The long term objective of this proposal is to test the hypothesis that the introduction of highly charged anionic polyelectrolytes into lung airway fluid as a result of inflammation and cell death contributes to the pathology of cystic fibrosis. Specifically the work will focus on the polyelectrolyte properties of F-actin and DNA as they relate to the abilities of these polymers to 1) increase the elasticity of CF sputum; 2) inactivate cationic antimicrobial proteins and peptides in the lung epithelium, and 3) interfere with gene therapy strategies that employ DNA-cationic lipid complexes for gene delivery into the epithelium. The experimental work will employ light scattering, fluorescence and polarization microscopy, rheology, and bacterial growth and lysis assays to study purified systems in which F-actin and DNA are caused to form bundles by lysozyme, defensins, histories and other cationic peptides released by epithelial cells and leukocytes. These studies will be expanded into determining how similar effects occur in sputum. Theories developed for polyelectrolyte condensation will be exploited to design soluble, polyvalent anions that independently or in combination with DNA- or actin-directed proteins such as DNAse and gelsolin can dissociate filament bundles and liberate antimicrobial function. The long term outcome of this work would be to determine if strategies based on depolymerizing DNA and cytoskeletal fibers or on dissociating bundles of these filaments by manipulating the ionic environment of airway fluid have potential to alleviate the pathology of CF.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL067286-02
Application #
6537988
Study Section
Lung Biology and Pathology Study Section (LBPA)
Program Officer
Banks-Schlegel, Susan P
Project Start
2001-06-01
Project End
2006-05-30
Budget Start
2002-05-31
Budget End
2003-05-30
Support Year
2
Fiscal Year
2002
Total Cost
$315,675
Indirect Cost
Name
University of Pennsylvania
Department
Physiology
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Wang, Yu-Hsiu; Bucki, Robert; Janmey, Paul A (2016) Cholesterol-Dependent Phase-Demixing in Lipid Bilayers as a Switch for the Activity of the Phosphoinositide-Binding Cytoskeletal Protein Gelsolin. Biochemistry 55:3361-9
Ku?akowska, Alina; Byfield, Fitzroy J; Zendzian-Piotrowska, Ma?gorzata et al. (2014) Increased levels of sphingosine-1-phosphate in cerebrospinal fluid of patients diagnosed with tick-borne encephalitis. J Neuroinflammation 11:193
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
Wang, Yu-Hsiu; Collins, Agnieszka; Guo, Lin et al. (2012) Divalent cation-induced cluster formation by polyphosphoinositides in model membranes. J Am Chem Soc 134:3387-95
Huisman, Elisabeth M; Wen, Qi; Wang, Yu-Hsiu et al. (2011) Gelation of semiflexible polyelectrolytes by multivalent counterions. Soft Matter 7:7257-7261
Cohen, Taylor S; Bucki, Robert; Byfield, Fitzroy J et al. (2011) Therapeutic potential of plasma gelsolin administration in a rat model of sepsis. Cytokine 54:235-8
Leszczynska, K; Namiot, A; Cruz, K et al. (2011) Potential of ceragenin CSA-13 and its mixture with pluronic F-127 as treatment of topical bacterial infections. J Appl Microbiol 110:229-38
Byfield, Fitzroy J; Wen, Qi; Leszczynska, Katarzyna et al. (2011) Cathelicidin LL-37 peptide regulates endothelial cell stiffness and endothelial barrier permeability. Am J Physiol Cell Physiol 300:C105-12
Byfield, Fitzroy J; Kowalski, Marek; Cruz, Katrina et al. (2011) Cathelicidin LL-37 increases lung epithelial cell stiffness, decreases transepithelial permeability, and prevents epithelial invasion by Pseudomonas aeruginosa. J Immunol 187:6402-9
Ellenbroek, Wouter G; Wang, Yu-Hsiu; Christian, David A et al. (2011) Divalent cation-dependent formation of electrostatic PIP2 clusters in lipid monolayers. Biophys J 101:2178-84

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