title: Secretion of a TNFR:Fc fusion protein following pulmonary administration of pseudotyped adeno-associated virus vectors.
abstract: This study evaluated and compared delivery of the tumor necrosis factor alpha receptor (TNFR)-immunoglobulin G1 (IgG1) Fc fusion (TNFR:Fc) gene to the lung by single and repeat administrations of multiple pseudotyped adeno-associated virus (AAV) vectors as a means for achieving systemic distribution of the soluble TNFR:Fc protein. A single endotracheal administration of AAV[2/5]cytomegalovirus (CMV)-TNFR:Fc vector (containing the AAV2 inverted terminal repeats and AAV5 capsid) to the rat lung resulted in long-term, high levels of serum TNFR:Fc protein that gradually declined over a period of 8 months. Endotracheal delivery of AAV[2/1]CMV-TNFR:Fc resulted in serum TNFR:Fc protein levels that were detectable for at least 4 months but were 10-fold lower than that of the AAV[2/5] vector. In contrast, secretion of the TNFR:Fc protein following pulmonary delivery of AAV[2/2]CMV-TNFR:Fc vector was very inefficient, and the protein was detected in the blood only when an airway epithelial cell-specific promoter, CC10, was substituted for the CMV enhancer/promoter to control transgene expression. In the context of AAV[2/5], the CC10 promoter was as efficient as CMV enhancer/promoter in generating similar levels of systemic TNFR:Fc protein, suggesting that this protein is secreted primarily from the airway epithelium. In mice, comparable long-term secretion of TNFR:Fc protein was demonstrated after AAV[2/2] and AAV[2/5] delivery, although the kinetics of transduction appeared to be different. All pseudotyped AAV vectors elicited serum anti-AAV capsid-neutralizing antibody responses, but these did not prevent lung transduction and efficient secretion of TNFR:Fc protein to the circulation following readministration with AAV[2/5]. These results highlight the potential utility of AAV vectors containing serotype 5 capsid to deliver and redeliver genes of secreted proteins to the lung to achieve long-term systemic protein expression.
authors.affiliation: Targeted Genetics Corporation, 1100 Olive Way, Suite 100, Seattle, WA 98101-1844, USA.
authors.forename: Ziv
authors.initials: Z
authors.lastname: Sandalon
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Bruckheimer
authors.forename: Kurt H
authors.initials: KH
authors.lastname: Lustig
authors.forename: Linda C
authors.initials: LC
authors.lastname: Rogers
authors.forename: Richard W
authors.initials: RW
authors.lastname: Peluso
authors.forename: Haim
authors.initials: H
authors.lastname: Burstein
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics* / Q000276:immunology; D005260:Female; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000096:biosynthesis* / Q000097:blood; D008168:Lung / Q000378:metabolism*; D051381:Rats; D011917:Rats, Inbred Lew; D018124:Receptors, Tumor Necrosis Factor / Q000096:biosynthesis* / Q000097:blood; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000097:blood; D014161:Transduction, Genetic
chemical_list: D000914:Antibodies, Viral; D007141:Immunoglobulin Fc Fragments; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins