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Cerebral Cavernous Malformations (CCM)

Cerebral Cavernous Malformations (CCM)

Methods and Protocols

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CHF 223.00
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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.


Part I: Overview of Cerebral Cavernous Malformations

1. 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 Malformations

Giovanni G. Vercelli, Fabio Cofano, Filippo Veneziani Santonio, Francesca Vincitorio, Francesco Zenga, and Diego Garbossa


Part II Diagnosis and Treatment of Cerebral Cavernous Malformations

4. 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) Disease

Valerio 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 Severity

Hélène Choquet and Helen Kim


7. Clinical Imaging of Cerebral Cavernous Malformations: Computed Tomography and Magnetic Resonance Imaging

Marc Mabray and Blaine Hart


8. Neuroradiology: Differential Diagnosis, Follow-Up, and Reporting

Alessandra Splendiani, Federico Bruno, and Alfonso Cerase


9. Surgical Management of Brain Cavernous Malformations

Marco 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 Malformations

10. 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 Disease

Preston 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 Disease

Luca Goitre, Claudia Fornelli, Alessia Zotta, Andrea Perrelli, and Saverio Francesco Retta


13. CRISPR/Cas9-mediated Generation of Human Endothelial Cell Knockout Models of CCM Disease

Konrad 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 Model

Alondra B. Schweizer Burguete and Amin S. Ghabrial


15. Generation and Analysis of CCM Phenotypes in C. elegans

Evelyn 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 Vasculature

Stefan Donat, and Salim Abdelilah-Seyfried


17. Vertebrate Models to Investigate CCM Pathogenesis: The Zebrafish and Mouse Model

Johnathan Abou-Fadel and Jun Zhang


18. Generation of Cerebral Cavernous Malformation in Neonatal Mouse Models using Inducible Cre-LoxP Strategy

Jaesung P. Choi and Xiangjian Zheng


19. Isolation of Cerebral Endothelial Cells from CCM1/KRIT1 Null Mouse Brain

Nicholas Nobiletti and Angela J. Glading


Part IV: Cellular and Molecular Mechanisms of Cerebral Cavernous Malformations

20. Identification of the KRIT1 Protein by LexA-based Yeast Two-Hybrid System

Ilya G. Serebriiskii, Mohamed Elmekawy, and Erica A. Golemis


21. Crystallographic Studies of the Cerebral Cavernous Malformations Proteins

Oriana S. Fisher, Xiaofeng Li, Weizhi Liu, Rong Zhang, and Titus J. Boggon


22. Microscopy Techniques to Investigate CCM Pathogenesis

Johnathan Abou-Fadel and Jun Zhang


23. Preparation and Analysis of Protein Extracts to Investigate CCM Pathogenesis

Johnathan Abou-Fadel and Jun Zhang


24. Systems Wide Analysis of CCM Signaling Complex (CSC) Alterations in CCM Deficient Models using Omics Approaches

Johnathan Abou-Fadel and Jun Zhang


25. Study of Molecular Interactions of CCM Proteins by using a GAL4-based Yeast Two-hybrid Screening

Federica Finetti and Lorenza Trabalzini


26. Study of CCM Microvascular Endothelial Phenotype by an in vitro Tubule Differentiation Model

Simona Delle Monache and Saverio Francesco Retta


27. Bidimentional in vitro Angiogenic Assays to Study CCM Pathogenesis: Endothelial Cell Proliferation and Migration

Federica Finetti and Lorenza Trabalzini


28. Measurement of Endothelial Barrier Function in Mouse Models of Cerebral Cavernous Malformations using Intravital Microscopy

Angela 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 Cells

Sandra 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 Immunofluorescence

Saverio Marchi, Saverio Francesco Retta, and Paolo Pinton


31. Notch Signaling in Familial Cerebral Cavernous Malformations and Immunohistochemical Detection of cleaved Notch1 Intracellular Domain

Sana S. Hasan and Andreas Fischer


32. Measuring the Kinase Activity of GCKIII Proteins in vitro

Juan 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) Disease

Cinzia Antognelli, Vincenzo Nicola Talesa, and Saverio Francesco Retta


34. Fluorescence Analysis of Reactive Oxygen Species (ROS) in Cellular Models of Cerebral Cavernous Malformation Disease

Andrea Perrelli and Saverio Francesco Retta


35. Small RNA Library Preparation for Profiling Small Non-coding RNAs in Patients Affected by Cavernous Malformations

Souvik Kar, Robert Geffers, Amir Samii, and Helmut Bertalanffy


36. Affinity Purification and Preparation of Peptides for Mass Spectrometry from C. elegans

Evelyn Popiel and William Brent Derry

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