About Us Significance Core Capabilities Tapping the RBL

1. The Regional Biocontainment Laboratory at the University of Louisville: A Shared Resource to Promote Basic and Translational Research

2. Merging Basic & Clinical Research • About Us • Significance • Core Capabilities Tapping the RBL

3. The RBL: About Us • History • Design & Construction: Dec. 2005 - Mar. 2009 • Commissioning: Mar. 2009 -July 2010 • BSL-3 Certification: Aug. 2010 – Oct. 2010 • Select Agent Approval: Nov. 2010 • Expansion Award (GLP): June 2010 • Estimation of Expansion completion: Jan 2014 Facility Specifications Total gross space: 38,135 sq. ft. Total laboratory space: 21,773 sq. ft. Total BSL-2 laboratory space: 2069 sq. ft. Total BSL-3 laboratory space: 2395 sq. ft. Total ABSL-3 laboratory space: 4893 sq. ft. Total ABSL-3 laboratory space: 2183 sq. ft. .

4. UofL RBL is Poised to Promote and Support Translational Research of CPM CPM Mission….to improve human health by conducting basic research that translates to the development of diagnostic markers, vaccines and therapeutics for pathogens requiring biocontainment. *The RBL will make itself available in the case of a national emergency.

5. Merging Basic & Clinical Research • About Us • Significance • Core Capabilities Tapping the RBL Service Center

6. Merging Basic & Clinical Research Merging Basic & Clinical Research The Perpetual Challenge ofInfectious Diseases Over the next hour alone, at least 1,500 people will die from an infectious disease - over half of them children under five (WHO). For children under five, pneumonia and diarrheal diseases are the two biggest killers, accounting for 18% and 15% of all deaths respectively in 2008.

7. Merging Basic & Clinical Research • About Us • Significance • Core Capabilities Tapping the RBL Service Center

8. From Basic to Translation Pathogenesis Efficacy Diagnosis Patient Outcome BIO-Imaging HT Biology Small Animal Models Microbiology Immunology

9. Merging Basic & Clinical Research HT Biology Core HT Biology Core Mission: To accelerate the discovery of small molecule for treatment of infections. Small Molecule Discovery Target Discovery

10. Merging Basic & Clinical Research HTB & Discovery Expertise Core Capabilities With our current state-of-the-art expertise and capabilities in Drug Discovery, our team can quickly identify putative small molecules or targets once an assay has been validated for screening. The HTB Core can provide primary and confirmatory screening and follow-on guidance for mechanism of action to confirm molecules as a viable probe to initiate a hit-to-lead discovery program as a potential new therapeutic or a tool for biological research. • Representative Publications & Patents 1. Discovery of novel benzoquinazolinones and thiazoloimidazoles, inhibitors of influenza H5N1 and H1N1 viruses, from a cell-based high-throughput screen, Journal of Biomolecular Screening 16, no. 1 (January 2011): 73-81. 2. HTS-Driven Discovery of New Chemotypes with West Nile Virus Inhibitory Activity, Molecules 15, no. 3 (March 2010): 1690-1704. 3. A Cell Based Assay for the Identification of Lead Compounds with Anti-Viral Activity Against West Nile Virus, NIH Bookshelf. NBK50698. 4. High-Throughput Screening of a 100,000 Compound Library for Inhibitors of Influenza A virus (H3N2), Journal of Biomolecular Screening 13, no. 9 (October 2008): 879-887. 5. A cell-based luminescence assay is effective for high-throughput screening of potential influenza antivirals, Antiviral Research 73, no. 1 (January 2007): 50-59. 6. Development and Validation of a High-Throughput Screen for Inhibitors of SARS CoV and Its Application in Screening of a 100,000-Compound Library, Journal of Biomolecular Screening 12(1); 2007:33-40.

11. Merging Basic & Clinical Research Animal Models Core Animal Core Mission: To determine how pathogens cause diseases & the efficacy of new therapeutics. Small Animal Capabilities Mouse Hamster Rat Guinea Pig Ferret Rabbit

12. Merging Basic & Clinical Research Microbiology Core Microbiology Core Mission: To determine how pathogens cause diseases & the efficacy of new therapeutics in vitro. To support the analyses of animal models of infection and treatment. • BSL2 Agents • Influenza Strains • Venezuelan encephalitis • virus vaccine strains • Respiratory Syncytial Virus • Yellow fever Virus • Dengue Virus • Chikungunya Virus vaccine • Yersina pseudotuberculosis • Klebsiella pneumoniae • BSL3 Agents • Arenaviruses • SARS CoV • Hantaviruses • West Nile Virus • H2N2 Influenza virus • BSL3 Select Agents • Influenza H5N1 strains • Yersinia pestis • Burkholderia pseudomallei • Burkholderia mallei • Venezuelan equine encephalitis

13. Merging Basic & Clinical Research Immunology Core Immunology Core Mission: To determine how hosts immune system respond to pathogens. • Integration • The Immunology Core offers expertise in full integration with the: • Vivarium Core to carefully isolate a variety of tissues from animal models for analysis. • Microbiology Core to process samples using IFA and cryotome sectioning for IHC and staining. • High-Throughput Biology Core to design assays using novel cytometric techniques. • ImagingCore to design molecular markers and cellular tracers to investigate disease progression with invivo and in vitro models. • Comprehensive Research Support • Our facilities and expertise allow the opportunity for thorough analyses of immune response to infectious diseases, including: • Hematology • Antibody response • Cell-cycle analyses • Lymphoproliferation • Live cell sorting • Multi-color flow cytometry • Multiplexed cytokine assays

14. Merging Basic & Clinical Research BIO-Imaging Core BIO-Imaging Core Mission: To determine how hosts immune system respond to pathogens. To Create New Probes for Diagnostic Imaging of Infection Whole Animal Imaging: Siemens Trimodal Whole Animal Imaging: IVIS Spectrum Cell-based Imaging: Zeiss LSM 710Duo Live

15. New technologies for Diagnostic Optical Imaging Jonathan Warawa, Ph.D. The bioluminescence lux operon was stably engineered into the chromosome of DD503 wild type B. pseudomallei and the capsule operon mutant Mice infected intranasally develop severe rhinitis and fatal pneumonia which can be monitored over time semi-quantitatively Y-intercept noise: 3.1 +/- 2.1x103 p/s CFU at 1 SD (JW280): 5.7x106 CFU CFU at 1 SD (JW280 Dwcb): 2.1x106 CFU

16. Discovery of Current Clinical tools for Imaging Pulmonary Inflammation in Infection FDG-PET/CT with H1N1 Infection, Colleen Jonsson Day 1 Day 3 Day 6 Jonsson CB, Camp JV, Wu A, Zheng H, Kraenzle JL, Biller AE, Vanover CD, Chu YK, Ng CK, Poctor M, Sherwood L, Steffen MC, Mollura DJ: Molecular Imaging Reveals a Progressive Pulmonary Inflammation in Lower Airways in Ferrets Infected with 2009 H1N1 Pandemic Influenza Virus. PLoS ONE. 2012. 7(7) e40094.

17. Merging Basic & Clinical Research • About Us • Significance • Core Capabilities • Tapping the RBL • Service Center

18. Merging Basic & Clinical Research RBL Welcomes External Users • NIH Funded Academic Research/Faculty • NIH Government Contracts (e.g. NIH IDIQ) • Academic Research/Faculty • Commercial/Biotech Research

19. Merging Basic & Clinical Research Possible Types of Usage • Collaboration with CPM or UofL Faculty • Work with a UofL faculty in collaboration for joint publication (grant, contract) • Fee for Service • CPM staff provide service, regulatory support • External User Access • Scientists directly access facility and publish findings independently with UofL regulatory oversight

20. Merging Basic & Clinical Research Process for Direct Use • Complete Agreements for Use • Facility use agreements • University Staff and Students • Visiting Scientists • DOJ Clearance • For direct access only • Complete Costing Agreement Prior to Use • Program Coordinator, Business Manager • Complete Training Program • BSL-2 • BSL-3 • Select Agent

21. CPM & Collaborators Support TEAM Director, CPM Colleen B. Jonsson, Ph.D. Director, Shared Resources William Severson, Ph.D. (On Site RBL Director) Shared Resources Program Coordinator E. Pete Womack, M.S. CPM Business Office Neshela Kulenovic, MBA, UBM Patricia Moss, Administrative Asst. High-throughput Biology Core Donghoon Chung, Ph.D., (faculty manger) Julie Sotsky, B.S. , (Technical Support) BioImaging Core Haixun Guo, Ph.D. (faculty manager) Chin Ng, Ph.D. (faculty) Jonathan Warawa (faculty manager) Huaiyu Zheng, Ph.D. (radiology) Colleen B. Jonsson, Ph.D. (faculty manager) Microbiology Core Yong-Kyu Chu, Ph.D. (Manager) Scott Adcock, (Technical Support) Immunology Core Daniel Cramer, Ph.D., (Manager) Animal Models Core (RRF personnel) Carol Vanover, R.L.A.T. Jennifer Kraenzle, R.L.A.T.g Ashley Biller, R.L.A.T. Tanner Hughes Colleen B. Jonsson, Ph.D. (faculty manager) CPM Facility Operations (VPBA personnel) Marlene Steffen, B.S. Troy Thompson UofL RRF Mary Proctor, D.V.M. Leslie Sherwood, D.V.M. Karen Powell, Ph.D., D.V.M. Environmental Health & Safety Carol Whetstone, Ph.D. RO Thomas Cremer, Ph.D. Clarissa Cowan, B.S.

22. Get to know us! Tours available by appointment UofL Regional Biocontainment Laboratory http://centerforpredictivemedicine.org/