PROJECT SUMMARY Funds are requested to purchase a Thermo Fisher Scientific Orbitrap IQ-X Mass Spectrometer System with Vanquish Flex UHPLC System (IQ-X) to establish a unique omics capability at the University of South Florida (USF), with analytical performance that is not currently available as a shared resource, for detailed, high-resolution structural analysis of low-level small molecules in complex biological matrices. Specifically, we have developed Nucleic Acid Adductomics (NAA) which aims to describe the totality of modifications in the genome i.e., DNA, and RNA products plus other significant classes of adducts e.g., DNA-DNA, RNA-RNA, DNA-protein, and RNA- protein crosslinks, all of which are implicated in pathogenesis. The requested instrument will be the only one of its kind outside of Taiwan performing NAA, and will support efforts to better understand the role of NA modifications in health and disease. Over twelve NIH-funded principal investigators (major users, representing 24 awards) across USF/Moffitt Comprehensive Cancer Center need the analytical capabilities provided by the IQ-X. Specifically, they need to identify the diverse range of NA modification types, and an unprecedented number of individual adducts, through ultra-high mass measurement accuracy and resolution, and unique scanning features (e.g., neutral loss-driven MS2 or MSn analysis for structure elucidation), while maintaining a respectable sample analysis time. An accompanying need is to perform targeted adductomics for specific nucleic acid-derived biomarkers in complex matrices with significant chemical noise. For analytical and user convenience, the instrument will be housed within the Proteomics Facility located in the Florida Center for Excellence for Drug Discovery and Innovation, at USF. The instrument will support NIH-funded investigators from multiple departments across major USF colleges (e.g., Arts and Sciences, Medicine, Public Health) and Moffitt. The IQ-X will be the first instrument of its kind at USF, and will be dedicated to performing non-targeted and targeted NAA, with options to focus on cellular, nuclear, or mitochondrial targets, and extracellular matrices, such as urine. The instrument will be used to enhance basic and translational, NIH-funded research programs at USF/Moffitt that cover a wide range of topics e.g., infectious disease, genome instability, carcinogenesis, respiratory, cardiovascular, and neurological disorders. This cutting- edge technology will accelerate new and important discoveries for multiple projects, as well as opening new directions of research in the biomedical sciences, and improving our understanding of the mechanisms underlying pathogenesis.

PROJECT NARRATIVE This application seeks funds for a state-of-the art, hybrid quadrupole-Orbitrap (IQ-X) high resolution mass spectrometer to be housed within an existing core facility at the University of South Florida (USF). The instrument will be dedicated to establishing a Nucleic Acid Adductomics capability as a campus-wide shared resource to support multiple NIH-funded projects across USF and Moffitt Comprehensive Cancer Center. This will provide a unique approach to characterizing the widest possible range of nucleic acid modifications, and hence advancing our knowledge of their role in health and disease.