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P2‐382: Targeting proinflammatory cytokine up‐regulation with a novel anti‐cytokine therapeutic, MW‐151, attenuates pathology in an APP/PS1 knock‐in mouse model
Author(s) -
Van Eldik Linda,
Bachstetter Adam,
Sompol Pradoldej,
St. Clair Daret,
Watterson D. Martin
Publication year - 2012
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2012.05.1092
Subject(s) - proinflammatory cytokine , neuroinflammation , cytokine , medicine , microglia , genetically modified mouse , immunology , pathology , inflammation , neuroscience , transgene , biology , gene , biochemistry
this hairpin structure. Therefore, modulation of exon 10 splicing in the tau gene via stabilization of the hairpin could potentially be targeted to prevent tauopathies. Methods: To target the tau pre-mRNA hairpin, we designed bipartite antisense molecules (ASOs) connected via a linker region, which could interact simultaneously with the regions that flank either side of the tau hairpin structure. Tripartite compounds (dubbed “molecular clasps”) also include an MTX moiety connected to the linker region, potentially allowing simultaneous binding to the base and either side of the hairpin structure. We have synthesized and tested these compounds for their ability to bind and stabilize the tau pre-mRNA stem-loop and their efficiency in shifting the splicing of tau away from the 4R isoform.Results: Via gel shift and RNase protection assays, we have demonstrated that the bipartite ASOs and tripartite molecular clasps bind and simultaneously interact with the sequences that immediately flank the tau pre-mRNA hairpin. EC 50 values for binding of the tripartite molecules to the tau RNA demonstrated that the linked MTX moiety of the conjugate interacts near the base of the hairpin. In vitro splicing assays showed that the bipartite and tripartite compounds reduced exon 10 splicing in a concentration-dependent manner. Moreover, these molecules decreased the splicing of exon 10 in cells from a minigene as well as from endogenous tau. Conclusions: The bipartite ASOs and tripartite molecular clasps can effectively inhibit tau exon 10 splicing and reverse the effects of destabilizing disease-causing mutations both in vitro and in cell culture.