Cercosporamide inhibits bone morphogenetic protein receptor type I kinase activity in zebrafish

Abstract

Zebrafish models are well established tools for investigating underlying mechanisms of diseases. Here, we identified cercosporamide, a metabolite from the fungus Ascochyta aquiliqiae, as a potent bone morphogenetic protein receptor (BMPR) type I kinase inhibitor through a zebrafish embryo phenotypic screen. The developmental defects in zebrafish, including lack of the ventral fin induced by cercosporamide was strikingly similar as the phenotypes caused by renowned small molecule BMPR type I kinase inhibitors and inactivating mutations in zebrafish BMPRs. In mammalian cell-based assays, cercosporamide blocked BMP/SMAD-dependent transcriptional reporter activity and BMP-induced SMAD1/5-phosphorylation. Biochemical assays with a panel of purified recombinant kinases demonstrated that cercosporamide directly inhibited kinase activity of BMPRs type I (also called activin receptor-like kinases (ALKs)). In mammalian cells, cercosporamide selectively inhibited constitutively active BMPR type I-induced SMAD1/5 phosphorylation. Importantly, cercosporamide rescued the developmental defects caused by constitutively active Alk2 in zebrafish embryos. Taken together, we believe cercosporamide may be the first of a new class of molecules with potential to be developed further for clinical use against diseases that are causally linked to overactivation of BMPR signaling, including Fibrodysplasia ossificans progressiva and diffuse intrinsic pontine glioma.

Migration of Kupffer’s vesicle derived cells is essential for tail morphogenesis in zebrafish embryos

“…The bone morphogenetic protein (BMP) gradient is essential for dorsoventral patterning of the zebrafish tail (Row and Kimelman, 2009). BMP signaling inhibitors such as dorsomorphin and DMH-1 result in a dorsalized phenotype including loss of ventral fins, as does cercosporamide that we identified previously (Hao et al, 2010;Hoeksma et al, 2020;Yang and Thorpe, 2011;Yu et al, 2008). Similar dorsoventral patterning defects are observed in loss-of-function mutants of key genes of the BMPsignaling pathway, including activin receptor-like kinase 2 (alk2) (in zebrafish also known as alk8 or losta-fin) (Bauer et al, 2001), its ligand, bmp2 (Kishimoto et al, 1997) or intracellular factors such as smad5 (Kramer et al, 2002).…”

“…Previously, several other small molecules, such as para-coumaric acid methyl ester (pCAME) and BMP-inhibitors dorsomorphin, DMH-1 and cercosporamide were reported to induce ectopic tail tissue (Gebruers et al, 2013;Hoeksma et al, 2020;Yang and Thorpe, 2011). The effect of these compounds was compared by testing them in parallel to geraniol in our assay (Fig.…”

“…In addition, we found that 25.2% of the active fungal filtrates induced tail defects. In a follow-up study, we focused on this category and identified the fungal compound cercosporamide as a BMP-signaling inhibitor, based on the dorsalized phenotype it induces in zebrafish tails (Hoeksma et al, 2020), demonstrating that fungal filtrates interfering with tail morphogenesis harbor compounds with interesting biological activities.…”

Dibenzofuran Derivatives Inspired from Cercosporamide as Dual Inhibitors of Pim and CLK1 Kinases

“…In addition, inhibition of Mnk pathways by cercosporamide enhanced glioblastoma cell response to chemotherapy and radionuclide therapy [11]. Recent results in the zebrafish model, through the inhibition of bone morphogenetic protein receptor (BMPR) type I kinase, suggest that cercosporamide represents a potential therapeutic strategy to combat diseases with overactive BMPR signaling, including the rare genetic disorder fibrodysplasia ossificans progressiva (FOP) and the rare childhood brainstem tumor diffuse intrinsic pontine glioma (DIPG) [12].…”

Cercosporamide, a polyketide-derived fungal metabolite, serves as an antifungal agent against phytopathogenic fungi

“…Cercosporamide was also reported to block phosphorylation of MNK-eukaryotic initiation factor 4E (EIF4E), resulting in potent inhibitory effects on primitive leuke-mic progenitors (Altman et al, 2013) and human hepatocellular carcinoma (Liu et al, 2016). Recently, cercosporamide was shown to inhibit a bone morphogenetic protein receptor (BMPR) type I kinase activity in zebrafish embryos (Hoeksma et al, 2020).…”