Sulindac metabolites decrease cerebrovascular malformations in CCM3 -knockout mice

Abstract

Cerebral cavernous malformation (CCM) is a disease of the central nervous system causing hemorrhage-prone multiple lumen vascular malformations and very severe neurological consequences. At present, the only recommended treatment of CCM is surgical. Because surgery is often not applicable, pharmacological treatment would be highly desirable. We describe here a murine model of the disease that develops after endothelial-cell-selective ablation of the CCM3 gene. We report an early, cell-autonomous, Wnt-receptor-independent stimulation of β-catenin transcription activity in CCM3-deficient endothelial cells both in vitro and in vivo and a triggering of a β-catenin-driven transcription program that leads to endothelial-tomesenchymal transition. TGF-β/BMP signaling is then required for the progression of the disease. We also found that the anti-inflammatory drugs sulindac sulfide and sulindac sulfone, which attenuate β-catenin transcription activity, reduce vascular malformations in endothelial CCM3-deficient mice. This study opens previously unidentified perspectives for an effective pharmacological therapy of intracranial vascular cavernomas.cerebral cavernous malformation | endothelial cells | β-catenin | sulindac metabolites | vascular pathology T he vascular malformations that characterize the disease known as cerebral cavernous malformation (CCM) are concentrated in the central nervous system, and they typically show multiple lumens and vascular leakage (1). These abnormalities can result in severe neurological symptoms, including hemorrhagic stroke (2), and, to date, the only possible therapy is surgery (3). In humans, loss-of-function mutations in any one of three independent genes known as cerebral cavernous malformation 1, 2, and 3 (CCM1, CCM2, and CCM3) are the cause of the genetic form of CCM (4). Similarly, in murine models, the vascular phenotype can be reproduced by endothelium-specific loss-of-function mutations of any one of these three CCM-linked genes (5-7).We have recently reported (7) that TGF-β/BMP signaling is activated after ablation of CCM1, CCM2, or CCM3 and induces endothelial-to-mesenchymal transition (EndMT) that plays a crucial role in the development of vascular malformations. Nevertheless, the sequence of signaling responses elicited by ablation of CCM genes still remains to be defined. Inhibitors of the TGF-β/BMP-signaling pathways reduce the number and size of the malformations, but not completely (7), suggesting that other signaling pathways may be implicated.The Wnt/β-catenin pathway, in synergy with TGF-β signaling (8), is responsible for the EndMT switch of endothelial cells giving rise to the heart cushion in the embryo. In addition, the knockdown of CCM1 and CCM3 expression in cultured aortic and artery endothelial cells promotes β-catenin signaling (9, 10), although no direct link with the in vivo model of the disease has been made. Activation of canonical Wnt/β-catenin signaling is critical for brain vascularization and acquisition, by the microvasculature, of blood-brain barr...

Cerebral Cavernous Malformations: Review of the Genetic and Protein–Protein Interactions Resulting in Disease Pathogenesis

“…Another recent intriguing development in the study of CCMs is the discovery that PDCD10-deficient endothelial cells in CCMs undergo endothelial-to-mesenchymal transformation (48). This transformation is the result of the loss of PDCD10-mediated regulation and subsequent upregulation of β-catenin signaling.…”

“…This transformation is the result of the loss of PDCD10-mediated regulation and subsequent upregulation of β-catenin signaling. Bravi et al also found that once this change occurred in the endothelial cells of CCMs, TGF-β/BMP signaling was subsequently required for the progression of the disease (48). The authors also found that this endothelial-to-mesenchymal cell transformation occurred in sporadic CCM lesions in addition to the familial and animal model lesions (49).…”

Combined HMG-COA reductase and prenylation inhibition in treatment of CCM

“…Moreover, fasudil (but not simvastatin) reduced lesion burden and improved survival in sensitized Ccm1 (but not Ccm2) heterozygous mice (11). The nonsteroidal antiinflammatory drug sulindac sulfide constrained vascular lesions in Cdh5(PAC)CreERT2/Ccm3 cKO mice lacking endothelial CCM3 (28). Combined treatment with fluvastatin and zoledronate appears to have effects comparable to sulindac sulfide on survival; both approaches also reduced lesion burden, although slight differences in analyses preclude direct comparisons.…”

“…We next tested the effects of combined treatment in an endothelial cell-specific, acute model of CCM3 disease. We used inducible Cdh5(PAC)CreERT2/Ccm3 cKO mice, which develop vascular malformations in the cerebellum and retina (27) that respond to pharmacological manipulation (28). Newborn pups were induced with tamoxifen and then were injected daily with fluvastatin [15 mg·kg −1 ·d −1 intragastrically (i.g.)]…”

RHO binding to FAM65A regulates Golgi reorientation during cell migration

“…Mutations of the CCM3 gene have been linked to cerebral cavernous malformations -vascular abnormalities characterised by dilated leaky cerebral lesions that can lead to brain haemorrhage (Draheim et al, 2014). The exact mechanism by which cerebral cavernous malformations arise is still subject to debate, with deregulation of several signalling pathways such as RHO (Richardson et al, 2013;Stockton et al, 2010;Borikova et al, 2010;Whitehead et al, 2009), TGFβ (Maddaluno et al, 2013), β-catenin (Bravi et al, 2015) and MEKK3-KLF2 or MEKK3-KLF4 (Cuttano et al, 2016;Zhou et al, 2016;Renz et al, 2015) having been demonstrated to be involved in development and progression of the disease. Crucially, loss of the CCM3 interaction with GCKIII kinases seems to be the crucial feature of all disease-associated CCM3 mutations (Fidalgo et al, 2010).…”