Effect of ubiquitination on Parkinson’s Disease associated proteins alpha-synuclein and synphilin-1 in humanized yeast models

Jens Loncke
Dit project onderzocht de rol van ubiquitinatie op de toxiciteit en het aggregeren van alfa-synucleïne en synphilin-1 in gehumaniseerde gistmodellen voor de ziekte van Parkinson.

Gist als rijzende ster tegen de ziekte van Parkinson

De ziekte van Parkinson

De ziekte van Parkinson is één van de meest voorkomende neurodegeneratieve aandoeningen met wereldwijd 7 tot 10 miljoen patiënten. Dit is hoogstwaarschijnlijk een onderschatting van het echte totaal, want artsen schrijven vaak symptomen van de ziekte van Parkinson toe aan normale verouderingsprocessen. De ziekte komt vooral voor bij ouderen en komt twee keer meer voor bij mannen dan bij vrouwen. De maandelijkse kost voor de gezondheidszorg ligt tussen de €2600 en €10 000 per patiënt in Europese landen. Deze kosten zullen nog fel stijgen, want door de vergrijzing van de bevolking zal de ziekte vaker voor komen. De belangrijkste symptomen zijn beven in rust, traagheid van bewegingen en spierstijfheid.  Wanneer deze symptomen ernstiger worden kan de arts symptomatische geneesmiddelen voorschrijven zoals levodopa of dopamine receptor agonisten. Deze behandelingen kunnen de patiënt echter niet genezen en zijn louter bedoeld om de symtomen draaglijker te maken. Bovendien verliezen deze behandelingen hun effectiviteit naarmate de ziekte erger wordt en komen er ook heel wat bijwerkingen bij kijken.

Het verlies van dopamine

De exacte mechanismes die bijdragen tot het ontwikkelen van de ziekte van Parkinson zijn niet bekend. Er wordt vermoed dat genetische factoren en omgevingsfactoren een rol spelen. Wat wel geweten is, is dat een groot deel van de dopamine producerende zenuwcellen in de hersenen sterven afsterft. Dopamine is een belangrijke neurotransmitter in het teweegbrengen van gecontroleerde spierbewegingen. Een groot deel van de cellen die afsterven bij het ontwikkelen van de ziekte van Parkinson zijn gepigmenteerd. Om die reden zijn hersenen van patiënten met de ziekte van Parkinson gemakkelijk te onderscheiden van die van gezonde patiënten. Onderstaande figuur toont de middenhersenen van een gezonde patiënt (links) en een patiënt met de ziekte van Parkinson (rechts).

In de overlevende zenuwcellen worden opeenstapelingen van eiwitten teruggevonden die ‘Lewy bodies’ worden genoemd.  Het grootste deel van die Lewy bodies bestaat uit het eiwit alfa-synucleïne en de interactiepartner synphilin-1. De exacte functies van deze eiwitten zijn niet gekend. Er wordt vermoed dat post-translationele modificaties, zoals ubiquitinatie, een invloed zouden kunnen hebben op het samenklonteren en de toxiciteit van deze eiwitten.

image-20190727111626-1http://maplecarephysiotherapy.com/services/parkinsons-disease/

Transversale sectie van normale middenhersenen (links) en middenhersenen van een patiënt met de ziekte van Parkinson (rechts)

Gist als model voor de ziekte van Parkinson

In dit project worden deze eiwitten onderzocht in bakkersgist, door gebruik te maken van zogenaamde gehumaniseerde gistmodellen voor de Ziekte van Parkinson. Dit is mogelijk, omdat veel cellulaire processen in gist goed lijken op die in menselijke cellen. Alfa-synucleïne en synphilin-1 kunnen we tot expressie brengen in gist door gebruik te maken van een plasmide. Net zoals in menselijke zenuwcellen klonteren alfa-synucleïne en synphilin-1 ook samen in gistcellen. Een voorbeeld van het samenklonteren van alfa-synucleïne in een gistcel wordt hieronder getoond.

image 56

Fluorescente microscopieopname van een gistcel met samengeklonterd alfa-synucleïne. 

In deze studie onderzochten we de invloed van ubiquitinatie op de samenklontering en toxiciteit van alfa-synucleïne en synphilin-1 in gehumaniseerde gistmodellen. We gebruikten verschillende mutanten met verwijderde genen die belangrijk zijn in het ubiquitinatiesysteem van de cel. Onze resultaten suggereren dat een verlaagde vrije ubiquitine voorraad in de cel het samenklonteren van alfa-synucleïne in grotere, beschermende eiwitklonters zou kunnen bevorderen. Voor synphilin-1 was dit niet het geval. Verdere resultaten suggereerden dat dit initiële voordeel verdwijnt naarmate cellen ouder worden. Onze resultaten lijken een belangrijke rol toe te schrijven aan ubiquitinatie in het verwerken van alfa-synucleïne in de cel. Verder onderzoek zal echter noodzakelijk zijn om meer uitsluitsel te kunnen geven en de exacte mechanismen van de verwerking van alfa-synucleïne en synphilin-1 bloot te leggen. Het begrijpen van deze mechanismen kan er voor zorgen dat meer bekend wordt over hoe de ziekte van Parkinson ontwikkelt. Op die manier zou het in de toekomst mogelijk kunnen zijn om een genezende behandeling te ontwikkelen voor de ziekte van Parkinson. Een dergelijke behandeling zou de kosten voor de maatschappij behoorlijk kunnen drukken en veel lijden kunnen voorkomen.

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Universiteit of Hogeschool
Master in de biochemie en de biotechnologie
Publicatiejaar
2019
Promotor(en)
Prof. Dr. Joris Winderickx
Kernwoorden
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