The structure-function relationship of oncogenic LMTK3

Sci Adv. 2020 Nov 13;6(46):eabc3099. doi: 10.1126/sciadv.abc3099.

Angeliki Ditsiou ¹, Chiara Cilibrasi ¹, Nikiana Simigdala ², Athanasios Papakyriakou ³, Leanne Milton-Harris ¹, Viviana Vella ¹, Joanne E Nettleship ⁴ ⁵, Jae Ho Lo ⁶, Shivani Soni ⁶, Goar Smbatyan ⁶, Panagiota Ntavelou ², Teresa Gagliano ¹, Maria Chiara Iachini ¹, Sahir Khurshid ⁷, Thomas Simon ¹, Lihong Zhou ⁸, Storm Hassell-Hart ⁹, Philip Carter ¹⁰, Laurence H Pearl ⁸, Robin L Owen ¹¹, Raymond J Owens ⁴ ⁵ ¹², S Mark Roe ⁸, Naomi E Chayen ⁷, Heinz-Josef Lenz ⁶, John Spencer ⁹, Chrisostomos Prodromou ¹, Apostolos Klinakis ², Justin Stebbing ¹⁰, Georgios Giamas ¹³

Affiliations

1 Department of Biochemistry and Biomedicine, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QG, UK.
2 Center of Basic Research, Biomedical Research Foundation of the Academy of Athens, 11527 Athens, Greece.
3 Institute of Biosciences and Applications, National Centre for Scientific Research "Demokritos," 15341 Athens, Greece.
4 Division of Structural Biology, University of Oxford, The Wellcome Centre for Human Genetics Headington, Oxford OX3 7BN, UK.
5 Protein Production UK, Research Complex at Harwell, Harwell Science and Innovation Campus, Didcot OX11 0FA, UK.
6 Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
7 Faculty of Medicine, Division of Systems Medicine, Department of Metabolism, Digestion and Reproduction, Imperial College, Sir Alexander Fleming Building, South Kensington Campus, London SW7 2AZ, UK.
8 Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Science Park Road, Falmer, Brighton BN1 9RQ, UK.
9 Department of Chemistry, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QJ, UK.
10 Faculty of Medicine, Department of Surgery and Cancer, Imperial College, London W12 0NN, UK.
11 Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, UK.
12 The Rosalind Franklin Institute, Harwell Campus, Didcot OX11 0FA, UK.
13 Department of Biochemistry and Biomedicine, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QG, UK. g.giamas@sussex.ac.uk.

PMID: 33188023 DOI: 10.1126/sciadv.abc3099

Abstract

Elucidating signaling driven by lemur tyrosine kinase 3 (LMTK3) could help drug development. Here, we solve the crystal structure of LMTK3 kinase domain to 2.1Å resolution, determine its consensus motif and phosphoproteome, unveiling in vitro and in vivo LMTK3 substrates. Via high-throughput homogeneous time-resolved fluorescence screen coupled with biochemical, cellular, and biophysical assays, we identify a potent LMTK3 small-molecule inhibitor (C28). Functional and mechanistic studies reveal LMTK3 is a heat shock protein 90 (HSP90) client protein, requiring HSP90 for folding and stability, while C28 promotes proteasome-mediated degradation of LMTK3. Pharmacologic inhibition of LMTK3 decreases proliferation of Cancer cell lines in the NCI-60 panel, with a concomitant increase in Apoptosis in breast Cancer cells, recapitulating effects of LMTK3 gene silencing. Furthermore, LMTK3 inhibition reduces growth of xenograft and transgenic breast Cancer mouse models without displaying systemic toxicity at effective doses. Our data reinforce LMTK3 as a druggable target for Cancer therapy.

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