Leishmania donovani Aurora kinase: A promising therapeutic target against visceral leishmaniasis |
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Institution: | 1. Infectious Diseases and Immunology Division, Council of Scientific and Industrial Research (CSIR), Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Jadavpur, Kolkata 700032, West Bengal, India;2. Assam University, Silchar (A Central University), Dept. of Pharmaceutical Science, Division of Drug Discovery Lab, Silchar 788011, Assam, India;3. Council of Scientific and Industrial Research (CSIR), Centre for Cellular and Molecular Biology, Uppal Road, Habshiguda, Hyderabad 500007, Telangana, India |
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Abstract: | BackgroundAurora kinases are key mitotic kinases executing multiple aspects of eukaryotic cell-division. The apicomplexan homologs being essential for survival, suggest that the Leishmania homolog, annotated LdAIRK, may be equally important.MethodsBioinformatics, stage-specific immunofluorescence microscopy, immunoblotting, RT-PCR, molecular docking, in-vitro kinase assay, anti-leishmanial activity assays, flow cytometry, fluorescence microscopy.ResultsLdairk expression is seen to vary as the cell-cycle progresses from G1 through S and finally G2M and cytokinesis. Kinetic studies demonstrate their enzymatic activity exhibiting a Km and Vmax of 6.12 μM and 82.9pmoles·min− 1mg− 1 respectively against ATP using recombinant Leishmania donovani H3, its physiological substrate. Due to the failure of LdAIRK −/+ knock-out parasites to survive, we adopted a chemical knock-down approach. Based on the conservation of key active site residues, three mammalian Aurora kinase inhibitors were investigated to evaluate their potential as inhibitors of LdAIRK activity. Interestingly, the cell-cycle progressed unhindered, despite treatment with GSK-1070916 or Barasertib, inhibitors with greater potencies for the ATP-binding pocket compared to Hesperadin, which at nanomolar concentrations, severely compromised viability at IC50s 105.9 and 36.4 nM for promastigotes and amastigotes, respectively. Cell-cycle and morphological studies implicated their role in both mitosis and cytokinesis.ConclusionWe identified an Aurora kinase homolog in L. donovani implicated in cell-cycle progression, whose inhibition led to aberrant changes in cell-cycle progression and reduced viability.General significanceHuman homologs being actively pursued drug targets and the observations with LdAIRK in both promastigotes and amastigotes suggest their potential as therapeutic-targets. Importantly, our results encourage the exploration of other proteins identified herein as potential novel drug targets. |
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