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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{
  "DOI": "10.1016/j.celrep.2019.03.081", 
  "container_title": "Cell Reports", 
  "title": "Quantitative Characterization of \u03b1-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control", 
  "issued": {
    "date-parts": [
      [
        2019, 
        4, 
        16
      ]
    ]
  }, 
  "abstract": "<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>", 
  "author": [
    {
      "family": "Perrino, Giansimone"
    }, 
    {
      "family": "Wilson, Cathal"
    }, 
    {
      "family": "Santorelli, Marco"
    }, 
    {
      "family": "di Bernardo, Diego"
    }
  ], 
  "page": "916-927", 
  "volume": "27", 
  "type": "article-journal", 
  "issue": "3", 
  "id": "2668451"
}
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