Title DISC1 AND FUS: CO-AGGREGATION OF PROTEINS FROM MENTAL AND
NEURODEGENERATIVE ILLNESS
Title (croatian) DISC1 I FUS: KOAGREGACIJA PROTEINA POVEZANIH S MENTALNIM I
NEURODEGENERATIVNIM BOLESTIMA
Author Martina Jeremić
Mentor Nicholas James Bradshaw (mentor)
Committee member Ivana Ratkaj (predsjednik povjerenstva)
Committee member Jelena Ban (član povjerenstva)
Granter University of Rijeka (Faculty of Biotechnology and Drug Development) Rijeka
Defense date and country 2021-07-14, Croatia
Scientific / art field, discipline and subdiscipline BIOTECHNICAL SCIENCES Biotechnology
Abstract Chronic mental illnesses (CMI) such as schizophrenia, bipolar disorder or depression have overlapping genetic backgrounds and symptoms. These illnesses are manifested as disruptions of mental health (cognitive, emotional and behavioural) from various etiological factors. As current diagnostic methods are almost entirely dependent on psychiatric evaluation, and given the complexity of the background, it is hard to develop new approaches to research these illnesses.
Neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS) or Alzheimer’s disease are incurable conditions that result in progressive degeneration of nerve cells. These illnesses are well known to be associated with the occurrence of protein aggregation. Recent studies suggest that this same biological state, in which misfolded proteins aggregate, may be found in mental illnesses as well.
The Disrupted in Schizophrenia 1 (DISC1) protein has been identified in the insoluble fraction of the brain tissue of patients suffering from schizophrenia, bipolar disorder and depression, implying that DISC1 aggregated. The Fused in Sarcoma protein (FUS) has been found in in the insoluble fraction of the brain tissue of patients suffering from various neurodegenerative illnesses, including ALS and frontotemporal dementia (FTD). Mutations of FUS have been described as contributing to the progress of illnesses.
We investigated the interaction of DISC1 with FUS in protein aggregates. We compared wild-type and mutated FUS protein with DISC1 in cultured mammalian cells. This interaction provides information on the biological link between mental illnesses and neurodegenerative disorders.
Microscopic examination of cells overexpressing these two proteins showed the co-aggregation of mutant type FUS with DISC1. Additionally, co-aggregation of fragments of DISC1 with the mutant type (mutation on C-terminal tyrosine) FUS protein was examined to determine which domains of this protein are involved in co-aggregation. One of DISC1 region has been seen aggregating in previous researches. We confirmed the importance of this region for aggregation in a different cell line – human neuroblastoma cell line, because it is of greater biological relevance compared to previously seen results in human embryonic kidney cells. The S>C region is another region of DISC1, which showed no aggregates in cells. Combined with FUS protein, it has been seen that the D>I region does not co-aggregate with FUS protein, while the results of the S>C region suggest potential coaggregation, but for now this remains uncertain.
The results obtained here, and further research on these proteins could help explain biological connections between neurodegenerative disorders and mental illnesses, overlap in symptoms of these illnesses and the difficulty of making the diagnose.
Abstract (croatian) Kronične mentalne bolesti, kao što su shizofrenija, bipolarni poremećaj ili depresija imaju zajedničku genetsku pozadinu i simptome. Ova oboljenja manifestiraju se kao poremećaj mentalnog zdravlja (kognitivnog, emotivnog i bihevioralnog) zbog različitih etioloških faktora. Budući da su dijagnostičke metode gotovo u potpunosti ovisne o psihijatrijskoj evaluaciji, a sama pozadina bolesti je kompleksna, teško je razviti nove pristupe.
Neurodegenerativne bolesti, kao što su amiotrofična lateralna skleroza (ALS) i Alzheimerova bolest, neizlječiva su stanja koja rezultiraju progresivnom degeneracijom živčanih stanica. Ove bolesti poznate su po pojavi proteinskih agregata. Nedavne studije pretpostavljaju da se ovo isto biološko stanje, u kojem nepravilno formirani proteini agregiraju, može naći i u mentalnim bolestima.
Disrupted-in-Schizophrenia 1 (DISC1) je protein identificiran u netopivim frakcijama moždanih tkiva pacijenata oboljelih od shizofrenije, bipolarnog poremećaja i depresije, što implicira da DISC1, u ovim stanjima, stvara agregate. Fused in Sarcoma protein (FUS) je identificiran u netopivim frakcijama moždanih tkiva pacijenata oboljelih od različitih neurodegenerativnih bolesti poput ALS-a i frontotemporalne demencije (FTD). U ovom slučaju, mutacije FUS proteina povezane su napredovanjem bolesti.
Istraživali smo interakciju proteina DISC1 i FUS u proteinskim agregatima. Uspoređivali smo wild-type i FUS mutant sa DISC1 proteinom u kultiviranim stanicama. Ova interakcija nudi informacije o biološkoj poveznici između mentalnih i neurodegenerativnih bolesti.
Mikroskopska analiza stanica s povećanom ekpresijom ova dva proteina je rezultirala u koagregaciji DISC1 proteina i FUS mutanta. Uz to, ispitivali smo koagregaciju fragmenata DISC1 proteina sa FUS mutantom (mutacija na C-terminalnom tirozinu) kako bismo utvrdili koja domena DISC1 je uključena u koagregaciju. Jedna od ovih regija pokazala je agregaciju u prethodnim istraživanjima. Važnost ove regije za proces agregacije potvrdili smo u boljoj staničnoj liniji – humane stanice neuroblastoma, zbog veće biološke relevantnosti za rezultate u usporedbi s prethodno korištenim humanim embrionalnim stanicama bubrega. S>C regija nije do sada agregirala u stanicama. Zajedno sa FUS proteinom, vidjeli smo kako D>I regija ne koagregira sa FUS mutantom, dok rezultati za S>C regiju pretpostavljaju potencijalnu koagregaciju, no za sad, to ostaje nedokazano.
Rezultati koje smo do sada prikupili i buduća istraživanja na ovim proteinima, mogli bi objasniti biološku pozadinu između neurodegenerativnih i mentalnih bolesti, preklapanje u simptomima kao i poteškoće prilikom postavljanja dijagnoze.
Keywords
protein aggregation
schizophrenia
mental illness
neurodegenerative disorders
Keywords (croatian)
agregacija proteina
shizofrenija
mentalne bolesti
neurodegenerativne bolesti
Language croatian
URN:NBN urn:nbn:hr:193:154879
Study programme Title: Biotechnology and drug research Study programme type: university Study level: undergraduate Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) biotehnologije i istraživanja lijekova (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) biotehnologije i istraživanja lijekova)
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File origin Born digital
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Created on 2021-07-14 10:10:46