Programmed cell death of infected cells is an ancient pathway of host defense. We now understand that cells can undergo distinct forms of programmed cell death: in addition to apoptosis, necroptosis is a recently-described form of cell suicide that can be induced by viral infection. Necroptosis involves cellular swelling and rupture, and has been hypothesized to trigger inflammatory and immune responses when it occurs in vivo, making the necroptotic pathway a form of host-inducible tissue damage. While this may be an effective strategy in tissues with high regenerative potential and the ability to sustain high inflammatory burdens, it is likely to be detrimental in essential tissues lacking these features, such as the CNS. To test these ideas, we assessed the role of the necroptotic pathway in defense against the neurotropic flavivirus West Nile virus (WNV), and found that while the key necroptotic kinase RIPK3 is required for CNS-intrinsic defense against WNV, necroptotic cell death does not play a role in this pathway. Rather, RIPK3 coordinates a transcriptional response essential for chemokine production and recruitment of inflammatory leukocytes to the brain. Based on these preliminary data, we hypothesized that the CNS represents a tissue that is ?cell death privileged,? in which inflammatory forms of cell death do not represent a beneficial form of host defense. Further, we propose that in these tissue, the pathway of necroptosis has taken on non-death roles in coordinating innate immune defense. To assess this idea, we will take complimentary approaches, first by deleting and then by forcing activation of necroptotic pathway components in the brain.
Aim 1 : We will use a conditional RIPK3flox allele to delete this essential necroptotic kinase within specific cell types in the CNS, then assess both cell death and transcriptional responses to both innate immune stimulation and WNV infection.
Aim 2 : We will make use of a new animal model, developed in our lab, in which RIPK3 can be specifically activated using a ligand drug. We will activate RIPK3 in specific neuronal tissues, then measure resulting transcriptional responses, chemokine production, neuroinflammation and neuropathology. Together, the experiments described here will expand our understanding of the link between cell death and inflammation, and will also identify a novel axis of innate immune response to a neurotropic virus representing a human health threat.

Public Health Relevance

The death of infected cells is an important part of our bodies? defense against infection, but in essential tissues such as those of the central nervous system, cell death can be detrimental. This proposal will explore how the pathways that normally kill infected cells have been re-configured in the brain, using the neuroinvasive West Nile virus as a model pathogen. This work will identify a new, death-independent role of signaling pathways normally associated with cell death, and will also demonstrate an unexpected form of host defense to neuroinvasive viruses.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
Exploratory/Developmental Grants (R21)
Project #
Application #
Study Section
Cellular and Molecular Biology of Glia Study Section (CMBG)
Program Officer
Wong, May
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Washington
Schools of Medicine
United States
Zip Code
Brault, Michelle; Olsen, Tayla M; Martinez, Jennifer et al. (2018) Intracellular Nucleic Acid Sensing Triggers Necroptosis through Synergistic Type I IFN and TNF Signaling. J Immunol 200:2748-2756
Giampazolias, Evangelos; Zunino, Barbara; Dhayade, Sandeep et al. (2017) Mitochondrial permeabilization engages NF-?B-dependent anti-tumour activity under caspase deficiency. Nat Cell Biol 19:1116-1129
Kolb, Joseph P; Oguin 3rd, Thomas H; Oberst, Andrew et al. (2017) Programmed Cell Death and Inflammation: Winter Is Coming. Trends Immunol 38:705-718
Orozco, Susana; Oberst, Andrew (2017) RIPK3 in cell death and inflammation: the good, the bad, and the ugly. Immunol Rev 277:102-112
Daniels, Brian P; Snyder, Annelise G; Olsen, Tayla M et al. (2017) RIPK3 Restricts Viral Pathogenesis via Cell Death-Independent Neuroinflammation. Cell 169:301-313.e11