Rift Valley Fever is a clinically and agriculturally important illness caused by Rift Valley Fever Virus (RVFV), a member of the Bunyaviridae endemic primarily in sub-Saharan Africa. Because of the severe illness it causes in humans and because of recent instances of natural spread outside of Africa, it has been identified as emerging disease agent and/or a potential agent of bioterrorism. The symptoms of RVF in humans can include severe fever, malaise, neurological damage, liver damage, hemorrhagic events, and death. Currently there are no clinically proven treatments for RVFV infection. Development of antiviral strategies requires understanding of the viral replication cycle so that specific steps can be targeted. In the research proposed here, we will characterize an essential molecular interaction in the RVFV replication cycle wherein nucleocapsid protein must recognize and bind to viral RNA sequences. To achieve this goal, we will perform truncation analysis on viral RNA transcripts to determine the minimum requirements for efficient RNA-protein binding. We will also perform a combinatorial in vitro selection scheme (SELEX) to obtain a high affinity RNA aptamer(s) to nucleocapsid protein. Sequence analysis of the aptamers will shed light on nucleocapsid target recognition and will subsequently be used to construct a tool to screen drugs that inhibit the nucleocapsid-RNA interaction. Specifically, the aptamer will be ligated to an RNA module that emits a fluorescent signal when the aptamer is displaced from its nucleocapsid binding site. The drug screening tool will then be used to screen compounds at NSRB. Molecules giving promising results in the aptamer displacement assay will be tested against RVFV replication in cell culture. Owing to its central role in replication, nucleocapsid protein has been recognized as a good potential antiviral target. A novel class of drug targeted against nucleocapsid protein has recently been shown to be effective in inhibiting replication of respiratory syncytial virus, also a negative-sense RNA virus, strongly supporting the potential for developing nucleocapsid as a viable drug target in RVFV (Chapman et al, 2007). This research project fits within the RMRCE Integrated Research Focus on Viral Therapeutics, and will interact directly with PI Brian Gowen (RP 3.2) and the NSRB drug screening core.

Public Health Relevance

Rift Valley fever virus causes significant mortality and morbidity in Africa and the Arabian peninsula and poses a serious threat to other regions of the world due to changing habitats of its insect hosts and because it could be introduced intentionally as a part of a bioterrorism effort. The goal of the research proposed here is to biochemically characterize a specific molecular interaction in the virus replication cycle to exploit it as a drug target. We will screen libraries of small molecules to find drugs that interfere with this essential step.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54AI065357-09
Application #
8465812
Study Section
Special Emphasis Panel (ZAI1-DDS-M)
Project Start
Project End
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
9
Fiscal Year
2013
Total Cost
$259,072
Indirect Cost
$51,621
Name
Colorado State University-Fort Collins
Department
Type
DUNS #
785979618
City
Fort Collins
State
CO
Country
United States
Zip Code
80523
Gibson, Christopher C; Zhu, Weiquan; Davis, Chadwick T et al. (2015) Strategy for identifying repurposed drugs for the treatment of cerebral cavernous malformation. Circulation 131:289-99
Wang, Hong; Siddharthan, Venkatraman; Hall, Jeffery O et al. (2014) Autonomic deficit not the cause of death in West Nile virus neurological disease. Clin Auton Res 24:15-23
Scharton, Dionna; Bailey, Kevin W; Vest, Zachary et al. (2014) Favipiravir (T-705) protects against peracute Rift Valley fever virus infection and reduces delayed-onset neurologic disease observed with ribavirin treatment. Antiviral Res 104:84-92
Shives, Katherine D; Beatman, Erica L; Chamanian, Mastooreh et al. (2014) West nile virus-induced activation of mammalian target of rapamycin complex 1 supports viral growth and viral protein expression. J Virol 88:9458-71
Calvert, Amanda E; Dixon, Kandice L; Delorey, Mark J et al. (2014) Development of a small animal peripheral challenge model of Japanese encephalitis virus using interferon deficient AG129 mice and the SA14-14-2 vaccine virus strain. Vaccine 32:258-64
Richert, Laura E; Rynda-Apple, Agnieszka; Harmsen, Ann L et al. (2014) CD11cýýý cells primed with unrelated antigens facilitate an accelerated immune response to influenza virus in mice. Eur J Immunol 44:397-408
Soffler, Carl; Bosco-Lauth, Angela M; Aboellail, Tawfik A et al. (2014) Pathogenesis of percutaneous infection of goats with Burkholderia pseudomallei: clinical, pathologic, and immunological responses in chronic melioidosis. Int J Exp Pathol 95:101-19
Porta, Jason; Jose, Joyce; Roehrig, John T et al. (2014) Locking and blocking the viral landscape of an alphavirus with neutralizing antibodies. J Virol 88:9616-23
Jones-Carson, Jessica; Zweifel, Adrienne E; Tapscott, Timothy et al. (2014) Nitric oxide from IFN?-primed macrophages modulates the antimicrobial activity of ?-lactams against the intracellular pathogens Burkholderia pseudomallei and Nontyphoidal Salmonella. PLoS Negl Trop Dis 8:e3079
Phillips, Aaron T; Schountz, Tony; Toth, Ann M et al. (2014) Liposome-antigen-nucleic acid complexes protect mice from lethal challenge with western and eastern equine encephalitis viruses. J Virol 88:1771-80

Showing the most recent 10 out of 181 publications