Abstract
Cerebral cavernous malformations (CCM) or cavernomas are slow-flow capillary vascular malformations with a mulberry-like appearance, which are predominantly located in the central nervous system. CCM can occur in a sporadic or a familial form. The latter is inherited in an autosomal dominant manner, and in the majority of the fragile lesions, mutations in the genes CCM1 (KRIT1), CCM2 (OSM), or CCM3 (PDCD10) can be detected. Loss of these genes leads to numerous alterations in endothelial cell signaling resulting in a disturbed vessel architecture and function. Lower activity of Notch signaling occurs upon loss of CCM1, CCM3, or the CCM1-interacting protein ICAP1 in cell culture and animal models. Notch signaling in endothelial cells is an essential regulator of angiogenesis, arterial-venous differentiation, vascular permeability and stability, mural cell recruitment, and flux of metabolites across the vessel wall. The purpose of this chapter is to briefly summarize the current understanding of Notch signaling in familial CCM and to provide a protocol for detecting cleaved Notch1 receptor proteins on paraformaldehyde-fixed paraffin-embedded mouse tissue.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Fischer A, Zalvide J, Faurobert E, Albiges-Rizo C, Tournier-Lasserve E (2013) Cerebral cavernous malformations: from CCM genes to endothelial cell homeostasis. Trends Mol Med 19(5):302–308
Zafar A, Quadri SA, Farooqui M, Ikram A, Robinson M, Hart BL et al (2019) Familial cerebral cavernous malformations. Stroke 50(5):1294–1301
Whitehead KJ, Plummer NW, Adams JA, Marchuk DA, Li DY (2004) Ccm1 is required for arterial morphogenesis: implications for the etiology of human cavernous malformations. Development 131(6):1437–1448
Bray SJ (2016) Notch signalling in context. Nat Rev Mol Cell Biol 17(11):722–735
Potente M, Gerhardt H, Carmeliet P (2011) Basic and therapeutic aspects of angiogenesis. Cell 146(6):873–887
Red-Horse K, Siekmann AF (2019) Veins and arteries build hierarchical branching patterns differently: bottom-up versus top-down. BioEssays 41(3):e1800198
Gaengel K, Genove G, Armulik A, Betsholtz C (2009) Endothelial-mural cell signaling in vascular development and angiogenesis. Arterioscler Thromb Vasc Biol 29(5):630–638
Hill-Felberg S, Wu HH, Toms SA, Dehdashti AR (2015) Notch receptor expression in human brain arteriovenous malformations. J Cell Mol Med 19(8):1986–1993
Murphy PA, Kim TN, Huang L, Nielsen CM, Lawton MT, Adams RH et al (2014) Constitutively active Notch4 receptor elicits brain arteriovenous malformations through enlargement of capillary-like vessels. Proc Natl Acad Sci U S A 111(50):18007–18012
Jabs M, Rose AJ, Lehmann LH, Taylor J, Moll I, Sijmonsma TP et al (2018) Inhibition of endothelial notch signaling impairs fatty acid transport and leads to metabolic and vascular remodeling of the adult heart. Circulation 137(24):2592–2608
Wieland E, Rodriguez-Vita J, Liebler SS, Mogler C, Moll I, Herberich SE et al (2017) Endothelial Notch1 activity facilitates metastasis. Cancer Cell 31(3):355–367
Fischer A, Gessler M (2007) Delta-Notch--and then? Protein interactions and proposed modes of repression by Hes and Hey bHLH factors. Nucleic Acids Res 35(14):4583–4596
Itoh F, Itoh S, Goumans MJ, Valdimarsdottir G, Iso T, Dotto GP et al (2004) Synergy and antagonism between Notch and BMP receptor signaling pathways in endothelial cells. EMBO J 23(3):541–551
Woltje K, Jabs M, Fischer A (2015) Serum induces transcription of Hey1 and Hey2 genes by Alk1 but not Notch signaling in endothelial cells. PLoS One 10(3):e0120547
Kar S, Baisantry A, Nabavi A, Bertalanffy H (2016) Role of Delta-Notch signaling in cerebral cavernous malformations. Neurosurg Rev 39(4):581–589
Wustehube J, Bartol A, Liebler SS, Brutsch R, Zhu Y, Felbor U et al (2010) Cerebral cavernous malformation protein CCM1 inhibits sprouting angiogenesis by activating DELTA-NOTCH signaling. Proc Natl Acad Sci U S A 107(28):12640–12645
Brutsch R, Liebler SS, Wustehube J, Bartol A, Herberich SE, Adam MG et al (2010) Integrin cytoplasmic domain-associated protein-1 attenuates sprouting angiogenesis. Circ Res 107(5):592–601
Maddaluno L, Rudini N, Cuttano R, Bravi L, Giampietro C, Corada M et al (2013) EndMT contributes to the onset and progression of cerebral cavernous malformations. Nature 498(7455):492–496
Schulz GB, Wieland E, Wustehube-Lausch J, Boulday G, Moll I, Tournier-Lasserve E et al (2015) Cerebral cavernous Malformation-1 protein controls DLL4-Notch3 signaling between the endothelium and pericytes. Stroke 46(5):1337–1343
He Y, Zhang H, Yu L, Gunel M, Boggon TJ, Chen H et al (2010) Stabilization of VEGFR2 signaling by cerebral cavernous malformation 3 is critical for vascular development. Sci Signal 3(116):ra26
Schleider E, Stahl S, Wustehube J, Walter U, Fischer A, Felbor U (2011) Evidence for anti-angiogenic and pro-survival functions of the cerebral cavernous malformation protein 3. Neurogenetics 12(1):83–86
You C, Sandalcioglu IE, Dammann P, Felbor U, Sure U, Zhu Y (2013) Loss of CCM3 impairs DLL4-Notch signalling: implication in endothelial angiogenesis and in inherited cerebral cavernous malformations. J Cell Mol Med 17(3):407–418
Acknowledgments
This work was supported by Deutsche Forschungsgemeinschaft (DFG) SFB1366 project C4 to A.F.
We thank Iris Moll for establishing the staining method and Leonie Uhl for providing the images presented in this article.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Hasan, S.S., Fischer, A. (2020). Notch Signaling in Familial Cerebral Cavernous Malformations and Immunohistochemical Detection of Cleaved Notch1 Intracellular Domain. In: Trabalzini, L., Finetti, F., Retta, S. (eds) Cerebral Cavernous Malformations (CCM) . Methods in Molecular Biology, vol 2152. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0640-7_31
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0640-7_31
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0639-1
Online ISBN: 978-1-0716-0640-7
eBook Packages: Springer Protocols