Dataset Information

Identification of Macrophage Inhibitory Factor as the Parthanatos Dependent AIF Associate Nuclease

ABSTRACT: Inhibition or genetic deletion of poly(ADP-ribose) polymerase-1 (PARP-1) is profoundly protective against a number of toxic insults in many organ systems1,2. In the brain the molecular mechanisms underlying PARP-1-dependent cell death (parthanatos) involves mitochondrial apoptosis-inducing factor (AIF) release and translocation to the nucleus resulting in chromatinolysis3-5. However, it is not known how AIF induces chromatinolysis and neuronal death in parthanatos3. Here we show through an unbiased screen of a ~16K human protein microarray followed by a second siRNA-based screen for protection against PARP-1- dependent cell death, that macrophage migration inhibitory factor (MIF)6 is an AIF associated nuclease that is required for cell death following N-methyl-D-aspartate (NMDA) glutamate receptor excitotoxicity and N-methyl-N-nitroso-N- nitroguanidine (MNNG) toxicity in primary neuronal cultures and focal stroke in mice. MIF contains a central four stranded mixed β-sheet flanked by two α- helices on both sides with an αβββαβ topology which is the structural core of many proteins belonging to the PD-D/E(X)K nuclease-like superfamily. MIF possesses previously unknown Mg2+/Ca2+-dependent nuclease activity, cleaving genomic DNA into large 20-50 kb fragments. AIF recruits MIF to the nucleus after NMDA treatment to induce DNA damage and cell death. Disruption of the AIF-MIF interaction prevents MIF recruitment to the nucleus and protects neurons from NMDA excitotoxicity and ischemic injury. Mutation of Glu22 to Gln in the catalytic nuclease domain completely blocks MIF’s nuclease activity, which almost completely inhibits DNA damage and cell death in vitro and in vivo. These findings reveal that MIF is recruited by AIF to the nucleus where it functions as a parthanatos dependent AIF associated nuclease that induces chromatinolysis and neuronal death, providing a new therapeutic target for stroke and other disorders involving parthanatos.

ORGANISM(S): Homo sapiens

PROVIDER: GSE65110 | GEO | 2016/12/19

SECONDARY ACCESSION(S): PRJNA273007

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Primary cortical neurons were isolated from E15 mice and after 5 days in vitro were untreated or treated for 24 h with mesenchymal stem cell conditioned medium and then untreated or treated for a further 24 h with NMDA. Neuron gene expression was profiled and compared between the four different conditions (neurons, neurons+MSC cm, neurons+NMDA, neurons+MSC cm+NMDA) to investigate the molecular mechanisms of MSC neuroprotection. Mesenchymal stem cells (MSC) promote functional recovery in experimental models of central nervous system (CNS) pathology and are currently being tested in clinical trials for stroke, multiple sclerosis and CNS injury. Their beneficial effects are attributed to activation of endogenous CNS repair processes and immune regulation but their mechanisms of action are poorly understood. Here we investigated the neuroprotective effects of MSC in simplified MSC-neuron co-culture systems and in mice using models of glutamate excitotoxicity. MSC protected primary cortical neurons against glutamate (NMDA) receptor-induced death and conditioned medium from MSC (MSC cm), but not control NIH3T3 cells, was sufficient for this effect. MSC cm neuroprotection in mouse cortical neurons was reduced by neutralizing antibodies to bFGF and associated with altered gene expression in neurons towards an immature phenotype as well as reduced neuronal Grin1, Grin2a and Grin2b mRNA levels in response to NMDA stimulation. Further, MSC cm neuroprotection in rat retinal ganglion cells was associated with absence of glutamate-induced calcium influx. Adoptive transfer of EGFP+MSC in a mouse kainic acid seizure model reduced CA3 neuron damage and hippocampal astrocytosis and resulted in the increased expression of neuronal genes that are upregulated by MSC cm, Bmi1, Ddx4, Ezh1, in the hippocampus. These results show that MSC mediate direct neuroprotection against glutamate excitotoxicity by secreting bFGF, reducing glutamate receptor expression and function and altering neuron gene expression towards an immature pattern, and provide evidence for a link between the therapeutic effects of MSC and the activation of endogenous repair processes following CNS injury. In vitro cultures primary cortical neurons from mice were protected from glutamate excitotoxicity when pre-treated with MSC cm. Global gene expression changes induced in neurons before and after treatment with MSC cm and/or NMDA were investigated using a cDNA spotted macroarray filter. Four samples were analysed in duplicate: neurons alone (untreated), neurons+MSC cm, neurons+NMDA, neurons+MSC cm+NMDA.

Dataset Information

Identification of Macrophage Inhibitory Factor as the Parthanatos Dependent AIF Associate Nuclease

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets