Achieving the full potential of intraarterial, intravenous, or pulmonary gene therapy requires quantitative bioengineering analysis of extracellular and intracellular barriers. With respect to the many viral and non-viral vectors tested, the field of gene transfer has been hampered by strong inflammatory and immunological responses during gene transfer as well as poor efficiency of gene transfer. Often, the readministration of the same virus serotype has proven extremely difficult due to neutralizing antibodies. We will investigate novel cationic glucocorticoids that deliver viral vectors and develop these formulations for anti-bacterial and anti-inflammatory activity. The following specific aims are proposed:
Aim 1 : Optimization of cationic steroids for improved delivery of Adeno- associated virus (AAV) serotypes to mouse lung. Delivery to normal or Pseudomonas aeurginosa infected mouse lung will be tested. We will measure the anti-inflammatory and bactericidal activity of cationic steroids in conjunction with gene expression and improved targeting to airway epithelium. We will test the ability of cationic steroids to facilitate vector readministration of the same AAV serotype.
Aim 2 : Enhancing gene transfer through chemical and siRNA screening to elucidate mechanisms and barriers of AAV gene transfer. We have conducted siRNA screening of over 5000 genes to identify a number of genes whose knockdown leads to either enhanced or reduced transgene expression following viral transduction. Both chemical and siRNA screening will be directed at understanding intracellular limits to viral transduction as a means for enhancing transgene expression and reducing viral doses during therapy. The overall project is focused on reducing side effects related to gene delivery, improving efficacy during gene delivery (even in the presence of local infection), and facilitating readministration of AAV vector to eliminate the need for biomanufacturing of multiple serotypes.

Public Health Relevance

The overall project is focused on reducing inflammatory and immunological side effects related to gene delivery so as to improve efficacy of adenoassociated virus gene delivery (even in the presence of local chronic infection). Novel cationic steroid delivery molecules will be tested for improved targeting and for allowing readministration of virus vector to eliminate the need for biomanufacturing of multiple serotypes.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL066565-12
Application #
8440757
Study Section
Bioengineering, Technology and Surgical Sciences Study Section (BTSS)
Program Officer
Mcdonald, Cheryl
Project Start
2000-12-15
Project End
2014-02-28
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
12
Fiscal Year
2013
Total Cost
$289,398
Indirect Cost
$100,902
Name
University of Pennsylvania
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Myint, Melissa; Limberis, Maria P; Bell, Peter et al. (2014) In vivo evaluation of adeno-associated virus gene transfer in airways of mice with acute or chronic respiratory infection. Hum Gene Ther 25:966-76
Leszczynska, Katarzyna; Namiot, Dorota; Byfield, Fitzroy J et al. (2013) Antibacterial activity of the human host defence peptide LL-37 and selected synthetic cationic lipids against bacteria associated with oral and upper respiratory tract infections. J Antimicrob Chemother 68:610-8
Wallen, Alexis J; Barker, Gregory A; Fein, David E et al. (2011) Enhancers of adeno-associated virus AAV2 transduction via high throughput siRNA screening. Mol Ther 19:1152-60
Fein, David E; Bucki, Robert; Byfield, Fitzroy et al. (2010) Novel cationic lipids with enhanced gene delivery and antimicrobial activity. Mol Pharmacol 78:402-10
Randazzo, R A S; Bucki, R; Janmey, P A et al. (2009) A series of cationic sterol lipids with gene transfer and bactericidal activity. Bioorg Med Chem 17:3257-65
Oh, Hana; Mohler 3rd, Emile R; Tian, Aiwei et al. (2009) Membrane cholesterol is a biomechanical regulator of neutrophil adhesion. Arterioscler Thromb Vasc Biol 29:1290-7
Barker, Gregory A; Diamond, Scott L (2008) RNA interference screen to identify pathways that enhance or reduce nonviral gene transfer during lipofection. Mol Ther 16:1602-8
Oh, Hana; Diamond, Scott L (2008) Ethanol enhances neutrophil membrane tether growth and slows rolling on P-selectin but reduces capture from flow and firm arrest on IL-1-treated endothelium. J Immunol 181:2472-82
Murphy, B R; Moayedpardazi, H S; Gewirtz, A M et al. (2007) Delivery and mechanistic considerations for the production of knock-in mice by single-stranded oligonucleotide gene targeting. Gene Ther 14:304-15
Handwerger, Rachel G; Diamond, Scott L (2007) Biotinylated photocleavable polyethylenimine: capture and triggered release of nucleic acids from solid supports. Bioconjug Chem 18:717-23

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