Protective effects of PJ34, a novel, potent inhibitor of poly(ADP-ribose) polymerase (PARP) in in vitro and in vivo models of stroke

Abdelkarim, Galaleldin E.; Gertz, Karen; Harms, Christoph; Katchanov, Juri; Dirnagl, Ulrich; Szabó, Csaba; Endres, Matthias

doi:10.3892/ijmm.7.3.255

Cited by 156 publications

(173 citation statements)

References 0 publications

Supporting

Mentioning

166

Contrasting

Unclassified

Order By: Relevance

“…We further tested the effect of blocking PARP-1 signaling with the pharmacological inhibitor PJ-34 (10 mg/kg intraperitonealy at MCAO onset, Jagtap et al, 2002;Abdelkarim et al, 2001) in ovary-intact WT female mice and obtained results consistent with those from female PARP-1-knockouts (Figure 9). Increased damage was observed in both cortex and striatum of PJ-34-treated mice with corresponding Physiological levels of E2 were restored to ovariectomized (OVX) PARPÀ/À and wild-type (WT) female mice.…”

Section: Pharmacological Inhibition Of Poly-adp Ribose Polymerase Alsmentioning

confidence: 78%

Ischemic Nitric Oxide and Poly (ADP-Ribose) Polymerase-1 in Cerebral Ischemia: Male Toxicity, Female Protection

McCullough

Zeng

Blizzard

et al. 2005

J Cereb Blood Flow Metab

297

326

View full text Add to dashboard Cite

It is well established that tissue damage and functional outcome after experimental or clinical stroke are shaped by biologic sex. We investigated the novel hypothesis that ischemic cell death from neuronally derived nitric oxide (NO) or poly-ADP ribose polymerase (PARP-1) activation is sexually dimorphic and that interruption of these molecular death pathways benefits only the male brain. Female neuronal nitric oxide synthase (nNOS) knockout (nNOSÀ/À) mice exhibited exacerbated histological injury after middle cerebral artery occlusion (MCAO) relative to wild-type (WT) females, unlike the protection observed in male nNOSÀ/À littermates. Similarly, treatment with the nNOS inhibitor (7-nitroindozole, 25 mg/kg) increased infarction in female C57Bl6 WT mice, but protected male mice. The mechanism for this sexually specific response is not mediated through changes in protein expression of endothelial NOS or inducible NOS, or differences in intraischemic cerebral blood flow. Unlike male PARP-1 knockouts (PARP1À/À), female PARP1À/À littermates sustained grossly increased ischemic damage relative to sex-matched WT mice. Treatment with a PARP inhibitor (PJ-34, 10 mg/kg) resulted in identical results. Loss of PARP-1 resulted in reversal of the neuroprotective activity by the female sex steroid, 17b estradiol. These data suggest that the previously described cell death pathways involving NO and PARP ischemic neurotoxicity may be operant solely in male brain and that the integrity of nNO/PARP-1 signaling is paradoxically protective in the female.

show abstract

Section: Pharmacological Inhibition Of Poly-adp Ribose Polymerase Alsmentioning

confidence: 78%

Ischemic Nitric Oxide and Poly (ADP-Ribose) Polymerase-1 in Cerebral Ischemia: Male Toxicity, Female Protection

McCullough

Zeng

Blizzard

et al. 2005

J Cereb Blood Flow Metab

297

326

View full text Add to dashboard Cite

show abstract

“…Treatment of J774 cells with 500 M H 2 O 2 induced a marked enhancement of ADP ribosylating activity, as indicated by the strong nuclear staining of H 2 O 2 -treated cells ( Figure 1C). Pretreatment of cells with the novel potent PARP inhibitor PJ34 (5 M) (Abdelkarim et al 2001;Soriano et al 2001) 30 min before H 2 O 2 exposure prevented enhancement of bio-NAD ϩ staining ( Figure 1D). Similar results were also obtained with other types of cells, including fibroblasts and human keratinocytes (data not shown).…”

Section: Resultsmentioning

confidence: 99%

Detection of Poly(ADP-ribose) Polymerase Activation in Oxidatively Stressed Cells and Tissues Using Biotinylated NAD Substrate

Bakondi

Bai

Szabó

et al. 2002

J Histochem Cytochem.

View full text Add to dashboard Cite

S U M M A R YPoly(ADP-ribose) polymerase (PARP) is a nuclear enzyme activated by DNA damage. Activated PARP cleaves NAD ϩ into nicotinamide and (ADP-ribose) and polymerizes the latter on nuclear acceptor proteins. Over-activation of PARP by reactive oxygen and nitrogen intermediates represents a pathogenetic factor in various forms of inflammation, shock, and reperfusion injury. Using a novel commercially available substrate, 6-biotin-17-nicotinamide-adenine-dinucleotide (bio-NAD ϩ ), we have developed three applications, enzyme cytochemistry, enzyme histochemistry, and cell ELISA, to detect the activation of PARP in oxidatively stressed cells and tissues. With the novel assay we were able to detect basal and hydrogen peroxide-induced PARP activity in J774 macrophages. We also observed that mitotic cells display remarkably elevated PARP activity. Hydrogen peroxide-induced PARP activation could also be detected in wild-type peritoneal macrophages but not in macrophages from PARP-deficient mice. Application of hydrogen peroxide to the skin of mice also induced bio-NAD ϩ incorporation in the keratinocyte nuclei. Hydrogen peroxide-induced PARP activation and its inhibition by pharmacological PARP inhibitors could be detected in J774 cells with the ELISA assay that showed good correlation with the traditional [ 3 H]-NAD incorporation method. The bio-NAD ϩ assays represent sensitive, specific, and non-radioactive alternatives for detection of PARP activation.

show abstract

“…PARP deficient mice are markedly resistant against stroke (21,23). Pharmacological inhibitors of PARP also provide neuroprotection (24)(25)(26)(27)(28)(29)(30), but the degree of protection tends to be less pronounced than the protection seen in PARP-1 deficient animals. Many PARP inhibitors appear to have a limited CNS uptake and cellular residence time (31).…”

Section: Discussionmentioning

confidence: 99%

Poly(ADP-ribose) polymerase inhibition protect neurons and the white matter and regulates the translocation of apoptosis-inducing factor in stroke

et al. 2004

View full text Add to dashboard Cite

Abstract. Focal cerebral ischemia activates the nuclear protein poly(ADP-ribose) polymerase (PARP). Apoptosis-inducing factor (AIF) is a flavoprotein that is normally confined to the mitochondria, but translocates to the nucleus, as shown by in vitro models of neuronal injury. Using INO-1001, a novel potent inhibitor of PARP, we determined the role of PARP activation in the process of AIF translocation in a rat model of focal cerebral ischemia. The potency of INO-1001 as a PARP inhibitor and its cytoprotective potential in oxidantchallenged human neuronal SK-N-MC cells was first confirmed in vitro. PARP inhibition markedly reduced infarct size and improved neurological status in both transient and permanent models of MCA occlusion in Sprague-Dawley rats, with a therapeutic window of 6 h and 2 h in the transient and permanent ischemia models, respectively. The PARP inhibitor reduced the accumulation of poly(ADP-ribose) in the ischemic/ reperfused hemisphere and reduced the accumulation of APP in the white matter of the affected hemisphere, consistently with protection against neuronal necrosis and axonal damage, respectively. Immunohistochemical analysis showed the appearance of AIF labeling in neuronal nuclei of the border zone ischemic area in the striatum after stroke. Cytoplasmatic (axonal) AIF staining was significantly diminished in the necrotic core of the striatum, while it was somewhat enhanced at the borderline ischemic territories of the white matter. Inhibition of PARP with INO-1001 reshifted the location of the apoptotic marker to the axons in the ipsilateral striatum. Thus, PARP inhibition is neuroprotective and regulates the ischemic nuclear translocation of AIF in stroke. IntroductionFocal cerebral ischemia activates the nuclear protein poly(ADP-ribose) polymerase (PARP) by single DNA strand breaks, which leads to energy depletion and cell death and significantly contributes to the pathogenesis of stroke (overviewed in ref. 1).Apoptosis-inducing factor (AIF) is a flavoprotein that is normally confined to the mitochondrial intermembrane space, yet translocates to the nucleus, as shown in experimental models of neuronal injury in vitro. Deletion or inhibition of PARP protects against ischemic brain injury and prevents AIF translocation in vitro (2).Here, using INO-1001, a novel potent inhibitor of PARP (3,4), we determined the role of PARP activation in the process of AIF translocation in stroke in vivo. Materials and methodsIn vitro PARP activity and cytoprotection studies. The potency of the isoindolinone-based novel PARP inhibitor INO-1001 (3,4) was determined in vitro using purified PARP enzyme as previously described using a commercially available PARS inhibition assay kit (Trevigen, Gaithersburg, MD) (3). The potency of INO-1001 was compared with that of PJ34 (5,6), a phenanthridinone-based potent PARP inhibitor, and with 3-aminobenzamide, a prototypical benchmark PARP inhibitor. The assay was carried out in 96-well ELISA plates following manufacturer's instructions. Briefly, wells were coat...

show abstract

Protective effects of PJ34, a novel, potent inhibitor of poly(ADP-ribose) polymerase (PARP) in in vitro and in vivo models of stroke

Cited by 156 publications

References 0 publications

Ischemic Nitric Oxide and Poly (ADP-Ribose) Polymerase-1 in Cerebral Ischemia: Male Toxicity, Female Protection

Ischemic Nitric Oxide and Poly (ADP-Ribose) Polymerase-1 in Cerebral Ischemia: Male Toxicity, Female Protection

Detection of Poly(ADP-ribose) Polymerase Activation in Oxidatively Stressed Cells and Tissues Using Biotinylated NAD Substrate

Poly(ADP-ribose) polymerase inhibition protect neurons and the white matter and regulates the translocation of apoptosis-inducing factor in stroke

Contact Info

Product

Resources

About