Top

Published in:

01-08-2009

Lysyl Oxidase (Lox) Gene Deficiency Affects Osteoblastic Phenotype

Authors: N. Pischon, J. M. Mäki, P. Weisshaupt, N. Heng, A. H. Palamakumbura, P. N’Guessan, A. Ding, R. Radlanski, H. Renz, T. A. L. J. J. Bronckers, J. Myllyharju, A. M. Kielbassa, B. M. Kleber, J.-P. Bernimoulin, P. C. Trackman

Published in: Calcified Tissue International | Issue 2/2009

Abstract

Lysyl oxidase (LOX) catalyzes cross-linking of elastin and collagen, which is essential for the structural integrity and function of bone tissue. The present study examined the role of Lox gene deficiency for the osteoblast phenotype in primary calvarial osteoblasts from E18.5 Lox knockout (Lox ^−/−) and wild type (wt) (C57BL/6) mice. Next to Lox gene depletion, mRNA expression of Lox isoforms, LOXL1–4, was significantly downregulated in Lox ^−/− bone tissue. A significant decrease of DNA synthesis of Lox ^−/− osteoblasts compared to wt was found. Early stages of osteoblastic apoptosis studied by annexin-V binding as well as later stages of DNA fragmentation were not affected. However, mineral nodule formation and osteoblastic differentiation were markedly decreased, as revealed by significant downregulation of osteoblastic markers, type I collagen, bone sialoprotein, and Runx2/Cbfa1.

Boskey AL (1996) Matrix protein and mineralization: an overview. Connect Tissue Res 35:357–363PubMedCrossRef

Fratzl P, Gupta HS, Paschalis EP, Roschger P (2004) Structure and mechanical quality of the collagen-mineral nano-composite in bone. J Mater Chem 14:2115–2123CrossRef

Kagan H, Trackman P (1991) Properties and function of lysyl oxidase. Am J Respir Cell Mol Biol 5:206–210PubMed

Prockop D, Kivirikko K (1995) Collagens: molecular biology, diseases, and potentials for therapy. Annu Rev Biochem 64:403–434PubMedCrossRef

Yamauchi M (1996) Collagen: the major matrix molecule in mineralized tissues. In: Anderson JJB, Garner S (eds) Calcium and phosphorus in health and disease. CRC Press, New York, pp 127–141

Knott L, Bailey A (1998) Collagen cross-links in mineralizing tissues: a review of their chemistry, function, and clinical relevance. Bone 22:181–187PubMedCrossRef

Kagan H (1986) Biology and regulation of extracellular matrix: a series. In: Mecham RP (ed) Regulation of matrix accumulation, vol 1. Academic Press, Orlando, pp 321–398

Mäki JM, Kivirikko KI (2001) Cloning and characterization of a fourth human lysyl oxidase isoenzyme. J Biochem 355:381–387CrossRef

Mäki JM, Tikkanen H, Kivirikko KI (2001) Cloning and characterization of a fifth human lysyl oxidase isoenzyme: the third member of the lysyl oxidase-related subfamily with four scavenger receptor cysteine-rich domains. Matrix Biol 20:493–496PubMedCrossRef

10.

Kenyon K, Modi WS, Contente S, Friedman RM (1993) A novel human cDNA with a predicted protein similar to lysyl oxidase maps to chromosome 15q24–q25. J Biol Chem 268:18435–18437PubMed

11.

Ito H, Akiyama H, Iguchi H, Iyama K, Miyamoto M, Ohsawa K et al (2001) Molecular cloning and biological activity of a novel lysyl oxidase-related gene expressed in cartilage. J Biol Chem 276:24023–24029PubMedCrossRef

12.

Jourdan-Le Saux C, Le Saux O, Donlon T, Boyd CD, Csiszar K (1998) The human lysyl oxidase-related gene (LOXL2) maps between markers D8S280 and D8S278 on chromosome 8p21.2–p21.3. Genomics 51:305–307PubMedCrossRef

13.

Huang Y, Dai J, Tang R, Zhao W, Zhou Z, Wang W et al (2001) Cloning and characterization of a human lysyl oxidase-like 3 gene (hLOXL3). Matrix Biol 20:153–157PubMedCrossRef

14.

Borel A, Eichenberger D, Farjanel J, Kessler E, Gleyzal C, Hulmes DJ et al (2001) Lysyl oxidase-like protein from bovine aorta. Isolation and maturation to an active form by bone morphogenetic protein-1. J Biol Chem 276:48944–48949PubMedCrossRef

15.

Kim MS, Kim SS, Jung ST, Park JY, Yoon HW, Ko J et al (2003) Expression and purification of enzymatically active forms of the human lysyl oxidase-like protein 4. J Biol Chem 278:52071–52074PubMedCrossRef

16.

Knott L, Whitehead C, Fleming R, Bailey A (1995) Biochemical changes in the collagenous matrix of osteoporotic avian bone. J Biochem 310:1045–1051

17.

Eyre DR, Dickson IR, Van Ness K (1988) Collagen crosslinking in human bone and articular cartilage: age-related changes in the content of mature hydroxypyridinium residues. J Biochem 252:495–500

18.

Geiger BJ, Steenbock H, Parsons HT (1933) Lathyrism in the rat. J Nutr 6:427–442

19.

Shim H, Harris ZL (2003) Genetic defects in copper metabolism. J Nutr 133:1527S–1531SPubMed

20.

Uzel M, Scott I, Babakhanlou-Chase H, Palamakumbura A, Pappano W, Hong H et al (2001) Multiple bone morphogenic protein-1 related mammal metalloproteinases process pro-lysyl oxidase at the correct physiological site and control lysyl oxidase activation in mouse embryo fibroblast cultures. J Biol Chem 276:22537–22543PubMedCrossRef

21.

Guo Y, Pischon N, Palamakumbura AH, Trackman P (2007) Intracellular distribution of the lysyl oxidase propeptide in osteoblastic cells. Am J Physiol Cell Physiol 292:2095–2102CrossRef

22.

Nellaiappan K, Risitano A, Liu G, Nicklas G, Kagan H (2000) Fully processed lysyl oxidase catalyst translocates from the extracellular space into nuclei of aortic smooth-muscle cells. J Biol Chem 79:576–582

23.

Li W, Nellaiappan K, Strassmaier T, Graham L, Thomas K, Kagan H (1997) Localization and activity of lysyl oxidase within nuclei of fibrogenic cells. Proc Natl Acad Sci USA 94:12817–12822PubMedCrossRef

24.

Palamakumbura A, Jeay S, Guo Y, Pischon N, Sommer P, Sonenshein G et al (2004) The propeptide domain of lysyl oxidase induces phenotypic reversion of ras-transformed cells. J Biol Chem 279:40593–40600PubMedCrossRef

25.

Jeay S, Pianetti S, Kagan H, Ge S (2003) Lysyl oxidase inhibits ras-mediated transformation by preventing activation of NF-kappa B. Mol Cell Biol 23:2251–2263PubMedCrossRef

26.

Giampuzzi M, Oleggini R, Di Donato A (2003) Demonstration of in vitro interaction between tumor suppressor lysyl oxidase and histones H1 and H2: definition of the regions involved. Biochim Biophys Acta 1647:245–251PubMed

27.

Min C, Kirsch KH, Zhao Y, Jeay S, Palamakumbura AH, Trackman PC et al (2007) The tumor suppressor activity of the lysyl oxidase propeptide reverses the invasive phenotype of Her-2/neu-driven breast cancer. Cancer Res 67:1105–1112PubMedCrossRef

28.

Wu M, Min C, Wang X, Yu Z, Kirsch K, Trackman PC et al (2007) Repression of BCL2 by the tumor suppressor activity of the lysyl oxidase propeptide inhibits transformed phenotype of lung and pancreatic cancer cells. Cancer Res 67:6278–6285PubMedCrossRef

29.

Hong HH, Pischon N, Santana RB, Palamakumbura AH, Chase HB, Gantz D et al (2004) A role for lysyl oxidase regulation in the control of normal collagen deposition in differentiating osteoblast cultures. J Cell Physiol 200:53–62PubMedCrossRef

30.

Mäki JM, Räsänen J, Tikkanen H, Sormunen R, Mäkikallio K, Kivirikko KI et al (2002) Inactivation of the lysyl oxidase gene Lox leads to aortic aneurysms, cardiovascular dysfunction, and perinatal death in mice. Circulation 106:2503–2509PubMedCrossRef

31.

Hornstra I, Birge S, Starcher B, Bailey A, Mecham R, Shapiro S (2003) Lysyl oxidase is required for vascular and diaphragmatic development in mice. J Biol Chem 278:14387–14393PubMedCrossRef

32.

Mäki J, Sormunen R, Lippo S, Kaarteenaho-Wiik R, Soininen R, Myllyharju J (2005) Lysyl oxidase is essential for normal development and function of the respiratory system and for the integrity of elastic and collagen fibers in various tissues. Am J Pathol 167:927–936PubMed

33.

Radlanski RJ, Renz H, Klarkowski MC (2003) Prenatal development of the human mandible. 3D reconstructions, morphometry and bone remodelling pattern, sizes 12–117 mm CRL. Anat Embryol 207:221–232PubMedCrossRef

34.

Bellows CG, Aubin JE, Heersche JNM, Antosz ME (1986) Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int 38:143–154PubMedCrossRef

35.

Ecarot-Charrier B, Glorieux FH, Van der Rest M, Pereira G (1983) Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture. J Cell Biol 96:639–643PubMedCrossRef

36.

Hippenstiel S, Schmeck B, N’Guessan PD, Seybold J, Krull M, Preissner K et al (2002) Rho protein inactivation induced apoptosis of cultured human endothelial cells. Am J Physiol Lung Cell Mol Physiol 283:L830–L838PubMed

37.

N’Guessan PD, Vigelahn M, Bachmann S, Zabel S, Opitz B, Schmeck B et al (2007) The UspA1 protein of Moraxella catarrhalis induces CEACAM-1-dependent apoptosis in alveolar epithelial cells. J Infect Dis 195:1651–1660PubMedCrossRef

38.

N’Guessan PD, Schmeck B, Ayim A, Hocke AC, Brell B, Hammerschmidt S et al (2005) Streptococcus pneumoniae R6x induced p38 MAPK and JNK-mediated caspase-dependent apoptosis in human endothelial cells. Thromb Haemost 94:295–303PubMed

39.

Livak KJ, Schnmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2^-∆∆C(T) method. Methods Enzymol 25:402–408CrossRef

40.

Stein G, Lian J, Stein J, Van Wijnen J, Montecino M (1996) Transcriptional control of osteoblast growth and differentiation. Physiol Rev 76:593–629PubMed

41.

Atsawasuwan P, Mochida Y, Parisuthiman D, Yamauchi M (2005) Expression of lysyl oxidase isoforms in MC3T3–E1 osteoblastic cells. Biochem Biophys Res Commun 327:1042–1046PubMedCrossRef

42.

Pischon N, Darbois LM, Palamakumbura AH, Kessler E, Trackman PC (2004) Regulation of collagen deposition and lysyl oxidase by tumor necrosis factor-alpha in osteoblasts. J Biol Chem 279:30060–30065PubMedCrossRef

43.

Turecek C, Fratzl-Zelman N, Rumpler M, Buchinger B, Spitzer S, Zoehrer R et al (2008) Collagen cross-linking influences osteoblastic differentiation. Calcif Tissue Int 82:392–400PubMedCrossRef

44.

Jordan MA (2002) Mechanism of action of antitumor drugs that interacts with microtubules and tubulin. Curr Med Chem Anti-Cancer Agents 2:1–17CrossRef

45.

Hock JM, Krishnan V, Onyia JE, Bidwell JP, Milas J, Stanislaus D (2001) Osteoblast apoptosis and bone turnover. J Bone Miner Res 16:975–984PubMedCrossRef

46.

Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89:747–754PubMedCrossRef

Title: Lysyl Oxidase (Lox) Gene Deficiency Affects Osteoblastic Phenotype
Authors: N. Pischon
J. M. Mäki
P. Weisshaupt
N. Heng
A. H. Palamakumbura
P. N’Guessan
A. Ding
R. Radlanski
H. Renz
T. A. L. J. J. Bronckers
J. Myllyharju
A. M. Kielbassa
B. M. Kleber
J.-P. Bernimoulin
P. C. Trackman
Publication date: 01-08-2009
Publisher: Springer-Verlag
Published in: Calcified Tissue International / Issue 2/2009
Print ISSN: 0171-967X
Electronic ISSN: 1432-0827
DOI: https://doi.org/10.1007/s00223-009-9252-8

Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin

Prof. Florian Limbourg

Prof. Anoop Chauhan

Developed by: Springer Medicine

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 2/2009

Abstracts Presented at the Bone Research Society Meeting, Jointly with the British Society for Matrix Biology

Formation of Tethers Linking the Epiphysis and Metaphysis Is Regulated by Vitamin D Receptor-Mediated Signaling

Influence of Drop-Landing Exercises on Bone Geometry and Biomechanical Properties in Prepubertal Girls: A Randomized Controlled Study

Association Analyses of RANKL/RANK/OPG Gene Polymorphisms with Femoral Neck Compression Strength Index Variation in Caucasians

Trabecular Microfracture Precedes Cortical Shell Failure in the Rat Caudal Vertebra Under Cyclic Overloading

Polymorphisms of the WNT10B Gene, Bone Mineral Density, and Fractures in Postmenopausal Women

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

At a glance: The STEP trials