2006 — 2008 |
Weiser, Douglas C |
F32Activity Code Description: To provide postdoctoral research training to individuals to broaden their scientific background and extend their potential for research in specified health-related areas. |
Regulation of Gastrulation by Gravin @ University of Washington
DESCRIPTION (provided by applicant): Proper control of cell migration and cell polarity is essential for proper vertebrate development. Inappropriate activation of developmental programs in an adult is linked to metastasis during tumor progression. Also, loss of proper cell migration during early development can lead to birth defect such as neural tube defects, including spina bifida. Thus, this proposal focuses on the establishment of cell polarity and cell migration during early development. A better understanding of the molecular control of cell migration during development, should lead to a better understanding of the root causes of birth defects and the mechanism of metastasis, and hopefully will aid in the prevention or treatment of these conditions.
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0.97 |
2017 |
Weiser, Douglas Conrad |
R15Activity Code Description: Supports small-scale research projects at educational institutions that provide baccalaureate or advanced degrees for a significant number of the Nation’s research scientists but that have not been major recipients of NIH support. The goals of the program are to (1) support meritorious research, (2) expose students to research, and (3) strengthen the research environment of the institution. Awards provide limited Direct Costs, plus applicable F&A costs, for periods not to exceed 36 months. This activity code uses multi-year funding authority; however, OER approval is NOT needed prior to an IC using this activity code. |
Regulation of the Unfolded Protein Response by Eif2 Alpha Phosphatases @ University of the Pacific-Stockton
Project Summary Eukaryotic cells must be able to maintain balance between the amount of protein entering the secretory pathway and the capacity to process, fold and traffic these proteins through the endoplasmic reticulum. Excessive protein synthesis can cause proteins to unfold and aggregate in the ER causing ER stress. To survive the cells launch a signal transduction cascade called the Unfolded Protein Response (UPR). The UPR leads to increases in protein folding capacity by upregulating chaperones, also prevents new proteins from entering the ER by lowering the rate of global translation, and the UPR also helps clear unfolded proteins from the ER by increase proteolysis and autophagy. Numerous human disease are linked to ER-stress including, Alzheimer?s and other neurodegenerative diseases, type II diabetes, fatty liver disease, spinal cord injury, among others. Multiple drugs have been developed that modulate UPR signaling; among the most promising are inhibitors of eIF2 alpha phosphatases (including salubrinal and sephin1). These drugs have shown great promise in delaying onset and prolonging survival in mouse models of Amyotrophic lateral sclerosis (ALS), while sephin1 has been recently approved as an orphan drug for treating Charcot-Marie-Tooth disease 1B (CMT). The eIF2 alpha phosphatase complexes are comprised of a catalytic subunit PP1 and either of two regulatory subunits, GADD34 and CReP, and are potent inhibitors of one branch of the UPR. This project uses the zebrafish model system to investigate the molecular mechanisms of ER-stress induced apoptosis. In aim 1 we will take a genetic approach and seek to understand how levels of GADD34 and CReP are regulated by ER stress. Our preliminary data indicates that GADD34 mRNA rapidly increases and remains elevated after ER-stress, while CReP transiently increases following ER stress and then declines to levels below baseline after prolonged stress. We will identify which of the branches of UPR signaling are required for this dynamic expression pattern. In addition, we will utilize CRIPSR-CAS9 genome editing for reverse-genetic analysis of the function of GADD34 and CReP. In aim 2 we will then focus on two major stress-induced phenotypes in zebrafish, ER-stress induced apoptosis in the developing caudal fin and ER-stress induced accumulation of fatty acids in the liver (steatosis). This will provide us with a highly tractable model of ER-stress induced cell death and ER-stress induced metabolic disorders. Our preliminary data indicates that inhibition of GADD34 and CReP protects fin epidermal cells from ER-stress induced apoptosis, but worsens fatty liver symptoms in the liver. In aim 3 we will examine the assembly of the active eIF2 alpha phosphatase by determining the amino acids required for in vivo function. In our preliminary data experiments we mapped the minimal PP1-binding domain of CReP and GADD34 and generated a suite of point mutants to map residues critical for function. Using zebrafish will allow us to combine reverse genetics with in vivo cell biology, and will provide an ideal model to test the function of GADD34 and CReP and shed light onto the mechanism of action of clinically relevant drugs that target these proteins.
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0.97 |