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Glyceryl triacetate for Canavan disease: A low-dose trial in infants and evaluation of a higher dose for toxicity in the tremor rat model

  • ORIGINAL ARTICLE
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Journal of Inherited Metabolic Disease

Summary

Canavan disease (CD) is a fatal dysmyelinating genetic disorder associated with aspartoacylase deficiency, resulting in decreased brain acetate levels and reduced myelin lipid synthesis in the developing brain. Here we tested tolerability of a potent acetate precursor, glyceryl triacetate (GTA), at low doses in two infants diagnosed with CD, aged 8 and 13 months. Much higher doses of GTA were evaluated for toxicity in the tremor rat model of CD. GTA was given orally to the infants for up to 4.5 and 6 months, starting at 25 mg/kg twice daily, doubling the dose weekly until a maximum of 250 mg/kg reached. Wild-type and tremor rat pups were given GTA orally twice daily, initially at a dose of 4.2 g/kg from postnatal days 7 through 14, and at 5.8 g/kg from day 15 through 23, and thereafter in food (7.5%) and water (5%). At the end of the trial (~90 to 120 days) sera and tissues from rats were analysed for changes in blood chemistry and histopathology. GTA treatment caused no detectable toxicity and the patients showed no deterioration in clinical status. In the high-dose animal studies, no significant differences in the mean blood chemistry values occurred between treated and untreated groups, and no lesions indicating toxicity were detectable in any of the tissues examined. Lack of GTA toxicity in two CD patients in low-dose trials, as well as in high-dose animal studies, suggests that higher, effective dose studies in human CD patients are warranted.

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Notes

  1. For weights outside the standard ranges, human equivalent dose is calculated using the formula: HED = animal dose in mg/kg × (animal weight in kg/human weight in kg)0.33.

Abbreviations

ASPA:

aspartoacylase

ASPAD:

ASPA deficiency

CD:

Canavan disease

GTA:

glyceryl triacetate

MRI:

magnetic resonance imaging

IP:

intraperitoneally

MRS:

magnetic resonance spectroscopy

NAA:

N-acetylaspartate

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Acknowledgements

C. N. Madhavarao was supported by the Rosalind Poss Rosen Clinical Research Training Fellowship sponsored by the American Academy of Neurology Foundation, St Paul, MN and Canavan Foundation, New York, NY. The study was supported by Jacob’s Cure Foundation, NTSAD, Boston, Samueli Institute and NIH R56 grant NS039387 for A. M. A. Namboodiri. GTA for the clinical trials was provided gratis by Cognis Oleo Chemicals, Germany. The authors thank the National Bio Resource Project for the Rat, Kyoto University, Kyoto, Japan, for providing breeding pairs of tremor rats.

Author information

Authors and Affiliations

  1. Department of Anatomy, Physiology and Genetics, Program in Neuroscience and Program in Molecular and Cell Biology, USUHS, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA

    C. N. Madhavarao, P. Arun, J. R. Moffett & A. M. A. Namboodiri

  2. Metabolic Disease Unit, Safra Children Hospital, Sheba Medical Center, Tel-Hashomer, and Sackler Medical School, Tel Aviv University, Tel Aviv, Israel

    Y. Anikster

  3. Division of Comparative Pathology, AFRRI, Bethesda, MD, USA

    S. R. Mog

  4. Medical Genetics Branch, NHGRI, NIH, Bethesda, MD, USA

    O. Staretz-Chacham & W. A. Gahl

  5. Department of Medical and Clinical Psychology, Program in Neuroscience, USUHS, Bethesda, MD, USA

    N. E. Grunberg

Authors
  1. C. N. Madhavarao
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  2. P. Arun
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  3. Y. Anikster
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  4. S. R. Mog
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  5. O. Staretz-Chacham
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  6. J. R. Moffett
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  7. N. E. Grunberg
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  8. W. A. Gahl
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  9. A. M. A. Namboodiri
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Corresponding author

Correspondence to A. M. A. Namboodiri.

Additional information

Communicating editor: Claude Bachmann

Competing interests: None declared

References to electronic databases: Canavan disease, ACY deficiency, ASPA deficiency: OMIM #271900. Gene map locus: 17pter-p13. Gene symbol: ASPA, aliases ASP and ACY2. Aspartoacylase (aminoacylase 2): EC 3.5.1.15. NCBI accession number for ASPA: NP_000040.1, GI:4557335

The authors C. N. Madhavarao, P. Arun, Y. Anikster contributed equally

Electronic supplementary material

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Supplementary Table 1a

Biochemical analyses of the blood serum samples of the GTA treated and untreated female tremor and untreated female wild type rats (DOC 178 KB)

Supplementary Figure S1

Comparative pictures of normal (a) and homozygous mutant (tremor, b) rat pups at 2 weeks of age. Note the thin coat, ring tail and curled whiskers in the tremor pup compared to the normal features in the wild-type pup (JPG 327 KB).

Supplementary Figure S2

Stomach sections from the wild-type female (a, b) and male (c, d) rats showing fundic glands (FG), muscularis mucosae (MM), submucosa (SM) and blood vessels (BV). No significant lesion was detected between GTA untreated (a, c) and treated (b, d) wild-type rats and the results were similar in tremor rats (JPG 2.57 MB).

Supplementary Figure S3

Small intestine (jejunum) sections from the wild-type female (a, b) and male (c, d) rats showing crypts of Lieberkuhn (CL), goblet cells (GC), muscularis mucosae (MM) and lumen (L). No significant lesion was detected between GTA untreated (a, c) and treated (b, d) wild-type rats and tremor rats (JPG 3.86 MB).

Supplementary Figure S4

Large intestine (colon) sections from the wild-type female (a, b) and male (c, d) rats showing crypts of Lieberkuhn (CL), goblet cells (GC), lumen (L), smooth muscle (SM) and blood vessels (BV). No significant lesion was detected between GTA untreated (a, c) and treated (b, d) wild-type rats and tremor rats (JPG 3.60 MB).

Supplementary Figure S5

Liver sections from the wild-type female (a, b) and male (c, d) rats showing the lobules of the liver with central vein (CV), sinusoids (Si), hepatocytes (H), plates of liver cells (PL) and portal vein (PV, in Fig. a). No significant lesion was detected between GTA untreated (a, c) and treated (b, d) rats except for diffuse minimal to mild hepatocellular vacuolar change of glycogen-type or lipid type (JPG 2.88 MB).

Supplementary Figure S6

Kidney cortical labyrinth sections from the wild-type female (a, b) and male (c, d) rats showing glomeruli. The renal capsules (RC) within the Bowman’s space (BS), proximal tubule (PT) and distal tubule (DT) are also shown. No significant lesion was detected among the GTA untreated (a, c) and treated (b, d) wild-type rats and tremor rats (JPG 3.46 MB).

Supplementary Figure S7

Spleen sections from the wild-type female (a, b) and male (c, d) rats showing the red pulp (RP) and white pulp (WP) regions. No significant lesion was detected among the GTA untreated (a, c) and treated (b, d) wild-type rats and tremor rats (JPG 4.57 MB).

Supplementary Figure S8

Heart sections from the wild-type female (a, b) and male (c, d) rats show the cardiac muscle cell nuclei (N), blood vessels (BV) and cardiac muscle fibers (CMF). No significant lesion was detected among the GTA untreated (a, c) and treated (b, d) wild-type rats and tremor rats (JPG 2.80 MB).

Supplementary Figure S9

Lung sections from the wild-type female (a, b) and male (c, d) rats showing the terminal bronchiole (TB), Clara cells (CC) and smooth muscle (SM) of the bronchiole. No significant lesion was detected among the GTA untreated (a, c) and treated (b, d) wild-type rats and tremor rats (JPG 2.90 MB).

Supplementary Figure S10

Forebrain sections from the wild-type female (a, b) and male (c, d) rats showing the white matter (WM) and gray matter (GM) regions. No significant lesion was detected among the GTA untreated (a, c) and treated (b, d) rats (JPG 2.79 MB).

Supplementary Figure S11

Cerebellum sections from the wild-type female (a, b) and male (c, d) rats showing the molecular layer (ML), granular layer (GRL) and Purkinje cells (PC). No significant lesion was detected among the GTA untreated (a, c) and treated (b, d) rats (JPG 3.17 MB).

Supplementary Figure S12

Spinal cord sections from the wild-type female (a, b) and male (c, d) rats showing the neurons (Nu), glial cells (Gl) and blood vessels (BV). No significant lesion was detected among the GTA untreated (a, c) and treated (b, d) rats (JPG 3.34 MB).

Supplementary Figure S13

Liver sections from the tremor female (a, b) and male (c, d) rats showing the lobules of the liver with central vein (PV), sinusoids (Si), hepatocytes (H) and plates of liver cells (PL). No significant lesion was detected between GTA untreated (a, c) and treated (b, d) rats except for diffuse minimal to mild hepatocellular vacuolar change of glycogen-type or lipid type seen with the treated rats more clearly than the untreated rats (JPG 2.85 MB).

Supplementary Figure S14

Forebrain sections from the tremor female (a, b) and male (c, d) rats showing the gray matter (GM), white matter (WM) and vacuoles seen in the white matter, typical of the mutant tremor rats. No significant lesion was detected that was attributable to GTA treatment among the GTA untreated (a, c) and treated (b, d) rats (JPG 2.83 MB).

Supplementary Figure S15

Cerebellum sections from the tremor female (a, b) and male (c, d) rats showing the molecular layer (ML), granular layer (GRL) and Purkinje cells (PC). No significant lesion was detected that was attributable to GTA treatment among the GTA untreated (a, c) and treated (b, d) rats (JPG 3.17 MB).

Supplementary Figure S16

Spinal cord sections from the tremor female (a, b) and male (c, d) rats showing the neurons (Nu), glial cells (Gl) and blood vessels (BV). Vacuoles are seen as typical of the mutant tremor rats. No significant lesion was detected that was attributable to GTA treatment among the GTA untreated (a, c) and treated (b, d) rats (JPG 3.09 MB).

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Madhavarao, C.N., Arun, P., Anikster, Y. et al. Glyceryl triacetate for Canavan disease: A low-dose trial in infants and evaluation of a higher dose for toxicity in the tremor rat model. J Inherit Metab Dis 32, 640–650 (2009). https://doi.org/10.1007/s10545-009-1155-3

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  • DOI: https://doi.org/10.1007/s10545-009-1155-3

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