Abstract
Background. Developing new treatments for glomerulonephritis makes the glomerulus a logical target for gene therapy. Microspheres may lodge in the glomerulus, and replication-deficient recombinant adenoviruses are potent mediators of gene transfer. We postulated that adenoviral-microsphere complexes could result in DNA transfer (transduction) into glomerular cells in vivo. Methods. Two adenoviruses, each one containing a luciferase or β-galactosidase (β-gal) transgene expression cassette, were complexed to polystyrene microspheres. To assess in vivo glomerular transduction with this tool, male Sprague-Dawley rats underwent aortic injections with adenovirus linked to 11 or 16 μm diameter microspheres. Results. After 48 hours, adenoviral-microsphere complexes resulted in transduction of up to 19% of glomeruli per kidney section. Endothelial and mesangial cells were transduced with this approach, and transprotein expression persisted for 21 days. Transduction efficiency was greater in the 16 μm group. For all rats, there was a strong correlation between kidney luciferase levels and the number of β-gal-positive glomeruli (r = 0.87), indicating that transgene expression was primarily glomerular in location. This was supported by reverse transcriptase in situ polymerase chain reaction, which demonstrated glomerular localization of the β-gal transgene. Conclusions. The aortic injection of adenoviral-microsphere complexes transduces the glomerulus in vivo and may be a useful tool in developing approaches to gene therapy of glomerular disease.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1500-1510 |
| Number of pages | 11 |
| Journal | Kidney International |
| Volume | 58 |
| Issue number | 4 |
| DOIs | |
| State | Published - Jan 1 2000 |
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Keywords
- Acute glomerulonephritis
- Chronic glomerular disease
- Gene therapy
- Glomerular gene transfer
- Mesangial cell vectors
- Nucleotide sequences
- Replication-deficient recombinant adenovirus
ASJC Scopus subject areas
- Nephrology
Cite this
Microsphere-adenoviral complexes target and transduce the glomerulus in vivo. / Nahman, N. Stanley; Sferra, Thomas J.; Kronenberger, Julie; Urban, Kathleen E.; Troike, Anne E.; Johnson, Amy; Holycross, Bethany J.; Nuovo, Gerard J.; Sedmak, Daniel D.
In: Kidney International, Vol. 58, No. 4, 01.01.2000, p. 1500-1510.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Microsphere-adenoviral complexes target and transduce the glomerulus in vivo
AU - Nahman, N. Stanley
AU - Sferra, Thomas J.
AU - Kronenberger, Julie
AU - Urban, Kathleen E.
AU - Troike, Anne E.
AU - Johnson, Amy
AU - Holycross, Bethany J.
AU - Nuovo, Gerard J.
AU - Sedmak, Daniel D.
PY - 2000/1/1
Y1 - 2000/1/1
N2 - Background. Developing new treatments for glomerulonephritis makes the glomerulus a logical target for gene therapy. Microspheres may lodge in the glomerulus, and replication-deficient recombinant adenoviruses are potent mediators of gene transfer. We postulated that adenoviral-microsphere complexes could result in DNA transfer (transduction) into glomerular cells in vivo. Methods. Two adenoviruses, each one containing a luciferase or β-galactosidase (β-gal) transgene expression cassette, were complexed to polystyrene microspheres. To assess in vivo glomerular transduction with this tool, male Sprague-Dawley rats underwent aortic injections with adenovirus linked to 11 or 16 μm diameter microspheres. Results. After 48 hours, adenoviral-microsphere complexes resulted in transduction of up to 19% of glomeruli per kidney section. Endothelial and mesangial cells were transduced with this approach, and transprotein expression persisted for 21 days. Transduction efficiency was greater in the 16 μm group. For all rats, there was a strong correlation between kidney luciferase levels and the number of β-gal-positive glomeruli (r = 0.87), indicating that transgene expression was primarily glomerular in location. This was supported by reverse transcriptase in situ polymerase chain reaction, which demonstrated glomerular localization of the β-gal transgene. Conclusions. The aortic injection of adenoviral-microsphere complexes transduces the glomerulus in vivo and may be a useful tool in developing approaches to gene therapy of glomerular disease.
AB - Background. Developing new treatments for glomerulonephritis makes the glomerulus a logical target for gene therapy. Microspheres may lodge in the glomerulus, and replication-deficient recombinant adenoviruses are potent mediators of gene transfer. We postulated that adenoviral-microsphere complexes could result in DNA transfer (transduction) into glomerular cells in vivo. Methods. Two adenoviruses, each one containing a luciferase or β-galactosidase (β-gal) transgene expression cassette, were complexed to polystyrene microspheres. To assess in vivo glomerular transduction with this tool, male Sprague-Dawley rats underwent aortic injections with adenovirus linked to 11 or 16 μm diameter microspheres. Results. After 48 hours, adenoviral-microsphere complexes resulted in transduction of up to 19% of glomeruli per kidney section. Endothelial and mesangial cells were transduced with this approach, and transprotein expression persisted for 21 days. Transduction efficiency was greater in the 16 μm group. For all rats, there was a strong correlation between kidney luciferase levels and the number of β-gal-positive glomeruli (r = 0.87), indicating that transgene expression was primarily glomerular in location. This was supported by reverse transcriptase in situ polymerase chain reaction, which demonstrated glomerular localization of the β-gal transgene. Conclusions. The aortic injection of adenoviral-microsphere complexes transduces the glomerulus in vivo and may be a useful tool in developing approaches to gene therapy of glomerular disease.
KW - Acute glomerulonephritis
KW - Chronic glomerular disease
KW - Gene therapy
KW - Glomerular gene transfer
KW - Mesangial cell vectors
KW - Nucleotide sequences
KW - Replication-deficient recombinant adenovirus
UR - http://www.scopus.com/inward/record.url?scp=0033801860&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033801860&partnerID=8YFLogxK
U2 - 10.1046/j.1523-1755.2000.00312.x
DO - 10.1046/j.1523-1755.2000.00312.x
M3 - Article
C2 - 11012885
AN - SCOPUS:0033801860
VL - 58
SP - 1500
EP - 1510
JO - Kidney International
JF - Kidney International
SN - 0085-2538
IS - 4
ER -