The migration of cranial neural crest (CNC) cells is controlled by proteins that regulate gene expression, cell proliferation, cell signaling and cell adhesion. Among these proteins the ADAM cell surface metalloproteases can regulate both cell signaling and cell adhesion. They do so by cleaving signaling and adhesion molecules from the cell surface. There is a fundamental gap in understanding how ADAM proteins control cell migration in general and CNC migration in particular. Our long-term goal is to understand how CNC migration is controlled in a developing embryo. The objective of this application is to determine how ADAM metalloproteases control CNC migration. The central hypothesis of this proposal, based on extensive preliminary data, is that ADAM metalloproteases control cell migration both by shedding cell adhesion molecules from the plasma membrane and by the independent activity of their cytoplasmic domains inside of the nucleus.
Aim 1 : To identify how ADAM cleavage of Cadherin-11 controls CNC migration. We have shown that Cad-11 is cleaved by ADAM13 during CNC migration and that the Cad-11 extracellular fragment containing the adhesive site is shed and can rescue migration in embryos lacking ADAM13 or overexpressing Cad-11. Our hypothesis is that this fragment binds to the surface of CNC via a receptor and promotes migration. We propose to determine how the extracellular fragment of Cad-11 promotes migration. We will determine, 1) what are the consequences of not cleaving Cad-11 on CNC specification and migration, 2) if the Cad-11 extracellular fragment stimulates directed cell migration and/or interfere with contact mediated inhibition, and 3) What are the signaling pathways involved in this process.
Aim 2 : To identify the mechanism by which the ADAM cytoplasmic domain controls CNC migration. We have shown that the ADAM13 cytoplasmic domain (C13) is cleaved and translocates into the nucleus. We showed that this is critical for gene expression and CNC migration in vivo. We also showed that one gene regulated by C13, Calpain8 is essential for CNC migration. Our hypothesis is that the ADAM13 cytoplasmic domain controls gene expression by modifying the function of a transcription factor to promote CNC migration. We will determine 1) the amino acid sequence(s) in C13 that are responsible for the control of gene expression and cell migration, and 2) the mechanism by which C13 regulates the expression of Calpain8. The approach is innovative, because it shows for the first time in any system that ADAM cytoplasmic domains function in the nucleus to regulate specific gene expression and cell migration. The proposed research is significant, because it is expected to vertically advance the field by identifying evolutionary conserved motifs in ADAM cytoplasmic domain that control the activity of transcription factors (e.g. FoxD3). In addition, the Cad-11 extracellular domain has been shown to increase cancer cell invasion, thus our results will shed the light on the mechanism by which this domain operates.

Public Health Relevance

The proposed project is relevant to public health because it increases our knowledge of craniofacial development in a vertebrate model organism amenable to experimental studies. The understanding of ADAM control of cell migration is also relevant as these proteins are often up-regulated and associated with invasion of cancer cells. The current study also shows that ADAM cytoplasmic domains control the expression of genes that are relevant to multiple human diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
2R01DE016289-06A1
Application #
8236674
Study Section
Intercellular Interactions (ICI)
Program Officer
Scholnick, Steven
Project Start
2005-01-08
Project End
2017-01-31
Budget Start
2012-03-08
Budget End
2013-01-31
Support Year
6
Fiscal Year
2012
Total Cost
$370,874
Indirect Cost
$120,874
Name
University of Massachusetts Amherst
Department
Veterinary Sciences
Type
Schools of Earth Sciences/Natur
DUNS #
153926712
City
Amherst
State
MA
Country
United States
Zip Code
01003
Abbruzzese, Genevieve; Cousin, Hélène; Salicioni, Ana Maria et al. (2014) GSK3 and Polo-like kinase regulate ADAM13 function during cranial neural crest cell migration. Mol Biol Cell 25:4072-82
Ji, Yon Ju; Hwang, Yoo-Seok; Mood, Kathleen et al. (2014) EphrinB2 affects apical constriction in Xenopus embryos and is regulated by ADAM10 and flotillin-1. Nat Commun 5:3516
Powder, Kara E; Cousin, Hélène; McLinden, Gretchen P et al. (2014) A nonsynonymous mutation in the transcriptional regulator lbh is associated with cichlid craniofacial adaptation and neural crest cell development. Mol Biol Evol 31:3113-24
Wang, L; Pawlak, E A; Johnson, P J et al. (2014) Expression and activity of collagenases in the digital laminae of horses with carbohydrate overload-induced acute laminitis. J Vet Intern Med 28:215-22
Wang, Le; Pawlak, Erica A; Johnson, Philip J et al. (2013) Impact of laminitis on the canonical Wnt signaling pathway in basal epithelial cells of the equine digital laminae. PLoS One 8:e56025
Cousin, Helene; Abbruzzese, Genevieve; Kerdavid, Erin et al. (2011) Translocation of the cytoplasmic domain of ADAM13 to the nucleus is essential for Calpain8-a expression and cranial neural crest cell migration. Dev Cell 20:256-63
Alfandari, Dominique; McCusker, Catherine; Cousin, Helene (2009) ADAM function in embryogenesis. Semin Cell Dev Biol 20:153-63
Coyne, Michael J; Cousin, Helene; Loftus, John P et al. (2009) Cloning and expression of ADAM-related metalloproteases in equine laminitis. Vet Immunol Immunopathol 129:231-41
Kashef, Jubin; Kohler, Almut; Kuriyama, Sei et al. (2009) Cadherin-11 regulates protrusive activity in Xenopus cranial neural crest cells upstream of Trio and the small GTPases. Genes Dev 23:1393-8
Hidalgo, Magdalena; Sirour, Cathy; Bello, Valerie et al. (2009) In vivo analyzes of dystroglycan function during somitogenesis in Xenopus laevis. Dev Dyn 238:1332-45

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