Dose- and route-dependent teratogenicity, toxicity, and pharmacokinetic profiles of the hedgehog signaling antagonist cyclopamine in the mouse.
The Hedgehog (Hh) signaling pathway is an essential regulator of embryonic development and appears to play important roles in postnatal repair and cancer progression and metastasis. The teratogenic Veratrum alkaloid cyclopamine is a potent Hh antagonist and is used experimentally both in vitro and in vivo to investigate the role of Hh signaling in diverse biological processes. Here, we set out to establish an administration regimen for cyclopamine-induced teratogenicity in the mouse. The dysmorphogenic concentration of cyclopamine was determined in vitro via mouse whole-embryo culture assays to be 2.0 microM. We administered cyclopamine to female C57BL/6J mice at varied doses by oral gavage, ip injection, or osmotic pump infusion and assessed toxicity and pharmacokinetic (PK) models. Bolus administration was limited by toxicity and rapid clearance. In vivo cyclopamine infusion at 160 mg/kg/day yielded a dam serum steady-state concentration of approximately 2 microM with a corresponding amniotic fluid concentration of approximately 1.5 microM. Gross facial defects were induced in 30% of cyclopamine-exposed litters, with affected embryos exhibiting cleft lip and palate. This is the first report describing the PKs and teratogenic potential of cyclopamine in the mouse and demonstrates that transient Hh signaling inhibition induces facial clefting anomalies in the mouse that mimic common human birth defects.
Lipinski
Robert J
RJ
Molecular and Environmental Toxicology Center, School of Medicine and Public Health, University of Wisconsin, Madison WI 53703, USA.
Hutson
Paul R
PR
Hannam
Paul W
PW
Nydza
Robert J
RJ
Washington
Ida M
IM
Moore
Robert W
RW
Girdaukas
Gary G
GG
Peterson
Richard E
RE
Bushman
Wade
W
D000653:Amniotic Fluid / Q000737:chemistry; D000818:Animals; D002971:Cleft Lip / Q000139:chemically induced*; D002972:Cleft Palate / Q000139:chemically induced*; D004305:Dose-Response Relationship, Drug; D004333:Drug Administration Routes; D004622:Embryo, Mammalian / Q000002:abnormalities / Q000187:drug effects; D005260:Female; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors*; D051379:Mice; D008810:Mice, Inbred C57BL; D011247:Pregnancy; D015398:Signal Transduction; D013723:Teratogens / Q000493:pharmacokinetics / Q000633:toxicity*; D014704:Veratrum Alkaloids / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics / Q000633:toxicity*
D053823:Hedgehog Proteins; D013723:Teratogens; D014704:Veratrum Alkaloids; C000541:cyclopamine