2668451
doi
10.1016/j.celrep.2019.03.081
oai:zenodo.org:2668451
user-cosy-bio
user-eu
Wilson, Cathal
Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy
Santorelli, Marco
Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy
di Bernardo, Diego
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
Quantitative Characterization of α-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control
Perrino, Giansimone
Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy
info:eu-repo/semantics/openAccess
Creative Commons Attribution 4.0 International
https://creativecommons.org/licenses/by/4.0/legalcode
Bioengineering
Microfluidics
Feedback control
Gene expression
Synuclein
Aggregation
<p><strong>Highlights</strong></p>
<p>• <em>In silico</em> feedback control enables regulation of α-synuclein expression in yeast</p>
<p>• α-Synuclein inclusion formation is strictly concentration, but not time, dependent</p>
<p>• The aggregation threshold of the α-synuclein A53T mutant is 56% of the wild-type</p>
<p>• Autophagy induction speeds up inclusion clearance in the A53T α-synuclein strain</p>
<p><strong>Summary</strong></p>
<p>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.</p>
Zenodo
2019-04-16
info:eu-repo/semantics/article
2668450
user-cosy-bio
user-eu
award_title=Control Engineering of Biological Systems for Reliable Synthetic Biology Applications; award_number=766840; award_identifiers_scheme=url; award_identifiers_identifier=https://cordis.europa.eu/projects/766840; funder_id=00k4n6c32; funder_name=European Commission;
1579532304.97796
4398468
md5:080ca2abef09a11ef8939ce14abab209
https://zenodo.org/records/2668451/files/Perrino et al Cell Reports 2019.pdf
public
Cell Reports
27
3
916-927
2019-04-16