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Author Notes:

Judith L Fridovich-Keil, Department of Human Genetics, Emory University School of Medicine, Rm. 325.2 Whitehead Bldg., 615 Michael St, Atlanta, GA 30322 TEL 404-727-3924, FAX 404-727-3949. Email: jfridov@emory.edu

Shauna Rasmussen helped to design and interpret experiments, performed or oversaw the majority of animal work performed, and participated in writing and editing of the manuscript. Jenna Daenzer helped to design and interpret experiments, performed or oversaw the majority of biochemical and metabolic work performed, and participated in writing and editing of the manuscript. Judith Fridovich-Keil coordinated the activities of all collaborators, helped to design and interpret experiments, and did most of the writing and editing of the manuscript.

We are grateful to numerous colleagues for their contributions to this project. We specifically thank Dr. Michael Castle for his priceless guidance on everything AAV, Drs. Oskar Laur and Xinping Huang and their colleagues in the Emory Integrated Cores for generating the AAV vector and viral stock used in this study, and Dr. Lyra Griffiths, also from the Emory Integrated Cores, for performing the quantitative PCR required to calculate viral genome copy number in tissue samples. We also thank all members of the Fridovich-Keil lab for constant support, and the many professionals working in the Emory Division of Animal Resources and the Emory IACUC without whom this work could not have been conducted.

All of the authors declare that they have no competing interests.

Subject:

Research Funding:

This work was supported in part by grants from the National Institutes of Health R01DK107900 and R21HD092785 (both to JLFK), and in part by the Emory Integrated Genomics Core (EIGC), which is subsidized by the Emory University School of Medicine and is one of the Emory Integrated Core Facilities.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Endocrinology & Metabolism
  • Genetics & Heredity
  • Medicine, Research & Experimental
  • Research & Experimental Medicine
  • AAV9
  • galactosemia
  • GALT
  • gene therapy
  • metabolite
  • rat
  • MICE

A pilot study of neonatalGALTgene replacement usingAAV9dramatically lowers galactose metabolites in blood, liver, and brain and minimizes cataracts inGALT-null rat pups

Journal Title:

JOURNAL OF INHERITED METABOLIC DISEASE

Volume:

Volume 44, Number 1

Publisher:

, Pages 272-281

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Classic galactosemia (CG) is a rare metabolic disorder that results from profound deficiency of galactose-1-P uridylyltransferase (GALT). Despite early detection by newborn screening and rapid and lifelong dietary restriction of galactose, which is the current standard of care, most patients grow to experience a broad constellation of long-term complications. The mechanisms underlying these complications remain unclear and likely differ by tissue. Here we conducted a pilot study testing the safety and efficacy of GALT gene replacement using our recently-described GALT-null rat model for CG. Specifically, we administered AAV9.CMV.HA-hGALT to seven GALT-null rat pups via tail vein injection on day 3 of life; eight GALT-null pups injected with PBS served as the negative control, and four GALT+ heterozygous pups injected with PBS served as the positive control. All pups were returned to their nursing mothers, weighed daily, and euthanized for tissue collection 2 weeks later. Among the AAV9-injected pups in this study, we achieved GALT levels in liver ranging from 64% to 595% wild-type, and in brain ranging from 3% to 42% wild-type. In liver, brain, and blood samples from these animals we also saw a striking drop in galactose, galactitol, and gal-1P. Finally, all treated GALT-null pups showed dramatic improvement in cataracts relative to their mock-treated counterparts. Combined, these results demonstrate that GALT restoration in both liver and brain of GALT-null rats by neonatal tail vein administration using AAV9 is not only attainable but effective.
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