• Users Online: 1370
  • Home
  • Print this page
  • Email this page

 Table of Contents  
Year : 2018  |  Volume : 13  |  Issue : 2  |  Page : 252-256

Dabrafenib, an inhibitor of RIP3 kinase-dependent necroptosis, reduces ischemic brain injury

1 Ottawa Hospital Research Institute; University of Ottawa, Brain and Mind Institute; Canadian Partnership for Storke Recovery, Ottawa, Canada
2 Ottawa Hospital Research Institute; Canadian Partnership for Storke Recovery, Ottawa, Canada
3 University of Ottawa Heart Institute, Ottawa, Canada

Date of Acceptance10-Nov-2017
Date of Web Publication16-Mar-2018

Correspondence Address:
Hsiao-Huei Chen
Ottawa Hospital Research Institute; University of Ottawa, Brain and Mind Institute; Canadian Partnership for Storke Recovery, Ottawa
Login to access the Email id

Source of Support: This study was supported by grants from the Heart and Stroke Foundation of Canada (HHC, AFRS), the Canadian Institutes of Health Research (to HHC and AFRS). HHC was also supported by a Mid-Career Investigator Award from the Heart and Stroke Foundation of Ontario, Conflict of Interest: None

DOI: 10.4103/1673-5374.226394

Rights and Permissions

Ischemic brain injury triggers neuronal cell death by apoptosis via caspase activation and by necroptosis through activation of the receptor-interacting protein kinases (RIPK) associated with the tumor necrosis factor-alpha (TNF-α)/death receptor. Recent evidence shows RIPK inhibitors are neuroprotective and alleviate ischemic brain injury in a number of animal models, however, most have not yet undergone clinical trials and safety in humans remains in question. Dabrafenib, originally identified as a B-raf inhibitor that is currently used to treat melanoma, was later revealed to be a potent RIPK3 inhibitor at micromolar concentrations. Here, we investigated whether Dabrafenib would show a similar neuroprotective effect in mice subjected to ischemic brain injury by photothrombosis. Dabrafenib administered intraperitoneally at 10 mg/kg one hour after photothrombosis-induced focal ischemic injury significantly reduced infarct lesion size in C57BL6 mice the following day, accompanied by a markedly attenuated upregulation of TNF-α. However, subsequent lower doses (5 mg/kg/day) failed to sustain this neuroprotective effect after 4 days. Dabrafenib blocked lipopolysaccharides-induced activation of TNF-α in bone marrow-derived macrophages, suggesting that Dabrafenib may attenuate TNF-α-induced necroptotic pathway after ischemic brain injury. Since Dabrafenib is already in clinical use for the treatment of melanoma, it might be repurposed for stroke therapy.

Keywords: ischemic brain injury; inflammation; macrophage; Dabrafenib; tumor necrosis factor-alpha; photothrombosis; receptor-interacting protein kinases; necroptosis; microglia; stroke; neural regeneration

How to cite this article:
Cruz SA, Qin Z, Stewart AF, Chen HH. Dabrafenib, an inhibitor of RIP3 kinase-dependent necroptosis, reduces ischemic brain injury. Neural Regen Res 2018;13:252-6

How to cite this URL:
Cruz SA, Qin Z, Stewart AF, Chen HH. Dabrafenib, an inhibitor of RIP3 kinase-dependent necroptosis, reduces ischemic brain injury. Neural Regen Res [serial online] 2018 [cited 2021 Jun 12];13:252-6. Available from: http://www.nrronline.org/text.asp?2018/13/2/252/226394

  Introduction Top

Stroke following ischemic brain injury is a leading cause of death worldwide. Ischemic brain injury resulting from disruption of blood supply to the brain triggers immediate cell death by necrosis, but also activates additional cellular mechanisms, including caspase-dependent apoptosis and caspase-independent necroptosis that propagate a wave of delayed cell death (Tovar-y-Romo et al., 2016).

Release of glutamate from dying neurons and astrocytes triggers excitotoxicity (Schock et al., 2008), a hyper-excitation of glutamatergic receptors that disrupts intracellular Ca2+ homeostasis, increases production of reactive oxygen species and release of cytochromce c from mitochondria that activates caspases (Degterev et al., 2003). Injured or dead neurons also activate local brain-resident macrophages (microglia) that secrete chemokines and inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), to recruit more immune cells in attempt for tissue repairs. However, inflammatory cytokines secreted from immune cells also induce cell death through the activation of the Fas/TNFR family of death-domain receptors (DRs).

Although activation of the caspase family of cysteine proteases is essential to mediate DR signal transduction and execution of apoptosis (Degterev et al., 2003), many studies have shown that caspase inhibitors, including the pan-caspase inhibitor zVAD.fmk, fail to block DR-induced cell death (Schulze-Osthoff et al., 1994; Kawahara et al., 1998; Vercammen et al., 1998; Khwaja and Tatton, 1999; Holler et al., 2000; Matsumura et al., 2000; Chan et al., 2003). Emerging evidence points to non-apoptotic caspase-independent cell death pathway called necroptosis, in which DRs activate receptor-interacting protein kinase 1 (RIPK1) and a signaling cascade involving the phosphorylation of RIPK3 by RIPK1 (Weinlich et al., 2017). Ischemic brain injury activates necroptotic cell death by activating RIPK3 (Vieira et al., 2014). The RIPK1 inhibitor necrostatin-1 limits experimentally induced ischemic brain injury in mice (Degterev et al., 2005; Degterev et al., 2008), but necrostatin-1 has not yet undergone phase 1 clinical trials for safety and tolerance in humans.

It has emerged that Dabrafenib, an inhibitor of B-raf currently approved for the treatment of melanoma (Hauschild et al., 2012), is also an effective high affinity inhibitor of RIPK3 that blocks TNF-α-induced necroptosis (Li et al., 2014b). In human vascular endothelial cells, Dabrafenib markedly attenuates the activation of TNF-α (Jung et al., 2016). Here, we tested whether Dabrafenib has a neuroprotective effect in an experimentally induced stroke model.

  Materials and Methods Top

Ischemic brain injury by photothrombosis in mice

C57BL6 adult male mice aged 2–3 months, weighing approximately 33 g, were fed with regular chow, and randomly assigned to experimental groups. All procedures were carried out with the approval of the animal care and use committee of the University of Ottawa (OGHRI-49), according to animal use guidelines of the Canadian Council on Animal Care (www.ccac.ca/en/standards/guidelines/). Photothrombosis-induced focal ischemic brain injury was carried out in two-month-old mice, using the protocol of Watson et al. (1985) with the modifications we previously described (Cruz et al., 2017). Mice were anesthesized in an induction chamber ventilated with 5% isoflurane gas (AbbVie Corporation, St-Laurent, Quebec, Canada), transferred to a stereotaxic frame and anesthesia was maintained with 1.5% isoflurane with a nose mask. A midline incision of the scalp exposed a 1 × 1 mm2 region of the skull. Rose Bengal (Sigma-Aldrich, Oakville, Ontario, Canada, 10 mg/mL in PBS, made fresh by vortexing and filtered) was administered by i.p. injection (100 mg/kg) and five minutes later, a laser was used to induce thrombosis. A collimated green laser (532 nm wavelength at 20 mW, Beta Instruments, Zaventem, Belgium) was placed 2 cm above the skull, positioned 0.7 mm anteroposterior and 2 mm mediolateral relative to the Bregma to target the sensorimotor cortex and used to illuminate the skull for 10 minutes. The skin wound was closed using cyanoacrylate glue, and mice were monitored until they regained consciousness. Sham operated mice received Rose Bengal and underwent surgery without laser illumination.

One hour after photothrombosis, Dabrafenib (10 mg/kg, Selleck, Houston, TX, USA) or vehicle (dimethyl sulfoxide, DMSO) was delivered by i.p. injection in 150 μL 80% DMSO in saline. Daily maintenance doses of Dabrafenib (5 mg/kg) were administered, prior to sacrifice on day 4.

Quantification of infarction

Mice were sacrificed and perfused with saline followed by 4% paraformaldehyde and brains were isolated and sectioned on a cryostatmicrotome for histology. Infarct volumes were determined by cresyl violet staining to reveal Nissl bodies of living neurons in 10 coronal sections (20 μm thick) sampled every 10 sections over a 2 mm distance overlapping the area of infarction, as previously described (Schock et al., 2008). Infarct volume was calculated by stacking infarct areas in serial sections using the ImageJ software (NIH, Bethesda, MD, USA). Lesion volumes were normalized to brain volumes of the corresponding sections to control for edema. Investigators were blinded to genotype.


Areas surrounding the infarction were visualized with primary antibodies incubated at 4°C overnight to the macrophage marker Iba1 (WAKO Chemicals, Richmond, VA, USA, #019-19741, rabbit anti-rat,1:500 dilution) and neuronal marker NeuN (Millipore Canada Ltd., Etobicoke, Ontario, Canada, MAB377, mouse anti-mouse, 1:500 dilution) by immunofluorescence as previously described (Chen et al., 2007). Cy2 and Cy3 conjugated secondary antibodies were obtained from Jackson Laboratories (Bar Harbour, ME, USA) and used at 1:1,000 dilution. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Iba1-positive cells were imaged on a Zeiss (Oberkoken, Germany) AxioImager Z1 fluorescence microscope and counted in three independent fields at 20× magnification from six sections per mouse.

Bone marrow-derived macrophage cultures

Bone marrow-derived macrophages (BMDM) were cultured for 6 days in HyClone™ Dulbecco's Modified Eagles Medium (GE Healthcare Life Sciences, Missassauga, Ontario, Canada) conditioned with medium from L926 cells, as we described (Chen et al., 2015) and treated with lipopolysaccharides (LPS; Sigma-Aldrich, Oakville, Ontario, Canada, 100 ng/mL) with or without Dabrafenib (10 μM) for 4 hours. RNA was isolated for quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis.


Total RNA from BMDM or frozen brain tissue was extracted using the Qiagen (Germantown, MD, USA) RNeasy Mini Kit (Hari et al., 2017). qRT-PCR was conducted as described previously (Pandey et al., 2013; Qin et al., 2015), and the results were normalized to beta-actin. In brief, 1 μg of RNA was used for cDNA synthesis (ABM 5x All-in-one RT MasterMix, #AG490), and 4 μL of 1:4 diluted cDNA was used for qPCR with the following primers: TNF-α: (F) 5′-CCA CCA CGC TCT TCT GTC TAC-3′, (R) 5′-AGG GTC TGG GCC ATA GAA CT-3′ and normalized to beta-actin: (F) 5′-CCT TCT GAC CCA TTC CCA CC, (R) 5′-GCT TCT TTG CAG CTC CTT CG-3′ for cultured BMDM ornormalized to GAPDH: (F) 5′-TGT TCC TAC CCC CAA TGT GT-3′, (R) 5′-TGT GAG GGA GAT GCT CAG TG-3′ for brain tissue.

Statistical analysis

All results are presented as the mean ± SEM. For infarct volumes, percentages were normalized by arcsine transformation. For between-group comparisons of fold changes in TNF-α expression, values were normalized by log transformation. Two-way analysis of variance (ANOVA) was used to compare the main effects of treatment (vehicle versus Dabrafenib) and time (1 day versus 4 days) and the interaction (treatment versus time) following photothrombosis on infarct volumes. For experiments in BMDM, two-way ANOVA was used to compare the effects of treatment (vehicle versus Dabrafenib) and immune stimulation (with or without LPS challenge) as well as the interaction on TNF-α mRNA levels. For post-hoc analysis, the Bonferroni correction was applied for multiple pairwise testing using two-tailed Student's t-test. Differences in means were considered significant at P < 0.05.

Results and Discussion

Dabrafenib (10 mg/kg) administered one hour after photothrombosis-induced focal ischemic injury significantly reduced infarct lesion size in C57Bl6 mice one day after infarction [Figure 1]. Two-factor ANOVA revealed a main effect of Dabrafenib (F = 16.458, P = 0.00036) and time (F = 10.131, P = 0.0035) to reduce infarction volume. Administration of a daily maintenance dose of Dabrafenib (5 mg/kg) for 3 days did not further reduce the infarction on day 4. Ischemic injury induced infiltration of Iba1-positive cells (microglia/macrophages), as revealed by immunofluorescence [Figure 2]A. Although Dabrafenib treatment did not result in a significant reduction in Iba1+ microglia recruitment to the site of injury [Figure 2]B, Dabrafenib treatment attenuated up-regulation of TNF-α mRNA levels one day after photothrombosis ([Figure 3]A, Dabrafenib effect: F = 5.479, P = 0.037; time effect: F = 15.412, P = 0.002). This result suggests that the effect of Dabrafenib on reduction of infarct volume is related to attenuated local inflammation after ischemic injury.
Figure 1: Dabrafenib attenuated ischemic brain injury in mice.
(A) Dabrafenib (Dab) was intraperitoneally injected after photothrombosis-induced ischemic brain injury as shown in the diagram. (B) Infarction volumes (white areas) were measured by Cresyl violet staining at one day and four days after infarction and expressed as a percentage of total brain volume (C). *P < 0.05. t-test was used with Bonferroni correction (n = 8 mice per group). Veh: Vehicle.

Click here to view
Figure 2: Immunofluorescence did not reveal an obvious difference in microglial recruitment between vehicle (Veh) and Dabrafenib (Dab)-treated mice one day after ischemic injury.
(A) The peri-infarct area shows Iba1-immunopositive (Iba1+) macrophages/microglia recruited to the area surrounding the ischemic core (IC). Scale bar: 100 μm. Sham-operated control shows few Iba+ cells in the same cortical region. (B) Iba1+ cell counts. n = 4 mice per group. NeuN: Neuronal marker. DAPI: 4′,6-diamidino-2-phenylindole, nuclei marker; Iba1: ionized calcium binding adaptor molecule 1.

Click here to view
Figure 3: Dabrafenib (Dab) attenuated tumor necrosis factor-alpha (TNF-α) activation in the brain.
(A)TNF-α mRNA levels in the brain measured by quantitative reverse transcription-polymerase chain reaction are elevated after photothrombosis. Dab treatment attenuated this effect 1 day after initial dose (10 mg/kg). n = 4 mice per group. (B) Dab (10 μM) blocked TNF-α mRNA expression in lipopolysaccharides (LPS)-stimulated bone marrow-derived macrophages. n = 4 independent cultures per group. *P < 0.05. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; Veh: Vehicle.

Click here to view

To test whether Dabrafenib affects TNF-α activation in macrophages, bone marrow-derived macrophages were stimulated with LPS in the absence or presence of Dabrafenib. Dabrafenib blocked LPS-induced activation of TNF-α expression in macrophages ([Figure 3]B; Dabrafenib effect: F = 642.54, P = 8.632E-12; LPS effect: 465.39, P = 5.735E-11; interaction: F = 220.49, P = 4.36E-9). This in vitro data may explain why markedly lower TNF-α mRNA level was detected one day after Dabrafenib treatment (10 mg/kg, 1 hour after photothrombosis) despite similar numbers of Iba1-positive microglia/macrophages recruited to the ischemic site. However, we are puzzled by the lack of a sustained effect of Dabrafenib 4 days after stroke. This suggests that the maintenance dose (5 mg/kg/day) we used was not sufficient. It is noteworthy that in humans, Dabrafenib is administered in two 150 mg doses daily (equivalent to 4 mg/kg body weight per day, given a 75 kg average weight) for the treatment of melanoma (Hauschild et al., 2012). On the other hand, in mice, daily oral administration of 10 mg/kg was effective at suppressing tumor growth (King et al., 2013), a dose we also found to be effective when administered intraperitoneally. Future studies are needed to test whether higher maintenance doses of Dabrafenib can further reduce infarct volumes and favor recovery.

Of note, vehicle treated mice showed regression of the infarction volume (expressed as a percentage of brain volume to correct for edema) at 4 days compared to day 1. Regression of infarction volume after photothrombosis has been reported by us (Cruz et al., 2017) and others (Li et al., 2014a). Local inflammation likely contributes to the size of infarction, since we previously showed that mice with sustained inflammation have delayed regression of infarction (Cruz et al., 2017).This would be consistent with the acute anti-inflammatory effect of the higher dose of Dabrafenib we observed 1 day after stroke.

Since the tumour necrosis factor receptor (TNFR) is a major death receptor involved in the activation of necroptosis, we propose a model whereby inhibition of TNF-α production from macrophages/microglia likely accounts for part of the protective effect by Dabrafenib to limit stroke injury (see diagram; [Figure 4]). Ischemic stroke results in an abrupt deprivation of nutrient supplies that quickly leads to irreversible damage in the core of the affected area. Damage-associated molecular patterns (DAMPS) released from dying cells are sensed by Toll-like receptors (TLRs). TLRs are a receptor family broadly expressed in many cell types including neurons and glia cells. Activation of TLRs triggers gene expression through several transcription factors. Interferon regulatory factors (IRFs) are the main targets of TLRs that initiate the innate immune response. Activation of TLRs/IRFs increases inflammatory cytokine expression, including TNF-α, leading to the recruitment of immune cells, like macrophages, to the site of injury for tissue repair (Kariko et al., 2004). On the other hand, TNF-α activates the TNF receptor on neurons and causes programmed cell death via caspase-dependent and independent pathways (Degterev et al., 2003).
Figure 4: Diagram of Dabrafenib action to block TNF-α-mediated activation of necroptosis induced after ischemic injury.
Dabrafenib is proposed to inhibit tumor necrosis factor-alpha (TNF-α) production from macrophages and microglia. At the same time, Dabrafenib can also inhibit RIPK3 in neurons and prevent necroptosis. TNFR: TNF-α-receptor; RIPK: receptor-interacting protein kinase; DAMP: damage-associated molecular patterns; TLR: toll-like receptors; IRF1: interferon regulatory factor-1; myd88: myeloid differentiation primary response gene 88.

Click here to view

TNF-α further activates IRF1 (Jaruga et al., 2004) that increases expression of RIPK proteins and amplifies TNF-α-induced RIPK3 activation and necroptosis (McComb et al., 2014). Consistent with the activation of TNF-α in ischemic brain injury, IRF1 expression is also elevated in ischemic neurons in mice after experimentally-induced stroke and in post-mortem neurons of stroke patients (Alexander et al., 2003). In line with these observations, mice that lack IRF1 have a smaller infarction after experimental focal brain ischemia (Iadecola et al., 1999).

Blocking necroptosis with the RIPK1 inhibitor necrostatin-1was found to be neuroprotective in ischemic brain injury (Degterev et al., 2005). However, necrostatin-1 and related RIPK1 inhibitors also exhibit some toxicity (Takahashi et al., 2012) and most have not undergone clinical trials for safety in humans. Recently, Dabrafenib was identified as a RIPK3 inhibitor (Li et al., 2014b). Importantly, Dabrafenib has already undergone clinical trials and is an FDA approved drug for the treatment of melanoma in humans (Hauschild et al., 2012). Thus, to the best of our knowledge, our study is the first to demonstrate that Dabrafenib attenuates stroke injury in a preclinical mouse model. Our finding is exciting because it suggests that Dabrafenib could readily be repurposed for stroke therapy.

Whether the effect of Dabrafenib is mediated by its inhibition of RIPK3 remains to be demonstrated. Another RIPK3 inhibitor was recently shown to protect cultured neurons against injury caused by oxygen and glucose deprivation (Fayaz et al., 2016). However, it should be pointed out that ablation of RIPK3 had no effect on brain injury caused by middle cerebral artery occlusion followed by reperfusion (Newton et al., 2016). The discrepancy between the RIPK3 knockout study and our findings could arise from compensatory effects of genetic ablation of RIPK3, or from a difference in stroke models, since photothrombosis does not involve reperfusion. We also cannot exclude that B-raf inhibition by Dabrafenib may itself be beneficial to stroke injury, as has been shown for other B-raf inhibitors (Ahnstedt et al., 2011). Lastly, additional behaviour studies will be required to test whether higher or more frequent doses of Dabrafenib can improve sensorimotor functional recovery after ischemic brain injury.[35]

Author contributions: SAC, ZQ, AFRS and HHC contributed to research design. SAC and ZQ conducted the experiments and acquired data. SAC, AFRS and HHC analyzed the data. AFRS and HHC wrote the manuscript. All authors reviewed and approved the final version of this paper.

Conflicts of interest: None declared.

Financial support: This study was supported by grants from the Heart and Stroke Foundation of Canada (HHC, AFRS), the Canadian Institutes of Health Research (to HHC and AFRS). HHC was also supported by a Mid-Career Investigator Award from the Heart and Stroke Foundation of Ontario.

Research ethics: All procedures were carried out with the approval of the animal care and use committee of the University of Ottawa (OGHRI-49), according to animal use guidelines of the Canadian Council on Animal Care (www.ccac.ca/en/standards/guidelines/).

Data sharing statement: Datasets analyzed during the current study are available from the corresponding author on reasonable request.

Plagiarism check: Checked twice by iThenticate.

Peer review: Externally peer reviewed.

Open peer reviewer: P Aguilera, National Institute of Neurology and Neurosurgery “Manuel Velasco Suárez”, Mexico.

Additional file: Open peer review report 1[Additional file 1].

Funding: This study was supported by grants from the Heart and Stroke Foundation of Canada (HHC, AFRS), the Canadian Institutes of Health Research (to HHC and AFRS). HHC was also supported by a Mid-Career Investigator Award from the Heart and Stroke Foundation of Ontario.

  References Top

Ahnstedt H, Saveland H, Nilsson O, Edvinsson L (2011) Human cerebrovascular contractile receptors are upregulated via a B-Raf/MEK/ERK-sensitive signaling pathway. BMC Neurosci 12:5.  Back to cited text no. 1
Alexander M, Forster C, Sugimoto K, Clark HB, Vogel S, Ross ME, Iadecola C (2003) Interferon regulatory factor-1 immunoreactivity in neurons and inflammatory cells following ischemic stroke in rodents and humans. Acta Neuropathol 105:420-424.  Back to cited text no. 2
Chan FK, Shisler J, Bixby JG, Felices M, Zheng L, Appel M, Orenstein J, Moss B, Lenardo MJ (2003) A role for tumor necrosis factor receptor-2 and receptor-interacting protein in programmed necrosis and antiviral responses. J Biol Chem 278:51613-51621.  Back to cited text no. 3
Chen HH, Schock SC, Xu J, Safarpour F, Thompson CS, Stewart AF (2007) Extracellular ATP-dependent upregulation of the transcription cofactor LMO4 promotes neuron survival from hypoxia. Exp Cell Res 313:3106-3116.  Back to cited text no. 4
Chen HH, Keyhanian K, Zhou X, Vilmundarson RO, Almontashiri NA, Cruz S, Pandey NR, Yap NL, Ho T, Stewart CA, Huang H, Hari A, Geoffrion M, McPherson R, Rayner KJ, Stewart AF (2015) IRF2BP2 reduces macrophage inflammation and susceptibility to atherosclerosis. Circ Res 117:671-683.  Back to cited text no. 5
Cruz SA, Hari A, Qin Z, Couture P, Huang H, Lagace DC, Stewart AFR, Chen HH (2017) Loss of IRF2BP2 in microglia increases inflammation and functional deficits after focal ischemic brain injury. Front Cell Neurosci 11:201.  Back to cited text no. 6
Degterev A, Boyce M, Yuan J (2003) A decade of caspases. Oncogene 22:8543-8567.  Back to cited text no. 7
Degterev A, Huang Z, Boyce M, Li Y, Jagtap P, Mizushima N, Cuny GD, Mitchison TJ, Moskowitz MA, Yuan J (2005) Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury. Nat Chem Biol 1:112-119.  Back to cited text no. 8
Degterev A, Hitomi J, Germscheid M, Ch'en IL, Korkina O, Teng X, Abbott D, Cuny GD, Yuan C, Wagner G, Hedrick SM, Gerber SA, Lugovskoy A, Yuan J (2008) Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol 4:313-321.  Back to cited text no. 9
Fayaz SM, Suvanish Kumar VS, Davis CK, Rajanikant GK (2016) Novel RIPK3 inhibitors discovered through a structure-based approach exert post-ischemic neuroprotection. Mol Divers 20:719-728.  Back to cited text no. 10
Hari A, Cruz SA, Qin Z, Couture P, Vilmundarson RO, Huang H, Stewart AFR, Chen HH (2017) IRF2BP2-deficient microglia block the anxiolytic effect of enhanced postnatal care. Sci Rep 7:9836.  Back to cited text no. 11
Hauschild A, Grob JJ, Demidov LV, Jouary T, Gutzmer R, Millward M, Rutkowski P, Blank CU, Miller WH Jr, Kaempgen E, Martín-Algarra S, Karaszewska B, Mauch C, Chiarion-Sileni V, Martin AM, Swann S, Haney P, Mirakhur B, Guckert ME, Goodman V, et al. (2012) Dabrafenib in BRAF-mutated metastatic melanoma: a multicentre, open-label, phase 3 randomised controlled trial. Lancet 380:358-365.  Back to cited text no. 12
Holler N, Zaru R, Micheau O, Thome M, Attinger A, Valitutti S, Bodmer JL, Schneider P, Seed B, Tschopp J (2000) Fas triggers an alternative, caspase-8-independent cell death pathway using the kinase RIP as effector molecule. Nat Immunol 1:489-495.  Back to cited text no. 13
Iadecola C, Salkowski CA, Zhang F, Aber T, Nagayama M, Vogel SN, Ross ME (1999) The transcription factor interferon regulatory factor 1 is expressed after cerebral ischemia and contributes to ischemic brain injury. J Exp Med 189:719-727.  Back to cited text no. 14
Jaruga B, Hong F, Kim WH, Gao B (2004) IFN-gamma/STAT1 acts as a proinflammatory signal in T cell-mediated hepatitis via induction of multiple chemokines and adhesion molecules: a critical role of IRF-1. Am J Physiol Gastrointest Liver Physiol 287:G1044-1052.  Back to cited text no. 15
Jung B, Kang H, Lee W, Noh HJ, Kim YS, Han MS, Baek MC, Kim J, Bae JS (2016) Anti-septic effects of dabrafenib on HMGB1-mediated inflammatory responses. BMB Rep 49:214-219.  Back to cited text no. 16
Kariko K, Weissman D, Welsh FA (2004) Inhibition of toll-like receptor and cytokine signaling--a unifying theme in ischemic tolerance. J Cereb Blood Flow Metab 24:1288-1304.  Back to cited text no. 17
Kawahara A, Ohsawa Y, Matsumura H, Uchiyama Y, Nagata S (1998) Caspase-independent cell killing by Fas-associated protein with death domain. J Cell Biol 143:1353-1360.  Back to cited text no. 18
Khwaja A, Tatton L (1999) Resistance to the cytotoxic effects of tumor necrosis factor alpha can be overcome by inhibition of a FADD/caspase-dependent signaling pathway. J Biol Chem 274:36817-36823.  Back to cited text no. 19
King AJ, Arnone MR, Bleam MR, Moss KG, Yang J, Fedorowicz KE, Smitheman KN, Erhardt JA, Hughes-Earle A, Kane-Carson LS, Sinnamon RH, Qi H, Rheault TR, Uehling DE, Laquerre SG (2013) Dabrafenib; preclinical characterization, increased efficacy when combined with trametinib, while BRAF/MEK tool combination reduced skin lesions. PLoS One 8:e67583.  Back to cited text no. 20
Li H, Zhang N, Lin HY, Yu Y, Cai QY, Ma L, Ding S (2014a) Histological, cellular and behavioral assessments of stroke outcomes after photothrombosis-induced ischemia in adult mice. BMC Neurosci 15:58.  Back to cited text no. 21
Li JX, Feng JM, Wang Y, Li XH, Chen XX, Su Y, Shen YY, Chen Y, Xiong B, Yang CH, Ding J, Miao ZH (2014b) The B-Raf(V600E) inhibitor dabrafenib selectively inhibits RIP3 and alleviates acetaminophen-induced liver injury. Cell Death Dis 5:e1278.  Back to cited text no. 22
Matsumura H, Shimizu Y, Ohsawa Y, Kawahara A, Uchiyama Y, Nagata S (2000) Necrotic death pathway in Fas receptor signaling. J Cell Biol 151:1247-1256.  Back to cited text no. 23
McComb S, Cessford E, Alturki NA, Joseph J, Shutinoski B, Startek JB, Gamero AM, Mossman KL, Sad S (2014) Type-I interferon signaling through ISGF3 complex is required for sustained Rip3 activation and necroptosis in macrophages. Proc Natl Acad Sci U S A 111:E3206-3213.  Back to cited text no. 24
Newton K, Dugger DL, Maltzman A, Greve JM, Hedehus M, Martin-McNulty B, Carano RA, Cao TC, van Bruggen N, Bernstein L, Lee WP, Wu X, DeVoss J, Zhang J, Jeet S, Peng I, McKenzie BS, Roose-Girma M, Caplazi P, Diehl L, et al. (2016) RIPK3 deficiency or catalytically inactive RIPK1 provides greater benefit than MLKL deficiency in mouse models of inflammation and tissue injury. Cell Death Differ 23:1565-1576.  Back to cited text no. 25
Pandey NR, Zhou X, Qin Z, Zaman T, Gomez-Smith M, Keyhanian K, Anisman H, Brunel JM, Stewart AF, Chen HH (2013) The LIM domain only 4 protein is a metabolic responsive inhibitor of protein tyrosine phosphatase 1B that controls hypothalamic leptin signaling. J Neurosci 33:12647-12655.  Back to cited text no. 26
Qin Z, Zhou X, Pandey NR, Vecchiarelli HA, Stewart CA, Zhang X, Lagace DC, Brunel JM, Beique JC, Stewart AF, Hill MN, Chen HH (2015) Chronic stress induces anxiety via an amygdalar intracellular cascade that impairs endocannabinoid signaling. Neuron 85:1319-1331.  Back to cited text no. 27
Schock SC, Xu J, Duquette PM, Qin Z, Lewandowski AJ, Rai PS, Thompson CS, Seifert EL, Harper ME, Chen HH (2008) Rescue of neurons from ischemic injury by peroxisome proliferator-activated receptor-gamma requires a novel essential cofactor LMO4. J Neurosci 28:12433-12444.  Back to cited text no. 28
Schulze-Osthoff K, Krammer PH, Droge W (1994) Divergent signalling via APO-1/Fas and the TNF receptor, two homologous molecules involved in physiological cell death. EMBO J 13:4587-4596.  Back to cited text no. 29
Takahashi N, Duprez L, Grootjans S, Cauwels A, Nerinckx W, DuHadaway JB, Goossens V, Roelandt R, Van Hauwermeiren F, Libert C, Declercq W, Callewaert N, Prendergast GC, Degterev A, Yuan J, Vandenabeele P (2012) Necrostatin-1 analogues: critical issues on the specificity, activity and in vivo use in experimental disease models. Cell Death Dis 3:e437.  Back to cited text no. 30
Tovar-y-Romo LB, Penagos-Puig A, Ramirez-Jarquin JO (2016) Endogenous recovery after brain damage: molecular mechanisms that balance neuronal life/death fate. J Neurochem 136:13-27.  Back to cited text no. 31
Vercammen D, Brouckaert G, Denecker G, Van de Craen M, Declercq W, Fiers W, Vandenabeele P (1998) Dual signaling of the Fas receptor: initiation of both apoptotic and necrotic cell death pathways. J Exp Med 188:919-930.  Back to cited text no. 32
Vieira M, Fernandes J, Carreto L, Anuncibay-Soto B, Santos M, Han J, Fernandez-Lopez A, Duarte CB, Carvalho AL, Santos AE (2014) Ischemic insults induce necroptotic cell death in hippocampal neurons through the up-regulation of endogenous RIP3. Neurobiol Dis 68:26-36.  Back to cited text no. 33
Watson BD, Dietrich WD, Busto R, Wachtel MS, Ginsberg MD (1985) Induction of reproducible brain infarction by photochemically initiated thrombosis. Ann Neurol 17:497-504.  Back to cited text no. 34
Weinlich R, Oberst A, Beere HM, Green DR (2017) Necroptosis in development, inflammation and disease. Nat Rev Mol Cell Biol 18:127-136.  Back to cited text no. 35


  [Figure 1], [Figure 2], [Figure 3], [Figure 4]

This article has been cited by
1 Ring Closure Strategy Leads to Potent RIPK3 Inhibitors
Shuwei Wu,Chen Xu,Kaijiang Xia,Yu Lin,Sheng Tian,Haikuo Ma,Yuting Ji,Fang Zhu,Sudan He,Xiaohu Zhang
European Journal of Medicinal Chemistry. 2021; : 113327
[Pubmed] | [DOI]
2 Neuronal protein-tyrosine phosphatase 1B hinders sensory-motor functional recovery and causes affective disorders in two different focal ischemic stroke models
ShellyA Cruz,Zhaohong Qin,KonradM Ricke,AlexandreF.R. Stewart,Hsiao-Huei Chen
Neural Regeneration Research. 2021; 16(1): 129
[Pubmed] | [DOI]
3 Cell death in head and neck cancer pathogenesis and treatment
Martina Raudenská,Jan Balvan,Michal Masarík
Cell Death & Disease. 2021; 12(2)
[Pubmed] | [DOI]
4 RIPK3: A New Player in Renal Fibrosis
Ying Shi,Xinming Chen,Chunling Huang,Carol Pollock
Frontiers in Cell and Developmental Biology. 2020; 8
[Pubmed] | [DOI]
5 Inhibitors Targeting RIPK1/RIPK3: Old and New Drugs
Sofie Martens,Sam Hofmans,Wim Declercq,Koen Augustyns,Peter Vandenabeele
Trends in Pharmacological Sciences. 2020; 41(3): 209
[Pubmed] | [DOI]
6 Targeting necroptosis in anticancer therapy: mechanisms and modulators
Ying Wu,Guoqiang Dong,Chunquan Sheng
Acta Pharmaceutica Sinica B. 2020;
[Pubmed] | [DOI]
7 RIP3/MLKL-mediated neuronal necroptosis induced by methamphetamine at 39°C
Li-Min Guo,Zhen Wang,Shi-Ping Li,Mi Wang,Wei-Tao Yan,Feng-Xia Liu,Chu-Dong Wang,Xu-Dong Zhang,Dan Chen,Jie Yan,Kun Xiong
Neural Regeneration Research. 2020; 15(5): 865
[Pubmed] | [DOI]
8 Emerging immune and cell death mechanisms in stroke: saponins as therapeutic candidates
Laura García-Pupo,Emily Van San,René Delgado-Hernández,Tom Vanden Berghe,Wim Vanden Berghe
Brain, Behavior, & Immunity - Health. 2020; : 100152
[Pubmed] | [DOI]
9 The Role of RIPK1 and RIPK3 in Cardiovascular Disease
Elise DeRoo,Ting Zhou,Bo Liu
International Journal of Molecular Sciences. 2020; 21(21): 8174
[Pubmed] | [DOI]
10 RSK3 mediates necroptosis by regulating phosphorylation of RIP3 in rat retinal ganglion cells
Mi Wang,Hao Wan,Shuchao Wang,Lvshuang Liao,Yanxia Huang,Limin Guo,Fengxia Liu,Lei Shang,Jufang Huang,Dan Ji,Xiaobo Xia,Bin Jiang,Dan Chen,Kun Xiong
Journal of Anatomy. 2020;
[Pubmed] | [DOI]
11 RIPK3 blockade attenuates tubulointerstitial fibrosis in a mouse model of diabetic nephropathy
Ying Shi,Chunling Huang,Yongli Zhao,Qinghua Cao,Hao Yi,Xinming Chen,Carol Pollock
Scientific Reports. 2020; 10(1)
[Pubmed] | [DOI]
12 Lytic Cell Death Mechanisms in Human Respiratory Syncytial Virus-Infected Macrophages: Roles of Pyroptosis and Necroptosis
Lori Bedient,Swechha Mainali Pokharel,Kim R. Chiok,Indira Mohanty,Sierra S. Beach,Tanya A. Miura,Santanu Bose
Viruses. 2020; 12(9): 932
[Pubmed] | [DOI]
13 RIPK3 blockade attenuates kidney fibrosis in a folic acid model of renal injury
Ying Shi,Chunling Huang,Hao Yi,Qinghua Cao,Yongli Zhao,Jason Chen,Xinming Chen,Carol Pollock
The FASEB Journal. 2020; 34(8): 10286
[Pubmed] | [DOI]
14 Non-Apoptotic Cell Death Signaling Pathways in Melanoma
Mariusz L. Hartman
International Journal of Molecular Sciences. 2020; 21(8): 2980
[Pubmed] | [DOI]
15 Protective effect of dabrafenib on renal ischemia-reperfusion injury in vivo and in vitro
Shu-su Liu,Yan-yi Chen,Shao-xia Wang,Qing Yu
Biochemical and Biophysical Research Communications. 2019;
[Pubmed] | [DOI]
16 Autophagy, anoikis, ferroptosis, necroptosis, and endoplasmic reticulum stress: Potential applications in melanoma therapy
Milad Ashrafizadeh,Reza Mohammadinejad,Shima Tavakol,Zahra Ahmadi,Sahar Roomiani,Majid Katebi
Journal of Cellular Physiology. 2019;
[Pubmed] | [DOI]
17 Therapeutic potential of a TrkB agonistic antibody for ischemic brain injury
Fang Han,Xiaoming Guan,Wei Guo,Bai Lu
Neurobiology of Disease. 2019; 127: 570
[Pubmed] | [DOI]
18 Serum and cerebrospinal fluid tau protein level as biomarkers for evaluating acute spinal cord injury severity and motor function outcome
Ying Tang,Hong-Liang Liu,Ling-Xia Min,Hao-Shi Yuan,Lei Guo,Peng-Bo Han,Yu-Xin Lu,Jian-Feng Zhong,Dong-Lin Wang
Neural Regeneration Research. 2019; 14(5): 896
[Pubmed] | [DOI]
19 Necroptosis: a crucial pathogenic mediator of human disease
Mary E. Choi,David R. Price,Stefan W. Ryter,Augustine M. K. Choi
JCI Insight. 2019; 4(15)
[Pubmed] | [DOI]
20 B-RAFV600E Inhibitor Dabrafenib Attenuates RIPK3-Mediated Necroptosis and Promotes Functional Recovery after Spinal Cord Injury
Takehiro Sugaya,Haruo Kanno,Michiharu Matsuda,Kyoichi Handa,Satoshi Tateda,Taishi Murakami,Hiroshi Ozawa,Eiji Itoi
Cells. 2019; 8(12): 1582
[Pubmed] | [DOI]
21 Current translational potential and underlying molecular mechanisms of necroptosis
Tamás Molnár,Anett Mázló,Vera Tslaf,Attila Gábor Szöllosi,Gabriella Emri,Gábor Koncz
Cell Death & Disease. 2019; 10(11)
[Pubmed] | [DOI]
22 Identification of a novel class of RIP1/RIP3 dual inhibitors that impede cell death and inflammation in mouse abdominal aortic aneurysm models
Ting Zhou,Qiwei Wang,Noel Phan,Jun Ren,Huan Yang,Conner C. Feldman,John B. Feltenberger,Zhengqing Ye,Scott A. Wildman,Weiping Tang,Bo Liu
Cell Death & Disease. 2019; 10(3)
[Pubmed] | [DOI]
23 Guidelines for evaluating myocardial cell death
Paras K Mishra,Adriana Adameová,Joseph A. Hill,Christopher P. Baines,Peter M. Kang,James Downey,Jagat Narula,Masafumi Takahashi,Antonio Abbate,Hande C. Piristine,Sumit Kar,Shi Su,Jason K Higa,Nicholas K. Kawasaki,Takashi Matsui
American Journal of Physiology-Heart and Circulatory Physiology. 2019;
[Pubmed] | [DOI]
24 RIP1/RIP3-regulated necroptosis as a target for multifaceted disease therapy (Review)
Yuping Liu,Ting Liu,Tiantian Lei,Dingding Zhang,Suya Du,Lea Girani,Dandan Qi,Chen Lin,Rongsheng Tong,Yi Wang
International Journal of Molecular Medicine. 2019;
[Pubmed] | [DOI]
25 Molecular Communication of a Dying Neuron in Stroke
Berta Puig,Santra Brenna,Tim Magnus
International Journal of Molecular Sciences. 2018; 19(9): 2834
[Pubmed] | [DOI]
26 The Pathogenesis of Necroptosis-Dependent Signaling Pathway in Cerebral Ischemic Disease
Yang Xu,Ji Zhang,Lingsong Ma,Shoucai Zhao,Shizun Li,Tingting Huang,Zhaohu Chu
Behavioural Neurology. 2018; 2018: 1
[Pubmed] | [DOI]


Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

  Materials and Me...
  In this article
Article Figures

 Article Access Statistics
    PDF Downloaded721    
    Comments [Add]    
    Cited by others 26    

Recommend this journal