A major limitation in the development of non-viral gene carriers for Cystic Fibrosis (CF) has been the sparse attention paid to effects of the mucosal barrier on stability of gene carriers and on their ability to access and efficiently enter, traffic within, and deliver cargo DNA to the nucleus of lung epithelial cells. Absorption of mucus components often destabilizes gene carriers and significantly changes important carrier physicochemical properties that affect gene delivery efficiency. However, modifications aimed at overcoming one barrier (e.g. mucin absorption) may create other significant barriers (e.g. reduced cell entry rate). Our overall hypothesis is that a more comprehensive and quantitative """"""""systems"""""""" approach to the identification of barriers to successful gene delivery for CF will allow our rational synthesis of novel gene carriers that resist mucus absorption and are capable of: (i) rapid transport through the mucosal barrier;(ii) facile entry into human CF bronchial epithelial cells following incubation in CF mucus;(iii) efficient and active accumulation around and in the cell nucleus within minutes of cell entry;and (iv) significantly improved gene expression in cell culture and in CF mouse models with gene carriers that have been pre-incubated in CF mucus. Specifically, starting with the synthesis and characterization of novel highly compacted (<22 nm in minor diameter) polymeric gene carriers (Aim 1), this proposal will utilize a number of powerful biophysical techniques to identify and quantify the rate limiting barriers to efficient gene carrier transport through human CF mucus (Aim 2) and through the cell to the nucleus (Aim 3). To ensure clinical relevance, gene carrier transport will be investigated in purulent/infected sputum and differentiated primary human bronchial epithelial cells grown at an air-interface, each freshly obtained from CF patients. Promising carriers will be tested for efficacy in a CF mouse model (Aim 4).The identification of important barriers in Aims 2-4 will guide the rational modification of the gene carrier physicochemical properties and surface chemistries (Aim 1 again) to potentially overcome the bottleneck. An interdisciplinary team, with expertise in bioengineering/biophysics, aerosol gene delivery, cellular trafficking/biology, and CF, has been assembled to investigate the hypothesis, with a long-term goal of safe and effective CF gene therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB003558-03
Application #
7619127
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Henderson, Lori
Project Start
2007-08-13
Project End
2011-04-30
Budget Start
2009-05-01
Budget End
2010-04-30
Support Year
3
Fiscal Year
2009
Total Cost
$356,474
Indirect Cost
Name
Johns Hopkins University
Department
Type
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Suk, Jung Soo; Kim, Anthony J; Trehan, Kanika et al. (2014) Lung gene therapy with highly compacted DNA nanoparticles that overcome the mucus barrier. J Control Release 178:8-17
Birket, Susan E; Chu, Kengyeh K; Liu, Linbo et al. (2014) A functional anatomic defect of the cystic fibrosis airway. Am J Respir Crit Care Med 190:421-32
Schuster, Benjamin S; Kim, Anthony J; Kays, Joshua C et al. (2014) Overcoming the cystic fibrosis sputum barrier to leading adeno-associated virus gene therapy vectors. Mol Ther 22:1484-1493
da Silva, Adriana L; Martini, Sabrina V; Abreu, Soraia C et al. (2014) DNA nanoparticle-mediated thymulin gene therapy prevents airway remodeling in experimental allergic asthma. J Control Release 180:125-33
Schuster, Benjamin S; Suk, Jung Soo; Woodworth, Graeme F et al. (2013) Nanoparticle diffusion in respiratory mucus from humans without lung disease. Biomaterials 34:3439-46
Kim, Anthony J; Boylan, Nicholas J; Suk, Jung Soo et al. (2013) Use of single-site-functionalized PEG dendrons to prepare gene vectors that penetrate human mucus barriers. Angew Chem Int Ed Engl 52:3985-8
Burke, Caitlin W; Suk, Jung Soo; Kim, Anthony J et al. (2012) Markedly enhanced skeletal muscle transfection achieved by the ultrasound-targeted delivery of non-viral gene nanocarriers with microbubbles. J Control Release 162:414-21
Ensign, Laura M; Schneider, Craig; Suk, Jung Soo et al. (2012) Mucus penetrating nanoparticles: biophysical tool and method of drug and gene delivery. Adv Mater 24:3887-94
Kim, Anthony J; Boylan, Nicholas J; Suk, Jung Soo et al. (2012) Non-degradative intracellular trafficking of highly compacted polymeric DNA nanoparticles. J Control Release 158:102-7
Kim, Anthony J; Hanes, Justin (2012) The emergence of multiple particle tracking in intracellular trafficking of nanomedicines. Biophys Rev 4:83-92

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