Rett syndrome is a devastating neurological disorder, almost exclusively affecting girls. Rett is predominantly caused by mutations in an X-linked gene encoding methyl-CpG-binding protein (MECP)2. Until recently, the etiology of the disease was assumed to be purely neuronal. However, the role of glia in the pathology of Rett syndrome (and of other CNS diseases) has now been recognized. Expression of wild type Mecp2 in astrocytes of Mecp2-null hosts has been shown to dramatically ameliorate disease pathology. Microglia were also recently suggested to play a role in Rett pathophysiology; Mecp2-null microglia were reported to be toxic to neurons through production of high levels of glutamate. Along these lines, it is generally well accepted by now that non-neuronal cells of the CNS are critically important for brain function. Our preliminary data that serves as a basis for this proposal, demonstrates the unique role myeloid cells play in arrest of Rett pathology. Transplantation of wild type bone marrow into irradiation-conditioned Mecp2-null hosts resulted in engraftment of brain parenchyma by bone marrow-derived myeloid cells of microglial phenotype, and arrest of disease development. However, when cranial irradiation was blocked by lead shield, and microglial engraftment was prevented, disease was not arrested. Similarly, targeted expression of Mecp2 in myeloid cells, driven by Lysmcre on an Mecp2-null background, dramatically attenuated disease symptoms. In irradiated mice, newly- engrafted brain mononuclear phagocytes produced high levels of insulin-like growth factor 1 (IGF-1), unlike resident mutant microglia, possibly one underlying molecular mechanism of rescue. Interestingly, however, inhibition of phagocytic activity using annexin V that blocks phosphatydilserine residues on apoptotic targets and prevents their recognition and engulfment by tissue-resident phagocytes, abolished disease arrest. Based on our preliminary results, we hypothesize that Mecp2-null microglia are incapable of providing neurotrophic support and are insufficient to the task of debris clearance (including synaptic pruning, elimination of dead cells etc.), thus contributing to the ongoing pathophysiology seen in Rett syndrome. Our data implicate microglia as major players in Rett pathophysiology, and suggest that bone marrow transplantation might offer a feasible therapeutic approach for this devastating disorder. However, a further understanding of the role played by microglia in disease pathology and repair on a molecular level is crucial for efficient translation of these studies to clinic. This proposal is aimed to study the underlying mechanism of microglia-mediated arrest of Rett pathology by (1) establishing the role of microglia in Rett pathology and repair using genetic and pharmacological approaches; (2) addressing the relative contribution of peripheral immune-derived IGF-1 and microglia-derived IGF-1 in arrest of Rett disease, using bone marrow transplantation and genetic crosses; and (3) testing the hypothesis that the deficiency in phagocytic clearance by microglia may underlie, in part, the pathophysiology of Rett using mice deficient in macrophage phagocytic activity as bone marrow donors.

Public Health Relevance

Rett syndrome is a devastating neurological disorder, almost exclusively affecting girls. Rett is predominantly caused by mutations in an X-linked gene encoding methyl-CpG- binding protein (MeCP)2. Until recently, the etiology of the disease was assumed to be purely neuronal. However, the role of glia in the pathology of Rett syndrome has now been recognized. Our results show that bone marrow transplantation into mouse model of Rett disease (Mecp2-null mice) arrests disease progression. Bone marrow transplantation could, therefore, be developed into a promising therapeutic approach for Rett disorder but an underlying mechanism needs to be better understood. This grant is aimed to understand the role played by myeloid cells in Rett arrest after bone marrow transplantation or expression of wild type Mecp2 in myeloid cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
4R01NS081026-05
Application #
9067524
Study Section
Developmental Brain Disorders Study Section (DBD)
Program Officer
Mamounas, Laura
Project Start
2012-09-01
Project End
2017-05-31
Budget Start
2016-06-01
Budget End
2017-05-31
Support Year
5
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Virginia
Department
Neurosciences
Type
Schools of Medicine
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Cronk, James C; Filiano, Anthony J; Louveau, Antoine et al. (2018) Peripherally derived macrophages can engraft the brain independent of irradiation and maintain an identity distinct from microglia. J Exp Med 215:1627-1647
Louveau, Antoine; Filiano, Anthony J; Kipnis, Jonathan (2018) Meningeal whole mount preparation and characterization of neural cells by flow cytometry. Curr Protoc Immunol 121:
Cronk, James C; Herz, Jasmin; Kim, Taeg S et al. (2017) Influenza A induces dysfunctional immunity and death in MeCP2-overexpressing mice. JCI Insight 2:e88257
Gadani, Sachin P; Smirnov, Igor; Smith, Ashtyn T et al. (2017) Characterization of meningeal type 2 innate lymphocytes and their response to CNS injury. J Exp Med 214:285-296
Cronk, James C; Derecki, Noel C; Litvak, Vladimir et al. (2016) Unexpected cellular players in Rett syndrome pathology. Neurobiol Dis 92:64-71
Filiano, Anthony J; Xu, Yang; Tustison, Nicholas J et al. (2016) Unexpected role of interferon-? in regulating neuronal connectivity and social behaviour. Nature 535:425-9
Filiano, Anthony J; Gadani, Sachin P; Kipnis, Jonathan (2015) Interactions of innate and adaptive immunity in brain development and function. Brain Res 1617:18-27
Cronk, James C; Derecki, Noël C; Ji, Emily et al. (2015) Methyl-CpG Binding Protein 2 Regulates Microglia and Macrophage Gene Expression in Response to Inflammatory Stimuli. Immunity 42:679-91
Derecki, Noël C; Katzmarski, Natalie; Kipnis, Jonathan et al. (2014) Microglia as a critical player in both developmental and late-life CNS pathologies. Acta Neuropathol 128:333-45
Schwartz, Michal; Kipnis, Jonathan; Rivest, Serge et al. (2013) How do immune cells support and shape the brain in health, disease, and aging? J Neurosci 33:17587-96

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