl, Claes-Göran ; Jensen, Ole Nørregaard ; Marko-Varga, György ; Westergren-Thorsson, Gunilla. / Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation : A feasibility study. I: Proteomics. 2002 ; Bind 2, Nr. 4. s. 394-404.
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title = "Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation: A feasibility study",
abstract = "The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients with asthmatic-like disorders. We isolated viable cells from 10 out of the 12 biopsies. The fibroblast-like cells were positive for the biomarker a-smooth muscle actin, indicating that the cells were in an activated state. Two different types of fibroblast-like cells were observed from human pulmonary connective tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most likely is of a synthetic phenotype. This is the first evidence of alterations in the proteoglycan expression pattern of versican, perlecan, biglycan and decorin which can be linked to the pathophysiological state of asthmatics proven by a combination of solid-phase extraction by reversed phase and by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Protein expression analysis using two-dimensional electrophoresis was interfaced to miniaturized sample preparation techniques using microcapillary extraction. Four protein groups were identified; cytoskeletal, adhesion, scavenger and metabolic proteins. These patient's proteomes showed a high degree of heterogeneity between patients but larger homogeneity within biopsies derived from different locations of the same patient.",
author = "Johan Malmstr{\"o}m and Kristoffer Larsen and Lennart Hansson and Claes-G{\"o}ran L{\"o}fdahl and Jensen, {Ole N{\o}rregaard} and Gy{\"o}rgy Marko-Varga and Gunilla Westergren-Thorsson",
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doi = "10.1002/1615-9861(200204)2:4<394::AID-PROT394>3.0.CO;2-6",
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Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation : A feasibility study. / Malmström, Johan; Larsen, Kristoffer; Hansson, Lennart; Löfdahl, Claes-Göran; Jensen, Ole Nørregaard; Marko-Varga, György; Westergren-Thorsson, Gunilla.

I: Proteomics, Bind 2, Nr. 4, 2002, s. 394-404.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation

T2 - A feasibility study

AU - Malmström, Johan

AU - Larsen, Kristoffer

AU - Hansson, Lennart

AU - Löfdahl, Claes-Göran

AU - Jensen, Ole Nørregaard

AU - Marko-Varga, György

AU - Westergren-Thorsson, Gunilla

PY - 2002

Y1 - 2002

N2 - The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients with asthmatic-like disorders. We isolated viable cells from 10 out of the 12 biopsies. The fibroblast-like cells were positive for the biomarker a-smooth muscle actin, indicating that the cells were in an activated state. Two different types of fibroblast-like cells were observed from human pulmonary connective tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most likely is of a synthetic phenotype. This is the first evidence of alterations in the proteoglycan expression pattern of versican, perlecan, biglycan and decorin which can be linked to the pathophysiological state of asthmatics proven by a combination of solid-phase extraction by reversed phase and by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Protein expression analysis using two-dimensional electrophoresis was interfaced to miniaturized sample preparation techniques using microcapillary extraction. Four protein groups were identified; cytoskeletal, adhesion, scavenger and metabolic proteins. These patient's proteomes showed a high degree of heterogeneity between patients but larger homogeneity within biopsies derived from different locations of the same patient.

AB - The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients with asthmatic-like disorders. We isolated viable cells from 10 out of the 12 biopsies. The fibroblast-like cells were positive for the biomarker a-smooth muscle actin, indicating that the cells were in an activated state. Two different types of fibroblast-like cells were observed from human pulmonary connective tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most likely is of a synthetic phenotype. This is the first evidence of alterations in the proteoglycan expression pattern of versican, perlecan, biglycan and decorin which can be linked to the pathophysiological state of asthmatics proven by a combination of solid-phase extraction by reversed phase and by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Protein expression analysis using two-dimensional electrophoresis was interfaced to miniaturized sample preparation techniques using microcapillary extraction. Four protein groups were identified; cytoskeletal, adhesion, scavenger and metabolic proteins. These patient's proteomes showed a high degree of heterogeneity between patients but larger homogeneity within biopsies derived from different locations of the same patient.

U2 - 10.1002/1615-9861(200204)2:4<394::AID-PROT394>3.0.CO;2-6

DO - 10.1002/1615-9861(200204)2:4<394::AID-PROT394>3.0.CO;2-6

M3 - Journal article

C2 - 12164698

VL - 2

SP - 394

EP - 404

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 4

ER -