Journal article Open Access

Quantitative Characterization of α-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control

Perrino, Giansimone; Wilson, Cathal; Santorelli, Marco; di Bernardo, Diego


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{
  "description": "<p><strong>Highlights</strong></p>\n\n<p>&bull;&nbsp;<em>In silico</em>&nbsp;feedback control enables regulation of &alpha;-synuclein expression in yeast</p>\n\n<p>&bull;&nbsp;&alpha;-Synuclein inclusion formation is strictly concentration, but not time, dependent</p>\n\n<p>&bull;&nbsp;The aggregation threshold of the &alpha;-synuclein A53T mutant is 56% of the wild-type</p>\n\n<p>&bull;&nbsp;Autophagy induction speeds up inclusion clearance in the A53T &alpha;-synuclein strain</p>\n\n<p><strong>Summary</strong></p>\n\n<p>Aggregation of&nbsp;&alpha;-synuclein&nbsp;and formation of inclusions are hallmarks of Parkinson&rsquo;s disease (PD). Aggregate formation is affected by cellular environment, but it has been studied almost exclusively in cell-free systems. We quantitatively analyzed &alpha;-synuclein inclusion formation and clearance in a yeast cell model of PD expressing either&nbsp;wild-type&nbsp;(WT) &alpha;-synuclein or the disease-associated A53T mutant from the&nbsp;galactose&nbsp;(Gal)-inducible promoter. A computer-controlled microfluidics device regulated &alpha;-synuclein in cells by means of closed-loop feedback control. We demonstrated that inclusion formation is strictly concentration dependent and that the aggregation threshold of the A53T mutant is about half of the WT &alpha;-synuclein (56%). We chemically modulated the proteasomal&nbsp;and autophagic pathways and demonstrated that&nbsp;autophagy&nbsp;is the main determinant of A53T &alpha;-synuclein inclusions&rsquo; clearance. In addition to proposing a technology to overcome current limitations in dynamically regulating&nbsp;protein expression&nbsp;levels, our results contribute to the&nbsp;biology&nbsp;of PD and have relevance for therapeutic applications.</p>", 
  "license": "https://creativecommons.org/licenses/by/4.0/legalcode", 
  "creator": [
    {
      "affiliation": "Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy", 
      "@id": "https://orcid.org/0000-0001-8095-9139", 
      "@type": "Person", 
      "name": "Perrino, Giansimone"
    }, 
    {
      "affiliation": "Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy", 
      "@id": "https://orcid.org/0000-0002-0637-8608", 
      "@type": "Person", 
      "name": "Wilson, Cathal"
    }, 
    {
      "affiliation": "Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy", 
      "@id": "https://orcid.org/0000-0002-9633-7825", 
      "@type": "Person", 
      "name": "Santorelli, Marco"
    }, 
    {
      "affiliation": "1. Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy; 2. Department of Chemical, Materials and Industrial Production Engineering, University of Naples Federico II, Piazzale Tecchio 80, 80125 Naples, Italy", 
      "@id": "https://orcid.org/0000-0002-1911-7407", 
      "@type": "Person", 
      "name": "di Bernardo, Diego"
    }
  ], 
  "headline": "Quantitative Characterization of \u03b1-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control", 
  "image": "https://zenodo.org/static/img/logos/zenodo-gradient-round.svg", 
  "datePublished": "2019-04-16", 
  "url": "https://zenodo.org/record/2668451", 
  "keywords": [
    "Bioengineering", 
    "Microfluidics", 
    "Feedback control", 
    "Gene expression", 
    "Synuclein", 
    "Aggregation"
  ], 
  "@context": "https://schema.org/", 
  "identifier": "https://doi.org/10.1016/j.celrep.2019.03.081", 
  "@id": "https://doi.org/10.1016/j.celrep.2019.03.081", 
  "@type": "ScholarlyArticle", 
  "name": "Quantitative Characterization of \u03b1-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control"
}
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