title: Development and characterization of polo-like kinase 2 loaded nanoparticles-A novel strategy for (serine-129) phosphorylation of alpha-synuclein.
abstract: Polo like kinase 2 (PLK2), a serine/threonine serum inducible kinase, has been proposed to be the major factor responsible for phosphorylating alpha-synuclein (α-syn) at Serine-129 (Ser-129) in Parkinson's disease (PD). A suitable strategy to gain insights into PLK2's biological effects might be to increase PLK2 intracellular levels with the aim of reproducing the slow progressive neuronal changes that occur in PD. The goal of this study was to develop and characterize a novel drug delivery system (DDS) for PLK2 cytosolic delivery using Total recirculating one machine system (TROMS), a technique capable of encapsulating fragile molecules while maintaining their native properties. A protocol for nanoparticle (NP) preparation using TROMS was set up. NPs showed a mean diameter of 257±15.61nm and zeta potential of -16±2mV, suitable for cell internalization. TEM and SEM images showed individual, spherical, dispersed NPs. The drug entrapment efficacy was 61.86±3.9%. PLK2-NPs were able to enter SH-SY5Y cells and phosphorylate α-syn at Ser-129, demonstrating that the enzyme retained its activity after the NP manufacturing process. This is the first study to develop a DDS for continuous intracellular delivery of PLK2. These promising results indicate that this novel nanotechnology approach could be used to elucidate the biological effects of PLK2 on dopaminergic neurons.
authors.affiliation: Pharmacy and Pharmaceutical Technology Department, University of Navarra, Pamplona, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Rodríguez-Nogales
authors.affiliation: Pharmacy and Pharmaceutical Technology Department, University of Navarra, Pamplona, Spain; Instituto de Investigación Sanitaria de Navarra (IDISNA), Pamplona, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Garbayo
authors.affiliation: Department of Neurology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Martínez-Valbuena
authors.affiliation: Chemical & Environmental Engineering Department & Nanoscience Institute of Aragon, University of Zaragoza, Zaragoza, Spain; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Madrid, Spain.
authors.forename: V
authors.initials: V
authors.lastname: Sebastián
authors.affiliation: Instituto de Investigación Sanitaria de Navarra (IDISNA), Pamplona, Spain; Department of Neurology, Clínica Universidad de Navarra, Pamplona, Spain.
authors.forename: M R
authors.initials: MR
authors.lastname: Luquin
authors.affiliation: Pharmacy and Pharmaceutical Technology Department, University of Navarra, Pamplona, Spain; Instituto de Investigación Sanitaria de Navarra (IDISNA), Pamplona, Spain. Electronic address: mjblanco@unav.es.
authors.forename: M J
authors.initials: MJ
authors.lastname: Blanco-Prieto
mesh_terms: D002460:Cell Line; D059290:Dopaminergic Neurons / Q000187:drug effects; D016503:Drug Delivery Systems / Q000379:methods; D006801:Humans; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D010300:Parkinson Disease / Q000188:drug therapy; D010766:Phosphorylation / Q000187:drug effects*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000494:pharmacology; D012694:Serine / Q000378:metabolism*; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein; D012694:Serine; C496126:PLK2 protein, human; D017346:Protein Serine-Threonine Kinases; C444640:serum-inducible kinase
keywords: Alpha-synuclein; Nanoparticles; PLGA; Phosphorylation; Polo-like kinase 2; TROMS