Project Summary/Abstract The goal of this K99/R00 application is to support and facilitate the transition of Dr. Robert Jackson’s mentored postdoctoral research project to an independent research program focused on spatiotemporal modeling of cancer-causing human papillomavirus (HPV) infection in human oral epithelia. HPVs are responsible for 5% of all cancers worldwide. HPV causes genital cancers, but also an increasing number of oral cancers, most often in the tonsils and caused by HPV16. In the United States, the rate of HPV-associated oral cancers has surpassed cervical cancers and is considered an epidemic. HPV16 can transform host oral epithelial cells and lead to oral cancer, most often arising in the tonsillar crypt, but its normal viral lifecycle is reliant on epithelial differentiation to yield a productive infection. To address the oral health burden caused by HPV16, Dr. Jackson proposes three specific aims to assess and characterize HPV16 transforming vs productive infections in oral epithelia. Dr. Jackson’s preliminary work, using three-dimensional oral epithelial culture and single-cell transcriptomics, demonstrates that tonsillar crypt and surface epithelial subtypes have unique differentiation phenotypes and host regulator activity. In the K99 mentored phase, Dr. Jackson will assess HPV16’s lifecycle in tonsil epithelial subtypes and test the hypothesis that a host transcription factor, Peroxisome Proliferator- Activated Receptor Alpha (PPARα), regulates viral productivity and tumorigenesis. In Specific Aim 1, he will quantify HPV16 lifecycle stages and tumourigenicity in tonsillar crypt and surface epithelia. In Specific Aim 2, he will modulate PPARα activity to determine if it controls viral productivity and tumorigenicity. In the R00 independent phase, in Specific Aim 3, Dr. Jackson will establish his independent research program and define the spatiotemporal viral-host transcriptome across oropharyngeal tissues. The proposed mentored research will be performed at the University of Arizona, an ideal research and training environment, and will be coupled with a career development plan including structured professional development and new techniques training. Dr. Jackson will be supported by his primary mentor, Dr. Koenraad Van Doorslaer, as well as an advisory committee that will ensure his successful transition to a tenure-track faculty position for the independent phase. Dr. Jackson’s proposed research aligns with the NIDCR’s strategic priority to integrate oral and general health, with an objective to advance prevention, early detection, and treatments for head and neck cancers using the tactic of characterizing HPV+ oropharyngeal cancers. Dr. Jackson’s research is relevant as it intersects with notices of special interest including single-cell level spatiotemporal mapping of oral tissue and incorporates computational approaches.

Project Narrative Human papillomavirus (HPV) causes 5% of cancers worldwide, including a growing number of oral cancers. In the United States, HPV-associated oral cancers, most often arising in the tonsils, are considered an epidemic but are poorly understood. To contribute to fundamental knowledge and address the oral health burden caused by HPV, this project will determine the host factors regulating oral HPV infection and carcinogenesis using three-dimensional epithelial models and computational approaches.