Main description:

This volume provides detailed, up-to-date methods used in research on Atherosclerosis. Chapters guide readers through an overview of the pathogenesis of atherosclerosis and model systems together with in vitro, ex vivo, in vivo and emerging methods in atherosclerosis research. 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, Atherosclerosis: Methods and Protocols serves as an invaluable resource for those engaging in research on atherosclerosis and cardiovascular disease, as well as for researchers who are new to the field and students at both the undergraduate and postgraduate level.

Contents:

PART I: Atherosclerosis: Pathogenesis, model systems and emerging concepts.

1. Atherosclerosis: Pathogenesis and key cellular processes, current and emerging therapies, key challenges and future research directions

Yee-Hung Chan and Dipak P. Ramji

2. Key roles of inflammation in atherosclerosis: Mediators involved in orchestrating the inflammatory response and its resolution in the disease along with therapeutic avenues targeting inflammation

Yee-Hung Chan and Dipak P. Ramji

3. Survey of in vitro model systems for investigation of key cellular processes associated with atherosclerosis

Dipak P. Ramji, Alaa Ismail, Jing Chen, Fahad Alradi, and Sulaiman Al Alawi

4. Survey of approaches for investigation of atherosclerosis in vivo

Dipak P. Ramji, Yee-Hung Chan, Alaa Alahmadi, Reem Alotibi, and Nouf Alshehri

5. Exploring the associations between clonal hematopoiesis of indeterminate potential, myeloid malignancy and atherosclerosis

Juan Bautista Menendez-Gonzalez, and Neil P. Rodrigues

6. Peptides as therapeutic agents for atherosclerosis

C. Roger White, Mayakonda Palgunachari, Paul Wolkowicz, and G. M. Anantharamaiah

PART II: In vitro and ex vivo model systems for atherosclerosis research.

7. Isolation and culturing of primary mouse and human macrophages

Alba de Juan and Begona Lavin

8. Effective transfection and gene silencing of primary murine macrophages with small interfering RNA

Kim van Kuijk, Jan Nagenborg, and Judith C. Sluimer

9. Monitoring celular proliferation, migration and apoptosis associated with atherosclerosis plaques in vitro

Kerry S. Wadey, Alexandros Somos, Stephen J. Cross, Lien M. Reolizo, Jason L. Johnson, and Sarah J. George

10. Monocyte adhesion assays for detecting endothelial cell activation in vascular inflammation and aterosclerosis

Keman Xu, Fatma Saaoud, Stephanie Yu, Charles Drummer IV, Ying Shao, Yu Sun, Yifan Lu, Jianxin Sun, Jun Yu, Xiaohua Jiang, Hong Wang, and Xiaofeng Yang

11. Chick embryo ex vivo aortic sprouting assays for cardiovascular research

Selvaraj Vimalraj and Anuradha Dhanasekaran

12. Purification and analysis of circulating lipid particles

Barnaby W. R. Roper, Basmah Al-Sayejh, Ahmed Al-Aufi, Gary A. Cuthbert, Katie Lacey, Shervanthi Homer-Vanniasinkam, Michael A. Harrison, Darren C. Tomlinson, Ramzi Ajjan, and Sreenivasan Ponnambalam

13. Preparation of LDL, oxidation, methods of detection and applications in atherosclerosis research

Chandrakala Aluganti Narasimhulu and Sampath Parthasarathy

14. Monitoring modified lipoprotein uptake and macropinocytosis associated with macrophage foam cell formation

Alaa A. Alahmadi and Dipak P. Ramji

15. A fluorescent based in vitro method to assess cholesterol efflux. Sara Fernandez-Castillejo, Anna Pedret, Ursula Catalan, and Rosa Sola

16. Monitoring the anisotropy and fluidity of the HDL monolayer as

surrogates of HDL functionality

Sara Fernandez-Castillejo, Anna Pedret, Ursula Catalan, Claude Motta, and Rosa Sola

17. Assessment of ex-vivo antioxidative potential of murine HDL in atherosclerosis

Andrea Rivas-Urbina, Noemi Rotllan, David Santos, Josep Julve, Joan Carles Escola-Gil, and Jose Luis Sanchez-Quesada

18. Dual-fluorescence in vitro efferocytosis assay

Arif Yurdagul Jr

19. Investigation of mitochondrial bioenergetic profile and dysfunction in atherosclerosis

Yee-Hung Chan, and Dipak P. Ramji

20. Probing inflammasome activation in atherosclerosis

Yee-Hung Chan and Dipak P. Ramji

21. Investigation of platelet aggregation in atherosclerosis

Ronan Lordan, Alexandros Tsoupras, and Ioannis Zabetakis

22. Extracellular vesicles in atherosclerosis research

Jessica O. Williams, Cass Whelan, Jamie Nash, and Philip E. James

23. The use of wire myography to investigate vascular tone and function

Kayleigh Griffiths and Melanie Madhani

PART III: In vivo model systems for atherosclerosis research.

24. Pig and mouse models of hyperlipidemia and atherosclerosis

Godfrey S. Getz and Catherine A. Reardon

25. Use of rabbit models to study atherosclerosis

Jianglin Fan, Manabu Nimimi, Yajie Chen, Ritsuko Suzuki, and Enqi Liu

26. Genetically engineered hamster models of dyslipidemia and atherosclerosis

Xunde Xian, Yuhui Wang, and George Liu

27. Atherosclerosis induced by adeno-associated virus encoding gain-of-function PCSK9

Martin Maeng Bjorklund, Juan A. Bernal, and Jacob F. Bentzon

28. Monitoring leukocyte migration during atherosclerosis in vivo

Keiichiro Matoba

29. Evaluation of plaque burden and lipid content in atherosclerotic plaques Yee-Hung Chan, Alaa Alahmadi, Reem Alotibi, and Dipak P. Ramji

30. Monitoring cellularity and expression of key markers in atherosclerotic plaques. Yee-Hung Chan, Reem Alotibi, Alaa Alahmadi, and Dipak P. Ramji

31. Monitoring cellular proliferation and apoptosis in atherosclerosis plaques and intimal thickenings

Kerry S. Wadey, Jason L. Johnson, and Sarah J. George

32. Investigation of atherosclerotic plaque vulnerability

Sarah J. George and Jason L. Johnson

33. Use of mouse carotid artery ligation model of intimal thickening to probe vascular smooth muscle cell remodelling and function in atherosclerosis

Helen Williams, Bethan A. Brown, Jason L. Johnson, and Sarah J. George

34. Assessing efferocytosis in atherosclerotic lesions in situ

Arif Yurdagul Jr

35. Monitoring atheroprotective macrophage cholesterol efflux in vivo

Andrea Rivas-Urbina, Noemi Rotllan, David Santos, Josep Julve, Jose Luis Sanchez-Quesada, and Joan Carles Escola-Gil

36. Flow cytometry analysis of hematopoietic stem/progenitor cells and mature blood cell subsets in atherosclerosis

Alhomidi Almotiri, Ali Abdelfattah, and Neil P. Rodrigues

37. Whole mount preparation of mouse aorta for confocal microscopy studies of the intima

Ana Barettino, Ignacio Benedicto, and Vicente Andres

38. Isolation of mouse aortic RNA for transcriptomics

Rosa M. Nevado, Magda R. Hamczyk, and Vicente Andres

39. Redox proteomics analysis of atherosclerotic aortas: Application of the "OxICAT" method

Manousos Makridakis and Antonia Vlahou

40. Intravital microscopy in atherosclerosis research

Georg Wissmeyer, Mohamad B. Kassab, Yoichiro Kawamura, Aaron D. Aguirre, and Farouc A. Jaffer

41. Localization of long non-coding RNA in formalin-fixed-paraffin embedded vascular tissue using in-situ hybridization

Jessica P. Scanlon, Andrew H. Baker, and Judith C. Sluimer

PART IV: Emerging techniques in atherosclerosis research.

42. In vivo gene editing in lipid and atherosclerosis research

Marco De Giorgi, Kelsey E. Jarrett, Thomas Q. de Aguiar Vallim, and William R. Lagor

43. Laser capture microdissection-based mRNA expression microarrays and single-cell RNA sequencing in atherosclerosis research

Xi Zhang, Zhihua Wang, Chuankai Zhang, Yutao Li, Shu Lu, Sabine Steffens, Sarajo Mohanta, Christian Weber, Andreas Habenicht, and Changjun Yin

44. Combined single cell RNA and single cell / T cell receptor sequencing of the arterial wall in atherosclerosis

Zhihua Wang, Xi Zhang, Chuankai Zhang, Yutao Li, Shu Lu, Sarajo Mohanta, Christian Weber, Andreas Habenicht, and Changjun Yin

45. Tissue clearing approaches in atherosclerosis

Ting Sun, Yuanfang Li, Benjamin Foerstera, Karen Stanic, Shu Lu, Sabine Steffens, Changjun Yin, Ali Erturk, Remco T. A. Megens, Christian Weber, Andreas Habenicht, and Sarajo K. Mohanta

46. Single cell antibody sequencing in atherosclerosis research

Ryosuke Saigusa, Christopher P. Durant, Vasantika Suryawanshi, and Klaus Ley

47. Flow cytometry and mass cytometry for measuring the immune cell infiltrate in atherosclerotic arteries

Marco Orecchioni, Melissa A. Meyer, Catherine C. Hedrick and Klaus Ley

48. Contrast-enhanced ultrasound molecular imaging in atherosclerosis

The Anh Nguyen and Jonathan R. Lindner

49. Nanoprobes for computed tomography and magnetic resonance imaging in atherosclerosis research

Ketan B. Ghaghada, Rohan Bhavane, Andrew Badachhape, Eric Tanifum, and Ananth Annapragada

50. Positron emission tomography in atherosclerosis research

Anne Roivainen, Mia Stahle, and Antti Saraste

51. Three-dimensional visualization of atherosclerotic vessels by tissue clearing and light sheet fluorescence microscopy

Tobias Becher, Dario F. Riascos-Bernal, Jingyi Chi, Paul Cohen, and Nicholas E. S. Sibinga

52. Intravascular fluorescence molecular imaging of atherosclerosis

Mohammed M. Chowdhury, Zhonglie Piao, Mazen S. Albaghdadi, Patrick A. Coughlin, James H.F. Rudd, Guillermo J. Tearney, and Farouc A. Jaffer