Project Summary/Abstract: HPV is a necessary cause of 10% of all cancers of women globally, including 99% of cervical cancers. Rates of cervical cancer vary markedly by geographic region and level of economic development, with >85% occurring in developing countries. Successful prophylactic vaccination against hrHPV types can prevent dysplasia and thus cervical cancer. Global estimates of HPV vaccination delivery and cervical screening by region and income level show virtually no significant delivery to many poorer populations of women worldwide. There are 13 hrHPV types plus a group of 11 possibly carcinogenic types. Licensed HPV vaccines all target HPV16 and HPV18, and one also targets the next five most common types in cervical cancer. Since they do not target all cancer-associated HPV types, screening must be maintained even in vaccinated women, adding to medical costs. In addition, the stringent cold-chain requirements of HPV vaccines provide a serious impediment to their delivery in developing countries. Our overall goal is to develop an affordable HPV vaccine that both extends the breadth of protection to all cancer-associated HPV types, and is stable at ambient temperature. Here we address this goal with two innovations: 1) displaying an L2-based protective epitope (RG1) that is conserved for HPVs in a unique position on the surface of HPV16 L1 VLPs, producing a single antigen, broadly protective HPV vaccine technology, RG1-VLP, and 2) the RG1-VLP vaccine is formulated on alum and embedded in glassy organic matrices formed by adjusting lyophilization and formulation parameters in order to control ice crystal nucleation rates, glass transition temperatures, and other material properties and thereby protect against degradation during processing, shipping and storage. HYPOTHESIS 1: Cold chain storage properties are a significant barrier to the distribution of current HPV vaccines, and the development of a powdered RG1-VLP vaccine including adjuvant that is thermostable will address this need. Specific Aim 1: Develop a GLP freeze-dry protocol for a powder formulation of RG1-VLPs in alum and study its in vitro temperature stability, and in murine models its immunogenicity and protective efficacy in comparison to Gardasil9. HYPOTHESIS 2: RG1-VLP vaccination is safe and well tolerated. Specific Aim 2: To perform a Dose Escalation Phase I Trial of the Safety and Immunogenicity of thermostable RG1-VLP in 36 healthy female volunteers with the inclusion of a control Gardasil9 arm in the study. HYPOTHESIS 3: RG1-VLP vaccination of healthy women induces broadly protective serum antibody. Specific Aim 3: To analyze the levels of protective antibodies in the serum of patients from the phase I study induced by RG1-VLP vaccination or Gardasil9. We will utilize the passive transfer assay to measure p...