The overall goal of this project is to determine whether the induction of the inflammatory response involving activation of caspase-1 and processing of interleukin-12 (IL-12) following moderate cervical spinal cord injury (SCI) in the rat is mediated by the formation of multiprotein inflammatory caspase-activating platforms termed inflammasomes. Our recent work has shown that the NLRP1 inflammasome is expressed in rat spinal cord neurons and regulates IL-12 and IL-18 production. We have found that in SCI neutralization of the adaptor protein apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (ASC) reduced inflammation and resulted in significant tissue sparing and functional improvement.
The Specific Aims of this proposal will extend these results to determine the cellular and molecular mechanisms regulating inflammasome activation in glia.
Aim 1 will identify the structural components and protein associations of inflammasomes in primary cultures of neurons, astrocytes, and microglia damaged by stretch injury. These studies will delineate which cell types express inflammasomes and will provide insight into the mechanisms leading to cytokine secretion caused by caspase-1 after injury.
Aim 2 will identify key inflammasome signaling components in distinct CNS cell types using lentiviral short hairpin (shRNA) vectors that target inflammasome genes and proteins. Related experiments will determine the cellular distribution and temporal sequence of protein expression of inflammasome proteins in spinal cords of normal and injured rats. Experiments in Specific Aim 4 will determine if disruption of inflammasome signaling by antibody neutralization of inflammasome components or lentiviral shRNA silencing of inflammasome genes results in inhibition of inflammasome signaling, thus leading to improved histopathological and behavioral outcomes. These studies will provide critical information about temporal and spatial activation patterns of the inflammasome and identify relevant therapeutic targets to control inflammation following SCI and other neurodegenerative disorders.

Public Health Relevance

Spinal cord injury (SCI) occurs throughout the world with an annual incidence of 15 to 40 million people per year and results in lifelong disabilities. The main cause of SCI in young males is motor-vehicle accidents and most spinal cord traumas occur at the cervical spinal cord level. The proposed research seeks to reduce cell death by inflammation that occurs within days of SCI, and to devise therapies to treat SCI and help in long-term recovery of function. The development of new treatments that can be given in the first hours after SCI may prevent or reverse much of the spinal cord damage resulting from SCI with a goal of restoring normal functioning in behavior.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS059836-03
Application #
8231379
Study Section
Clinical Neuroplasticity and Neurotransmitters Study Section (CNNT)
Program Officer
Hicks, Ramona R
Project Start
2010-04-01
Project End
2013-09-30
Budget Start
2012-04-01
Budget End
2013-09-30
Support Year
3
Fiscal Year
2012
Total Cost
$299,852
Indirect Cost
$103,852
Name
University of Miami School of Medicine
Department
Physiology
Type
Schools of Medicine
DUNS #
052780918
City
Coral Gables
State
FL
Country
United States
Zip Code
33146
Kigerl, Kristina A; de Rivero Vaccari, Juan Pablo; Dietrich, W Dalton et al. (2014) Pattern recognition receptors and central nervous system repair. Exp Neurol 258:5-16
de Rivero Vaccari, Juan Pablo; Dietrich, W Dalton; Keane, Robert W (2014) Activation and regulation of cellular inflammasomes: gaps in our knowledge for central nervous system injury. J Cereb Blood Flow Metab 34:369-75
de Rivero Vaccari, Juan Pablo; Brand 3rd, Frank J; Sedaghat, Christina et al. (2014) RIG-1 receptor expression in the pathology of Alzheimer's disease. J Neuroinflammation 11:67
Minkiewicz, Julia; de Rivero Vaccari, Juan Pablo; Keane, Robert W (2013) Human astrocytes express a novel NLRP2 inflammasome. Glia 61:1113-21
Stojadinovic, Olivera; Minkiewicz, Julia; Sawaya, Andrew et al. (2013) Deep tissue injury in development of pressure ulcers: a decrease of inflammasome activation and changes in human skin morphology in response to aging and mechanical load. PLoS One 8:e69223
de Rivero Vaccari, Juan Pablo; Bastien, Dominic; Yurcisin, Geoffrey et al. (2012) P2X4 receptors influence inflammasome activation after spinal cord injury. J Neurosci 32:3058-66
de Rivero Vaccari, Juan Pablo; Minkiewicz, Julia; Wang, Xiaoliang et al. (2012) Astrogliosis involves activation of retinoic acid-inducible gene-like signaling in the innate immune response after spinal cord injury. Glia 60:414-21
Wang, Xiaoliang; Vaccari, Juan Pablo de Rivero; Wang, Handong et al. (2012) Activation of the nuclear factor E2-related factor 2/antioxidant response element pathway is neuroprotective after spinal cord injury. J Neurotrauma 29:936-45