Abstract
Grant Number:
1 X01 MH077608-01
PI Name:
PAVITT, GRAHAM
PI Email:
PI Title:
SENIOR LECTURER
Project Title:
High-throughput screen for a therapeutic agent for elF2B-related disorders
Abstract:
DESCRIPTION (provided by applicant): 'eIF2B-related disorders' are genetically inheriteddiseases caused by mutations in the general translation initiation factor eIF2B. Although eIF2B functions inall cells, affected individuals have tissue-specific disease. The most serious disease is a dysfunction in brainmyelin. Progressive demyelination causes severe disability. The impairment leads to an increasingly poorquality of life and early death for affected individuals. 'Classical' onset is in children aged 2-5, but severeforms of disease present from as early as birth and milder forms do not present until early adulthood. Milderforms are also associated with germ cell defects. There is currently no therapy or cure available.Experimental work suggests that disease results from a moderate impairment of eIF2B function in many if not all cell types. As control of eIF2B function is important for cellular stress responses, reduced eIF2Bfunction correlates with the observed stress-induced (fever, trauma) deterioration/ degeneration. In theory achemical intervention that can boost the remaining function of eIF2B should be able to prevent or slowfurther decline in affected individuals. Alternatively, a compound that is not directly beneficial to patientsmay also be a useful research tool to probe eIF2B structure and function. As eIF2B is a universallyconserved protein among eukaryotic organisms, we have developed a cellular assay system using yeastcells, Saccharomyces cerevisiae. Assays applicable to high throughput screening have been optimized. Thefirst assay is a simple growth assay, where absorbance increase is measured. eIF2B mutations cause cells togrow at a slower rate than normal cells. Compounds that rescue this defect are sort. This assay was validatedusing a small library of 2000 compounds. The second assay is a reporter gene assay using a luminescentsubstrate. GCN4 expression is inversely correlated with eIF2B activity in cells. Consequently cells bearingeIF2B mutations have aberrantly high GCN4 expression. Compounds that can correct (lower) this are sort.A program of post-screening analysis of mutations in a variety of human, mammal and yeast model systemsis outlined. Post-screening studies involve expertise within collaborating laboratories. Lay Summary-relevance for Human Health: Leukodystrophies are degenerative brain disorders that affect children of allracial groups, including Native Americans. Affected individuals have increasingly poor life quality and dieprematurely. Recent work has discovered that one common cause of leukodystrophy is genetically inheritedmutations of a protein called eIF2B. eIF2B is known to be an important protein involved in normal growthand development. A reduction in eIF2B function rather than a loss of its function is responsible for thedisease. There is currently no therapy or cure available for eIF2B related leukodystrophies. We havedeveloped cell based assays that we wish to employ to screen a large library of compounds, to determine if any may be useful therapeutic agents or research tools for this disease. The aim is to identify a chemical thatmay specifically boost the remaining activity of eIF2B. If successful in cellular models of the disease thismay ultimately result in the slowing or halting of disease progression.
Thesaurus Terms:
High throughput screening, eIF2B, demyelination, Saccharomyces cerevisiae, GCN4 expression, Leukodystrophies, degenerative brain disorders
of 00
National Institutes of Health: Abstract Pavitt 1 X01 MH077608-01
80 Reads
Info and Rating
| Category: | Uncategorized. |
| Rating: | |
| Upload Date: | 01/19/2008 |
| Copyright: | Attribution Non-commercial |
| Tags: |
Leave a Comment