Effects of VLA-4 antagonists in rat whole embryo culture

Teratology. 2002 Jan;65(1):26-37. doi: 10.1002/tera.1095.

Abstract

Background: Pharmacological antagonism of VLA-4 (Very Late Antigen 4, alpha(4)beta(1) integrin) has become an attractive target for the treatment of predominantly eosinophil mediated disease states such as asthma, allergic rhinitis, multiple sclerosis, rheumatoid arthritis, diabetes, and inflammatory bowel disease. Gene knockouts of the alpha(4)-integrin subunit of VLA-4 or its cell surface ligand, VCAM-1, however, have been shown to result in embryo-lethality in homozygous null mice due to defects in chorio-allantoic or epi-myocardial fusion. Although gene knockout phenotypes are not always manifested by pharmacological antagonism, those studies suggested that VLA-4 antagonists might cause embryo-lethality or drug-induced malformations.

Methods: To test these concepts, early neurulating rat embryos were cultured by the methods of New ('78) after intra-coelomic microinjection of a VLA-4 blocking antibody or in the presence of small molecule VLA-4 antagonists.

Results: Defects in chorio-allantoic fusion were induced after microinjection of VLA4 blocking antibody and after continuous exposure to small molecule antagonists. In a minority of affected embryos chorio-allantoic fusion was completely blocked whereas the majority of affected embryos had only superficial chorio-allantoic fusion and the allantois was enlarged and edematous. Although the allantoic mesoderm covered the trophoblasts of the chorionic plate and contained blood vessels there was only minimal invasion of the trophoblasts by the allantoic mesoderm. The lowest observed effect level generally correlated with the IC(approximately 95), as determined in 90% plasma.

Discussion: Based on these data, VLA-4 antagonism might represent a significant risk to the developing embryo/fetus. In vitro exposure, however, is "constant" and does not take into account the elimination phase of these xenobiotics in vivo. Given the high concentrations required to elicit an effect, therapeutic blood levels in vivo may be several fold lower than those that affect the conceptus, depending on the tissue penetration of the compound and the route of administration. VLA-4 also exists in a range of conformations and activation states in vivo and the gene KOs and present studies do not define whether these developmental processes are dependent upon a particular activation state of VLA-4. Therefore, state-selective antagonists may have an improved embryonic safety profile. Additional studies will be required to determine potential effects of VLA-4 antagonists on embryo/fetal development in vivo.

MeSH terms

  • Allantois / abnormalities
  • Allantois / embryology
  • Animal Husbandry
  • Animals
  • Antibodies / immunology
  • Antibodies / pharmacology
  • Culture Techniques
  • Embryo, Mammalian / drug effects*
  • Female
  • Integrin alpha4beta1
  • Integrins / antagonists & inhibitors*
  • Male
  • Mice
  • Placenta / abnormalities
  • Placenta / embryology
  • Placentation / drug effects*
  • Placentation / physiology
  • Pregnancy
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Lymphocyte Homing / antagonists & inhibitors*

Substances

  • Antibodies
  • Integrin alpha4beta1
  • Integrins
  • Receptors, Lymphocyte Homing