The long-term objectives of this proposal are to elucidate the role of epigenetic regulation of gene expression during mammalian orofacial development. Formation of the mammalian orofacial tissue is a multifaceted, synchronized developmental process involving cell migration, proliferation, differentiation, apoptosis, and synthesis of extracellular matrix. All these critical cellular processes are under the control of a variety of genes encoding growth and differentiation factors, signaling mediators, transcription factors and extracellular matrix proteins, whose expression is controlled, in turn, by epigenetically-modulated regulatory elements. Disruption of any one of these processes can lead to orofacial clefting. Emerging evidence strongly supports the importance of epigenetic mechanisms controlling the expression of key genes critical for orofacial ontogenesis. Recent studies from our lab have demonstrated that precise patterns of differential gene methylation and expression occur in embryonic orofacial tissue. Preliminary data revealed several epigenetically regulated genes which have documented critical roles in orofacial morphogenesis. We thus propose to test the hypothesis that regulation of differentially-expressed genes via differentially methylated regions, is essential for proper development of the embryonic orofacial region. Three hypothesis-driven specific aims will be pursued: 1) Differentially expressed genes encoding mRNAs are epigenetically regulated during orofacial development. 2) Differentially expressed and epigenetically regulated gene transcripts are essential for the regulation of specific biological processes as well as phenotypic outcomes associated with orofacial development, and 3) Selected differentially-expressed target genes are epigenetically regulated via differentially methylated regions during orofacial development.

Public Health Relevance

Orofacial clefts are amongst the most prevalent birth defects in humans, which can lead to difficulties in infant feeding, breathing and speech impairment. Studies proposed in the current application are designed to advance our understanding of the complex mechanisms that control development of the orofacial region. This research proposal will adopt a novel bioinformatic approach and will utilize a mouse model that mimics human orofacial development, to investigate the role of epigenetic regulation of gene expression during mammalian orofacial development. Successful completion of the proposed studies will provide a better insight of the specific control mechanisms that are important in orofacial development and will ultimately reveal potential mechanisms of clefting.

National Institute of Health (NIH)
National Institute of Dental & Craniofacial Research (NIDCR)
Small Research Grants (R03)
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Special Emphasis Panel (ZDE1-JR (11))
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Harris, Emily L
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University of Louisville
Schools of Dentistry
United States
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Seelan, Ratnam S; Mukhopadhyay, Partha; Warner, Dennis R et al. (2017) Determinants of orofacial clefting II: Effects of 5-Aza-2'-deoxycytidine on gene methylation during development of the first branchial arch. Reprod Toxicol 67:100-110
Mukhopadhyay, Partha; Seelan, Ratnam S; Rezzoug, Francine et al. (2017) Determinants of orofacial clefting I: Effects of 5-Aza-2'-deoxycytidine on cellular processes and gene expression during development of the first branchial arch. Reprod Toxicol 67:85-99
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Seelan, Ratnam S; Appana, Savitri N; Mukhopadhyay, Partha et al. (2013) Developmental profiles of the murine palatal methylome. Birth Defects Res A Clin Mol Teratol 97:171-86
Seelan, Ratnam S; Warner, Dennis R; Mukhopadhyay, Partha M et al. (2013) Epigenetic analysis of laser capture microdissected fetal epithelia. Anal Biochem 442:68-74
Seelan, Ratnam S; Mukhopadhyay, Partha; Warner, Dennis R et al. (2013) Epigenetic regulation of Sox4 during palate development. Epigenomics 5:131-46
Seelan, Ratnam S; Mukhopadhyay, Partha; Pisano, M Michele et al. (2012) Developmental epigenetics of the murine secondary palate. ILAR J 53:240-52