CHILDREN'S HOSPITAL MEDICAL CENTER
Grant: $7,500 - National Institutes of Health - Jun. 8, 2009
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Award Description: This study founded by this award provides a summer research opportunity for an undergraduate student that will engage the interest in biomedical science. The study addresses the following hypotheses: 1) Prototype competitive inhibitors, termed pharmacologic chaperones, can increase selected mutant enzymes’ activity/function and be therapeutic in vivo. This is important for the CNS variants of Gaucher disease, since enzyme therapies (ET) or gene therapies are not currently avail¬able for them. 2) Such chaperones could be used to enhance ET by improving the administered en¬zyme’s activity and/or delivery. Insufficient activity of acid ?-glucosidase (GCase) initiates the patho¬logical pro¬cesses by decreasing the substrate—glucosylceramide (GC) or, more importantly for CNS variants, glucosylsphingosine (GS)—flux in the organs. Normalization of substrate flux in tissues is essential in halting the progression of the disease and restoring health. The objective of this proposal is to evaluate the ex vivo and in vivo effects of pharmacological chaperones in correcting the GC and GS metab¬olism using the newly created GC synthase overexpressing GCase point-mutated mouse model that presents GC and GS accumulation. Tissue and cellular substrate levels in this model will be char¬acterized and the role of chaperones in normalizing GSL metabolism in visceral and CNS tissues will be evaluated. The results will have implications for developing new therapeutic treatments for Gaucher disease. This short-term project will enable a student working in the lab in summer 2009 to take part in biomedical research and will be of value to those who are interested in pursuing a career in public health.
Project Description: To create a rapidly progressive Gaucher disease mouse model, transgenic mice over expressing the glucosylceramide-synthesizing enzyme, UDP-glucose:ceramide glucosyltransferase (GCS) were created that were then bred into a GCase D409V(9V)/null background. GCS knockout (GCS-/-) in mice is an embryonic lethal. This lethal phenotype was rescued through expression of the GCS transgene (tg/GCS-/-) under the control of the ubiquitous ROSA promoter; thus demonstrating a functional transgene in vivo. Skin fibroblast cells from tg/GCS-/- mice were used to characterize expression levels and activity of the transgene, and lipid accumulation compared to fibroblasts expressing endogenous GCS (9V/null). Western analyses of tg/GCS-/- skin fibroblasts showed GCS protein levels similar to WT levels. Activity of transgenic GCS was ~40% of WT activity in fibroblast. Localization studies using an antibody specific for mouse GCS showed transgenic GCS was expressed in the Golgi of fibroblasts and liver hepatocytes. 9v/null variants expressing Tg showed increased size of storage cells and 2-fold increases of GC levels in lung. The use of several other GCase mutant mice with this GCS Tg should be useful in understanding the complexity of Gaucher disease and for determining the threshold level of substrate flux that influences phenotypic development.
Jobs Summary: With the support of the Recovery Act Fund, we hired one college student in our medical research laboratory for 10 weeks. The student participated in the project addressing basic biological and medical questions directly related to human inherited diseases. The student learn and performed experiments with DNA, cells, tissues, and animal models. Those experiences provided student with an excellent foundation in medical research and will assist them in making informed career choices in the biomedical sciences.This funding not only provide an opportunity for students to explore basic medical research but also promote job and help economic recovery. (Total jobs reported: 1)
Project Status: Completed
This award's data was last updated on Jun. 8, 2009. Help expand these official descriptions using the wiki below.
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