Intratracheal gene transfer of tissue factor pathway inhibitor attenuates pulmonary fibrosis

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Abstract

Activation of the coagulation system and increased expression of tissue factor (TF) in pulmonary fibrosis associated with acute and chronic lung injury have been previously documented. In the present study, we evaluated the effect of TF inhibition with intratracheal gene transfer of tissue factor pathway inhibitor (TFPI), a potent and highly specific endogenous inhibitor of TF-dependent coagulation activation, in a rat model of bleomycin-induced lung fibrosis. Significant lung fibrotic changes as assessed by histologic findings and hydroxyproline content, and increased procoagulant activity and thrombin generation in bronchoalveolar lavage fluid were detected in rats after intratracheal injection of bleomycin. Intratracheal administration of an adenovirus vector expressing TFPI significantly decreased bleomycin-induced procoagulant and thrombin generation resulting in a strong inhibition of pulmonary fibrosis. TFPI-overexpression in the lung was associated with a significant reduction in gene expression of the connective tissue growth factor, a potent profibrotic growth factor. This is the first report showing that direct inhibition of TF-mediated coagulation activation abrogates bleomycin-induced pulmonary fibrosis.

Section snippets

Materials and methods

Animal treatment. Pulmonary fibrosis was induced in male Wistar rats weighing 280–320 g by a single intratracheal instillation of bleomycin (2 U/kg, Nippon Kayaku, Tokyo, Japan) diluted in sterile endotoxin-free phosphate-buffered saline (PBS), pH 7.4, in a final volume of 300 μl. After anesthesia with pentobarbital sodium (60 mg/kg, intraperitoneal injection), rats received intratracheal instillation of PBS (control), bleomycin or bleomycin + AdTFPI (the recombinant replication-defective adenovirus

Suppression of procoagulant activity and thrombin generation by TFPI overexpression in bleomycin-treated lungs

As previously reported [10], [11], [33], a single intratracheal injection of bleomycin caused remarkable increases in procoagulant activity and thrombin generation in BALF, which began to increase on day 3 and peaked on day 7 (Fig. 1). It is worth noting that while thrombin activity sharply returned to baseline values (Fig. 1C) procoagulant activity remained high on day 14, returning to baseline values on day 21 (Fig. 1B). The explanation for this discrepancy is not clear but it may depend on

Discussion

In the present study, we assessed the pathophysiological implication of TF and the therapeutic value of adenovirus-mediated intratracheal gene transfer of TFPI in bleomycin-associated lung fibrosis [28]. This study showed that TFPI-overexpression almost completely inhibited procoagulant activity and thrombin generation in bleomycin-induced lung injury, thus providing evidence that coagulation activation in this model occurs due to excessive TF expression. The main finding of this study is that

References (39)

  • H. Takeya et al.

    Synergistic effect of sphingosine 1-phosphate on thrombin-induced tissue factor expression in endothelial cells

    Blood

    (2003)
  • U.R. Pendurthi et al.

    Factor VIIa and thrombin induce the expression of Cyr61 and connective tissue growth factor, extracellular matrix signaling proteins that could act as possible downstream mediators in factor VIIa × tissue factor-induced signal transduction

    J. Biol. Chem.

    (2000)
  • M. Gharaee-Kermani et al.

    CC-chemokine receptor 2 required for bleomycin-induced pulmonary fibrosis

    Cytokine

    (2003)
  • H.A. Chapman

    Disorders of lung matrix remodeling

    J. Clin. Invest.

    (2004)
  • R.C. Chambers et al.

    Coagulation cascade proteases and tissue fibrosis

    Biochem. Soc. Trans.

    (2002)
  • D.C. Howell et al.

    Role of thrombin and its major cellular receptor, protease-activated receptor-1, in pulmonary fibrosis

    Biochem. Soc. Trans.

    (2002)
  • S. Idell

    Coagulation, fibrinolysis, and fibrin deposition in acute lung injury

    Crit. Care Med.

    (2003)
  • C. Kuhn et al.

    An immunohistochemical study of architectural remodeling and connective tissue synthesis in pulmonary fibrosis

    Am. Rev. Respir. Dis.

    (1989)
  • A. Gunther et al.

    Prevention of bleomycin-induced lung fibrosis by aerosolization of heparin or urokinase in rabbits

    Am. J. Respir. Crit. Care Med.

    (2003)
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