Reactivation of latent human cytomegalovirus (HCMV) remains a significant cause of morbidity and mortality in transplant recipients, despite the use of antiviral drugs. Therefore, new approaches are required to reduce the complications from this pathogen. Due to the species specificity of HCMV, we have developed a novel transplant model using the highly related murine CMV (MCMV) as a model to study CMV latency and reactivation in the context of organ transplantation. In this model, MCMV latently infected kidneys are transplanted into immunocompromised NOD.Cg-PrkdcscidIL2rgtm1Wjl/Szj (NSG) mice, which are deficient in T, B, and NK cells. Transplantation of latently infected kidneys into NSG recipients results in reactivation of latent virus in the donor kidney, which disseminates systemically. Reactivation of HCMV is associated with various inflammatory insults. Our preliminary data shows that IL-6, soluble CD40L, IL-18, and LIF are elevated in the plasma of recipient mice with timing consistent with a causal role in reactivation. IL-6 induces reactivation of HCMV in hematopoietic cell models. The central hypothesis of this proposal is that the inflammatory response elicited by the transplanted kidney results in the release of mediators in the recipient mice. The resulting signaling cascade stimulates epigenetic reprogramming of latent viral genomes, transcriptional reactivation of viral immediate early (IE) gene expression, and in immunocompromised recipients, re-entry of latent virus into the lytic replication program. To test this hypothesis, in Aim 1 we will investigate the requirement for candidate factors identified in our preliminary studies, and their downstream signaling intermediates, in inducing reactivation in the NSG model. Our previous studies and those of others show that lytic viral gene expression is repressed in CMV latency due to heterochromatinization of viral genomes. In addition, we have shown that transplant-induced reactivation of MCMV immediate early gene expression is associated with epigenetic reprogramming. Previous studies by our collaborator, Dr. Thomas Kristie, have identified epigenetic inhibitors that suppress reactivation of latent herpes simplex virus.
In Aim 2 we will investigate these promising therapeutic interventions for their ability to prevent reactivation of latent MCMV in the NSG model. Finally, in Aim 3 we will investigate new in vitro models for MCMV latency and differentiation-induced reactivation in murine hematopoietic progenitor cells. This complementary in vitro model would be exceptionally useful for defining molecular mechanisms in the absence of the complexity of the organism. Upon completion, our studies will have identified signaling pathways that lead to epigenetic reprogramming of viral chromatin to reactivate latent MCMV in response to transplantation and potential therapeutic targets.

Public Health Relevance

Cytomegalovirus (CMV) is a ubiquitous herpesvirus which establishes a lifelong latent infection in which the viral DNA is present, but no virus is produced Reactivation of latent virus can cause significant morbidity or mortality in immunocompromised patients. The goal of this proposal is to understand the molecular mechanisms by which the virus reactivates from latency in transplant recipients. These studies may lead to development of new therapies to prevent reactivation of latent CMV, and thus, to prevent CMV disease due to reactivation from latency.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
1R01AI112911-01
Application #
8752165
Study Section
Virology - B Study Section (VIRB)
Program Officer
Beisel, Christopher E
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Northwestern University at Chicago
Department
Surgery
Type
Schools of Medicine
DUNS #
City
Chicago
State
IL
Country
United States
Zip Code
60611