Title Mehanizmi antitumorskog djelovanja biološki aktivnih sastavnica ljekovitih gljiva na modelu kolorektalnog karcenoma
Title (english) Antitumor Mechanisms of Biologically Compounds from Medicinal Mushrooms on the Colorectal Cancer Model
Author Boris Jakopović
Mentor Nada Oršolić (mentor)
Mentor Sandra Kraljević Pavelić (komentor)
Committee member Marina Ćetković-Cvrlje (predsjednik povjerenstva)
Committee member Hrvoje Jakovac (član povjerenstva)
Committee member Draženka Komes (član povjerenstva)
Committee member Nada Oršolić (član povjerenstva)
Committee member Sandra Kraljević Pavelić (član povjerenstva)
Granter University of Rijeka (Faculty of Biotechnology and Drug Development) Rijeka
Defense date and country 2021, Croatia
Scientific / art field, discipline and subdiscipline BIOTECHNICAL SCIENCES Biotechnology
Universal decimal classification (UDC ) 577 - Biochemistry. Molecular biology. Biophysics
Abstract Kolorektalni karcinom (CRC) treća je najčešća vrsta karcinoma kod muškaraca i žena, s oko 35% pacijenata kojima je dijagnosticirana metastatska bolest u IV stadiju. Unatoč napretku u liječenju, očekivano trajanje života u bolesnika s etastatskom bolešću još uvijek nije zadovoljavajuće. Zbog česte rezistencije na lijekove tijekom konvencionalnih i ciljanih tretmana karcinoma, razvoj i ispitivanje terapija koje djeluju na više ciljeva važno je polje istraživanja. Izolirani spojevi specifični za ljekovite gljive, kao i složene smjese ekstrakata, detaljno su proučavani, a mnoge vrste gljiva dokazano su netoksični inhibitori raznih specifičnih onkogenih putova, kao i snažni imunomodulatori. Utvrdili smo značajan antiproliferativni i proapoptotički učinak ljekovitih gljiva pripravaka AGARIKON.1 i AGARIKON PLUS na ne-kolorektalne (A549, MCF-7, MIA PaCa2, HepG2) i kolorektalne (HCT-116, SW620) tumorske stanične linije, dok je učinak na staničnu liniju ljudskih fibroblasta (WI-38) bio proliferativan. Dalje smo istraživali učinak ljekovitih gljivičnih pripravaka AGARIKON.1 i AGARIKON PLUS u različitim kombinacijama s 5- fluorouracilom na modelu kolorektalnog karcinoma CT26.WT. Budući da je liječenje započelo tek nakon 14 dana nakon inokulacije tumora, ovo predstavlja vrlo kasni tumorski model uznapredovalog metastatskog karcinoma. Rezultati pokazuju da AGARIKON.1 i AGARIKON PLUS pokazuju imunostimulacijska i antiangiogena svojstva koja različito utječu na preživljenje: AGARIKON PLUS ima najslabiji učinak na povećanje ukupnog preživljenja (engl. overall survival) (OS = 22,2%), u kombinaciji sa 5-FU značajno ga produžuje u odnosu na negativnu kontrolu (OS = 87,5%), ali ne statistički značajno više u odnosu na 5-FU (OS = 77,7%) samostalno. Antitumorski učinci AGARIKON.1 i AGARIKON PLUS, sa ili bez 5-fluorouracila, temelje se na povećanju polarizacije M1 makrofaga, inhibiciji polarizacije M2 i TAM, učincima na profile citokina Th1/Th2/Th17, inhibiciji vaskularnih endotelnih čimbenika rasta (VEGF) i modulaciji metaloproteinaza 2 i 9 (MMP-2 i MMP-9). Neki od ovih učinaka mogu se pripisati antioksidativnoj sposobnosti fenolnih i drugih sastavnica prisutnih u ljekovitim gljivama. Primjena AGARIKON.1 i AGARIKON PLUS nije pokazala genotoksičnost niti bilo kakav štetan učinak na kliničke hematološke ili biokemijske parametre. Nadalje, proteomsko profiliranje tumorskih tkiva otkrilo je da AGARIKON.1 i AGARIKON PLUS sami ili u kombinaciji s 5-fluorouracilom ostvaruju antitumorske učinke utječući na temeljne procese u progresiji raka debelog crijeva (CRC), poput translacije, odgovor na stres nesmotanih proteina (UPR) kao i ciklus lipida i trikarboksilne kiseline (TCA). Ovi podaci pružaju dobru osnovu za proširenu translacijsku studiju.
Abstract (english) Colorectal cancer (CRC) is the third most frequent cancer type in both males and females, with about 35% of patients being diagnosed in stage IV metastatic disease. Despite advancements in treatment, life expectancy in patients with metastatic disease is still not satisfying. Due to frequent drug resistance during conventional and targeted cancer treatments, the development and testing of multi-target therapies is an important research field. Medicinal mushrooms specific isolated compounds as well as complex extract mixtures have been studied in depth, and many mushroom
species have been proven to be non-toxic multi-target inhibitors of specific oncogenic pathways, as well as potent immunomodulators. We established a significant antiproliferative and proapoptotic effect of medicinal mushroom preparations AGARIKON.1 and AGARIKON PLUS on non-colorectal (A549, MCF-7, MIA PaCa-2, HepG2) and colorectal (HCT-116, SW620) tumor cell lines, while the effect on human fibroblast cell line (WI-38) was proliferative. We further
investigated the effect of medicinal mushroom preparations AGARIKON.1 and AGARIKON PLUS in various combinations with 5-fluorouracil in CT26.WT mouse model. Since the treatment started only after 14 days after tumor inoculation, this represents a very late tumor model of advanced metastatic cancer. The results show that AGARIKON.1 and AGARIKON PLUS exhibit immunostimulatory, antiangiogenic properties which affect survival differently: AGARIKON PLUS has the weakest effect on increasing overall survival (OS = 22.2%), in combination with 5- FU significantly prolongs it in relation to the negative control (OS = 87.5%), but not statistically significantly more in relation to 5-FU (OS = 77.7%) alone. The antitumor effects of AGARIKON.1 and AGARIKON PLUS, with or without 5-fluorouracil, are based on enhancing M1 macrophage polarization, inhibition of M2 and TAM polarization, effects on Th1/Th2/Th17 citokine profiles, direct inhibition of CT26.WT tumor growth, inhibition of vascular endothelial growth factors (VEGF) and metalloproteinases 2 and 9 (MMP-2 and MMP-9) modulation. Some of these effects can be attributed to the antioxidant ability of phenolic and other components present in medicinal mushrooms. The administration of AGARIKON.1 and AGARIKON PLUS did not show
genotoxicity nor any adverse effect on clinical hematological or biochemical parameters. Furthermore, proteomic profiling of tumor tissues revealed that AGARIKON.1 and AGARIKON PLUS alone, or in combination with 5-fluorouracil, exert their antitumor effects by affecting fundamental processes in colorectal cancer (CRC) progression, such as translation, unfolded
protein response (UPR) as well as lipid and tricarboxylic acid cycle (TCA). This data provides good basis for an expanded translational study.
Keywords
ljekovite gljive
kolorektalni karcinom
kombinirana terapija
5-fluorouracil
preživljenje
angiogeneza
polarizacija makrofaga
translacija
stres nesmotanih proteina
tumorski metabolizam
proteomika
Keywords (english)
medicinal mushrooms
colorectal cancer
combination therapy
5-fluorouracil
survival
angiogenesis
macrophage polarization
translation
unfolded protein stress
tumor metabolism
proteomics
Language croatian
URN:NBN urn:nbn:hr:193:257710
Promotion 2022
Study programme Title: Medicinal chemistry Study programme type: university Study level: postgraduate Academic / professional title: doktor/doktorica znanosti, interdisciplinarna područja znanosti, polje biotehnologija u biomedicini (doktor/doktorica znanosti, interdisciplinarna područja znanosti, polje biotehnologija u biomedicini)
Type of resource Text
File origin Born digital
Access conditions Closed access
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Created on 2022-03-14 09:59:11