The overall goal of this proposal is to engineer (R21) and evaluate (R33) the next generation of a novel probiotic designed to prevent or treat infection by the diarrheagenic pathogen Clostridium difficile and, thereby, limit reliance on antibiotic therapy.
Three Specific Aims are proposed for the R21 phase.
In Aim 1, we will genetically engineer complete bio-containment into our existing first-generation probiotic, so that it will be lost from the gastrointestinal tract over 3-5 days unless continually dosed. Further, its anti-C. difficile infection (CDI) property will be expressed only upon concomitant sugar-substitute supplementation.
In Aim 2, we will assess in vitro properties of the second-generation probiotic.
In Aim 3, we will determine colonization efficacy, and also perform proof-of-concept anti-CDI efficacy studies. Quantifiable transition milestones will be available in order to progress to the R33 phase, where two Specific Aims are proposed.
In Aim 4, we will determine dosing, delivery and biological containment efficacy.
In Aim 5, we will comprehensively evaluate ability of the biotherapeutic agent to prevent or treat CDI (including primary infection and relapse), elicit adaptive immune response(s) to specific antigens, and impact endogenous microbiota recovery following antibiotic suppression or CDI-specific dysbiosis. Thus, the immediate impact of this work will be to verify pre-clinical readiness of our agent, via comprehensive interrogation of its disease-preventing modalities.

Public Health Relevance

Hundreds of thousands of cases of Clostridium difficile infection (CDI) occur annually in the USA, imposing a burden of >$3 billion on the healthcare system. CDI now surpasses all other bacterial infections in many US hospitals. The studies proposed in this application are focused on a novel, engineered probiotic designed to prevent or treat CDI, thereby limiting the problems associated with antibiotic use. This probiotic competitively prevents C. difficile establishment in the mammalian gastrointestinal (GI) tract. In work proposed in this application, the agent will be further refined to be completely biologically containable such that (a) it will be lost from the GI tract unless continually dosed, and (b) its unique anti-CDI property manifest only upon specific, simple, sugar supplementation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants Phase II (R33)
Project #
4R33AI121590-03
Application #
9591094
Study Section
Special Emphasis Panel (ZAI1)
Program Officer
Ranallo, Ryan
Project Start
2015-12-01
Project End
2020-11-30
Budget Start
2017-12-01
Budget End
2018-11-30
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Arizona
Department
Veterinary Sciences
Type
Earth Sciences/Resources
DUNS #
806345617
City
Tucson
State
AZ
Country
United States
Zip Code
85721
Vedantam, Gayatri; Kochanowsky, Joshua; Lindsey, Jason et al. (2018) An Engineered Synthetic Biologic Protects Against Clostridium difficile Infection. Front Microbiol 9:2080