Nonencapsulated (nontypeable) Haemophilus influenzae (NTHi) are human-adapted commensal organisms that can also cause chronic mucosal infections, particularly otitis media. Since NTHi are not susceptible to the routinely-administered Hib vaccine, these infections are usually treated with antibiotics. Many children exhibit recurrent otitis media caused by NTHi despite antibiotic treatment, and these episodes can lead to deafness and language development delays in early life. Although the ability to form biofilms has been implicated in persistent infections, the exact molecular mechanism by which NTHi causes chronic disease is not fully understood. NTHi contain a set of four highly conserved gene pairs termed toxin-antitoxin (TA) loci. Our preliminary data shows that the deletion of two of these gene pairs significantly reduces the ability of the organism to cause long-term infections in a primary human tissue model. Our hypothesis is that these four TA loci are involved in protease-mediated metabolic regulation that allows the organism to enter a state of growth arrest precipitated by stress (such as antibiotic therapy or nutrient limitation), and this enhances their ability to survive under these suboptimal conditions. Once conditions improve (e.g. cessation of antibiotic treatment, or nutrient upshift), bacterial growth resumes. Since most antibiotics target essential biosynthetic pathways that are not utilized during growth arrest, this mechanism could explain the observed clinical failure in children with recurrent otitis media, and result in new targets for the treatment of chronic infections. To investigate this, we aim to a) progressively delete all four TA loci from two different NTHi strains;b) assess the ability of these mutants to survive in long-term infections of a primary human tissue model grown at the air-liquid interface, and c) analyze the mutants that were found to be significantly attenuated for persistence in the chinchilla model of otitis media. Because TA loci appear in hundreds of different bacterial species, in addition to discovering novel targets for immunological or pharmacological intervention for those patients that exhibit chronic disease, these studies have implications for the treatment of many other organisms that persist within a host or in the environment.

Public Health Relevance

antibiotic treatment failure in children that results in recurrent middle ear infections (otitis media). These episodes can lead to deafness as well as language development delays. Our research could lead to the identification of new targets for vaccines or pharmaceuticals that would synergize with existing treatments for those patients that exhibit chronic otitis media.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
7R01DC010187-04
Application #
8277800
Study Section
Clinical Research and Field Studies of Infectious Diseases Study Section (CRFS)
Program Officer
Watson, Bracie
Project Start
2009-07-09
Project End
2014-06-30
Budget Start
2012-08-28
Budget End
2014-06-30
Support Year
4
Fiscal Year
2012
Total Cost
$307,870
Indirect Cost
$92,248
Name
Old Dominion University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
041448465
City
Norfolk
State
VA
Country
United States
Zip Code
23508
Ren, Dabin; Kordis, Alexis A; Sonenshine, Daniel E et al. (2014) The ToxAvapA toxin-antitoxin locus contributes to the survival of nontypeable Haemophilus influenzae during infection. PLoS One 9:e91523
Cline, Susan D; Saleem, Sehresh; Daines, Dayle A (2012) Regulation of the vapBC-1 toxin-antitoxin locus in nontypeable Haemophilus influenzae. PLoS One 7:e32199
Ren, Dabin; Daines, Dayle A (2011) Use of the EpiAirway model for characterizing long-term host-pathogen interactions. J Vis Exp :e3261