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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  <dc:creator>Perrino, Giansimone</dc:creator>
  <dc:creator>Wilson, Cathal</dc:creator>
  <dc:creator>Santorelli, Marco</dc:creator>
  <dc:creator>di Bernardo, Diego</dc:creator>
  <dc:date>2019-04-16</dc:date>
  <dc:description>Highlights

• In silico feedback control enables regulation of α-synuclein expression in yeast

• α-Synuclein inclusion formation is strictly concentration, but not time, dependent

• The aggregation threshold of the α-synuclein A53T mutant is 56% of the wild-type

• Autophagy induction speeds up inclusion clearance in the A53T α-synuclein strain

Summary

Aggregation of α-synuclein and formation of inclusions are hallmarks of Parkinson’s disease (PD). Aggregate formation is affected by cellular environment, but it has been studied almost exclusively in cell-free systems. We quantitatively analyzed α-synuclein inclusion formation and clearance in a yeast cell model of PD expressing either wild-type (WT) α-synuclein or the disease-associated A53T mutant from the galactose (Gal)-inducible promoter. A computer-controlled microfluidics device regulated α-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 α-synuclein (56%). We chemically modulated the proteasomal and autophagic pathways and demonstrated that autophagy is the main determinant of A53T α-synuclein inclusions’ clearance. In addition to proposing a technology to overcome current limitations in dynamically regulating protein expression levels, our results contribute to the biology of PD and have relevance for therapeutic applications.</dc:description>
  <dc:identifier>https://zenodo.org/record/2668451</dc:identifier>
  <dc:identifier>10.1016/j.celrep.2019.03.081</dc:identifier>
  <dc:identifier>oai:zenodo.org:2668451</dc:identifier>
  <dc:relation>info:eu-repo/grantAgreement/EC/H2020/766840/</dc:relation>
  <dc:relation>url:https://zenodo.org/communities/cosy-bio</dc:relation>
  <dc:rights>info:eu-repo/semantics/openAccess</dc:rights>
  <dc:rights>https://creativecommons.org/licenses/by/4.0/legalcode</dc:rights>
  <dc:source>Cell Reports 27(3) 916-927</dc:source>
  <dc:subject>Bioengineering</dc:subject>
  <dc:subject>Microfluidics</dc:subject>
  <dc:subject>Feedback control</dc:subject>
  <dc:subject>Gene expression</dc:subject>
  <dc:subject>Synuclein</dc:subject>
  <dc:subject>Aggregation</dc:subject>
  <dc:title>Quantitative Characterization of α-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control</dc:title>
  <dc:type>info:eu-repo/semantics/article</dc:type>
  <dc:type>publication-article</dc:type>
</oai_dc:dc>
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