Spinal cord injury (SCI) triggers a neuroinflammatory reaction that can aggravate tissue injury (e.g., neuronal death, axonal injury, demyelination) and promote repair (e.g., axon regeneration, remyelination, revascularization). Recently, functionally distinct subsets of macrophages, i.e., """"""""M1"""""""" (pro-inflammatory) and """"""""M2"""""""" (anti-inflammatory) cells have been identified at sites of SCI which may underlie the functional dichotomy. M1 macrophages are neurotoxic and dominate the injured spinal cord for several weeks post-injury. In contrast, M2 macrophages promote axon growth, even in the presence of inhibitory molecules (e.g. CSPG and myelin), without concomitant neurotoxicity. Unfortunately, M2 macrophages populate the injury site for only a few days, eventually becoming overwhelmed by M1 macrophages. Thus, the high M1:M2 ratio might explain why repair of the injured CNS is slow and inefficient relative to tissues in the periphery. We predict that the efficiency and magnitude of spinal cord repair will be improved by modulating the phenotype and function of macrophages that respond to the injury. We will test this hypothesis using genetic loss-of-function (knockout) and gain-of-function (lentiviral) techniques to manipulate expression of the triggering receptor expressed on myeloid cells-2 (TREM2) on resident microglia and myeloid precursor cells, i.e., cells that give rise to monocyte-derived macrophages (MDMs), since overexpressing TREM2 in macrophages induces an M2 phenotype.
In Aim 1 we will examine how TREM2 overexpression or selective knockout on microglia vs. MDMs effects inflammation, motor recovery and anatomical indices of repair after contusive spinal cord injury. By using lentiviral constructs to overexpress TREM2 on MDMs in a model of dorsal spinal hemisection injury in Aim 2, we will determine if TREM2 manipulation influences macrophage effects on myelin/axon phagocytosis, axon regeneration and axonal retraction or """"""""die-back"""""""" of injured axons. Using a model of focal intraspinal demyelination (lysolecithin) in conjunction with the gain of function protocols, we will determine if manipulating macrophage TREM2 affects OPC differentiation and remyelination within the spinal cord in Aim 3. The current proposal outlines proof-of-principle experiments that will advance our understanding of how a distinct molecular signaling pathway, i.e., TREM2, influences the natural course of CNS macrophage function after SCI. Importantly, if data from these studies indicate that manipulating TREM2 confers anatomical or functional benefits with minimal or no adverse effects on CNS structure or function, then it should be feasible to develop similar protocols for human clinical trials. Indeed, intravenous delivery of bone marrow cells (the primary technique to be used in the proposed studies) has already been tried in SCI patients and without adverse effects. Moreover, enzyme replacement therapies, using lentiviral transduction of autologous peripheral blood mononuclear cells, were shown to be safe and effective in human subjects.

Public Health Relevance

The injured spinal cord does not heal properly resulting in chronic paralysis and expensive health care costs for patients with spinal cord injuries. The goal of this proposal is to alter the response to injury to promote increased regeneration, reduced tissue loss, and ultimately increased functional recovery by genetically engineering inflammatory cells that normally respond to injury. Similar therapeutic approaches are being used to cure a variety of neurological diseases;therefore data obtained from our studies could provide the groundwork for a reparative therapy to treat spinal cord injury.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS072304-03
Application #
8488503
Study Section
Clinical Neuroplasticity and Neurotransmitters Study Section (CNNT)
Program Officer
Hicks, Ramona R
Project Start
2011-08-15
Project End
2014-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
3
Fiscal Year
2013
Total Cost
$294,325
Indirect Cost
$101,325
Name
Ohio State University
Department
Neurosciences
Type
Schools of Medicine
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Goldstein, Evan Z; Church, Jamie S; Hesp, Zoe C et al. (2016) A silver lining of neuroinflammation: Beneficial effects on myelination. Exp Neurol 283:550-9
Carpenter, Randall S; Kigerl, Kristina A; Marbourg, Jessica M et al. (2015) Traumatic spinal cord injury in mice with human immune systems. Exp Neurol 271:432-44
Gaudet, Andrew D; Sweet, David R; Polinski, Nicole K et al. (2015) Galectin-1 in injured rat spinal cord: implications for macrophage phagocytosis and neural repair. Mol Cell Neurosci 64:84-94
Puga, Denise A; Tovar, C Amy; Guan, Zhen et al. (2015) Stress exacerbates neuron loss and microglia proliferation in a rat model of excitotoxic lower motor neuron injury. Brain Behav Immun 49:246-54
Gaudet, Andrew D; Popovich, Phillip G (2014) Extracellular matrix regulation of inflammation in the healthy and injured spinal cord. Exp Neurol 258:24-34
Lerch, Jessica K; Puga, Denise A; Bloom, Ona et al. (2014) Glucocorticoids and macrophage migration inhibitory factor (MIF) are neuroendocrine modulators of inflammation and neuropathic pain after spinal cord injury. Semin Immunol 26:409-14
Gensel, John C; Kigerl, Kristina A; Mandrekar-Colucci, Shweta S et al. (2012) Achieving CNS axon regeneration by manipulating convergent neuro-immune signaling. Cell Tissue Res 349:201-13