Microsphere-adenoviral complexes target and transduce the glomerulus in vivo

N. Stanley Nahman, Thomas J. Sferra, Julie Kronenberger, Kathleen E. Urban, Anne E. Troike, Amy Johnson, Bethany J. Holycross, Gerard J. Nuovo, Daniel D. Sedmak

Research output: Contribution to journalArticle

18 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)1500-1510
Number of pages11
JournalKidney International
Volume58
Issue number4
DOIs
StatePublished - Jan 1 2000

Fingerprint

Microspheres
Transgenes
Adenoviridae
Luciferases
Genetic Therapy
Kidney Glomerulus
Galactosidases
Injections
Mesangial Cells
RNA-Directed DNA Polymerase
Polystyrenes
Glomerulonephritis
Sprague Dawley Rats
Endothelial Cells
Kidney
Polymerase Chain Reaction
DNA
Genes

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

Nahman, N. S., Sferra, T. J., Kronenberger, J., Urban, K. E., Troike, A. E., Johnson, A., ... Sedmak, D. D. (2000). Microsphere-adenoviral complexes target and transduce the glomerulus in vivo. Kidney International, 58(4), 1500-1510. https://doi.org/10.1046/j.1523-1755.2000.00312.x

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 journalArticle

Nahman, NS, Sferra, TJ, Kronenberger, J, Urban, KE, Troike, AE, Johnson, A, Holycross, BJ, Nuovo, GJ & Sedmak, DD 2000, 'Microsphere-adenoviral complexes target and transduce the glomerulus in vivo', Kidney International, vol. 58, no. 4, pp. 1500-1510. https://doi.org/10.1046/j.1523-1755.2000.00312.x
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. / Microsphere-adenoviral complexes target and transduce the glomerulus in vivo. In: Kidney International. 2000 ; Vol. 58, No. 4. pp. 1500-1510.
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AU - Urban, Kathleen E.

AU - Troike, Anne E.

AU - Johnson, Amy

AU - Holycross, Bethany J.

AU - Nuovo, Gerard J.

AU - Sedmak, Daniel D.

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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.

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