title: Recombinant pre-miR-29b for Alzheimer´s disease therapeutics.
abstract: MicroRNAs are arising as the next generation of diagnostic and therapeutic tools for gene silencing. Studies demonstrated that the miR-29 expression is decreased in Alzheimer's disease (AD) patients displaying high levels of human β-secretase (hBACE1). Recent advances toward an effective therapy for AD intend to employ miR-29 to suppress hBACE1 expression and subsequent Amyloid-β (Aβ) peptide. However, delivery of mature miRNA has demonstrated modest efficacy in vitro; therefore, the preparation of highly pure and biologically active pre-miRNA arises as one of the most important challenges in the development of these therapeutic strategies. Recently, we described a new strategy based arginine-affinity chromatography to specifically purify the recombinant pre-miR-29b. Following this strategy, the purified pre-miR-29b was successfully encapsulated into polyplexes that were further delivered in cytoplasm. It was verified that Chitosan/pre-miR-29b and Polyethylenimine/pre-miR-29b systems efficiently delivered pre-miR-29b to N2a695 cells, thus reducing the hBACE1 protein expression (around 78% and 86%, respectively) and Aβ42 levels (approximately 44% and 47%, respectively). Furthermore, pre-miR-29b downregulates the hBACE1 mRNA expression in 80%. Overall, it was demonstrated that the recombinant pre-miR-29b using polyplexes allowed to decrease the hBACE1 and Aβ42 expression levels, improving the currently available methodologies of miRNA-based therapeutics.
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Avenida Infante D. Henrique, Covilhã, 6200-506, Portugal.
authors.forename: Patrícia A
authors.initials: PA
authors.lastname: Pereira
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Avenida Infante D. Henrique, Covilhã, 6200-506, Portugal.
authors.forename: Joana F
authors.initials: JF
authors.lastname: Tomás
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Avenida Infante D. Henrique, Covilhã, 6200-506, Portugal.
authors.forename: João A
authors.initials: JA
authors.lastname: Queiroz
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Avenida Infante D. Henrique, Covilhã, 6200-506, Portugal.|CNC - Center of Neuroscience and Cell Biology, University of Coimbra, Largo Marquês de Pombal, Coimbra, 3004-517, Portugal.
authors.forename: Ana R
authors.initials: AR
authors.lastname: Figueiras
authors.affiliation: CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Avenida Infante D. Henrique, Covilhã, 6200-506, Portugal.
authors.forename: Fani
authors.initials: F
authors.lastname: Sousa
mesh_terms: D000544:Alzheimer Disease / Q000235:genetics* / Q000628:therapy; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics; D016229:Amyloid beta-Peptides / Q000378:metabolism; D016282:Aspartic Acid Endopeptidases / Q000235:genetics; D045744:Cell Line, Tumor; D004339:Drug Compounding; D015870:Gene Expression; D005786:Gene Expression Regulation; D015316:Genetic Therapy; D006801:Humans; D035683:MicroRNAs / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D009474:Neurons / Q000378:metabolism; D034622:RNA Interference; D012322:RNA Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D016229:Amyloid beta-Peptides; C532045:MIRN29a microRNA, human; D035683:MicroRNAs; D012322:RNA Precursors; D012333:RNA, Messenger; D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human
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