title: The alpha-synuclein 5'untranslated region targeted translation blockers: anti-alpha synuclein efficacy of cardiac glycosides and Posiphen.
abstract: Increased brain α-synuclein (SNCA) protein expression resulting from gene duplication and triplication can cause a familial form of Parkinson's disease (PD). Dopaminergic neurons exhibit elevated iron levels that can accelerate toxic SNCA fibril formation. Examinations of human post mortem brain have shown that while mRNA levels for SNCA in PD have been shown to be either unchanged or decreased with respect to healthy controls, higher levels of insoluble protein occurs during PD progression. We show evidence that SNCA can be regulated via the 5'untranslated region (5'UTR) of its transcript, which we modeled to fold into a unique RNA stem loop with a CAGUGN apical loop similar to that encoded in the canonical iron-responsive element (IRE) of L- and H-ferritin mRNAs. The SNCA IRE-like stem loop spans the two exons that encode its 5'UTR, whereas, by contrast, the H-ferritin 5'UTR is encoded by a single first exon. We screened a library of 720 natural products (NPs) for their capacity to inhibit SNCA 5'UTR driven luciferase expression. This screen identified several classes of NPs, including the plant cardiac glycosides, mycophenolic acid (an immunosuppressant and Fe chelator), and, additionally, posiphen was identified to repress SNCA 5'UTR conferred translation. Western blotting confirmed that Posiphen and the cardiac glycoside, strophanthidine, selectively blocked SNCA expression (~1 μM IC(50)) in neural cells. For Posiphen this inhibition was accelerated in the presence of iron, thus providing a known APP-directed lead with potential for use as a SNCA blocker for PD therapy. These are candidate drugs with the potential to limit toxic SNCA expression in the brains of PD patients and animal models in vivo.
authors.affiliation: Neurochemistry Laboratory, Psychiatry-Neuroscience, Massachusetts General Hospital, Charlestown, MA 02129, USA. Jrogers@partners.org
authors.forename: Jack T
authors.initials: JT
authors.lastname: Rogers
authors.forename: Sohan
authors.initials: S
authors.lastname: Mikkilineni
authors.forename: Ippolita
authors.initials: I
authors.lastname: Cantuti-Castelvetri
authors.forename: Deborah H
authors.initials: DH
authors.lastname: Smith
authors.forename: Xudong
authors.initials: X
authors.lastname: Huang
authors.forename: Sanghamitra
authors.initials: S
authors.lastname: Bandyopadhyay
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Cahill
authors.forename: Maria L
authors.initials: ML
authors.lastname: Maccecchini
authors.forename: Debomoy K
authors.initials: DK
authors.lastname: Lahiri
authors.forename: Nigel H
authors.initials: NH
authors.lastname: Greig
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D002298:Cardenolides / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004009:Dicloxacillin / Q000494:pharmacology*; D006801:Humans; D007501:Iron / Q000378:metabolism*; D016874:Neurofibrillary Tangles / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D014176:Protein Biosynthesis / Q000187:drug effects / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013327:Strophanthidin / Q000494:pharmacology*; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
chemical_list: D020121:5' Untranslated Regions; D000900:Anti-Bacterial Agents; D002298:Cardenolides; D012333:RNA, Messenger; C497604:SNCA protein, human; D051844:alpha-Synuclein; D013327:Strophanthidin; D004009:Dicloxacillin; D007501:Iron; C007846:gitoxigenin