Identification of inhibitors for the transmembranei Trypanosoma/ii cruzi/i eIF2α kinase relevant for parasite proliferation

0583467 - BC 2024 RIV NL eng J - Journal Article
Marcelino, T.P. - Fala, A.M. - da Silva, M.M. - Souza-Melo, N. - Malvezzi, A.M. - Klippel, A.H. - Zoltner, M. - Padilla-Mejia, N.E. - Kosto, S. - Field, Mark Christian - Burle-Caldas, G.A. - Teixeira, S.M.R. - Counago, R.M. - Massirer, K.B. - Schenkman, S.
Identification of inhibitors for the transmembranei Trypanosoma/ii cruzi/i eIF2α kinase relevant for parasite proliferation.
Journal of Biological Chemistry. Roč. 299, č. 7 (2023), č. článku 104857. ISSN 0021-9258. E-ISSN 1083-351X
R&D Projects: GA MŠMT(CZ) EF16_019/0000759
Institutional support: RVO:60077344
Keywords : chemical biology * mechanism * target * complex * stress * brucei * domain * dna
OECD category: Microbiology
Impact factor: 4.8, year: 2022
Method of publishing: Open access
https://www.sciencedirect.com/science/article/pii/S0021925823018859?via%3Dihub

The TcK2 protein kinase of Trypanosoma cruzi, the causative agent of Chagas disease, is structurally similar to the human kinase PERK, which phosphorylates the initiation factor eIF2 alpha and, in turn, inhibits translation initiation. We have previously shown that absence of TcK2 kinase impairs parasite proliferation within mammalian cells, positioning it as a potential target for treatment of Chagas disease. To better understand its role in the parasite, here we initially confirmed the importance of TcK2 in parasite proliferation by generating CRISPR/Cas9 TcK2-null cells, albeit they more efficiently differentiate into infective forms. Proteomics indicates that the TcK2 knockout of proliferative forms expresses proteins including trans-sialidases, normally restricted to infective and nonproliferative trypomastigotes explaining decreased proliferation and better differentiation. TcK2 knockout cells lost phosphorylation of eukaryotic initiation factor 3 and cyclic AMP responsive-like element, recognized to promote growth, likely explaining both decreased proliferation and augmented differentiation. To identify specific inhibitors, a library of 379 kinase inhibitors was screened by differential scanning fluo- rimetry using a recombinant TcK2 encompassing the kinase domain and selected molecules were tested for kinase inhibition. Only Dasatinib and PF477736, inhibitors of Src/Abl and ChK1 kinases, showed inhibitory activity with IC50 of 0.2 +/- 0.02 mM and 0.8 +/- 0.1, respectively. In infected cells Dasatinib inhibited growth of parental amastigotes (IC50 = 0.6 +/- 0.2 mM) but not TcK2 of depleted parasites (IC50 > 34 mM) identifying Dasatinib as a potential lead for development of therapeutics for Chagas disease targeting TcK2.
Permanent Link: https://hdl.handle.net/11104/0351436