The signal transduction pathways driving morphogenesis and migration are still not well understood, yet defects in these processes underlie a wide spectrum of human diseases and syndromes, including mental retardation, chronic obstructive pulmonary disease (COPD), and cancer. The long term objectives of this proposal are to delineate the cellular mechanisms that control morphogenesis and migration in mammalian epithelial cells. The approaches used here involve the analysis of: (i) collective epithelial cell migration and (ii) the establishment of cell-cell junctions and apical/basal polarity in epithelial cells. The key hypothesis is that despite being very different biological processes, morphogenesis and migration share many of the same molecular components and signaling pathways and that Rho GTPases play a central role in both. In work funded by the current grant, we identified a number of regulators of Rho signaling pathways required for the collective migration of a human bronchial epithelial cell line, 16HBE, and developed time-lapse ex vivo imaging to show that Rac1 is required for the collective migration of anterior visceral endoderm (AVE) cells in embryonic day 5.5 mouse embryos.
In Aim 1, micro-patterned surfaces will be used to identify the contribution of Rho pathways and polarity proteins to the collective behavior of migrating 16HBE cells, and microinjection techniques coupled with real time imaging will be used to identify molecular players in the collective migration of AVE cells in e5.5 mouse embryos. In other work funded by the current grant, we identified three kinases downstream of Rho and Cdc42 required for cell-cell junction assembly in 16HBE cells, and identified several additional Rho family target proteins required for 3D morphogenesis (polarized cyst formation) of the colorectal cell line Caco-2.
In Aim 2, we will identify the kinase substrates involved in junction assembly in 16HBE cells, identify the molecular basis of apical/basal polarity establishment during the first cell division of Caco-2 cells grown in 3D, and delineate the contribution of Rho family targets to Caco-2 morphogenesis. This program of research will lead to a significant advance in understanding the control of migration and morphogenesis and provide greater insight into the underlying cell biology associated with a wide range of human disorders.

Public Health Relevance

In the embryo, cells must migrate to appropriate locations and adopt specialized shapes to form organized tissues and organs. These structures then need to be maintained throughout adult life. The biochemical mechanisms controlling these processes are poorly understood, but defects contribute to a wide range of congenital and acquired human disorders, ranging from mental retardation to cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM081435-07
Application #
8537934
Study Section
Intercellular Interactions (ICI)
Program Officer
Nie, Zhongzhen
Project Start
2007-09-28
Project End
2015-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
7
Fiscal Year
2013
Total Cost
$653,050
Indirect Cost
$295,997
Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065
Durgan, Joanne; Tao, Guangbo; Walters, Matthew S et al. (2015) SOS1 and Ras regulate epithelial tight junction formation in the human airway through EMP1. EMBO Rep 16:87-96
Jin, Dan; Durgan, Joanne; Hall, Alan (2015) Functional cross-talk between Cdc42 and two downstream targets, Par6B and PAK4. Biochem J 467:293-302
Omelchenko, Tatiana; Rabadan, M Angeles; Hernández-Martínez, Rocío et al. (2014) β-Pix directs collective migration of anterior visceral endoderm cells in the early mouse embryo. Genes Dev 28:2764-77
Xu, Xiaojian; Jin, Dan; Durgan, Joanne et al. (2013) LKB1 controls human bronchial epithelial morphogenesis through p114RhoGEF-dependent RhoA activation. Mol Cell Biol 33:2671-82
Omelchenko, Tatiana (2012) Regulation of collective cell migration by RhoGAP myosin IXA. Small GTPases 3:213-8
Omelchenko, Tatiana; Hall, Alan (2012) Myosin-IXA regulates collective epithelial cell migration by targeting RhoGAP activity to cell-cell junctions. Curr Biol 22:278-88
Magudia, Kirti; Lahoz, Aurelia; Hall, Alan (2012) K-Ras and B-Raf oncogenes inhibit colon epithelial polarity establishment through up-regulation of c-myc. J Cell Biol 198:185-94
Durgan, Joanne; Kaji, Noriko; Jin, Dan et al. (2011) Par6B and atypical PKC regulate mitotic spindle orientation during epithelial morphogenesis. J Biol Chem 286:12461-74
Wallace, Sean W; Magalhaes, Ana; Hall, Alan (2011) The Rho target PRK2 regulates apical junction formation in human bronchial epithelial cells. Mol Cell Biol 31:81-91
Pulvirenti, Teodoro; Van Der Heijden, Maartje; Droms, Leif A et al. (2011) Dishevelled 2 signaling promotes self-renewal and tumorigenicity in human gliomas. Cancer Res 71:7280-90

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