Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Intra-cisterna magna delivery of an AAV vector with the GLUT1 promoter in a pig recapitulates the physiological expression of SLC2A1

Gene Therapy volume 28, pages 329–338 (2021)Cite this article

Subjects

Abstract

Glucose transporter 1 deficiency syndrome (GLUT1DS) is caused by haplo-insufficiency of SLC2A1, which encodes GLUT1, resulting in impaired hexose transport into the brain. Previously, we generated a tyrosine-mutant AAV9/3 vector in which SLC2A1 was expressed under the control of the endogenous GLUT1 promoter (AAV-GLUT1), and confirmed the improved motor function and cerebrospinal fluid glucose levels of Glut1-deficient mice after cerebroventricular injection of AAV-GLUT1. In preparation for clinical application, we examined the expression of transgenes after intra-cisterna magna injection of AAV-GFP (tyrosine-mutant AAV9/3-GFP with the CMV promoter) and AAV-GLUT1. We injected AAV-GFP or AAV-GLUT1 (1.63 × 1012 vector genomes/kg) into the cisterna magna of pigs to compare differential promoter activity. After AAV-GFP injection, exogenous GFP was expressed in broad areas of the brain and peripheral organs. After AAV-GLUT1 injection, exogenous GLUT1 was expressed predominantly in the brain. At the cellular level, exogenous GLUT1 was mainly expressed in the endothelium, followed by glia and neurons, which was contrasted with the neuronal-predominant expression of GFP by the CMV promotor. We consider intra-cisterna magna injection of AAV-GLUT1 to be a feasible approach for gene therapy of GLUT1DS.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Structure of AAV-GFP and AAV-GLUT1.
Fig. 2: Intra-cisterna magna injection of the pig with AAV vectors.
Fig. 3: Vector genome levels of the brain, spinal cord, and other organs after AAV-GLUT1 injection.
Fig. 4: Exogenous GLUT1 mRNA expression after AAV-GLUT1 injection.
Fig. 5: Immunohistostaining of the pig brain with an anti-myc-tag antibody.
Fig. 6: Immunohistostaining of the spinal cord of the pig with an anti-myc-tag antibody.
Fig. 7: Types of GLUT1-expressing cells in the lumbar spinal cord after AAV-GLUT1 injection.
Fig. 8: Immunohistostaining outside the central nervous system with an anti-myc-tag antibody.

Similar content being viewed by others

References

  1. Maher F, Vannucci SJ, Simpson IA. Glucose transporter proteins in brain. FASEB J. 1994;8:1003–11.

    Article  CAS  Google Scholar 

  2. Vannucci SJ, Maher F, Simpson IA. Glucose transporter proteins in brain: delivery of glucose to neurons and glia. Glia. 1997;21:2–21.

    Article  CAS  Google Scholar 

  3. Simpson IA, Carruthers A, Vannucci SJ. Supply and demand in cerebral energy metabolism: the role of nutrient transporters. J Cereb Blood Flow Metab. 2007;27:1766–91.

    Article  CAS  Google Scholar 

  4. Choeiri C, Staines W, Messier C. Immunohistochemical localization and quantification of glucose transporters in the mouse brain. Neuroscience. 2002;111:19–34.

    Article  CAS  Google Scholar 

  5. Hildebrand MS, Damiano JA, Mullen SA, Bellows ST, Oliver KL, Dahl HH, et al. Glucose metabolism transporters and epilepsy: only GLUT1 has an established role. Epilepsia. 2014;55:e18–21.

    Article  CAS  Google Scholar 

  6. Klepper J, Leiendecker B. GLUT1 deficiency syndrome-2007 update. Dev Med Child Neurol. 2007;49:707–16.

    Article  Google Scholar 

  7. De Giorgis V, Veggiotti P. GLUT1 deficiency syndrome 2013: current state of the art. Seizure. 2013;22:803–11.

    Article  Google Scholar 

  8. Ito Y, Takahashi S, Kagitani-Shimono K, Natsume J, Yanagihara K, Fujii T, et al. Nationwide survey of glucose transporter-1 deficiency syndrome (GLUT-1DS) in Japan. Brain Dev. 2015;37:780–9.

    Article  Google Scholar 

  9. Leen WG, Wevers RA, Kamsteeg EJ, Scheffer H, Verbeek MM, Willemsen MA. Cerebrospinal fluid analysis in the workup of GLUT1 deficiency syndrome: a systematic review. JAMA Neurol. 2013;70:1440–4.

    Article  Google Scholar 

  10. Leen WG, Taher M, Verbeek MM, Kamsteeg EJ, van de Warrenburg BP, Willemsen MA. GLUT1 deficiency syndrome into adulthood: a follow-up study. J Neurol. 2014;261:589–99.

    Article  CAS  Google Scholar 

  11. Klepper J. Glucose transporter deficiency syndrome (GLUT1DS) and the ketogenic diet. Epilepsia. 2008;49(Suppl 8):46–49.

    Article  Google Scholar 

  12. Fujii T, Ito Y, Takahashi S, Shimono K, Natsume J, Yanagihara K, et al. Outcome of ketogenic diets in GLUT1 deficiency syndrome in Japan: a nationwide survey. Brain Dev. 2016;38:628–37.

    Article  Google Scholar 

  13. Ito Y, Oguni H, Ito S, Oguni M, Osawa M. A modified Atkins diet is promising as a treatment for glucose transporter type 1 deficiency syndrome. Dev Med Child Neurol. 2011;53:658–63.

    Article  Google Scholar 

  14. Kwiterovich PO Jr, Vining EP, Pyzik P, Skolasky R Jr, Freeman JM. Effect of a high-fat ketogenic diet on plasma levels of lipids, lipoproteins, and apolipoproteins in children. JAMA. 2003;290:912–20.

    Article  CAS  Google Scholar 

  15. Nakamura S, Muramatsu SI, Takino N, Ito M, Jimbo EF, Shimazaki K, et al. Gene therapy for Glut1-deficient mouse using an adeno-associated virus vector with the human intrinsic GLUT1 promoter. J Gene Med. 2018;20:e3013.

    Article  Google Scholar 

  16. Nakamura S, Osaka H, Muramatsu SI, Takino N, Ito M, Aoki S, et al. Gene therapy for a mouse model of glucose transporter-1 deficiency syndrome. Mol Genet Metab Rep. 2017;10:67–74.

    Article  CAS  Google Scholar 

  17. Tang M, Gao G, Rueda CB, Yu H, Thibodeaux DN, Awano T, et al. Brain microvasculature defects and Glut1 deficiency syndrome averted by early repletion of the glucose transporter-1 protein. Nat Commun. 2017;8:14152.

    Article  CAS  Google Scholar 

  18. Iida A, Takino N, Miyauchi H, Shimazaki K, Muramatsu S. Systemic delivery of tyrosine-mutant AAV vectors results in robust transduction of neurons in adult mice. Biomed Res Int. 2013;2013:974819.

    Article  Google Scholar 

  19. Hordeaux J, Hinderer C, Goode T, Buza EL, Bell P, Calcedo R, et al. Toxicology study of intra-cisterna magna adeno-associated virus 9 expressing iduronate-2-sulfatase in rhesus macaques. Mol Ther Methods Clin Dev. 2018;10:68–78.

    Article  CAS  Google Scholar 

  20. Huda F, Konno A, Matsuzaki Y, Goenawan H, Miyake K, Shimada T, et al. Distinct transduction profiles in the CNS via three injection routes of AAV9 and the application to generation of a neurodegenerative mouse model. Mol Ther Methods Clin Dev. 2014;1:14032.

    Article  Google Scholar 

  21. Evers MM, Miniarikova J, Juhas S, Vallès A, Bohuslavova B, Juhasova J, et al. AAV5-miHTT gene therapy demonstrates broad distribution and strong human mutant huntingtin lowering in a Huntington’s disease minipig model. Mol Ther. 2018;26:2163–77.

    Article  CAS  Google Scholar 

  22. Hinderer C, Bell P, Katz N, Vite CH, Louboutin JP, Bote E, et al. Evaluation of intrathecal routes of administration for adeno-associated viral vectors in large animals. Hum Gene Ther. 2018;29:15–24.

    Article  CAS  Google Scholar 

  23. Hinderer C, Bell P, Vite CH, Louboutin JP, Grant R, Bote E, et al. Widespread gene transfer in the central nervous system of cynomolgus macaques following delivery of AAV9 into the cisterna magna. Mol Ther Methods Clin Dev. 2014;1:14051.

    Article  Google Scholar 

  24. Hinderer C, Katz N, Dyer C, Goode T, Johansson J, Bell P, et al. Translational feasibility of lumbar puncture for intrathecal AAV administration. Mol Ther Methods Clin Dev. 2020;17:969–74.

    Article  CAS  Google Scholar 

  25. Sorrentino NC, Maffia V, Strollo S, Cacace V, Romagnoli N, Manfredi A, et al. A comprehensive map of CNS transduction by eight recombinant adeno-associated virus serotypes upon cerebrospinal fluid administration in pigs. Mol Ther. 2016;24:276–86.

    Article  CAS  Google Scholar 

  26. Bailey RM, Rozenberg A, Gray SJ. Comparison of high-dose intracisterna magna and lumbar puncture intrathecal delivery of AAV9 in mice to treat neuropathies. Brain Res. 2020;1739:146832.

    Article  CAS  Google Scholar 

  27. Taghian T, Marosfoi MG, Puri AS, Cataltepe OI, King RM, Diffie EB, et al. A safe and reliable technique for CNS delivery of AAV vectors in the cisterna magna. Mol Ther. 2020;28:411–21.

    Article  CAS  Google Scholar 

  28. Bey K, Deniaud J, Dubreil L, Joussemet B, Cristini J, Ciron C, et al. Intra-CSF AAV9 and AAVrh10 administration in nonhuman primates: promising routes and vectors for which neurological diseases? Mol Ther Methods Clin Dev. 2020;17:771–84.

    Article  CAS  Google Scholar 

  29. Albright AL, Ferson SS. Intrathecal baclofen therapy in children. Neurosurg Focus. 2006;21:e3.

    Article  Google Scholar 

  30. Saraiva J, Nobre RJ, Pereira de Almeida L. Gene therapy for the CNS using AAVs: the impact of systemic delivery by AAV9. J Control Rel. 2016;241:94–109.

    Article  CAS  Google Scholar 

  31. Gray SJ, Matagne V, Bachaboina L, Yadav S, Ojeda SR, Samulski RJ. Preclinical differences of intravascular AAV9 delivery to neurons and glia: a comparative study of adult mice and nonhuman primates. Mol Ther. 2011;19:1058–69.

    Article  CAS  Google Scholar 

  32. Gray SJ, Nagabhushan Kalburgi S, McCown TJ, Jude, Samulski R. Global CNS gene delivery and evasion of anti-AAV-neutralizing antibodies by intrathecal AAV administration in non-human primates. Gene Ther. 2013;20:450–9.

    Article  CAS  Google Scholar 

  33. Olson AL, Pessin JE. Structure, function, and regulation of the mammalian facilitative glucose transporter gene family. Annu Rev Nutr. 1996;16:235–56.

    Article  CAS  Google Scholar 

  34. Hordeaux J, Hinderer C, Goode T, Katz N, Buza EL, Bell P, et al. Toxicology study of intra-cisterna magna adeno-associated virus 9 expressing human alpha-L-iduronidase in rhesus macaques. Mol Ther Methods Clin Dev. 2018;10:79–88.

    Article  CAS  Google Scholar 

  35. Hwang DY, Ismail-Beigi F. Stimulation of GLUT-1 glucose transporter expression in response to hyperosmolarity. Am J Physiol Cell Physiol. 2001;281:C1365–1372.

    Article  CAS  Google Scholar 

  36. Hwang DY, Ismail-Beigi F. Control of Glut1 promoter activity under basal conditions and in response to hyperosmolarity: role of Sp1. Am J Physiol Cell Physiol. 2006;290:C337–344.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Japan Society for the Promotion of Science, JSPS KAKENHI (19K17374), grants for the Project for Health Research on Infants, Children, Adolescents, and Young Adults from the Agency of Medical Research and Development, and a grant from the Japan Agency for Medical Research and Development (AMED) under Grant Number 19lm0203049h0002 and JP20ae0201007.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Jichi Medical University, Tochigi, Japan

    Sachie Nakamura, Hitoshi Osaka, Eriko F. Jimbo, Chika Watanabe & Takanori Yamagata

  2. Division of Neurological Gene Therapy, Jichi Medical University, Tochigi, Japan

    Shin-ichi Muramatsu, Naomi Takino & Mika Ito

  3. Center for Gene and Cell Therapy, The Institute of Medical Science, The University of Tokyo, Bunkyo City, Tokyo, Japan

    Shin-ichi Muramatsu

  4. Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan

    Shuji Hishikawa

  5. Department of Neurosurgery, Jichi Medical University, Tochigi, Japan

    Takeshi Nakajima

Authors
  1. Sachie Nakamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hitoshi Osaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shin-ichi Muramatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Naomi Takino
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mika Ito
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Eriko F. Jimbo
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Chika Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shuji Hishikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Takeshi Nakajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Takanori Yamagata
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hitoshi Osaka.

Ethics declarations

Conflict of Interest

SM owns equity in a gene therapy company (Gene Therapy Research Institution) that commercializes the use of AAV vectors for gene therapy applications. To the extent that the work in this manuscript may increase the value of these commercial holdings, SM has a potential conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nakamura, S., Osaka, H., Muramatsu, Si. et al. Intra-cisterna magna delivery of an AAV vector with the GLUT1 promoter in a pig recapitulates the physiological expression of SLC2A1. Gene Ther 28, 329–338 (2021). https://doi.org/10.1038/s41434-020-00203-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41434-020-00203-z

This article is cited by

Search

Advanced search

Quick links