The long-term objective of this research project is to develop a technique of somatic gene therapy in which endothelial cells that have undergone gene transfer ex vivo are transplanted into the capillary bed of skeletal muscle. This technique of somatic gene therapy could be used to achieve a regional or systemic effect by co-or repopulating a vascular bed with endothelial cells which constitutively or inducibly express a protective recombinant protein such as tissue plasminogen activator or Factor IX. Thus, the development of such a technique of somatic gene therapy could have a wide application to both surgical and genetic human deceases.
The Specific Aims of the project are:
Aim I : To identify and characterize the molecular mechanism mediating the adhesion of endothelial cells seeded onto quiescent, confluent endothelial cell monolayers.
Aim II; To optimize the adhesion and incorporation of genetically transduced endothelial cells transplanted into the capillary bed of skeletal muscle.
Aim III; To document long-term recombinant gene expression by endothelial cells transplanted into the capillary bed of skeletal muscle. The investigators have considerable experience with an in vitro model to study endothelial cell-endothelial cell adhesion as well as with an in vivo rt model to study transplantation of genetically modified endothelial cells into skeletal muscle capillary beds. The proposed experiments to achieve th above Aims include identification and characterization of specific adhesion molecules mediating endothelial cell-endothelial cell adhesion. These experiments would reveal fundamental new knowledge concerning endothelial cell-endothelial cell interactions. This knowledge could also be used to enhance adhesion and incorporation of transduced endothelium into the capillary wall. In a rat model the adhesion and incorporation of genetically transduced endothelium transplanted into the capillary bed will be optimized by angiogenesis induced with injection of basic fibroblast growth factor or stimulation f skeletal muscle regeneration. Finally, the optimal techniques of transplantation of transduced endothelium into host capillary beds, determined in the aforenoted studies, will be used to document the efficacy of this technique to deliver recombinant proteins into the circulation. At the completion of these studies, this technique of somatic gene therapy will be ready for evaluation in a primate model in preparations for its use in humans.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL051184-01
Application #
2227764
Study Section
Surgery and Bioengineering Study Section (SB)
Project Start
1994-04-09
Project End
1997-03-31
Budget Start
1994-04-09
Budget End
1995-03-31
Support Year
1
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Surgery
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Chang, David S; Su, Hua; Tang, Gale L et al. (2003) Adeno-associated viral vector-mediated gene transfer of VEGF normalizes skeletal muscle oxygen tension and induces arteriogenesis in ischemic rat hindlimb. Mol Ther 7:44-51
Brevetti, Lucy S; Chang, David S; Tang, Gale L et al. (2003) Overexpression of endothelial nitric oxide synthase increases skeletal muscle blood flow and oxygenation in severe rat hind limb ischemia. J Vasc Surg 38:820-6
Messina, Louis M; Brevetti, Lucy S; Chang, David S et al. (2002) Therapeutic angiogenesis for critical limb ischemia: invited commentary. J Control Release 78:285-94
Paek, Robert; Chang, David S; Brevetti, Lucy S et al. (2002) Correlation of a simple direct measurement of muscle pO(2) to a clinical ischemia index and histology in a rat model of chronic severe hindlimb ischemia. J Vasc Surg 36:172-9
Brevetti, Lucy S; Chang, David S; Sarkar, Rajabrata et al. (2002) Effect of adenoviral titer and instillation pressure on gene transfer efficiency to arterial and venous grafts ex-vivo. J Vasc Surg 36:263-70
Brevetti, L S; Paek, R; Brady, S E et al. (2001) Exercise-induced hyperemia unmasks regional blood flow deficit in experimental hindlimb ischemia. J Surg Res 98:21-6
Brevetti, L S; Sarkar, R; Chang, D S et al. (2001) Administration of adenoviral vectors induces gangrene in acutely ischemic rat hindlimbs: role of capsid protein-induced inflammation. J Vasc Surg 34:489-96
Furukawa, L; Brevetti, L S; Brady, S E et al. (2000) Adenoviral-mediated gene transfer of ICP47 inhibits major histocompatibility complex class I expression on vascular cells in vitro. J Vasc Surg 31:558-66
Zelenock, J A; Welling, T H; Sarkar, R et al. (1997) Improved retroviral transduction efficiency of vascular cells in vitro and in vivo during clinically relevant incubation periods using centrifugation to increase viral titers. J Vasc Surg 26:119-27
Ford, J W; Welling 3rd, T H; Stanley, J C et al. (1996) PKH26 and 125I-PKH95: characterization and efficacy as labels for in vitro and in vivo endothelial cell localization and tracking. J Surg Res 62:23-8

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