Nicotine imparts its effects on cellular function through interaction with neuronal nicotinic acetylcholine receptors (nAChR). Some of these receptors, especially those composed of a4b2 subunits, respond to sustained ligand exposure through the process of upregulation as defined by a substantial increase in the density of high affinity nicotine binding sites. The physiological consequences of upregulation have been linked to many diverse processes ranging from addiction to pathophysiological processes contributing to early phases of Alzheimers disease. An exciting and relatively new role for nAChRs is emerging with regard to the interaction with pro-inflammatory cytokines. In fact, the anti-inflammatory properties of nicotine, acting through various nAChRs, is now widely reported to influence many diseases of inflammatory etiology. However, relatively little is known about the mechanisms controlling the inflammatory:nAChR interaction. The overall goal of this proposal is to define intracellular mechanisms that regulate the interactions between key pro- inflammatory cytokines and defined combinations of nAChR subtypes.
SPECIFIC AIM 1. Hypothesis: TNFa-activation of the p38MAPK pathway and/or IL-1b activation of PKA pathways enhance a4b2 expression and these are antagonized by pathways involving PI3K/Akt signaling. In this Aim we will pursue the definition of intracellular pathways initiated by TNFa and/or IL-1b to enhance nicotine-mediated upregulation of nAChRa4b2.
SPECIFIC AIM 2. Hypothesis: Direct phosphorylation of a4 and the presence of other nAChR subunits modify the pro-inflammatory cytokine signals mediating enhanced upregulation of a4b2.
This Aim will determine the impact of: 1) modifying PKA phosphorylation sites in a4;2) introducing (or deleting) other nAChRs (specifically a7 or a5);and 3) determining effects of TNFa or IL-1b intracellular signaling leading to enhanced a4b2 upregulation in cultured neurons.
SPECIFIC AIM 3. Hypothesis: Interactions between cells of the CNS such as neurons and microglia (Mg) direct the outcome of the overall inflammatory:nAChR response. Neurons and microglia will be co-cultured in different ratios to dissect the relative contribution of paracrine, autocrine or juxtacrine interactions in modulating the TNFa and IL-1b-mediated enhancement of a4b2-upregulation in a mixed cell system.

Public Health Relevance

We are interested in understanding the interaction between nicotine and inflammation. We will determine the signal transduction pathways cytokines use to promote or inhibit enhanced upregulation of nicotinic receptors. Two major cytokines, tumor necrosis factor alpha (TNFa) and interleukin-1b (IL-1b) enhance nicotine-induced upregulation of the high affinity nicotine receptor termed nAChRa4b2. The upregulation response is correlated with addiction behaviors and multiple pathologies of the brain such as Alzheimer's disease. Understanding how the inflammatory system and response to nicotine interact will impact upon how we approach these problems with therapeutic agents.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
5R01DA025057-04
Application #
8310245
Study Section
Molecular Neuropharmacology and Signaling Study Section (MNPS)
Program Officer
Wu, Da-Yu
Project Start
2009-09-18
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
4
Fiscal Year
2012
Total Cost
$358,912
Indirect Cost
$118,837
Name
University of Utah
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Gahring, Lorise C; Myers, Elizabeth; Palumbos, Sierra et al. (2014) Nicotinic receptor Alpha7 expression during mouse adrenal gland development. PLoS One 9:e103861
Gahring, Lorise C; Enioutina, Elena Y; Myers, Elizabeth J et al. (2013) Nicotinic receptor alpha7 expression identifies a novel hematopoietic progenitor lineage. PLoS One 8:e57481
Rogers, Scott W; Tvrdik, Petr; Capecchi, Mario R et al. (2012) Prenatal ablation of nicotinic receptor alpha7 cell lineages produces lumbosacral spina bifida the severity of which is modified by choline and nicotine exposure. Am J Med Genet A 158A:1135-44
Gahring, Lorise C; Vasquez-Opazo, Gustavo A; Rogers, Scott W (2010) Choline promotes nicotinic receptor alpha4 + beta2 up-regulation. J Biol Chem 285:19793-801
Gahring, Lorise C; Rogers, Scott W (2010) Nicotinic receptor subunit alpha5 modifies assembly, up-regulation, and response to pro-inflammatory cytokines. J Biol Chem 285:26049-57
Gahring, Lorise C; Osborne, Amber V; Reed, Michelle et al. (2010) Neuronal nicotinic alpha7 receptors modulate early neutrophil infiltration to sites of skin inflammation. J Neuroinflammation 7:38