Publisher Description

This volume provides experimental approaches aimed to characterize the Cerebral Cavernous Malformations (CCM) disease and to define the cellular and molecular mechanisms underlying this pathology. Chapters are divided into four sections providing a general overview of the natural history, epidemiology, and pathogenetic mechanisms of CCM disease, describing methods currently used for diagnosis and treatment, production and analysis of distinct cellular and animal models in which CCM can be studied, and different methodological approaches to study the mechanisms underlying the CCM onset and progression. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and cutting-edge, Cerebral Cavernous Malformations (CCM): Methods and Protocols  aims to ensure successful results in the further study of this complex disease and its pathophysiologic correlates.

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This volume provides experimental approaches aimed to characterize the Cerebral Cavernous Malformations (CCM) disease and to define the cellular and molecular mechanisms underlying this pathology. Chapters are divided into four sections providing a general overview of the natural history, epidemiology, and pathogenetic mechanisms of CCM disease, describing methods currently used for diagnosis and treatment, production and analysis of distinct cellular and animal models in which CCM can be studied, and different methodological approaches to study the mechanisms underlying the CCM onset and progression. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Cerebral Cavernous Malformations (CCM): Methods and Protocols aims to ensure successful results in the further study of this complex disease and its pathophysiologic correlates.

Table of Contents

Part I: Overview of Cerebral Cavernous Malformations1. From Genes and Mechanisms to Molecular-targeted Therapies: the Long Climb to the Cure of Cerebral Cavernous Malformation (CCM) Disease Saverio Francesco Retta, Federica Finetti, Andrea Perrelli, and Lorenza Trabalzini 2. Incidence, Prevalence, and Clinical Presentation of Cerebral Cavernous Malformations Kelly D. Flemming 3. Natural History, Clinical, and Surgical Management of Cavernous MalformationsGiovanni G. Vercelli, Fabio Cofano, Filippo Veneziani Santonio, Francesca Vincitorio, Francesco Zenga, and Diego Garbossa  Part II Diagnosis and Treatment of Cerebral Cavernous Malformations4. Molecular Genetics Screening of CCM patients, An Overview Elisabeth Tournier-Lasserve 5. Next Generation Sequecing (NGS) Strategies for Genetic Testing and Diagnosis of Cerebral Cavernous Malformation (CCM) DiseaseValerio Benedetti, Elisa Pellegrino, Alfredo Brusco, Roberto Piva, and Saverio Francesco Retta 6. Genome-wide Genotyping of Cerebral Cavernous Malformation Type 1 Individuals to Identify Genetic Modifiers of Disease SeverityHélène Choquet and Helen Kim 7. Clinical Imaging of Cerebral Cavernous Malformations: Computed Tomography and Magnetic Resonance ImagingMarc Mabray and Blaine Hart  8. Neuroradiology: Differential Diagnosis, Follow-Up, and ReportingAlessandra Splendiani, Federico Bruno, and Alfonso Cerase 9. Surgical Management of Brain Cavernous MalformationsMarco M. Fontanella, Luca Zanin, Alessandro Fiorindi, Giannantonio Spena, Federico Nicolosi, Francesco Belotti, Pierpaolo Panciani, Claudio Cornali, and Francesco Doglietto Part III: Animal and Cellular Models of Cebral Cavernous Malformations10. Generation of CCM Phenotype by a Human Microvascular Endothelial Model Simona Delle Monache and Saverio Francesco Retta 11. Isolation and Purification of Mouse Brain Endothelial Cells to Study Cerebral Cavernous Malformation DiseasePreston Hale, Shady Ibrahim Soliman, Hao Sun, and Miguel Alejandro Lopez-Ramirez 12. Production of KRIT1-knockout and KRIT1-knockin Mouse Embryonic Fibroblasts as Cellular Models of CCM DiseaseLuca Goitre, Claudia Fornelli, Alessia Zotta, Andrea Perrelli, and Saverio Francesco Retta 13. CRISPR/Cas9-mediated Generation of Human Endothelial Cell Knockout Models of CCM DiseaseKonrad Schwefel, Stefanie Spiegler, Christiane D. Much, Ute Felbor, and Matthias Rath 14. Dissection of the Role of CCM Genes in Tubulogenesis using the Drosophila Tracheal System as a ModelAlondra B. Schweizer Burguete and Amin S. Ghabrial 15. Generation and Analysis of CCM Phenotypes in C. elegansEvelyn Popiel and William Brent Derry 16. Generation of Transgenic Lines of Zebrafish Expressing Fluorescently-tagged CCM Proteins to Study Their Function and Subcellular Localization within the VasculatureStefan Donat, and Salim Abdelilah-Seyfried 17. Vertebrate Models to Investigate CCM Pathogenesis: The Zebrafish and Mouse ModelJohnathan Abou-Fadel and Jun Zhang 18. Generation of Cerebral Cavernous Malformation in Neonatal Mouse Models using Inducible Cre-LoxP StrategyJaesung P. Choi and Xiangjian Zheng 19. Isolation of Cerebral Endothelial Cells from CCM1/KRIT1 Null Mouse BrainNicholas Nobiletti and Angela J. Glading Part IV: Cellular and Molecular Mechanisms of Cerebral Cavernous Malformations20. Identification of the KRIT1 Protein by LexA-based Yeast Two-Hybrid SystemIlya G. Serebriiskii, Mohamed Elmekawy, and Erica A. Golemis 21. Crystallographic Studies of the Cerebral Cavernous Malformations ProteinsOriana S. Fisher, Xiaofeng Li, Weizhi Liu, Rong Zhang, and Titus J. Boggon 22. Microscopy Techniques to Investigate CCM PathogenesisJohnathan Abou-Fadel and Jun Zhang 23. Preparation and Analysis of Protein Extracts to Investigate CCM PathogenesisJohnathan Abou-Fadel and Jun Zhang 24. Systems Wide Analysis of CCM Signaling Complex (CSC) Alterations in CCM Deficient Models using Omics ApproachesJohnathan Abou-Fadel and Jun Zhang 25. Study of Molecular Interactions of CCM Proteins by using a GAL4-based Yeast Two-hybrid ScreeningFederica Finetti and Lorenza Trabalzini 26. Study of CCM Microvascular Endothelial Phenotype by an in vitro Tubule Differentiation ModelSimona Delle Monache and Saverio Francesco Retta 27. Bidimentional in vitro Angiogenic Assays to Study CCM Pathogenesis: Endothelial Cell Proliferation and MigrationFederica Finetti and Lorenza Trabalzini 28. Measurement of Endothelial Barrier Function in Mouse Models of Cerebral Cavernous Malformations using Intravital MicroscopyAngela J. Glading 29. Immunofluorescence of Cell-Cell and Cell-Extracellular Matrix Adhesive Defects in in vitro Endothelial CCM Model. Juxtacrine Role of Mutant Extracellular Matrix on Wild-Type Endothelial CellsSandra Manet, Daphné Vannier, Anne-Pascale Bouin, Justyna Lisowska, Corinne Albiges-Rizo, and Eva Faurobert 30. Detection of p62/SQSTM1 Aggregates in Cellular Models of CCM Disease by ImmunofluorescenceSaverio Marchi, Saverio Francesco Retta, and Paolo Pinton 31. Notch Signaling in Familial Cerebral Cavernous Malformations and Immunohistochemical Detection of cleaved Notch1 Intracellular DomainSana S. Hasan and Andreas Fischer 32. Measuring the Kinase Activity of GCKIII Proteins in vitroJuan Zalvide, Cristina Almengló, Sara Vázquez, Mar García-Colomer, Miriam Sartages, and Celia M. Pombo 33. Spectrophotometric Method for Determining Glyoxalase 1 Activity in Cerebral Cavernous Malformation (CCM) DiseaseCinzia Antognelli, Vincenzo Nicola Talesa, and Saverio Francesco Retta 34. Fluorescence Analysis of Reactive Oxygen Species (ROS) in Cellular Models of Cerebral Cavernous Malformation DiseaseAndrea Perrelli and Saverio Francesco Retta 35. Small RNA Library Preparation for Profiling Small Non-coding RNAs in Patients Affected by Cavernous MalformationsSouvik Kar, Robert Geffers, Amir Samii, and Helmut Bertalanffy 36. Affinity Purification and Preparation of Peptides for Mass Spectrometry from C. elegansEvelyn Popiel and William Brent Derry

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Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

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