Lung Fibroblasts from Patients with Emphysema Show Markers of Senescence in Vitro

Overview

Journal Respir Res

Specialty Pulmonary Medicine

Date 2006 Mar 1

PMID 16504044

Citations 74

Authors

K-C Muller

L Welker

K Paasch

B Feindt

V J Erpenbeck

J M Hohlfeld

N Krug

M Nakashima

D Branscheid

H Magnussen

R A Jorres

O Holz

Affiliations

Soon will be listed here.

Abstract

Background: The loss of alveolar walls is a hallmark of emphysema. As fibroblasts play an important role in the maintenance of alveolar structure, a change in fibroblast phenotype could be involved in the pathogenesis of this disease. In a previous study we found a reduced in vitro proliferation rate and number of population doublings of parenchymal lung fibroblasts from patients with emphysema and we hypothesized that these findings could be related to a premature cellular aging of these cells. In this study, we therefore compared cellular senescence markers and expression of respective genes between lung fibroblasts from patients with emphysema and control patients without COPD.

Methods: Primary lung fibroblasts were obtained from 13 patients with moderate to severe lung emphysema (E) and 15 controls (C) undergoing surgery for lung tumor resection or volume reduction (n = 2). Fibroblasts (8E/9C) were stained for senescence-associated beta-galactosidase (SA-beta-Gal). In independent cultures, DNA from lung fibroblasts (7E/8C) was assessed for mean telomere length. Two exploratory 12 k cDNA microarrays were used to assess gene expression in pooled fibroblasts (3E/3C). Subsequently, expression of selected genes was evaluated by quantitative PCR (qPCR) in fibroblasts of individual patients (10E/9C) and protein concentration was analyzed in the cell culture supernatant.

Results: The median (quartiles) percentage of fibroblasts positive for SA-beta-Gal was 4.4 (3.2;4.7) % in controls and 16.0 (10.0;24.8) % in emphysema (p = 0.001), while telomere length was not different. Among the candidates for differentially expressed genes in the array (factor > or = 3), 15 were upregulated and 121 downregulated in emphysema. qPCR confirmed the upregulation of insulin-like growth factor-binding protein (IGFBP)-3 and IGFBP-rP1 (p = 0.029, p = 0.0002), while expression of IGFBP-5, -rP2 (CTGF), -rP4 (Cyr61), FOSL1, LOXL2, OAZ1 and CDK4 was not different between groups. In line with the gene expression we found increased cell culture supernatant concentrations of IGFBP-3 (p = 0.006) in emphysema.

Conclusion: These data support the hypothesis that premature aging of lung fibroblasts occurs in emphysema, via a telomere-independent mechanism. The upregulation of the senescence-associated IGFBP-3 and -rP1 in emphysema suggests that inhibition of the action of insulin and insulin-like growth factors could be involved in the reduced in vitro-proliferation rate.

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References

Hwa V, Oh Y, Rosenfeld R . The insulin-like growth factor-binding protein (IGFBP) superfamily. Endocr Rev. 1999; 20(6):761-87. DOI: 10.1210/edrv.20.6.0382. View

Valdes A, Andrew T, Gardner J, Kimura M, Oelsner E, Cherkas L . Obesity, cigarette smoking, and telomere length in women. Lancet. 2005; 366(9486):662-4. DOI: 10.1016/S0140-6736(05)66630-5. View

Kurz D, Decary S, Hong Y, Erusalimsky J . Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells. J Cell Sci. 2000; 113 ( Pt 20):3613-22. DOI: 10.1242/jcs.113.20.3613. View

Csiszar K . Lysyl oxidases: a novel multifunctional amine oxidase family. Prog Nucleic Acid Res Mol Biol. 2001; 70:1-32. DOI: 10.1016/s0079-6603(01)70012-8. View

Balin A, Fisher A, Anzelone M, Leong I, Allen R . Effects of establishing cell cultures and cell culture conditions on the proliferative life span of human fibroblasts isolated from different tissues and donors of different ages. Exp Cell Res. 2002; 274(2):275-87. DOI: 10.1006/excr.2002.5485. View

Wilson H, Birnbaum R, Poot M, Quinn L, Swisshelm K . Insulin-like growth factor binding protein-related protein 1 inhibits proliferation of MCF-7 breast cancer cells via a senescence-like mechanism. Cell Growth Differ. 2002; 13(5):205-13. View

Nobukuni S, Watanabe K, Inoue J, Wen F, Tamaru N, Yoshida M . Cigarette smoke inhibits the growth of lung fibroblasts from patients with pulmonary emphysema. Respirology. 2002; 7(3):217-23. DOI: 10.1046/j.1440-1843.2002.00400.x. View

Noordhoek J, Postma D, Chong L, Vos J, Kauffman H, Timens W . Different proliferative capacity of lung fibroblasts obtained from control subjects and patients with emphysema. Exp Lung Res. 2003; 29(5):291-302. DOI: 10.1080/01902140303789. View

Erpenbeck V, Hohlfeld J, Volkmann B, Hagenberg A, Geldmacher H, Braun A . Segmental allergen challenge in patients with atopic asthma leads to increased IL-9 expression in bronchoalveolar lavage fluid lymphocytes. J Allergy Clin Immunol. 2003; 111(6):1319-27. DOI: 10.1067/mai.2003.1485. View

10.

Bird J, Ostler E, Faragher R . Can we say that senescent cells cause ageing?. Exp Gerontol. 2003; 38(11-12):1319-26. DOI: 10.1016/j.exger.2003.09.011. View

11.

Newman R, Mobascher A, Mangold U, Koike C, Diah S, Schmidt M . Antizyme targets cyclin D1 for degradation. A novel mechanism for cell growth repression. J Biol Chem. 2004; 279(40):41504-11. DOI: 10.1074/jbc.M407349200. View

12.

Holz O, Zuhlke I, Jaksztat E, Muller K, Welker L, Nakashima M . Lung fibroblasts from patients with emphysema show a reduced proliferation rate in culture. Eur Respir J. 2004; 24(4):575-9. DOI: 10.1183/09031936.04.00143703. View

13.

Ning W, Li C, Kaminski N, Feghali-Bostwick C, Alber S, Di Y . Comprehensive gene expression profiles reveal pathways related to the pathogenesis of chronic obstructive pulmonary disease. Proc Natl Acad Sci U S A. 2004; 101(41):14895-900. PMC: 522001. DOI: 10.1073/pnas.0401168101. View

14.

King G, Kahn C, Rechler M, NISSLEY S . Direct demonstration of separate receptors for growth and metabolic activities of insulin and multiplication-stimulating activity (an insulinlike growth factor) using antibodies to the insulin receptor. J Clin Invest. 1980; 66(1):130-40. PMC: 371514. DOI: 10.1172/JCI109826. View

15.

Smith P, Krohn R, Hermanson G, Mallia A, Gartner F, Provenzano M . Measurement of protein using bicinchoninic acid. Anal Biochem. 1985; 150(1):76-85. DOI: 10.1016/0003-2697(85)90442-7. View

16.

Goldstein R, Poliks C, Pilch P, Smith B, Fine A . Stimulation of collagen formation by insulin and insulin-like growth factor I in cultures of human lung fibroblasts. Endocrinology. 1989; 124(2):964-70. DOI: 10.1210/endo-124-2-964. View

17.

Harley C, Futcher A, Greider C . Telomeres shorten during ageing of human fibroblasts. Nature. 1990; 345(6274):458-60. DOI: 10.1038/345458a0. View

18.

Goldstein S, Moerman E, Jones R, BAXTER R . Insulin-like growth factor binding protein 3 accumulates to high levels in culture medium of senescent and quiescent human fibroblasts. Proc Natl Acad Sci U S A. 1991; 88(21):9680-4. PMC: 52782. DOI: 10.1073/pnas.88.21.9680. View

19.

Harley C . Telomere loss: mitotic clock or genetic time bomb?. Mutat Res. 1991; 256(2-6):271-82. DOI: 10.1016/0921-8734(91)90018-7. View

20.

Allsopp R, Vaziri H, Patterson C, Goldstein S, Younglai E, Futcher A . Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci U S A. 1992; 89(21):10114-8. PMC: 50288. DOI: 10.1073/pnas.89.21.10114. View