Project Details
Projekt
Gene expression changes during Merkel Cell Polyomavirus productive and persistent infection in skin organoids
Applicant
Professorin Dr. Nicole Brigitte Fischer
Subject Area
Virology
Term
since 2021
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 443644894
Human polyomaviruses (hPyVs) are prevalent pathogens that establish lifelong persistence in their host. These viruses are opportunistic pathogens and, under immunosuppression, contribute to uncontrolled replication and pathogenesis by reactivating persistence. A major challenge in understanding these viruses and how to combat them is the restricted cell tropism and the associated limitation in infection and pathogenicity models. Thus, the cell types of persistence and the mechanisms contributing to persistence for MCPyV remain to be elucidated. The primary objectives of this project are to elucidate the molecular mechanisms that regulate productive Merkel cell polyomavirus (MCPyV) infection and to identify the factors responsible for the persistence of MCPyV. To achieve this objective, an MCPyV 3D tissue infection model was established, which is based on a previously published iPSC-derived skin organoid model and which represents the first infection model for this human tumor virus. In this model, previously identified central molecular switches of viral infection will be analyzed We will focus on viral mutants of sT, of ALTO, a gene product of an alternative open reading frame, and the viral miRNA, miR-M1, all of which were identified in overexpression experiments as playing a role in maintaining persistence. Of importance is the manipulation of crucial cellular components, namely stimulator of interferon genes (STING) and Interferon gamma inducible protein 16 (IFI16), which we will target with inhibitors or genetic manipulation, e.g. with an auxin-inducible DEGRON system in the organoids. We will employ: single-cell RNA sequencing (scRNA-seq), single-cell ATAC sequencing (scATAC-seq), spatial transcriptomics, and advanced imaging methods available in DEEP-DV. The resulting data, as well as time-resolved data from hPyV in primary cell lines, will be incorporated into the DEEP-DV transcriptional atlas. This will facilitate the identification and understanding of factors and mechanisms of infection and persistence within polyomaviruses and across other human DNA viruses.
DFG Programme
Research Units
