izvor podataka: poirot

Naziv

Hiperfosforilacija, agregacija i transsinaptički prijenos tau proteina u Alzheimerovoj bolesti: analiza likvora i ispitivanje potencijalnih neuroprotektivnih spojeva

Tau protein hyperphosphorylation, aggregation and trans-synaptic transfer in Alzheimer's disease: cerebrospinal fluid analysis and assessment of potential neuroprotective compounds

Opis projekta

Za Alzheimerovu bolest (AB) ne postoji uzročno liječenje jer su brojni dosadašnji pokušaji intervencije temeljeni na amiloidnoj kaskadnoj hipotezi bili neuspješni. Usporedno s napretkom razumijevanja uloge tau proteina u patogenezi AB, metabolizam tau proteina je postao i važan terapijski cilj. Naime, smatra se da je u AB ravnoteža između kinaza i fosfataza tau proteina narušena, što dovodi do njegove hiperfosforilacije. Posljedično se povećava i frakcija tau proteina nevezanih za mikrotubule, pa se hiperfosforilirani tau monomeri vežu jedan za drugoga proizvodeći oligomere. Spajanje tau oligomera dovodi do oblikovanja sparenih uzvojitih filamenata, od kojih se sastoje sve neurofibrilarne promjene za koje je dokumentirano da predstavljaju najraniji neuropatološki događaj (tzv. ''pre-tangle'' stadij) od ključnog značaja za patogenezu AB. Ciljevi našeg projekta su: 1) bolje definirati odnos između bioloških biljega ranih stadija AB: ukupnih i fosforiliranih tau proteina, kao i amiloida ß u cerebrospinalnoj tekućini (likvoru) bolesnika s AD, MCI (blagi spoznajni poremećaj) i kontrolnih ispitanika. Uporabom inovativnog ELISA postupka i modificiranog eseja pomoću tioflavina S, u uzorcima likvora odrediti ćemo omjer između monomera i oligomera (agregata) tau proteina. Ti će novi podatci biti uspoređeni s kliničkim, neuropsihološkim i MRI nalazima; 2) testirati ćemo neke poznate i potencijalno neuroprotektivne spojeve (donepezil, klorgilin, nekoliko različitih flavonoida, te PF9601N i ASS234) na fosforilaciju tau proteina izazvanu okadaičnom kiselinom i nokodazolom u kulturama SH-SY5Y i P19 stanica. Epitope tau proteina fosforilirane na Thr181, Ser199 i Thr231 analizirati ćemo ELISA postupkom, a one na epitopima Thr202, Thr205, Ser396 i Ser404 pomoću westernske razložbe; i 3) istražiti ćemo biokemijski i imunocitokemijski transsinaptičko širenje neurofibrilarne degeneracije i razinu izraženosti tau mRNA nakon injekcije tau oligomera u entorinalnu moždanu koru miša.

There is no effective treatment for Alzheimer’s disease (AD). As many drugs discovery attempts based on the amyloid cascade hypothesis have not been successful, tau protein has become an increasingly important therapeutic target owing to advances in understanding of its role in AD pathogenesis. In AD, the balance of tau kinase and phosphatase activity is shifted, creating a highly phosphorylated species of tau. This raises the fraction of unbound tau, which is no longer attached to microtubules, allowing for monomeric hyperphosphorylated tau to bind to one another to produce oligomers. Fusion of these oligomeric species leads to the formation of paired helical filaments, the primary constituent of neurofibrillary changes, which have been documented to be an early and pivotal event (so-called „pre-tangle“ stage) in the pathogenesis of AD. Our aims are: 1) to define better the relationship between the early AD biomarkers total and phospho-tau proteins, as well as amyloid ß in cerebrospinal fluid (CSF) of AD, MCI (mild cognitive impairment), and control subjects. By using innovative ELISA and modified thioflavin S binding assay on CSF samples, we will also assess the ratio between monomer and oligomer (aggregated) tau species. These new data will be correlated to clinical, neuropsychological, and MRI findings; 2) to test the effect of several known and potentially neuroprotective compounds (donepezil, clorgyline, several flavonoids, PF9601N and ASS234, among others) on tau phosphorylation induced by okadaic acid and nocodazole in SH-SY5Y and P19 cell cultures. Tau epitopes phosphorylated at Thr181, Ser199 and Thr231 residues will be analyzed by ELISA, while p-tau epitopes at Thr202, Thr205, Ser396 and Ser404 will be analyzed by Western blot; and 3) to investigate biochemically and immunocytochemically the trans-synaptic spreading of neurofibrillary degeneration and tau mRNA expression after the injection of tau oligomers in the mouse entorhinal cortex.

Ključne riječi

Alzheimer's disease, cerebrospinal fluid, ELISA, neurofibrillary degeneration, neuroprotective compounds, tau protein, thioflavine S

Znanstveno-istraživački projekti

ALZTAUPROTECT

IP-2014-09-9730

12.10.2015

11.10.2019

nije evidentirano

HRK 1.000.000

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Podaci o institucijama

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