Voditelj	Prof. dr. sc. Dora Višnjić
Istraživači	Prof.dr.sc. Drago Batinić, Medicinski fakultet Sveučilišta u Zagrebu, KBC Zagreb Prof.dr.sc. Antonio Bedalov, Fred Hutchinson Cancer Research Center, SAD dr. sc. Hrvoje Lalić, Medicinski fakultet Sveučilišta u Zagrebu dr. sc. Josip Batinić, KBC Zagreb dr. sc. Vilma Dembitz, Medicinski fakultet Sveučilišta u Zagrebu dipl. ing. Klara Dubravčić, KBC Zagreb
Iznos potpore	1.000.000,00 kuna
Početak projekta	15.04.2017.
Kraj projekta	14.04.2021.

SAŽETAK PROJEKTA

Akutna mijeloična leukemija (AML) je heterogena skupina bolesti koja je obilježena nekontroliranom proliferacijom blasta koji su zakočeni u ranoj fazi diferencijacije. Najuspješnije farmakološko liječenje AML je diferencijacijsko liječenje pomoću sve-trans-retinske kiseline (ATRA, prema engl. all-trans retinoic acid). Međutim, ATRA se rabi isključivo u liječenju akutne promijelocitne leukemije (APL),  podvrste AML koja sadrži translokaciju t(15;17). Svi ostali oblici AML liječe se intenzivnom kemoterapijom koja nije značajno produljila trajanje remisije ili opće preživljenje. Naše nedavno istraživanje pokazalo je da 5-aminoimidazol-4-karboksamid ribonukleozid (AIKAR, akadezin) potiče apoptozu i pospješuje diferencijaciju staničnih linija AML koje nisu APL, ali je mehanizam djelovanja AIKAR-a još uvijek nepoznat. Stoga je svrha predloženih istraživanja određivanje mehanizama koji su odgovorni za povoljno djelovanje AIKAR-a u stanicama non-APL AML te otkrivanje signalnih mehanizama i metaboličkih promjena koje su odgovorne za monocitnu i granulocitnu diferencijaciju stanica AML. U istraživanju ćemo rabiti komercijalne stanične linije AML da bismo opisali: a) ulogu i mehanizam autofagije u monocitno/makrofagnoj i granulocitnoj diferencijaciji, b) promjene u metabolizmu tijekom diferencijacije i prolaska kroz stanični ciklus, te c) ulogu deacetilaza Sirt u monocitno/makrofagnoj i granulocitnoj diferencijaciji staničnih linija AML. Dodatno, ispitat ćemo učinke AIKAR-a in vitro na uzorcima blasta izoliranih iz oboljelih od AML. Istraživanja signalnih mehanizama odgovornih za diferencirajući učinak u leukemijskim stanicama mogu u konačnici pridonijeti uspješnijem liječenju oboljelih od AML.

SUMMARY 

Acute myeloid leukemia (AML) is a heterogeneous group of diseases characterized by clonal proliferation of blasts that are blocked at an early stage of differentiation. The most successful pharmacological therapy of AML is differentiation therapy with all-trans-retinoic acid (ATRA); however, ATRA-based therapy is restricted to acute promyelocytic leukemia (APL), a particular subtype that carries t(15;17) translocation. All other AMLs are treated with intensive chemotherapy, which have not significantly improved the disease-free or overall survival. Our recent study demonstrated that 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR, acadesine) induces apoptosis and enhances differentiation of non-APL AML cell lines, but the mechanism is still unknown. Therefore, proposed studies are aimed to determine the mechanism responsible for beneficial effects of AICAR in non-APL AML cells and to further elucidate signaling mechanisms and metabolic changes responsible for monocytic and granulocytic differentiation of AML cells. We will use commercially available AML cell lines to define: a) the role and the mechanism of autophagy in monocytic/macrophage and granulocytic differentiation, b) the changes in metabolism during differentiation and cell cycle progression, and to determine c) the role of deacetylases Sirt in monocytic/macrophage and granulocytic differentiation of AML cell lines. In addition, in vitro profiling of the sensitivity of primary AML samples to AICAR will be performed. The proposed project aims to define signaling mechanisms responsible for differentiation of leukemia cells, which may eventually lead to the development of an improved therapy, thus contributing to well-being of AML patients.

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PUBLIKACIJE VODITELJA I ISTRAŽIVAČA KOJE SU PROIZAŠLE IZ PROJEKTA

1. Dembitz V, Lalic H, Visnjic D. 5-Aminoimidazole-4-carboxamide ribonucleoside-induced autophagy flux during differentiation of monocytic leukemia cells. Cell Death Discov. 2017 Oct 2;3:17066. doi: 10.1038/cddiscovery.2017.66. eCollection 2017.

Konferencijska izlaganja

1. Dembitz V, Lalić H, Kodvanj I, Višnjić D. Glycolysis and autophagy flux during differentiation along the monocytic lineage“, Annual Meeting of the CIS with EFIS on Tour. Zagreb 2017

2. Dembitz V, Lalic H, Visnjic D. Autophagy in AICAR-mediated differentiation of acute myeloid leukemia cells U937. EMBO Conference - Autophagy, Dubrovnik-Cavtat, 2017. Poster presentation

3. Dembitz V, Lalic H, Visnjic D. 5-aminoimidazole-4-carboxamide ribonucleoside induces autophagy in U937 cells. 4th Congress of Croatian Physiological Society (CPS) and 2nd Regional Congress of the Physiological Societies, Dubrovnik 2017. Poster presentation

SUMMARY

Acute myeloid leukemia (AML) is a heterogeneous group of diseases characterized by clonal proliferation of blasts that are blocked at an early stage of differentiation. The most successful pharmacological therapy of AML is differentiation therapy with all-trans-retinoic acid (ATRA); however, ATRA-based therapy is restricted to acute promyelocytic leukemia (APL), a particular subtype that carries t(15;17) translocation. All other AMLs are treated with intensive chemotherapy, which have not significantly improved the disease-free or overall survival. Our recent study demonstrated that 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR, acadesine) induces apoptosis and enhances differentiation of non-APL AML cell lines, but the mechanism is still unknown. Therefore, proposed studies are aimed to determine the mechanism responsible for beneficial effects of AICAR in non-APL AML cells and to further elucidate signaling mechanisms and metabolic changes responsible for monocytic and granulocytic differentiation of AML cells. We will use commercially available AML cell lines to define: a) the role and the mechanism of autophagy in monocytic/macrophage and granulocytic differentiation, b) the changes in metabolism during differentiation and cell cycle progression, and to determine c) the role of deacetylases Sirt in monocytic/macrophage and granulocytic differentiation of AML cell lines. In addition, in vitro profiling of the sensitivity of primary AML samples to AICAR will be performed. The proposed project aims to define signaling mechanisms responsible for differentiation of leukemia cells, which may eventually lead to the development of an improved therapy, thus contributing to well-being of AML patients.