Efficient Gene Delivery Admitted by small Metabolites Specifically Targeting Astrocytes in the Mouse Brain.

in Molecular therapy : the journal of the American Society of Gene Therapy by Haibin Zhou, Jiajing Dai, Dong Li, Luyao Wang, Meng Ye, Xiaoling Hu, Joseph LoTurco, Ji Hu, Wenzhi Sun

TLDR

  • The study introduces a gene delivery approach called gDAM, which targets astrocytes in the adult mouse brain and shows promise for treating gliomas.
  • gDAM combines DNA with small metabolites to achieve targeted delivery of naked DNA, leading to transient and robust gene expression.

Abstract

The development of efficient and targeted methods for delivering DNA in vivo has long been a major focus of research. In this study, we introduce a gene Delivery approach Admitted by small Metabolites, named gDAM, for the efficient and targeted delivery of naked DNA into astrocytes in the adult brains of mice. gDAM utilizes a straightforward combination of DNA and small metabolites, including glycine, L-proline, L-serine, L-histidine, D-alanine, Gly-Gly, and Gly-Gly-Gly, to achieve astrocyte-specific delivery of naked DNA, resulting in transient and robust gene expression in these cells. Using gDAM, we successfully co-deliver the PiggyBac (PB) transposon and the CRISPR/Cas9 system to induce long-term overexpression of the oncogene EGFRvIII and knockout of tumor suppressor genes Nf1, Pten, and Trp53 in astrocytes, leading to the development of astrocyte-derived gliomas in immunocompetent mice. Furthermore, gDAM facilitates the delivery of naked DNA to peripheral glioma astrocytes. The overexpression of interferon-β (IFN-β) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in these peripheral glioma astrocytes significantly prolongs the overall survival of mice bearing 73C glioma cells. This approach offers a new perspective on developing gene delivery systems that specifically target astrocytes to meet the varied needs of both research and gene therapy. The innovative strategy behind gDAM is expected to provide fresh inspiration in the quest for DNA delivery to other tissues, such as skeletal muscle and skin.

Overview

  • The study introduces a gene delivery approach called gDAM, which efficiently and specifically delivers naked DNA to astrocytes in the adult mouse brain.
  • gDAM combines DNA with small metabolites, including amino acids, to achieve targeted delivery of naked DNA to astrocytes.
  • The primary objective is to develop a gene delivery system that specifically targets astrocytes for research and gene therapy applications.

Comparative Analysis & Findings

  • gDAM successfully delivers DNA to astrocytes in the adult mouse brain, resulting in transient and robust gene expression.
  • gDAM co-delivers the PiggyBac transposon and CRISPR/Cas9 system to induce long-term overexpression of EGFRvIII and knockout of tumor suppressor genes Nf1, Pten, and Trp53.
  • The delivery of naked DNA to peripheral glioma astrocytes using gDAM leads to the overexpression of IFN-β and GM-CSF, significantly prolonging the overall survival of mice bearing 73C glioma cells.

Implications and Future Directions

  • The innovative strategy behind gDAM provides new inspiration for developing gene delivery systems that target other tissues, such as skeletal muscle and skin.
  • The study highlights the potential of gDAM for meeting the varied needs of both research and gene therapy applications.
  • Future studies can explore the application of gDAM in different disease models and modalities to further expand its potential.
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