Title: Anti-EDAR Antibody Treatment Resulted Secondary Palate Closure in Pax9 Deficient Mice
Shihai Jia, University of Utah
Jing zhou, university of Utah
Yinshen Wee, University of Utah
Marja Mikkola, University of Helsinki
Pascal Schneider, University of Lausanne
Rena D'Souza (Presenter)
University of Utah
Objectives: PAX9 is known to be associated with human craniofacial abnormalities such as cleft palate and tooth agenesis. Homozygous mice lacking Pax9 gene die at birth with cleft palate. The objective of this study is to use this mouse model to explore a novel therapeutic treatment and the molecular relationship of Pax9 and Eda/Edar signaling pathway in palate development. Our analyses provides new molecular mechanism of Pax9-Wnt-Eda signaling in regulating palate formation and proposes candidates for the therapeutic treatment of patients with craniofacial defects.
Methods: To increase Eda/Edar signaling activities, the agonist anti-EDAR antibody was injected into pregnant Pax9+/- mice which had been mated with Pax9+/- males for Pax9-/- embryos. The agonist was injected through the tail vein of Pax9+/- pregnant mice at embryonic day E10.5 and E12.5 with dosage of 16 mg/kg, which covered the early developmental stages of palate formation. Further the Eda antagonist anti-EDA antibody together with Wnt agonist was administrated to Pax9-/- embryos during palate development stages. The phenotypes were analyzed at E18.5 via whole mount view and HE stained sections.
Results: The agonist anti-Edar antibody treatment rescued cleft palate defects in Pax9-/- embryos with a success ratio of 90%. However, correction of the palatal defects did not prevent postnatal death of Pax9-/- pups. Further the co-treatment of anti-EDA antibody and Wnt agonist to Pax9-/- embryos reduced the rescue efficiency of Wnt agonist treatment from 100% to 20%.
Conclusions: Our data uncover a unique relationship between Pax9 and the Eda/Edar signaling pathway that can be further exploited for the development of noninvasive, safe, and effective therapies for the treatment of cleft palate conditions and other single-gene disorders affecting the craniofacial complex.