http___www.irb.hr_Istrazivanja_Zavodi_Zavod-za-molekularnu-medicinu_Laboratorij-za-istrazivanje-neurodegenerativnih-bolesti_Projekti_Molekularni-mehanizam-neurodegeneracije-u-Niemann-Pickovoj-bolesti-tip-C_1.xml
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Ispitivanje uloge BACE1 za razvoj liječenja protiv bolesti Niemann-Pick tipa C The B-secretase, known as B-site amyloid precursor protein cleavage enzyme 1 (BACE1), plays a central role in Alzheimer’s disease (AD) pathogenesis as it initiates the production of the toxic amyloid-B peptides (AB) that accumulate in the brains of AD patients. BACE1 has become a prime therapeutic target for lowering AB, and clinical development of BACE1 inhibitors is being intensely pursued as avenue for treatment of AD. Interestingly, a rare inherited - yet untreatable - lysosomal storage disorder Niemann-Pick type C (NPC) and AD have several key features in common, including accumulation of AB and the B-secretase generated C-terminal fragments of the B-amyloid precursor protein (APP) - CTFB, hyperphosphorylation of tau, neurodegeneration, and altered cholesterol metabolism. In light with the recently discovered similarities between AD and NPC, the goal of this project is to elucidate the role of BACE1 in the pathogenesis of NPC and whether BACE1 may represent a novel target for treating/ameliorating NPC disease. Furthermore, NPC can serve as an innovative model in which understanding of a defect caused by a single gene/protein (NPC1/NPC2) may help us to dissect molecular details of a more complex Alzheimer’s disease. However, these two neurodegenerative disorders show different vulnerabilities of the brain regions such as hippocampus, which is the most affected in AD and spared in NPC disease, and the cerebellum, which is spared in AD but is primarily affected in NPC. Further studies of similarities and differences between AD and NPC are encouraged to increase our understanding of the molecular pathogenesis of these conditions, as well as our chances of finding disease-modifying therapies. Based on our recent finding that B-secretase (BACE1) cleavage of APP is enhanced in NPC, in this project we will analyze whether enhanced BACE1-proteolysis in NPC is specific for APP or could also be observed for other BACE1 substrates such as seizure protein 6 (Sez6) and seizure 6-like protein 1 (Sez6L1). Both proteins seem to be involved in synaptic (dys)function and neurodegeneration – characteristic features of both AD and NPC disease. We will also test whether enhanced BACE1-cleavage in NPC correlates with distinct regional/neuronal vulnerabilities, accumulation of AB and CTFB, tau hyperphosphorylation, activation of astrocytes and microglia. Lastly, using BACE1-inhibitor we will determine whether targeting BACE1 could be considered as an alternative approach for treating/ameliorating NPC disease. These studies may indicate a novel role of BACE1 in NPC pathogenesis and the potential of BACE1-inhibition as a novel strategy for treating NPC. - Glavni istraživač / voditelj - Ostali suradnici Martina Malnar - Nemate ovlasti za pregled ovog sadržaja. DZNE - Munchen, Njemačka Sabina Tahirović https://www.dzne.de/en/sites/munich/research-groups/tahirovic.html Alessio Colombo Anna Daria
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Molekularni mehanizam neurodegeneracije u Niemann-Pickovoj bolesti tip C Bolest Niemann-Pick tipa C (NPC) je rijetka nasljedna lizosomska bolest nakupljanja koju karakterizira nakupljanje kolesterola u lizosomima što dovodi do progresivne neurodegeneracije. Zanimljivo, ova rijetka bolest (uzrokovana mutacijama u genu NPC1ili NPC2) pokazuje niz značajki kompleksne Alzheimerove bolesti (AB) koje uključuju nakupljanje peptida amiloid-B (AB) i hiperfosforilaciju proteina tau. Međutim, ove neurodegenerativne bolesti pokazuju različitu „ranjivost“ regija mozga; hipokampus je primarno oštećen u AB, dok je pošteđen u bolesti NPC, a cerebelum je pošteđen u AB, dok je primarno zahvaćen u bolesti NPC gdje dolazi do značajnog gubitka Purkinje neurona. U ovom projektu ćemo primijeniti saznanja opsežnih istraživanja najčešće neurodegenerativne bolesti AB s ciljem razumijevanja molekularnog mehanizam neurodegeneracije u rijetkoj bolesti NPC. Naši rezultati pokazuju da nakupljanje kolesterola u staničnom modelu bolesti NPC potiče povećano cijepanje proteina prekursora peptida amiloid-B (APP) proteazom BACE1 i uzrokuje povećan nastanak peptida AB. Nedavno su, uz protein APP, otkriveni i brojni drugi supstrati proteaze BACE1 u mozgu čime je pokazana izrazito važna uloga ove proteaze u mozgu. Potaknuti tim otkrićima, ispitat ćemo ulogu proteolize enzimom BACE1 u patogenezi bolesti NPC i može li BACE1 predstavljati novu ciljnu molekulu za tretiranje/ublažavanje bolesti NPC. U hipokampusu i cerebelumu NPC1- i wt-miševa ispitat ćemo proteolizu i distribuciju APP kao i dva, nedavno otkrivena, isključiva supstrata za BACE1. Primarne kulture neurona hipokampusa i Purkinjeovih neurona cerebeluma ćemo upotrijebiti za otkrivanje mehanizma učinka BACE1 na patološke značajke NPC bolesti. Također, razvit ćemo ex vivo model bolesti NPC korištenjem organotipskih kultura moždanih rezova hipokampusa i cerebeluma te ćemo pratiti utjecaj inhibicije BACE1 na prevenciju/ublažavanje patoloških značajki bolesti NPC. Konačno, generirat ćemo mišji model NPC1 u kojem ćemo deletirati gen BACE1 za potvrdu dobivenih rezultatain vivo. Ova istraživanja će ukazati na ulogu BACE1 u patogenezi NPC bolesti i na potencijalno ciljanje enzima BACE1 kao alternativnog pristupa liječenja ove još uvijek neizlječive bolesti. - Glavni istraživač / voditelj - Ostali suradnici Kristina Dominko, mag.biol.mol., istraživač DZNE - Munchen, Njemačka Sabina Tahirović https://www.dzne.de/en/sites/munich/research-groups/tahirovic.html Alessio Colombo Stefan Lichtenthaler https://www.dzne.de/en/sites/munich/research-groups/lichtenthaler.html - Napomena - instalacija Olympus SZ61 Stereo Mikroskopa 14. prosinca 2017. - NPC1 mišji model stigao na IRB Kristina Dominko na djelu ... - Otvoreni dani IRBa - predstavljanje projekta u okviru punkta "BašTIna kulture stanica" - voditelj punkta: mag.biol.mol. Kristina Dominko - Kristina je sa svojim kolegicama predstavila različite stanične linije, primarne kulture neurona kao i bojanje proteina u stanicama i u rezovima mišjih mozgova metodom imunofluorescencije