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": {
      "id": "CC-BY-4.0"
    }, 
    "title": "Quantitative Characterization of \u03b1-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control", 
    "journal": {
      "volume": "27", 
      "issue": "3", 
      "pages": "916-927", 
      "title": "Cell Reports"
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        "program": "H2020", 
        "funder": {
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          "name": "European Commission", 
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    "keywords": [
      "Bioengineering", 
      "Microfluidics", 
      "Feedback control", 
      "Gene expression", 
      "Synuclein", 
      "Aggregation"
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    "publication_date": "2019-04-16", 
    "creators": [
      {
        "orcid": "0000-0001-8095-9139", 
        "affiliation": "Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy", 
        "name": "Perrino, Giansimone"
      }, 
      {
        "orcid": "0000-0002-0637-8608", 
        "affiliation": "Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy", 
        "name": "Wilson, Cathal"
      }, 
      {
        "orcid": "0000-0002-9633-7825", 
        "affiliation": "Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy", 
        "name": "Santorelli, Marco"
      }, 
      {
        "orcid": "0000-0002-1911-7407", 
        "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", 
        "name": "di Bernardo, Diego"
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