Nonsegmented negative strand RNA viruses: viral RNA cap methylation and potential applications as an anticancer therapy

1 online resource (177 pages) : PDF

2012

University of North Carolina at Charlotte

The viruses of the order Mononegavirales include important human, animal, and plant pathogens and additionally, have great potential as vaccine, oncolytic and gene therapy vectors. This dissertation focuses on two prototypic Mononegavirales, vesicular stomatitis virus (VSV) and Sendai virus (SeV), their virus-encoded cap methylation function, and potential applications as an anticancer therapy. The L protein of Mononegavirales has six conserved domains postulated to constitute the specific enzymatic activities of this multifunctional protein. We conducted a comprehensive mutational analysis by targeting the entire SeV L protein domain VI, creating twenty-four infectious L mutants. Our analysis identified several residues required for successful cap methylation and virus replication. This study confirms structural and functional similarity of this domain across different families of the order Mononegavirales. Additionally, the oncolytic potential of VSV was analyzed for the first time in a panel of human pancreatic ductal adenocarcinoma (PDA) cell lines and compared to other oncolytic viruses. VSV showed superior oncolytic abilities; however, cells were heterogeneous in their susceptibility to virus-induced oncolysis and several cell lines were resistant to all tested viruses. Four cell lines that varied in their permissiveness to VSV were tested in mice, and in vivo results closely mimicked those in vitro. While our results demonstrate VSV is a promising oncolytic agent against PDA, further studies are needed to better understand the molecular mechanisms of resistance to oncolytic virotherapy.

doctoral dissertations

Virology
Biology

Ph.D.

5' Cap Methylation
Oncolytic Virotherapy
Sendai Virus
Vesicular Stomatitis Virus

Biology

Grdzelishvili, Valery

Marriott, Ian
Mukherjee, Pinku
Schrum, Laura
Turner, Michael

Thesis (Ph.D.)--University of North Carolina at Charlotte, 2012.

This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). For additional information, see http://rightsstatements.org/page/InC/1.0/.

Copyright is held by the author unless otherwise indicated.

Murphy_uncc_0694D_10318

http://hdl.handle.net/20.500.13093/etd:1272