In vivo protein transduction to the CNS

L. T. Loftus, H. F. Li, A. J. Gray, C. Hirata-Fukae, B. A. Stoica, Junichiro Futami, H. Yamada, P. S. Aisen, Y. Matsuoka

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Proteins and peptides are useful research and therapeutic tools, however applications are limited because delivery to the desired location is not easily achievable. There are two hurdles in protein/peptide delivery to the brain: the blood-brain barrier and intracellular penetration. Penetration to both brain and the intracellular space can be achieved by adjusting hydrophilicity, and small molecule pharmacological agents have been successfully developed using this approach. But with proteins and peptides, it is difficult to modify the hydrophilicity without influencing biological functions. Trans-acting factor protein from the human immunodeficiency virus contains a highly conserved cationic peptide sequence necessary for transduction across the cell membrane. While trans-acting factor peptide has been used for in vitro protein transduction, its in vivo application is very limited because it is rapidly degraded by proteolysis. Polyethylenimine is a chemically synthesized small molecule cationization agent; the charge density is greater than a peptide-based cationic cluster such as trans-acting factor, and it is resistant to proteolysis in vivo. We first tested intracellular protein transduction following direct brain injection in mice using polyethylenimine-conjugated green fluorescence protein and β-galactosidase (molecular weights 29 and 540 kDa, respectively). Polyethylenimine-conjugates penetrated to the intracellular space immediately surrounding the injection site within one hour. We further tested polyethylenimine-mediated protein transduction following intranasal administration, which bypasses the blood-brain barrier. Polyethylenimine-conjugates in pH 7.5 solution did not reach the brain, probably because the polyethylenimine-conjugates penetrated into the intracellular space where first exposed to the tissue, i.e. at the nasal mucosae. We temporarily reduced the electrostatic interaction between cationized polyethylenimine-conjugates and cellular surfaces by adjusting the pH to 4.5; solution rapidly reached the brain and penetrated to the intracellular space. This study suggests that polyethylenimine is a useful protein transduction agent in the brain in vivo, and adjusting cationic charge interaction can determine the extent of brain penetration.

Original languageEnglish
Pages (from-to)1061-1067
Number of pages7
JournalNeuroscience
Volume139
Issue number3
DOIs
Publication statusPublished - 2006

Fingerprint

Polyethyleneimine
Intracellular Space
Peptides
Trans-Activators
Brain
Proteins
Blood-Brain Barrier
Hydrophobic and Hydrophilic Interactions
Proteolysis
Galactosidases
Therapeutic Human Experimentation
Human Immunodeficiency Virus Proteins
Intranasal Administration
Injections
Nasal Mucosa
Static Electricity
Molecular Weight
Fluorescence
Cell Membrane
Pharmacology

Keywords

  • blood-brain barrier
  • drug delivery
  • intranasal delivery
  • polyethylenimine
  • protein transduction
  • trans-acting factor (Tat)

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Loftus, L. T., Li, H. F., Gray, A. J., Hirata-Fukae, C., Stoica, B. A., Futami, J., ... Matsuoka, Y. (2006). In vivo protein transduction to the CNS. Neuroscience, 139(3), 1061-1067. https://doi.org/10.1016/j.neuroscience.2006.01.041

In vivo protein transduction to the CNS. / Loftus, L. T.; Li, H. F.; Gray, A. J.; Hirata-Fukae, C.; Stoica, B. A.; Futami, Junichiro; Yamada, H.; Aisen, P. S.; Matsuoka, Y.

In: Neuroscience, Vol. 139, No. 3, 2006, p. 1061-1067.

Research output: Contribution to journalArticle

Loftus, LT, Li, HF, Gray, AJ, Hirata-Fukae, C, Stoica, BA, Futami, J, Yamada, H, Aisen, PS & Matsuoka, Y 2006, 'In vivo protein transduction to the CNS', Neuroscience, vol. 139, no. 3, pp. 1061-1067. https://doi.org/10.1016/j.neuroscience.2006.01.041
Loftus LT, Li HF, Gray AJ, Hirata-Fukae C, Stoica BA, Futami J et al. In vivo protein transduction to the CNS. Neuroscience. 2006;139(3):1061-1067. https://doi.org/10.1016/j.neuroscience.2006.01.041
Loftus, L. T. ; Li, H. F. ; Gray, A. J. ; Hirata-Fukae, C. ; Stoica, B. A. ; Futami, Junichiro ; Yamada, H. ; Aisen, P. S. ; Matsuoka, Y. / In vivo protein transduction to the CNS. In: Neuroscience. 2006 ; Vol. 139, No. 3. pp. 1061-1067.
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