Upgrading infectious disease research facilities at University of Louisville RBL

The University of Louisville Regional Biocontainment Laboratory (RBL) is an operational unit of the University's Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases (CPM). The CPM operates the only Biosafety Level 3 laboratories and animal research facilities available to researchers in the Commonwealth of Kentucky. As such we are a regional resource for two Carnegie Research I (highly research intensive) Universities-the University of Louisville and the University of Kentucky-as well as for the local biotechnology industry and other universities and non-profit organizations. The University of Louisville has been an outstanding steward of the RBL, paying meticulous attention to the facility needs. However, after a decade, we have encountered some challenges, primarily with obsolescence of control systems, and wear and tear on some of the building air handling systems. The necessary HVAC upgrades are already being addressed by the University, but we request support from NIAID through this G20 for additional facility related upgrades and modernization of research equipment SPECIFIC AIMS We propose to address specific facility operational needs in both the fixed building infrastructure and in upgrading and modernizing our in vivo and in vitro research capabilities. We propose incorporating modern state-of-the-art instrumentation to build solid foundations for innovations in biodefense and emerging infectious disease research. The building systems upgrades that directly affect operations of the containment space will be scheduled during the annual preventative maintenance shutdown periods to be conducted in November/December 2021 and 2022. Similarly, new in vivo and in vitro research equipment shall be installed during the annual shutdown to allow for service professionals to install and commission the equipment without needing to address biosafety training and Select Agent-specific standard operating procedures. Aim 1: Repair, renovate and modernize RBL building systems to enhance functions and operations of the existing research facilities. We propose to address specific facility operational needs in the fixed building infrastructure. We shall upgrade critical facility systems that are facing imminent obsolescence and rectify one major facility design defect that placed some inconveniences and limitations on research operations: the BSL-3- ABSL-3 passthrough boxes. The long-term commitment of the CPM is to support basic and translational research that leads to the development of countermeasures for biodefense and emerging infectious agents. These upgrades will secure our ability to maintain our BSL-3 research program and continue to develop innovative countermeasures against biodefense and emerging infectious disease threats. Aim 2: Enhance and modernize the CPM RBL in vivo research capabilities. The COVID-19 pandemic validated the significance and need for ABSL-3 facilities such as we have at the CPM. Over the past 14 months our in vivo research facilities were heavily utilized to support development of novel COVID-19 therapeutics, vaccines and other prophylactic strategies. Until COVID-19 the vast majority of our in vivo research utilized murine models, with sporadic use of ferrets for influenza studies. Recently, we have made extensive use of the Golden Syrian Hamster (GSH) model. Consequently, we have a practical need to expand our caging for medium sized rodent models. Moreover, emerging data on post-COVID human pathologies impacting different organ systems-lung, cardiovascular, neural and renal in particular-raise the need for more sophisticated tools for assessing physiological function in research models. To this end, we propose to elevate our in vivo research facilities by adding Promethion metabolic isocaging systems that record animal metabolic and behavioral data and allow us to integrate research studies with our existing gnotobiotic rodent core. We shall also add telemetry capabilities to precisely record body temperature, heart rate and blood pressure metrics. For lung function assessments, we will add rodent plethysmography capabilities. These research infrastructure enhancements will build new collaborations with our colleagues in centers of excellence in environmental health, cardiovascular disease, diabetes and obesity. We will also consolidate existing collaborations with our Functional Microbiomics, Inflammation and Pathogenicity COBRE by facilitating use of gnotobiotic animals to assess the impact of microbiome communities in pathogenesis of emerging infectious diseases. In vivo imaging capability is an extremely useful adjunct to physiological and most-mortem pathology assessments. Our existing IVIS optical imaging system is nearing obsolescence, and needs upgrading to secure ongoing research studies, particularly in Yersinia pestis and Burkholderia pseudomallei infection models. Aim 3: Enhance and modernize the CPM RBL in vitro research capabilities. The CPM RBL is a wellequipped facility, but the vast majority of the research instrumentation was installed as the University cost-share contribution to the RBL construction between 2007 and 2010. We have a meticulous equipment maintenance and service program coordinated by Marlene Steffen and funded by the Executive Vice President for Research annual budget to the CPM, which has kept all of our equipment fit for purpose. Over the past decade, technology Contact PD/PI: PALMER, KENNETH E Program Narrative Page 7 for analysis of immune cell function by flow cytometry has become significantly more sophisticated. We propose to enhance and modernize our flow cytometry analytical capabilities by adding a Becton Dickinson FACSymphony analyzer with 4 laser/16 color capabilities and high throughput system that will accommodate 96/384-well plates. This will significantly expand the speed, throughput and sophistication of the data acquisition, complementing our existing 11-year-old FACSAria II cell sorter. We have existing high throughput screening capabilities in our High Throughput Biology Core. Most of the HTPB equipment is also a decade old. We propose that our biodefense and emerging infectious disease research programs would benefit immensely by acquisition of the Cytation C10 high throughput confocal microscopy instrument that allows rapid collection of high quality and high-resolution quantitative data under normal cell and tissue incubation conditions. Several of our faculty have critical need for this instrument to be located in our RBL BSL-3 laboratories to facilitate their research on emerging viruses and bacterial pathogens.