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Chapter 3. The adenoviral vector angiogenesis/lymphangiogenesis assay.

Janice A Nagy1, Shou-Ching Shih, Wendy H Wong

  • 1Department of Pathology, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA.

Methods in Enzymology
|November 15, 2008
PubMed
Summary

Adenoviral vectors expressing vascular endothelial growth factor (VEGF-A(164)) create diverse abnormal blood and lymphatic vessels. This assay aids in studying pathological angiogenesis and evaluating therapies.

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Area of Science:

  • Vascular biology
  • Molecular medicine
  • Regenerative medicine

Background:

  • Pathological angiogenesis and lymphangiogenesis are key in diseases like cancer.
  • Understanding the formation of abnormal vasculature is crucial for developing targeted therapies.
  • Adenoviral vectors offer a method to deliver genetic material for studying these processes.

Purpose of the Study:

  • To investigate the utility of adenoviral vectors expressing VEGF-A(164) in modeling pathological angiogenesis and lymphangiogenesis.
  • To characterize the distinct types of abnormal blood and lymphatic vessels induced by Ad-VEGF-A(164).
  • To establish a screening assay for evaluating pro- and anti-angiogenic therapies.

Main Methods:

  • Injection of adenoviral vectors expressing VEGF-A(164) (Ad-VEGF-A(164)) into various tissues of nude mice and rats.
  • Visual monitoring of angiogenic and lymphangiogenic responses, particularly in ear injection sites.
  • Characterization of induced vessel types including mother vessels, capillaries, glomeruloid microvascular proliferations, vascular malformations, feeding arteries, and draining veins.

Main Results:

  • Ad-VEGF-A(164) induced six distinct types of abnormal blood vessels and "giant" lymphatics.
  • The assay allowed elucidation of the mechanisms and steps of new vessel formation.
  • Ear injection sites provided a convenient model for visual, real-time assessment of therapeutic efficacy and toxicity.

Conclusions:

  • Adenoviral vectors expressing VEGF-A(164) provide a robust model for studying pathological angiogenesis and lymphangiogenesis.
  • The Ad-VEGF-A(164) assay is a valuable tool for screening anti- and pro-angiogenic agents.
  • This methodology can be extended to study other growth factors and cytokines involved in neovascularization.