There have been tremendous recent advances in our understanding of the pathogenesis of otitis media (OM) due to nontypeable Haemophilus influenzae (NTHI). Significantly, we?ve come to appreciate the role that biofilms play in both post-tympanostomy tube otorrhea as well as in the chronicity and recurrence of NTHI- induced OM. Bacteria dwelling within a biofilm present a formidable obstacle to effectors of immunity and antibiotic therapies. Thereby, in order to design novel and effective strategies to better treat and/or prevent otorrhea and OM, it is necessary to understand both the biology of biofilms, including their unique biochemical and proteomic composition, as well as how one might undermine these structures to mediate a therapeutic ?cure?. Through the successful conduct of a series of three highly integrated Specific Aims, we will expand upon a solid foundation of a growing body of literature from our laboratories which show that we can target a family of bacterial DNA-binding proteins (IHF and HU) that are critical to stabilize extracellular DNA (eDNA) present within an NTHI biofilm to reduce or eradicate that structure both in vitro and in vivo. Here, we will now identify and characterize the best therapeutic antigenic targets within both IHF and HU, with a focus on specific functional domains. We will also determine both the kinetics of release and the availability of IHF and HU subunits within the NTHI eDNA-DNABII dependent extracellular matrix in order to identify an optimal therapeutic window for intervention. Moreover, we will determine the therapeutic value of using biofilm-focused, epitope-targeted monoclonal antibodies and cocktails thereof to resolve biofilms associated with chronic otorrhea and otitis media. These latter studies will first be conducted in vitro using both monospecies biofilms, as well as those of mixed otopathogen etiology, to better reflect the clinical condition. Finally, we will determine the relative therapeutic efficacy of this approach in vivo, using a chinchilla model of experimental OM. The chinchilla represents an anatomically appropriate, robust, reproducible and extensively studied animal model of this highly prevalent pediatric biofilm disease in which to conduct these essential pre-clinical studies. Importantly, whereas our primary target for disease intervention is OM, because NTHI causes many biofilm-associated diseases of the human respiratory tract, the studies proposed here will also likely provide a basis for the design of better practices to treat disease in other parts of the airway such as the sinuses (chronic rhinosinusitis), the tonsils and adenoids (tonsillitis, adenoiditis), the bronchus (chronic cough and bronchitis) and the lung (COPD, early stage CF, community acquired pneumonia).
Our Specific Aims are perfectly aligned with both Healthy People 2020 (Hearing and Other Sensory or Communication Disorders, ENT-VSL-2: ?Reduce otitis media in children and adolescents?), and the NIDCD strategic plan FY 2012-2016, wherein Priority Research Area 3 ? Improving Diagnosis, Treatment and Prevention, declares a mandate to ?Develop therapies to prevent and treat biofilms.?

Public Health Relevance

It is estimated that 709 million new cases of acute OM (AOM) and 31 million new episodes of chronic suppurative OM (CSOM) occur each year worldwide, making the clinical spectrum of diseases known collectively as ?otitis media? the number one reason for antibiotic use. Moreover, worldwide it is reported that between 65 and 330 million children have existing CSOM, 60% of which have an associated hearing loss from this disease state that is characterized by chronically draining ears that can extend over a period of many years. The most cost-effective way to manage this highly prevalent pediatric disease, and have a transformational effect on the health of children globally, is through the design of innovative vaccines to prevent OM, as well as the discovery of novel methods to better treat existing OM; continued development and optimization of a highly innovative and effective therapeutic for treatment of children with CSOM and post- tympanostomy tube otorrhea (PTTO) by targeting the biofilms associated with these conditions is the focus of this application and represents one of the long-standing and overarching goals of the collaborative research program of the Bakaletz and Goodman laboratories.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
5R01DC011818-08
Application #
9534041
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Watson, Bracie
Project Start
2011-07-20
Project End
2021-08-31
Budget Start
2018-09-01
Budget End
2019-08-31
Support Year
8
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Nationwide Children's Hospital
Department
Type
DUNS #
147212963
City
Columbus
State
OH
Country
United States
Zip Code
43205
Novotny, Laura A; Clements, John D; Goodman, Steven D et al. (2017) Transcutaneous Immunization with a Band-Aid Prevents Experimental Otitis Media in a Polymicrobial Model. Clin Vaccine Immunol 24:
Freire, M O; Devaraj, A; Young, A et al. (2017) A bacterial-biofilm-induced oral osteolytic infection can be successfully treated by immuno-targeting an extracellular nucleoid-associated protein. Mol Oral Microbiol 32:74-88
Rocco, C J; Davey, M E; Bakaletz, L O et al. (2017) Natural antigenic differences in the functionally equivalent extracellular DNABII proteins of bacterial biofilms provide a means for targeted biofilm therapeutics. Mol Oral Microbiol 32:118-130
Jurcisek, Joseph A; Brockman, Kenneth L; Novotny, Laura A et al. (2017) Nontypeable Haemophilus influenzae releases DNA and DNABII proteins via a T4SS-like complex and ComE of the type IV pilus machinery. Proc Natl Acad Sci U S A 114:E6632-E6641
Novotny, Laura A; Jurcisek, Joseph A; Goodman, Steven D et al. (2016) Monoclonal antibodies against DNA-binding tips of DNABII proteins disrupt biofilms in vitro and induce bacterial clearance in vivo. EBioMedicine 10:33-44
Idicula, Winslo K; Jurcisek, Joseph A; Cass, Nathan D et al. (2016) Identification of biofilms in post-tympanostomy tube otorrhea. Laryngoscope 126:1946-51
Devaraj, Aishwarya; Justice, Sheryl S; Bakaletz, Lauren O et al. (2015) DNABII proteins play a central role in UPEC biofilm structure. Mol Microbiol 96:1119-35
Novotny, Laura A; Jurcisek, Joseph A; Ward Jr, Michael O et al. (2015) Antibodies against the majority subunit of type IV Pili disperse nontypeable Haemophilus influenzae biofilms in a LuxS-dependent manner and confer therapeutic resolution of experimental otitis media. Mol Microbiol 96:276-92
Brockson, M Elizabeth; Novotny, Laura A; Mokrzan, Elaine M et al. (2014) Evaluation of the kinetics and mechanism of action of anti-integration host factor-mediated disruption of bacterial biofilms. Mol Microbiol 93:1246-58
Novotny, Laura A; Amer, Amal O; Brockson, M Elizabeth et al. (2013) Structural stability of Burkholderia cenocepacia biofilms is reliant on eDNA structure and presence of a bacterial nucleic acid binding protein. PLoS One 8:e67629

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