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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2004;24:1646.)
© 2004 American Heart Association, Inc.

Vascular Biology

Functional Analysis of Genomic DNA, cDNA, and Nucleotide Sequence of the Mature C-Type Natriuretic Peptide Gene in Vascular Cells

Jaroslav Pelisek; Andreas Kuehnl; Pierre H. Rolland; Choukri Mekkaoui; Alexandra Fuchs; Greg F. Walker; Manfred Ogris; Ernst Wagner; Sigrid Nikol

From the Department of Cardiology and Angiology (J.P., A.K., S.N.), University Hospital, Westfaelische Wilhelm University, Muenster, Germany; the Department of Pharmacy (J.P., G.F.W., M.O., E.W.), Center of Drug Research, Pharmaceutical Biology-Biotechnology; Paediatric Cardiology (A.F.), Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany; and Laboratoire Cardiovasculaire, Faculté de Médécine, Université de Marseille, France.

Correspondence to Sigrid Nikol, MD, Associate Professor for Molecular Cardiology, Medical Clinic C, Cardiology and Angiology, University of Münster, Albert-Schweitzer-Str. 33, 48129 Münster, Germany. E-mail nikol{at}uni-muenster.de

Objective— The aim of this study was to investigate the effect of various C-type natriuretic peptide (CNP) sequences (genomic DNA [CNPDNA], cDNA derived from mRNA [CNPcDNA], and sequence coding for 22 amino acids of the mature CNP [CNP22aa]) on the growth of primary porcine vascular cells.

Methods and Results— Gene transfer was performed with cationic lipid DOCSPER or linear polyethylenimine. All 3 CNP sequences led to significant inhibition of smooth muscle cell (SMC) proliferation. In contrast, significant stimulation of cell growth was observed in endothelial cells (ECs) using CNPDNA or CNPcDNA but not CNP22aa. In a porcine restenosis model, a significant reduction in neointima hyperplasia was found 3 months after application of the CNPcDNA vector compared with the control transfection.

Conclusions— The results demonstrate that the first intron in the CNP sequence does not contain any additional enhancer-binding sites. However, the signal sequence is indispensable for secretion of CNP and its appropriate physiological function. Furthermore, the results show for the first time the therapeutic effect of CNP using liposome-mediated gene transfer and local adventitial delivery. Advantages of the CNP gene are its dual effects with inhibition of SMC proliferation and simultaneous promotion of EC growth.

Functional analysis of various C-type natriuretic peptide (CNP) sequences on growth of vascular cells. For the first time, dual therapeutic effects of CNP with inhibition of smooth muscle cell proliferation and stimulation of re-endothelialization were demonstrated in a pig restenosis model using liposome-mediated gene transfer and local adventitial delivery.

Key Words: C-type natriuretic peptide • vascular cells • gene transfer • proliferation • restenosis