PI- Phillip B. Hylemon, Ph.D. Title: ?Bile Acids and Clostridium scindens Inhibit C. difficile: Role of Secreted Antibacterial Compounds? Clostridium difficile, the cause of antibiotic associated diarrhea and colitis, is a growing health threat for patients taking broad spectrum antibiotics. It has been estimated that C. difficile may be responsible for almost a half a million infections per year and 29,000 deaths in the US at an annual cost of $4.8 billion dollars. Patients on broad-spectrum antibiotics markedly decrease the levels of protective gut microbiota and allows proliferation of C. difficile normally found in low levels in some individuals. Patients treated with antibiotics, especially in hospitals, are also at risk for colonization by C. difficile spores, which germinate in the GI tract (stimulated by specific bile acids) producing vegetative cells that secrete toxins A and B causing diarrhea and colitis. Patients with antibiotics associated diarrhea and/or colitis are routinely treated with either metronidazole or vancomycin to kill C. difficile vegetative cells colonizing the colon; however, up to 30% all patients successfully treated with these antibiotics will relapse following cessation of antibiotic therapy. Fecal transplants, using gut microbiota from healthy donors, have been highly successful in treating relapsing patients. Recent studies, published in Nature, were undertaken to determine which members of the gut microbiota are responsible for resistance to C. difficile infection. It was reported that Clostridium scindens, a human gut bacterium that converts the primary bile acids, cholic acid (CA) and chenodeoxycholic acid (CDCA) to secondary bile acids, deoxycholic acid (DCA) and lithocholic acid (LCA), respectively, is strongly associated with inhibition of C. difficile infections in both animal models and in human patients. This past year, our laboratory made two important discoveries that may explain how C. difficile is able to colonize the human GI tract, when patients are treated with antibiotics, and how C. scindens inhibits the growth and colonization by C. difficile.
Aim 1 : Confirm the structure and characterize the mechanism of action of a cyclic 6 amino acid antibiotic peptide secreted by C. difficile that inhibits the growth of C. scindens and other human gut bacteria.
Sub aim 1 a. Determine if clinical strains of C. difficile secrete the same compound.
Aim 2 : Purify, characterize, and determine the structure of a CA inducible antibacterial compound(s) secreted by C. scindens ATCC 35704 and VPI 12708 that inhibits the growth of C. difficile and other pathogenic bacteria in vitro.
Sub aim 2 a. Determine the spectrum of different bacteria inhibited by this compound(s).
Sub aim 2 b. Identify the bile acid inducible genes encoding the enzymes(s) involved in the synthesis of this antibacterial compound using RNAseq technology.
Sub aim 2 c. Characterize the mechanism of inhibition of C. difficile growth by this compound(s).
Aim 3 : Screen multiple strains of C. scindens for antibacterial compounds that inhibit C. difficile.
Sub aim 3 a. Develop a ?cocktail? of C. scindens strains producing different antibacterial compounds to use in the treatment of patients with infections.
Sub aim 3 b. Develop strains of C. scindens resistant to the 6 amino acid cyclic antibiotic peptide secreted by C. difficile. Long Term Objective: Find safe and effective ways to prevent and treat C. difficile infections using different strains of C. scindens and inhibitory compounds secreted by C. scindens. Determine how bile acids and antibacterial compounds, secreted by different species of the genus Clostridium, regulate the gut microbiome community structure.

Public Health Relevance

PI- Phillip B. Hylemon, Ph.D. Title: ?Bile Acids and Clostridium scindens Inhibit C. difficile: Role of Secreted Antibacterial Compounds? Clostridium difficile is the cause of antibiotic associated diarrhea and colitis (AAD&C). Patients on broad-spectrum antibiotics markedly decrease the levels of protective gut microbiota and allows proliferation of C. difficile. Patients with AAD&C are routinely treated with antibiotics but up to 30% relapse after therapy. Recent studies reported that Clostridium scindens, a bacterium that converts cholic acid (CA) to deoxycholic acid (DCA), was highly associated with resistance to C. difficile infection in animal models and patients. This past year, we discovered that C. scindens secretes antibacterial compound(s) that inhibits C. difficile. We also discovered that C. difficile also secretes a 6 amino acid cyclic antibiotic peptide that inhibits C. scindens and other Gram-positive gut bacteria. The studies proposed in this application are aimed at determining the interaction between C. difficile, C. scindens, bile acids and AAD&C.

Agency
National Institute of Health (NIH)
Institute
Veterans Affairs (VA)
Type
Non-HHS Research Projects (I01)
Project #
5I01BX001328-06
Application #
9412372
Study Section
Gastroenterology (GAST)
Project Start
2012-07-01
Project End
2020-12-31
Budget Start
2018-01-01
Budget End
2018-12-31
Support Year
6
Fiscal Year
2018
Total Cost
Indirect Cost
Name
VA Veterans Administration Hospital
Department
Type
DUNS #
146678115
City
Richmond
State
VA
Country
United States
Zip Code
23249
Bajaj, J S; Sterling, R K; Betrapally, N S et al. (2016) HCV eradication does not impact gut dysbiosis or systemic inflammation in cirrhotic patients. Aliment Pharmacol Ther 44:638-43
Bhowmik, Shiva; Chiu, Hsien-Po; Jones, David H et al. (2016) Structure and functional characterization of a bile acid 7? dehydratase BaiE in secondary bile acid synthesis. Proteins 84:316-31
Bajaj, Jasmohan S; Ahluwalia, Vishwadeep; Steinberg, Joel L et al. (2016) Elderly patients have an altered gut-brain axis regardless of the presence of cirrhosis. Sci Rep 6:38481
Kang, Dae J; Kakiyama, Genta; Betrapally, Naga S et al. (2016) Rifaximin Exerts Beneficial Effects Independent of its Ability to Alter Microbiota Composition. Clin Transl Gastroenterol 7:e187
Kang, Dae Joong; Betrapally, Naga S; Ghosh, Siddhartha A et al. (2016) Gut microbiota drive the development of neuroinflammatory response in cirrhosis in mice. Hepatology 64:1232-48