Targeting DYRK1A/B kinases to modulate p21‐cyclin D1‐p27 signalling and induce anti‐tumour activity in a model of human glioblastoma

Abstract

The dual‐specificity tyrosine‐regulated kinases DYRK1A and DYRK1B play a key role in controlling the quiescence‐proliferation switch in cancer cells. Serum reduction of U87MG 2D cultures or multi‐cellular tumour spheroids induced a quiescent like state characterized by increased DYRK1B and p27, and decreased pRb and cyclin D1. VER‐239353 is a potent, selective inhibitor of the DYRK1A and DYRK1B kinases identified through fragment and structure‐guided drug discovery. Inhibition of DYRK1A/B by VER‐239353 in quiescent U87MG cells increased pRb, DYRK1B and cyclin D1 but also increased the cell cycle inhibitors p21 and p27. This resulted in exit from G0 but subsequent arrest in G1. DYRK1A/B inhibition reduced the proliferation of U87MG cells in 2D and 3D culture with greater effects observed under reduced serum conditions. Paradoxically, the induced re‐expression of cell cycle proteins by DYRK1A/B inhibition further inhibited cell proliferation. Cell growth arrest induced in quiescent cells by DYRK1A/B inhibition was reversible through the addition of growth‐promoting factors. DYRK inhibition‐induced DNA damage and synergized with a CHK1 inhibitor in the U87MG spheroids. In vivo, DYRK1A/B inhibition‐induced tumour stasis in a U87MG tumour xenograft model. These results suggest that further evaluation of VER‐239353 as a treatment for glioblastoma is therefore warranted.

Comparative Efficacy and Selectivity of Pharmacological Inhibitors of DYRK and CLK Protein Kinases

“…VER-239353, a related product (#34 in ref 97), was investigated as a potential drug for glioblastoma (DYRK1A IC 50 = 76 nM). 99 Our results show that compounds 40 and 48 have potent activity toward not only DYRK1A (IC 50 = 8 and 5 nM, respectively), DYRK2 (5 and 3 nM), and DYRK2 (10 and 9 nM) but also CLK1 (16 and 8 nM) and CLK4 (10 and 16 nM). This potency was confirmed in the luminescence assay (Table 2).…”

Chemical, Biochemical, Cellular, and Physiological Characterization of Leucettinib-21, a Down Syndrome and Alzheimer’s Disease Drug Candidate

“…Given the literature supporting the link between DYRK1A inhibition and induction of glioblastoma cell death ,− (see the review in ref ) and given the relative brain permeability of Leucettinib-21 ( 4 ) (unpublished results), we tested this compound and its kinase-inactive isomer on patient-derived, Temozolomide-resistant glioblastoma stem-like cells (GSCs) (Figure ). Cells were exposed for 48 h to a range of concentrations of both compounds, and cell death was assessed using the Cell Titer-Glo luminescent ATP assay.…”

“…Leucettinib-21 (4) and iso-Leucettinib-21 (7) were next tested at 1 and 10 μM, respectively, in a panel of 413 kinases (radiometric assay, Eurofins-Cerep) (Table 5, Table S5). Thirteen kinases showed more than 85% inhibition by Leucettinib-21 (4), while not a single kinase was inhibited by iso-Leucettinib-21 (7) (Table 5A).…”

“…The IC 50 value for GSK-3β was 2 μM. Iso-Leucettinib-21 (7) was completely inactive on all kinases (IC 50 > 10 μM, the highest dose tested). These results were closely confirmed with the proximity assay (Table 7).…”

CCT2 enhances the proliferation and migration of head and neck squamous cell carcinoma via regulating cell cycle

“…GSK-3β is well known for its suppressive function on cyclin D1 and E1 protein level by downregulating mRNA transcription and protein stability [32]. Both p21 and p27 protein are members of the Cip/Kip family of cyclin-dependent kinase (CDK) inhibitors that can inhibit kinase activities of CDK2-cyclin E complex, CDK4/6-cyclin D complex, and other cyclins [33][34][35]. Enhancement of p27protein level may cause G1 arrest [36].…”