Liver GALT Activity Mediated by AAV8 Gene Therapy Público

Abstract

Classic galactosemia is a genetic disorder characterized by a deficiency of functional galactose 1-phosphate uridylyl transferase (GALT). The GALT enzyme catalyzes a critical reversible reaction within the Leloir pathway of galactose metabolism where galactose 1- phosphate (gal-1P) is reversibly converted into UDP-galactose. Failure to produce adequate levels of functional GALT results in the accumulation of multiple metabolites associated with the Leloir pathway in tissues throughout the body. This study aims to discern if liver specific GALT restoration can be achieved through the use of AAV8 gene therapy in a GALT-null rat model of CG. GALT activity was assessed using GALT assays in liver and brain tissues to determine if the transgene was expressed within these tissues. Enzyme assays were performed on tissues collected from rats at 14, 30, 45, and 60 days after the administration of treatment to measure the amount and duration of GALT activity in treated animals. The results of the assays showed GALT activity in the livers of treated GALT-null rats, while brain GALT activity remained below detectable limits. After GALT activity was detected in the livers of treated animals, galactose, galactitol, and gal-1P metabolite levels were quantified in liver and brain tissue, while galactose and galactitol levels were measured in plasma samples. Metabolite levels were examined to determine if liver specific GALT restoration could provide meaningful decreases to liver metabolite levels in addition to those of the brain and plasma. Results indicated that there may be a pattern of decreased metabolite accumulation within all tissues analyzed. Overall, this experiment demonstrates liver specific GALT restoration can be achieved using an AAV8 virus. In addition, metabolite data suggest liver GALT activity may provide some correction to metabolite levels in the liver as well as other tissues and blood components throughout the organism. 

Table of Contents

Table of Contents

Chapter 1: Background ................................................................................................................ 1

Chapter 2: Liver GALT Restoration by AAV8-Gene Therapy in GALT-null Rats .................... 9

Chapter 3: Metabolic Efficacy of Liver GALT Restoration by AAV8-Mediated Gene Therapy in GALT-null Rats ..................................................................................................... 27

Chapter 4: General Discussion ................................................................................................... 56

References ................................................................................................................................... 61 

About this Honors Thesis

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School	Emory College
Department	Chemistry
Degree	B.S.
Submission	Honors Thesis
Language	English
Research Field	Biology, Molecular Chemistry, Biochemistry Health Sciences, General
Palavra-chave	AAV Gene Therapy Galactosemia AAV8 Rat Model Classic Galactosemia Gene Therapy
Committee Chair / Thesis Advisor	Fridovich-Keil, Judith, Emory University
Committee Members	Lynn, David, Emory University Himes, Richard, Emory University

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