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ApoE-2 Brain-Targeted Gene Therapy Through Transferrin and Penetratin Tagged Liposomal Nanoparticles

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Abstract

Purpose

Apolipoprotein E2 (ApoE2) gene therapy is a potential disease-modifying therapy for Alzheimer’s disease (AD). We investigated the potential of plasmid encoding ApoE2 loaded brain-targeted functionalized-liposomes for treatment of AD. This was achieved via systemic administration of liposomes entrapping therapeutic gene targeting the brain of mice.

Methods

Targeting and transfection efficiency of designed liposomes were determined in bEnd.3, primary glial and primary neuronal cells. The ability of liposomal formulations to translocate across in vitro blood-brain barrier (BBB) and, thereafter, transfect primary neuronal cells was investigated using in vitro triple co-culture BBB model. We quantified ApoE expression in the brain of mice after single intravenous injection of brain-targeted liposomes loaded with plasmid ApoE2.

Results

Dual surface modification enhanced the in vitro transfection efficiency of designed liposomes. Successful delivery of therapeutic gene overcoming BBB by Transferrin-Penetratin- modified liposomes was demonstrated both in vitro and in vivo. Significant (p < 0.05) increase in ApoE levels in the brain of mice was observed after intravenous administration of Tf-Pen-liposomes encasing plasmid ApoE2.

Conclusion

The results indicate that dual-ligand based liposomal gene delivery systems had both enhanced brain targeting and gene delivery efficiencies. Transferrin-Penetratin modified liposomes for delivery of plasmid ApoE2 has great potential for AD treatment.

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Abbreviations

AD:

Alzheimer’s disease

BBB:

Blood brain barrier

CPP:

Cell-penetrating peptide

DOPE:

1,2-dioleoyl-sn-glycero-3-phosphoethanolamine

DOTAP:

1,2-dioleoyl-3-trimethylammonium-propane

FBS:

Fetal bovine serum

H&E:

Hematoxylin and eosin

LOAD:

Late onset form of Alzheimer’s disease

MTT:

(3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide)

p/s:

Penicillin and streptomycin

Pen:

Penetratin

TEER:

Transepithelial electrical resistance

Tf:

Transferrin

TfR:

Transferrin receptor

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Acknowledgments and Disclosures

This research was supported by National Institutes of Health (Grant R01AG051574). B.S.R. is supported by doctoral fellowship from The Brazilian National Council for Scientific and Technological Development (CNPq, Brazil) with a scholarship for B.S.R (Full Doctorate Fellowship (GDE): 221327/2014–2). The authors report no conflict of interest.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy, College of Health Professions, North Dakota State University, Fargo, North Dakota, 58105, USA

    Bruna dos Santos Rodrigues & Jagdish Singh

  2. Department of Neuroscience, Mayo Clinic, Jacksonville, Florida, 32224, USA

    Takahisa Kanekiyo

Authors
  1. Bruna dos Santos Rodrigues
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  2. Takahisa Kanekiyo
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  3. Jagdish Singh
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Correspondence to Jagdish Singh.

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dos Santos Rodrigues, B., Kanekiyo, T. & Singh, J. ApoE-2 Brain-Targeted Gene Therapy Through Transferrin and Penetratin Tagged Liposomal Nanoparticles. Pharm Res 36, 161 (2019). https://doi.org/10.1007/s11095-019-2691-7

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