The normal balance between coagulation and fibrinolysis, both intravascularly and extravascularly, is significantly altered during pulmonary injuries. The coagulation pathway is activated that is accompanied by an inhibition of fibrinolytic activity. This imbalance leads to fibrin deposition in the alveolar, interstitial and vascular spaces of the lung, thereby adversely affecting cardiovascular and respiratory functions. In addition to impaired blood perfusion and gas exchange due to clot formation, fibrin also promotes an acute cellular inflammatory response, contributes to surfactant dysfunction and provides a provisional matrix for collagen deposition in the lung. Thus, decreasing fibrin deposition within the lung is not only important for the improvement of hemodynamics and prevention of pulmonary damage at the early stages of pulmonary injury but also beneficial for the prevention of late-stage fibrosis. We propose in this application to develop a novel and safe nonviral vector for pulmonary uPA gene transfer for the treatment of pulmonary injury. Our basic hypothesis is that systemic delivery of uPA gene to the lung would upregulate the fibrinolytic activity within the lung and reduce collagen accumulation that follows pulmonary inflammation. Accordingly, specific aims of this comprehensive proposal are:
Aim 1 : To develop a novel polymer-based delivery system that is highly efficient in selective gene delivery to pulmonary circulation with minimal toxicity.
Aim 2 : To develop a mini/mutated uPA that fully retains its proteolytic activity but lacks pro-inflammatory activity.
Aim 3 : To study the biological consequences of expressed uPA following pulmonary uPA gene transfer.
Aim 4 : To examine the therapeutic efficacy of uPA gene transfer in a mouse model of bleomycin-induced pulmonary injury. Completion of the above studies will lead to development of a novel therapy that will be beneficial not only for the treatment of acute lung injury but also for the prevention and treatment of late-stage complications such as pulmonary fibrosis. The novel nonviral vectors developed in this application can also be readily employed for targeted delivery of different therapeutic genes for the treatment of other pulmonary diseases such as pulmonary hypertension.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL091828-04
Application #
8197275
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Eu, Jerry Pc
Project Start
2008-12-01
Project End
2013-11-30
Budget Start
2011-12-01
Budget End
2013-11-30
Support Year
4
Fiscal Year
2012
Total Cost
$374,963
Indirect Cost
$127,463
Name
University of Pittsburgh
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Zhang, Yifei; Ghazwani, Mohammed; Li, Jiang et al. (2014) MiR-29b inhibits collagen maturation in hepatic stellate cells through down-regulating the expression of HSP47 and lysyl oxidase. Biochem Biophys Res Commun 446:940-4
Zhang, Peng; Huang, Yixian; Makhov, Alexander M et al. (2013) Characterization of spherulites as a lipidic carrier for low and high molecular weight agents. Pharm Res 30:1525-35
Li, Jiang; Ghazwani, Mohammed; Zhang, Yifei et al. (2013) miR-122 regulates collagen production via targeting hepatic stellate cells and suppressing P4HA1 expression. J Hepatol 58:522-8
Li, Jiang; Zhang, Yifei; Kuruba, Ramalinga et al. (2011) Roles of microRNA-29a in the antifibrotic effect of farnesoid X receptor in hepatic stellate cells. Mol Pharmacol 80:191-200
Li, Jiang; Kuruba, Ramalinga; Wilson, Annette et al. (2010) Inhibition of endothelin-1-mediated contraction of hepatic stellate cells by FXR ligand. PLoS One 5:e13955
Gao, Xiang; Kuruba, Ramalinga; Damodaran, Krishnan et al. (2009) Polyhydroxylalkyleneamines: a class of hydrophilic cationic polymer-based gene transfer agents. J Control Release 137:38-45