ASO targeting RBM3 temperature-controlled poison exon splicing prevents neurodegeneration in vivo

Neurodegenerative diseases are increasingly prevalent in the aging population, yet no disease-modifying treatments are currently available. Increasing the expression of the cold-shock protein RBM3 through therapeutic hypothermia is remarkably neuroprotective. However, systemic cooling poses a health risk, strongly limiting its clinical application. Selective upregulation of RBM3 at normothermia thus holds immense therapeutic potential. Here we identify a poison exon within the RBM3 gene that is solely responsible for its cold-induced expression. Genetic removal or antisense oligonucleotide (ASO)-mediated manipulation of this exon yields high RBM3 levels independent of cooling. Notably, a single administration of ASO to exclude the poison exon, using FDA-approved chemistry, results in long-lasting increased RBM3 expression in mouse brains. In prion-diseased mice, this treatment leads to remarkable neuroprotection, with prevention of neuronal loss and spongiosis despite high levels of disease-associated prion protein. Our promising results in mice support the possibility that RBM3-inducing ASOs might also deliver neuroprotection in humans in conditions ranging from acute brain injury to Alzheimer's disease

Astrocyte Unfolded Protein Response Induces a Specific Reactivity State that Causes Non-Cell-Autonomous Neuronal Degeneration.

Recent interest in astrocyte activation states has raised the fundamental question of how these cells, normally essential for synapse and neuronal maintenance, become pathogenic. Here, we show that activation of the unfolded protein response (UPR), specifically phosphorylated protein kinase R-like endoplasmic reticulum (ER) kinase (PERK-P) signaling-a pathway that is widely dysregulated in neurodegenerative diseases-generates a distinct reactivity state in astrocytes that alters the astrocytic secretome, leading to loss of synaptogenic function in vitro. Further, we establish that the same PERK-P-dependent astrocyte reactivity state is harmful to neurons in vivo in mice with prion neurodegeneration. Critically, targeting this signaling exclusively in astrocytes during prion disease is alone sufficient to prevent neuronal loss and significantly prolongs survival. Thus, the astrocyte reactivity state resulting from UPR over-activation is a distinct pathogenic mechanism that can by itself be effectively targeted for neuroprotection

Feedback between deformation and magmatism in the Lloyds River Fault Zone : an example of episodic fault reactivation in an accretionary setting, Newfoundland Appalachians

Author Posting. © American Geophysical Union, 2006. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Tectonics 25 (2006): TC4004, doi:10.1029/2005TC001789.The Lloyds River Fault Zone is a 10–15 km wide amphibolite-grade shear zone that formed during the Ordovician Taconic Orogeny. It separates ophiolites and arc–back-arc complexes formed in Iapetus from a peri-Laurentian microcontinent (Dashwoods microcontinent). The Lloyds River Fault Zone comprises three high-strain zones, dominantly composed of mylonitic amphibolites, separated by less deformed plutonic rocks. Structural, age and metamorphic data suggest the Lloyds River Fault Zone accommodated sinistral-oblique underthrusting of ophiolites underneath the Dashwoods microcontinent prior to 471 ± 5 Ma at 800°C and 6 kbar. Plutonic rocks within the Lloyds River Fault Zone comprise two suites dated at 464 ± 2 plus 462 ± 2 and 459 ± 3 Ma, respectively. The younger age of the plutons with respect to some of the amphibolites, evidence for magmatic deformation, and the elongate nature of the plutons parallel to the Lloyds River Fault Zone suggest they were emplaced within the fault zone during deformation. Both intrusive episodes triggered renewed deformation at high temperatures (770–750°C), illustrating the positive feedback between deformation and magmatism. Offshoots of the plutons intruded undeformed ophiolitic gabbros outside the Lloyds River Fault Zone. Deformation localized within the intrusive sheets, coeval with static contact metamorphism of the host gabbros, leading to the development of new, small-scale shear zones. This illustrates that channeling of plutons into shear zones and nucleation of shear zones in melt-rich zones may occur simultaneously within the same fault system.This research is funded by a scholarship from the Faculty of Graduate and Postdoctoral Studies, University of Ottawa, to C.J.L. and a NSERC grant to C.v.S in his position as Adjunct Professor at the University of Ottawa

Timing and spatial distribution of deformation in the Newfoundland Appalachians: a "multi-stage collision" history

The Newfoundland Appalachians have been interpreted as an area where Lower Paleozoic plate convergence culminated in collision between an Ordovician volcanic chain and the North American craton hi Middle Ordovician times. Closure of the intervening proto-Atlantic (Iapetus) ocean was considered incomplete. Subsequent deformation gave rise to regional folding and faulting.Recent studies in the Newfoundland Dunnage zone have revealed that the deformation history is far more complex than previously recognized. Large-scale thrusting, folding and faulting occurred in Silurian-Devonian times. Furthermore, it has been suggested that the Dunnage zone is an allochthonous terrane underlain by dominantly continental crust rather than representing remnants of a "rooted" ocean basin.In view of these results a revision of tectonic scenarios and zonal subdivision is warranted and a "multi-stage collision" history will be discussed, with emphasis on the spatial distribution and significance of Silurian-Devonian deformation in central Newfoundland.Subduction in Lower Paleozoic times gave rise to the formation of a volcanic terrane; concurrently, to the southeast a marginal sea was formed (Mariana-type subduction). In Middle Ordovician times the volcanic terrane collided with the North American craton ("first-stage collision") and back-arc spreading terminated. Continued crustal shortening resulted in the formation of a Silurian accretionary terrane (telescoped marginal sea), and its subsequent deformation ("second-stage collision"). Devonian (-Carboniferous?) strike-slip faulting represents the third stage in the collision history.The model is applicable to large tracts of the Caledonian-Appalachian chain. Its main characteristics are: 1. (a) the revised zonal subdivision of the area is based on characteristics of Silurian and older rocks, rather than Middle Ordovician and older rocks only;2. (b) the central part of the orogen represents a telescoped marginal sea that formed to the southeast of the Ordovician volcanic chain, rather than a remnant of the incompletely closed Iapetus ocean;3. (c) the earliest deformation is progressively younger toward the southeast;4. (d) the Appalachian collision history is a result of the activity of a single deformation regime over a long period of at least 75 Ma.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26738/1/0000289.pd

TrkB signaling regulates the cold-shock protein RBM3-mediated neuroprotection.

Increasing levels of the cold-shock protein, RNA-binding motif 3 (RBM3), either through cooling or by ectopic over-expression, prevents synapse and neuronal loss in mouse models of neurodegeneration. To exploit this process therapeutically requires an understanding of mechanisms controlling cold-induced RBM3 expression. Here, we show that cooling increases RBM3 through activation of TrkB via PLCγ1 and pCREB signaling. RBM3, in turn, has a hitherto unrecognized negative feedback on TrkB-induced ERK activation through induction of its specific phosphatase, DUSP6. Thus, RBM3 mediates structural plasticity through a distinct, non-canonical activation of TrkB signaling, which is abolished in RBM3-null neurons. Both genetic reduction and pharmacological antagonism of TrkB and its downstream mediators abrogate cooling-induced RBM3 induction and prevent structural plasticity, whereas TrkB inhibition similarly prevents RBM3 induction and the neuroprotective effects of cooling in prion-diseased mice. Conversely, TrkB agonism induces RBM3 without cooling, preventing synapse loss and neurodegeneration. TrkB signaling is, therefore, necessary for the induction of RBM3 and related neuroprotective effects and provides a target by which RBM3-mediated synapse-regenerative therapies in neurodegenerative disorders can be used therapeutically without the need for inducing hypothermia

Trazodone rescues dysregulated synaptic and mitochondrial nascent proteomes in prion neurodegeneration

The unfolded protein response (UPR) is rapidly gaining momentum as a therapeutic target for protein misfolding neurodegenerative diseases, in which its overactivation results in sustained translational repression leading to synapse loss and neurodegeneration. In mouse models of these disorders, from Alzheimer’s to prion disease, modulation of the pathway—including by the licensed drug, trazodone—restores global protein synthesis rates with profound neuroprotective effects. However, the precise nature of the translational impairment, in particular the specific proteins affected in disease, and their response to therapeutic UPR modulation are poorly understood. We used non-canonical amino acid tagging (NCAT) to measure de novo protein synthesis in the brains of prion-diseased mice with and without trazodone treatment, in both whole hippocampus and cell-specifically. During disease the predominant nascent proteome changes occur in synaptic, cytoskeletal and mitochondrial proteins in both hippocampal neurons and astrocytes. Remarkably, trazodone treatment for just 2 weeks largely restored the whole disease nascent proteome in the hippocampus to that of healthy, uninfected mice, predominantly with recovery of proteins involved in synaptic and mitochondrial function. In parallel, trazodone treatment restored the disease-associated decline in synapses and mitochondria and their function to wild-type levels. In conclusion, this study increases our understanding of how translational repression contributes to neurodegeneration through synaptic and mitochondrial toxicity via depletion of key proteins essential for their function. Further, it provides new insights into the neuroprotective mechanisms of trazodone through reversal of this toxicity, relevant for the treatment of neurodegenerative diseases via translational modulation.Sección Deptal. de Bioquímica y Biología Molecular (Veterinaria)Fac. de VeterinariaTRUEpu

ASO targeting RBM3 temperature-controlled poison exon splicing prevents neurodegeneration in vivo.

Funder: Freie Universität Berlin; Id: http://dx.doi.org/10.13039/501100007537Funder: Cambridge Centre for Parkinson's PlusNeurodegenerative diseases are increasingly prevalent in the aging population, yet no disease-modifying treatments are currently available. Increasing the expression of the cold-shock protein RBM3 through therapeutic hypothermia is remarkably neuroprotective. However, systemic cooling poses a health risk, strongly limiting its clinical application. Selective upregulation of RBM3 at normothermia thus holds immense therapeutic potential. Here we identify a poison exon within the RBM3 gene that is solely responsible for its cold-induced expression. Genetic removal or antisense oligonucleotide (ASO)-mediated manipulation of this exon yields high RBM3 levels independent of cooling. Notably, a single administration of ASO to exclude the poison exon, using FDA-approved chemistry, results in long-lasting increased RBM3 expression in mouse brains. In prion-diseased mice, this treatment leads to remarkable neuroprotection, with prevention of neuronal loss and spongiosis despite high levels of disease-associated prion protein. Our promising results in mice support the possibility that RBM3-inducing ASOs might also deliver neuroprotection in humans in conditions ranging from acute brain injury to Alzheimer's disease