The goal of this proposal is to determine how membrane trafficking of critical Notch pathway components during asymmetric cell division controls a Notch-mediated cell fate switch in cells of the nervous system. Notch function is required in a variety of normal stem and progenitor cells, including neural stem cells, blood cell precursors, and gut progenitors in both Drosophila and vertebrates. Aberrant Notch regulation in stem cells leads to severe defects in developmental events and contributes to human diseases, such as CADASIL syndrome, and cancers, such T-cell acute lymphoblastic leukemia. To understand in detail the cell biological mechanisms that control Notch function, the proposal focuses on the sensory organ progenitor (SOP) cells of the Drosophila adult peripheral nervous system, where cell fate decisions depend on activation or inhibition of Notch activity. The work is based on the findings over the last five years that establish the functional role of a key regulato of Notch signaling in sensory organ cells, Sanpodo. In the previous funding cycle, studies demonstrated that sanpodo promotes Notch activity to confer correct cell fates after asymmetric cell division in the adult peripheral nervous system. Through biochemical studies, in vivo imaging, and rescue experiments, functional domains of Sanpodo have been identified that mediate two critical functions. First, Sanpodo associates with the g-secretase complex to promote Notch activation in the cell with high Notch activity. Second, Sanpodo controls Notch recycling at the plasma membrane in the cell with low Notch activity, acting as a repressor of Notch signaling in this context. This proposal aims to determine how these antagonistic functions are regulated at the level of vesicle trafficking and to define the role of conserved motifs that determine Sanpodo's interaction with Notch regulators such as Numb. The role of conserved vesicle trafficking regulators on controlling Notch levels in asymmetrically dividing neural progenitors will be assessed, using the unique ability to combine molecular modeling, biochemical analysis, and live cell imaging of progenitor cell behavior in the intact animal, to tet the hypothesis that a constitutive level of endocytosis establishes steady state levels of Notch in epithelial cells, whereas Sanpodo acts synergistically with Numb to deplete Notch from the plasma membrane in asymmetrically diving neural progenitors. This proposal will therefore provide a more complete understanding of the evolutionarily conserved mechanisms governing Notch trafficking in this context.

Public Health Relevance

This proposal seeks to reveal the mechanisms underlying how cell fate decisions are made during nervous system development in the fly, Drosophila. While the complexity of the nervous system in humans is greater than in flies, the genes and mechanisms that control nervous system development are largely the same. Understanding how neurons and glial cells are generated may provide critical insight to studies seeking to promote regeneration of nerves and may open up new therapeutic avenues for the treatment of cancers as well as developmental syndromes involving the nervous system.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
High Priority, Short Term Project Award (R56)
Project #
2R56NS059971-06A1
Application #
8928697
Study Section
Neurogenesis and Cell Fate Study Section (NCF)
Program Officer
Owens, David F
Project Start
2007-07-01
Project End
2015-08-31
Budget Start
2014-09-30
Budget End
2015-08-31
Support Year
6
Fiscal Year
2014
Total Cost
$446,250
Indirect Cost
$196,250
Name
Research Institute of Fox Chase Cancer Center
Department
Type
DUNS #
064367329
City
Philadelphia
State
PA
Country
United States
Zip Code
19111