Prion infection impairs the cellular response to oxidative stress

Milhavet, Ollivier; McMahon, Hilary E.M.; Rachidi, Walid; Nishida, Naoshi; Katamine, Shigeru; Mangé, Alain; Arlotto, Marie; Casanova, Danielle; Riondel, J; Favier, Alain; Lehmann, Sylvain

doi:10.1073/pnas.250289197

Cited by 206 publications

(161 citation statements)

References 41 publications

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“…1d), and by repeating the experiment we established that the effective concentration for 50% reduction of PrP Sc (IC 50 ) over 72 h was Ϸ1.35 M. The normal PrP C expression in uninfected cells was unaffected. A similar effect was confirmed by using other scrapie-infected GT cells (GTϩ22L and GTϩCh) (20), and also by using GTϩBSE cells stably infected with mouseadapted bovine spongiform encephalopathy (BSE) (Fig. 1e).…”

Section: Resultssupporting

confidence: 68%

“…We used a mouse neuronal cell culture uninfected (GT1-7) and persistently infected with human TSE agent (Fukuoka-1 strain), designated GTϩFK (20). Of the 59 compounds, we tested the 44 that were commercially available (see SI Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…The immortalized mouse neuronal cell line GT1-7 was cultured as described (20). GT1-7 cells stably infected with Fukuoka-1, 22L, or Chandler/RML (designated GTϩFK, GTϩ22L, or GTϩCh, respectively) were maintained for more than a year in our laboratory.…”

Section: Methodsmentioning

confidence: 99%

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Hot spots in prion protein for pathogenic conversion

Kuwata

Nishida

Matsumoto

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Prion proteins are key molecules in transmissible spongiform encephalopathies (TSEs), but the precise mechanism of the conversion from the cellular form (PrP C ) to the scrapie form (PrP Sc ) is still unknown. Here we discovered a chemical chaperone to stabilize the PrP C conformation and identified the hot spots to stop the pathogenic conversion. We conducted in silico screening to find compounds that fitted into a ''pocket'' created by residues undergoing the conformational rearrangements between the native and the sparsely populated high-energy states (PrP*) and that directly bind to those residues. Forty-four selected compounds were tested in a TSE-infected cell culture model, among which one, 2-pyrrolidin-1-yl-N-[4-[4-(2-pyrrolidin-1-yl-acetylamino)-benzyl]-phenyl]-acetamide, termed GN8, efficiently reduced PrP Sc . Subsequently, administration of GN8 was found to prolong the survival of TSE-infected mice. Heteronuclear NMR and computer simulation showed that the specific binding sites are the A-S2 loop (N159) and the region from helix B (V189, T192, and K194) to B-C loop (E196), indicating that the intercalation of these distant regions (hot spots) hampers the pathogenic conversion process. Dynamics-based drug discovery strategy, demonstrated here focusing on the hot spots of PrP C , will open the way to the development of novel anti-prion drugs.anti-prion compound ͉ binding sites ͉ chemical chaperone ͉ dynamicsbased drug discovery ͉ transmissible spongiform encephalopathy

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Section: Resultssupporting

confidence: 68%

Section: Resultsmentioning

confidence: 99%

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Hot spots in prion protein for pathogenic conversion

Kuwata

Nishida

Matsumoto

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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175

225

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“…Immortalized neuronal and neuroglial cell lines persistently infected by prions generally show no overt signs of cytotoxicity, although producing readily detectable amounts of PrP . Various phenotypical alterations have been reported (16,17), yet it remains uncertain whether similar dysfunctions may affect TSE-injured postmitotic neurons. This calls for renewed efforts toward the development of primary cell culture systems in which the individual contribution of various CNS cells to prion propagation and its effects could be evaluated.…”

mentioning

confidence: 99%

Prions can infect primary cultured neurons and astrocytes and promote neuronal cell death

Cronier

Laude

Peyrin

2004

Proc. Natl. Acad. Sci. U.S.A.

139

150

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Transmissible spongiform encephalopathies arise as a consequence of infection of the central nervous system by prions, where neurons and glial cells are regarded as primary targets. Neuronal loss and gliosis, associated with the accumulation of misfolded prion protein (PrP), are hallmarks of prion diseases; yet the mechanisms underlying such disorders remain unclear. Here we introduced a cell system based on primary cerebellar cultures established from transgenic mice expressing ovine PrP and then exposed to sheep scrapie agent. Upon exposure to low doses of infectious agent, such cultures, unlike cultures originating from PrP null mice, were found to accumulate de novo abnormal PrP and infectivity, as assessed by mouse bioassay. Importantly, using astrocyte and neuron͞astrocyte cocultures, both cell types were found capable of sustaining efficient prion propagation independently, leading to the production of proteinase K-resistant PrP of the same electrophoretic profile as in diseased brain. Moreover, contrasting with data obtained in chronically infected cell lines, late-occurring apoptosis was consistently demonstrated in the infected neuronal cultures. Our results provide evidence that primary cultured neural cells, including postmitotic neurons, are permissive to prion replication, thus establishing an approach to study the mechanisms involved in prion-triggered neurodegeneration at a cellular level.T ransmissible spongiform encephalopathies (TSE), which include Creutzfeldt-Jakob disease in humans, bovine spongiform encephalopathy in cattle, and scrapie in sheep, are fatal neurodegenerative disorders caused by prions, a class of unconventional agents that targets the CNS in mammals. A hallmark of prion diseases is the accumulation of abnormal prion protein (PrP Sc ), a misfolded form of the cellular PrP (PrP c ). Transmissibility is believed to stem from the ability of the prion isoform to promote the conformational transition from PrP c to PrP Sc . Biologically distinct prion strains can propagate in a same host, presumably through the perpetuation of different specific PrP Sc conformers (1-3).Although it seems clear that neuronal dysfunction must lie at the root of the clinical disorders observed in these diseases, it is still obscure what triggers neurodegeneration and what role nonneuronal cells may play in this process. There is ample evidence to support a primary role of the neurons in prion propagation and neuropathogenesis into the CNS. Intra-or perineuronal PrP Sc deposition, spongiform vacuolation involving cell soma and processes, and neuronal loss are typical histopathological changes observed in TSE-affected brain tissues (4, 5). Transgenic mice with PrP expression specifically targeted to neurons have been obtained that turned out to be fully susceptible to prion disease (6). More recently, it was shown that an acute neuron-targeted depletion of PrP in the brain of mice with ongoing infection is able to prevent neuronal loss and progression to disease and even to reverse early spongiform chang...

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“…For instance, infection with several murine prion strains impairs the cell response of GT1 and N2a cells to oxidative stress [89], presumably through a decrease in superoxide dismutase activity. RML infection was also shown to sensitise N2a and GT1 cells to pro-apoptotic stimuli such as endoplasmic reticulum stress or to mild proteasome inhibition [54,71].…”

Section: Mechanisms Of Neurodegenerationmentioning

confidence: 99%

Cell models of prion infection

Vilette

2007

Vet. Res.

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-Due to recent renewal of interest and concerns in prion diseases, a number of cell systems permissive to prion multiplication have been generated in the last years. These include established cell lines, neuronal stem cells and primary neuronal cultures. While most of these models are permissive to experimental, mouse-adapted strains of prions, the propagation of natural field isolates from sheep scrapie and chronic wasting disease has been recently achieved. These models have improved our knowledge on the molecular and cellular events controlling the conversion of the PrP C protein into abnormal isoforms and on the cell-to-cell spreading of prions. Infected cultured cells will also facilitate investigations on the molecular basis of strain identity and on the mechanisms that lead to neurodegeneration. The ongoing development of new cell models with improved characteristics will certainly be useful for a number of unanswered critical issues in the prion field.

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Prion infection impairs the cellular response to oxidative stress

Cited by 206 publications

References 41 publications

Hot spots in prion protein for pathogenic conversion

Hot spots in prion protein for pathogenic conversion

Prions can infect primary cultured neurons and astrocytes and promote neuronal cell death

Cell models of prion infection

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