Cheung, E. C. et al. Apoptosis-inducing factor is a key factor in neuronal cell death propagated by BAX-dependent and BAX-independent mechanisms. J. Neurosci. 25, 1324-1334

Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario, Canada
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience (Impact Factor: 6.34). 03/2005; 25(6):1324-34. DOI: 10.1523/JNEUROSCI.4261-04.2005
Source: PubMed


Mitochondria release proteins that propagate both caspase-dependent and caspase-independent cell death pathways. AIF (apoptosis-inducing factor) is an important caspase-independent death regulator in multiple neuronal injury pathways. Presently, there is considerable controversy as to whether AIF is neuroprotective or proapoptotic in neuronal injury, such as oxidative stress or excitotoxicity. To evaluate the role of AIF in BAX-dependent (DNA damage induced) and BAX-independent (excitotoxic) neuronal death, we used Harlequin (Hq) mice, which are hypomorphic for AIF. Neurons carrying double mutations for Hq/Apaf1-/- (apoptosis proteases-activating factor) are impaired in both caspase-dependent and AIF-mediated mitochondrial cell death pathways. These mutant cells exhibit extended neuroprotection against DNA damage, as well as glutamate-induced excitotoxicity. Specifically, AIF is involved in NMDA- and kainic acid- but not AMPA-induced excitotoxicity. In vivo excitotoxic studies using kainic acid-induced seizure showed that Hq mice had significantly less hippocampal damage than wild-type littermates. Our results demonstrate an important role for AIF in both BAX-dependent and BAX-independent mechanisms of neuronal injury.

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    • "Strikingly, we made the observation that MK-2206 exposure led to an induction of pro-apoptotic protein AIF and its translocation from mitochondria to the nucleus of the GEO cells (Figure  6A, C). It has been reported that AIF is responsible for caspase-independent death in ovarian cancer cells [30,36,37]. AIF is localized in the mitochondria but upon activation it translocates to the nucleus and causes DNA fragmentation [38]. "
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    ABSTRACT: There is extensive evidence for the role of aberrant cell survival signaling mechanisms in cancer progression and metastasis. Akt is a major component of cell survival-signaling mechanisms in several types of cancer. It has been shown that activated Akt stabilizes XIAP by S87 phosphorylation leading to survivin/XIAP complex formation, caspase inhibition and cytoprotection of cancer cells. We have reported that TGFbeta/PKA/PP2A-mediated tumor suppressor signaling regulates Akt phosphorylation in association with the dissociation of survivin/XIAP complexes leading to inhibition of stress-dependent induction of cell survival. IGF1R-dependent colon cancer cells (GEO and CBS) were used for the study. Effects on cell proliferation and cell death were determined in the presence of MK-2206. Xenograft studies were performed to determine the effect of MK-2206 on tumor volume. The effect on various cell death markers such as XIAP, survivin, AIF, Ezrin, pEzrin was determined by western blot analysis. Graph pad 5.0 was used for statistical analysis. P < 0.05 was considered significant. We characterized the mechanisms by which a novel Akt kinase inhibitor MK-2206 induced cell death in IGF1R-dependent colorectal cancer (CRC) cells with upregulated PI3K/Akt signaling in response to IGF1R activation. MK-2206 treatment generated a significant reduction in tumor growth in vivo and promoted cell death through two mechanisms. This is the first report demonstrating that Akt inactivation by MK-2206 leads to induction of and mitochondria-to-nuclear localization of the Apoptosis Inducing Factor (AIF), which is involved in caspase-independent cell death. We also observed that exposure to MK-2206 dephosphorylated Ezrin at the T567 site leading to the disruption of Akt-pEzrin-XIAP cell survival signaling. Ezrin phosphorylation at this site has been associated with malignant progression in solid tumors. The identification of these 2 novel mechanisms leading to induction of cell death indicates MK-2206 might be a potential clinical candidate for therapeutic targeting of the subset of IGF1R-dependent cancers in CRC.
    BMC Cancer 03/2014; 14(1):145. DOI:10.1186/1471-2407-14-145 · 3.36 Impact Factor
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    • "The autophagic nature of this event was tested by confocal analysis on MOI 50-infected hippocampal cultures at 12 h after double immunofluorescence with cytC (green) and LC3 (red), two markers used routinely to detect activation of mitophagy in neurons (Cai et al., 2012; Van Humbeeck et al., 2011). CytC is a dynamic mitochondrial marker localized to intermembrane space of these organelles and its cytosolic release is a very late event occurring during excitotoxic death (Cheung et al., 2005; Cregan et al., 2002), as previously reported to occur after the exogenous expression of NH 2 htau in neurons (Amadoro et al., 2006). LC3 (Microtubule-Associated Protein 1A/ 1B-light chain 3) is a known autophagic indicator that localizes in its lipidated form (LC3II) to autophagosomal membranes (Klionsky et al., 2008, 2012). "
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    ABSTRACT: Functional as well as structural alterations in mitochondria size, shape and distribution are precipitating, early events in progression of Alzheimer's Disease (AD). We reported that a 20–22 kDa NH2-tau fragment (aka NH2htau), mapping between 26 and 230 amino acids of the longest human tau isoform, is detected in cellular and animal AD models and is neurotoxic in hippocampal neurons. The NH2htau –but not the physiological full-length protein– interacts with Aβ at human AD synapses and cooperates with it in inhibiting the mitochondrial ANT-1-dependent ADP/ATP exchange. Here we show that the NH2htau also adversely affects the interplay between the mitochondria dynamics and their selective autophagic clearance. Fragmentation and perinuclear mislocalization of mitochondria with smaller size and density are early found in dying NH2htau-expressing neurons. The specific effect of NH2htau on quality control of mitochondria is accompanied by (i) net reduction in their mass in correlation with a general Parkin-mediated remodeling of membrane proteome; (ii) their extensive association with LC3 and LAMP1 autophagic markers; (iii) bioenergetic deficits and (iv) in vitro synaptic pathology. These results suggest that NH2htau can compromise the mitochondrial biology thereby contributing to AD synaptic deficits not only by ANT-1 inactivation but also, indirectly, by impairing the quality control mechanism of these organelles.
    Neurobiology of Disease 02/2014; 62:489–507. DOI:10.1016/j.nbd.2013.10.018 · 5.08 Impact Factor
    • "Although Aif-null mouse embryos show lethality, they do not seem to have increased cell numbers63. Cerebellar granule cells from Hq mice show partial resistance to cell death induced by serum-withdrawal5263. Cortical and hippocampal neurons from Hq mice are resistant to death induced by N-methyl-D-Aspartate (NMDA) and glutamate646566, which reflects the results of in vivo excitotoxic studies showing protection of hippocampal neurons of Hq mice from kainic acid-induced seizures64. Intracellular ROS levels possibly regulate the cleavage and release of Aif which is supported by an observation that treatment with an Manganese Superoxide Dismutase (MnSOD) mimetic attenuated nuclear translocation of Aif and ischaemic cell death in neurons67. "
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    ABSTRACT: Multiple checkpoints regulating finely balanced death-versus-survival decisions characterize both thymic development and peripheral homeostasis of T lymphocytes. While exploring the mechanisms of T cell death involved at various stages during the life of a T cell, we have observed and reported a variety of non-redundant roles for apoptosis inducing factor (Aif), a mitochondrial flavoprotein. Aif is ubiquitously expressed in all cell lineages and functions as an NADH oxidase in its mitochondrial location. It is released following the mitochondrial death signals, whereupon it translocates to the nucleus, binds to DNA and causes large-scale DNA fragmentation. During T cell development, Aif is important for developing thymocytes to navigate the double negative (DN)3 to DN4 transition (beta-selection), via its oxidoreductase property which protects the rapidly proliferating cells from death due to reactive oxygen species (ROS). In peripheral mature T cells, Aif deficiency leads to an increased susceptibility of T cell blasts to activation induced cell death (AICD), possibly mediated by its antioxidant function, and decreased sensitivity to neglect-induced death (NID). Thus, Aif seems to have pro-apoptotic and anti-apoptotic roles in the same lineage in different contexts and at different stages. Surprisingly, in the closely related B lymphocyte lineage, Aif deficiency does not result in any abnormality. These findings generate the possibility of specific T cell dysfunction in human disease caused by Aif deficiency, as well as in mitochondriopathies due to other causes. Also, these data raise questions regarding the basis of lineage-specific consequences of the dysfunction/deficiency of apparently ubiquitous molecules.
    The Indian Journal of Medical Research 11/2013; 138(5):577-90. · 1.40 Impact Factor
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