1. A collaborative journey down the rabbit hole Barbara Van Der Pol, PhD, MPH (HPER) David E Nelson, PhD (Biology)

3. Inflammatory response in the urethra causing discharge, pain on urination and burning/itching Leading cause of male healthcare office visits in US 69.5% of initial visits in Massachusetts HMO* <20% had identifiable infection *Ratelle, et al STD 2001

4. Pathogens?Host Response?

5. Clinical/Epidemiologic questions Why are some men at greater risk of urethritis? How does this impact STI control efforts? Need patient samples and data Molecular tools provide answers What microbial communities are related to these syndromes? How can we identify & quantify them?

6. Access to male volunteers $ to compensate for time Access to STI diagnostics $ for testing Sequencing technologies $!!! Input on study design Consider analysis a priori

7. Enrolled men from STD clinic population Collected paired samples, behavioral data, sexual history, urethral health and prostatitis history Ran STD diagnostic assays Samples sent in nucleic acid preservative to Dave Nelson’s lab DNA isolated/quantified PCR amplified using broad-range 16S primers Deep pyrosequenced specimens at the IU CGB Analysis

10. C. trachomatis N. gonorrhea Tenericutes

12. Which would YOU choose?

14. H+ L. crispatusN. gonnorhoeae

15. Have demonstrated utility of urine sampling Have shown differences among men based on STI status Have identified further differences among men WITH urethritis Have begun to identify pathogens similar to those seen in vaginal syndromes Have begun longitudinal analysis of microbiome in young men

16. R01 with 32 nd percentile (to be resubmitted Nov 2010). $3.7 million TC UH3 funded (Oct 2010-2013, $7 million TC) Manuscripts: Preliminary exploration of male urethral microbiome (under revision) Comparison of urethral swabs to urine (in preparation) Unique composition of microbiome in urethritis related to STI (in prep) Baseline analysis of young men: differences based on sexual behavior & circumcision status (in prep) Presentations: NIH Human Microbiome Symposium [2 posters, 1 oral presentation] ISSTDR London [1 poster, 1 oral presentation] Midwest Microbial Pathogenesis Conference [1 poster]

17. Through collaboration comes synergy Assistance with experimental design (with analysis in mind) is key Opportunities such as CTSI PDT can open MANY doors Long-term outcome still in-process!

18. CTSI PDT JD Fortenberry (IU Adolescent Medicine) Q Dong (UNT Bioinformatics) BP Katz(IU Biostatistics) Evelyn Toh(IU Biology) George Weinstock (WUStL Genomics)