Abstract
Astrocytes, the “star-shaped” glial cells, are appealing gene-delivery targets to treat neurological diseases due to their diverse roles in brain homeostasis and disease. Cationic polymers have successfully delivered genes to mammalian cells and hence present a viable, non-immunogenic alternative to widely used viral vectors. In this study, we investigated the gene delivery potential of a series of arginine- and polyethylene glycol-modified, siloxane-based polyethylenimine analogs in primary cultured human neural cells (neurons and astrocytes) and in mice. Plasmid DNAs encoding luciferase reporter were used to measure gene expression. We hypothesized that polyplexes with arginine would help in cellular transport of the DNA, including across the blood-brain barrier; polyethylene glycol will stabilize polyethylenimine and reduce its toxicity while maintaining its DNA-condensing ability. Polyplexes were non-toxic to human neural cells and red blood cells. Cellular uptake of polyplexes and sustained gene expression were seen in human astrocytes as well as in mouse brains post-intravenous-injections. The polyplexes also delivered and expressed genes driven by astrocyte-restricted glial fibrillary acidic protein promoters, which are weaker than viral promoters. To our knowledge, the presented work validates a biocompatible and effective polymer-facilitated gene-delivery system for both human brain cells and mice for the first time.
| Original language | English |
|---|---|
| Pages (from-to) | 645-657 |
| Number of pages | 13 |
| Journal | Molecular Therapy - Nucleic Acids |
| Volume | 12 |
| DOIs | |
| State | Published - 7 Sep 2018 |
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Keywords
- GFAP promoters
- blood-brain barrier
- gene therapy
- human neural cells
- intravenous gene delivery
- polyplexes
Cite this
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Reaching for the Stars in the Brain : Polymer-Mediated Gene Delivery to Human Astrocytes. / Joshi, Chaitanya R.; Raghavan, Vijay; Vijayaraghavalu, Sivakumar; Gao, Yue; Saraswathy, Manju; Labhasetwar, Vinod; Ghorpade, Anuja.
In: Molecular Therapy - Nucleic Acids, Vol. 12, 07.09.2018, p. 645-657.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Reaching for the Stars in the Brain
T2 - Polymer-Mediated Gene Delivery to Human Astrocytes
AU - Joshi, Chaitanya R.
AU - Raghavan, Vijay
AU - Vijayaraghavalu, Sivakumar
AU - Gao, Yue
AU - Saraswathy, Manju
AU - Labhasetwar, Vinod
AU - Ghorpade, Anuja
PY - 2018/9/7
Y1 - 2018/9/7
N2 - Astrocytes, the “star-shaped” glial cells, are appealing gene-delivery targets to treat neurological diseases due to their diverse roles in brain homeostasis and disease. Cationic polymers have successfully delivered genes to mammalian cells and hence present a viable, non-immunogenic alternative to widely used viral vectors. In this study, we investigated the gene delivery potential of a series of arginine- and polyethylene glycol-modified, siloxane-based polyethylenimine analogs in primary cultured human neural cells (neurons and astrocytes) and in mice. Plasmid DNAs encoding luciferase reporter were used to measure gene expression. We hypothesized that polyplexes with arginine would help in cellular transport of the DNA, including across the blood-brain barrier; polyethylene glycol will stabilize polyethylenimine and reduce its toxicity while maintaining its DNA-condensing ability. Polyplexes were non-toxic to human neural cells and red blood cells. Cellular uptake of polyplexes and sustained gene expression were seen in human astrocytes as well as in mouse brains post-intravenous-injections. The polyplexes also delivered and expressed genes driven by astrocyte-restricted glial fibrillary acidic protein promoters, which are weaker than viral promoters. To our knowledge, the presented work validates a biocompatible and effective polymer-facilitated gene-delivery system for both human brain cells and mice for the first time.
AB - Astrocytes, the “star-shaped” glial cells, are appealing gene-delivery targets to treat neurological diseases due to their diverse roles in brain homeostasis and disease. Cationic polymers have successfully delivered genes to mammalian cells and hence present a viable, non-immunogenic alternative to widely used viral vectors. In this study, we investigated the gene delivery potential of a series of arginine- and polyethylene glycol-modified, siloxane-based polyethylenimine analogs in primary cultured human neural cells (neurons and astrocytes) and in mice. Plasmid DNAs encoding luciferase reporter were used to measure gene expression. We hypothesized that polyplexes with arginine would help in cellular transport of the DNA, including across the blood-brain barrier; polyethylene glycol will stabilize polyethylenimine and reduce its toxicity while maintaining its DNA-condensing ability. Polyplexes were non-toxic to human neural cells and red blood cells. Cellular uptake of polyplexes and sustained gene expression were seen in human astrocytes as well as in mouse brains post-intravenous-injections. The polyplexes also delivered and expressed genes driven by astrocyte-restricted glial fibrillary acidic protein promoters, which are weaker than viral promoters. To our knowledge, the presented work validates a biocompatible and effective polymer-facilitated gene-delivery system for both human brain cells and mice for the first time.
KW - GFAP promoters
KW - blood-brain barrier
KW - gene therapy
KW - human neural cells
KW - intravenous gene delivery
KW - polyplexes
UR - http://www.scopus.com/inward/record.url?scp=85050865372&partnerID=8YFLogxK
U2 - 10.1016/j.omtn.2018.06.009
DO - 10.1016/j.omtn.2018.06.009
M3 - Article
VL - 12
SP - 645
EP - 657
JO - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
SN - 2162-2531
ER -