Existing therapies for autoimmune and inflammatory diseases have minimal effects on resolution and recovery from chronic injury. There is also a fundamental gap in understanding how macrophages regulate cellular phenotype to mediate either injury or repair. The long-term goal is to develop treatments that prevent and reverse chronic inflammatory injury. Using a combination of cellular, genomic, proteomic and in vivo approaches, a novel splice variant of the human sodium channel gene, SCN5A, has been identified as a central regulator of anti-inflammatory macrophage signaling. This splice variant encodes a novel, endosomal cation channel that couples intracellular calcium flux to downstream signaling and gene transcription. Because this splice variant is not expressed in mice, a transgenic knock-in model, the C57BL6cfms-hSCN5A mouse, was developed to study its in vivo function in macrophages. Transfer of bone marrow derived macrophages from these mice mediates clinical recovery in the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS). The objective of this study is to characterize the SCN5A signaling network in macrophages and determine how it regulates clinical recovery in the EAE model of chronic inflammatory disease. The central hypothesis is that persistent expression of SCN5A in macrophages initiates and maintains a cellular phenotype that enhances recovery from inflammatory injury. The rationale for this study is that understanding this mechanism will lead to innovative approaches to prevent and treat chronic inflammatory disease. The hypothesis will be tested in two specific aims: 1. Determine how the SCN5A variant cation channel regulates macrophage phenotype. 2. Analyze how SCN5A+ macrophages mediate recovery during EAE. Based on preliminary data, the working model is that endosomal channel activity mediates short-term biochemical signaling and long-term gene expression patterns that determine and maintain cellular phenotype.
In Aim 1, channel-dependent regulation of cytosolic calcium, cyclic AMP, and activating transcription factor 2 (ATF2) pathways will be examined. The experimental approaches will include a combination of existing biochemical, imaging and molecular techniques in mouse and human primary macrophages.
In Aim 2, the EAE model will be utilized to assess this pathway in vivo and determine how SCN5A+ macrophages mediate recovery from inflammatory injury. In vivo gene and protein expression will be analyzed and correlated with in vitro assessment of SCN5A regulation of macrophage vesicular secretion and intercellular communication. The proposed work is innovative because it represents a new approach to regulate macrophage function and treat chronic inflammatory disease. It is significant because promotion of recovery in MS and other chronic inflammatory diseases represents an unmet therapeutic need.
Multiple sclerosis (MS) is an autoimmune, inflammatory disease that is a common cause of disability in young adults. The VA provides care for approximately 16,000 MS patients a year, and 6,000 are service connected for this condition. The cost of caring for Veterans with MS is very high. Medications used to treat MS are only partially effective and very costly. In addition, they do not prevent progressive worsening of symptoms. The goals of our research are to identify ways to prevent tissue injury and enhance recovery in patients with MS and other inflammatory diseases. Using a mouse model of MS, we have identified a novel channel protein that induces a subtype of immune cells that can migrate to brain and spinal cord lesions, decrease inflammation, and promote recovery. The goals of this proposal are to examine how this channel activates anti-inflammatory pathways in immune cells and how this signaling mechanism works in our animal models.
| Lee, Cara; Jones, Alexis; Kainz, Danielle et al. (2015) A sodium channel variant in Aedes aegypti as a candidate pathogen sensor for viral-associated molecular patterns. Biochem Biophys Res Commun 463:1203-9 |
| Carrithers, Michael D (2014) Innate immune viral recognition: relevance to CNS infections. Handb Clin Neurol 123:215-23 |
| Jones, Alexis; Kainz, Danielle; Khan, Faatima et al. (2014) Human macrophage SCN5A activates an innate immune signaling pathway for antiviral host defense. J Biol Chem 289:35326-40 |
| Rahgozar, Kusha; Wright, Erik; Carrithers, Lisette M et al. (2013) Mediation of protection and recovery from experimental autoimmune encephalomyelitis by macrophages expressing the human voltage-gated sodium channel NaV1.5. J Neuropathol Exp Neurol 72:489-504 |
