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. 1994 Aug;94(2):497–505. doi: 10.1172/JCI117361

Gene transfer into the rat renal glomerulus via a mesangial cell vector: site-specific delivery, in situ amplification, and sustained expression of an exogenous gene in vivo.

M Kitamura 1, S Taylor 1, R Unwin 1, S Burton 1, F Shimizu 1, L G Fine 1
PMCID: PMC296122  PMID: 8040302

Abstract

To evaluate the pathophysiological function of specific molecules in the renal glomerulus, selective, sustained, and modifiable expression of such molecules will be required. Towards achieving this end, we devised a gene transfer system using the glomerular mesangial cell as a vector for gene delivery. A reporter gene which encodes bacterial beta-galactosidase was introduced into cultured rat mesangial cells, and the stable transfectants were transferred into the rat kidney via the renal artery, leading to selective entrapment within the glomeruli. In the normal kidney, the reporter cells populated into 57 +/- 13% of glomeruli site specifically, and the expression of beta-galactosidase was sustained for 4 wk and declined thereafter. Within the glomerulus, some of the reporter cells remained in the glomerular capillaries, while others repopulated the mesangial area and, in part, extended their cytoplasmic processes toward the surrounding capillaries. When the cells were transferred into glomeruli subjected to transient mesangiolysis induced by monoclonal antibody 1-22-3, in situ expression of beta-galactosidase was amplified 7-12-fold, and the enhanced level of expression continued for up to 8 wk. The mesangial cell vector system thus achieves site-specific delivery of an exogenous gene into the glomerulus and is amenable to in situ amplification and sustained expression by preconditioning of the target site.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bandara G., Mueller G. M., Galea-Lauri J., Tindal M. H., Georgescu H. I., Suchanek M. K., Hung G. L., Glorioso J. C., Robbins P. D., Evans C. H. Intraarticular expression of biologically active interleukin 1-receptor-antagonist protein by ex vivo gene transfer. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10764–10768. doi: 10.1073/pnas.90.22.10764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baricos W. H., Shah S. V. Proteolytic enzymes as mediators of glomerular injury. Kidney Int. 1991 Aug;40(2):161–173. doi: 10.1038/ki.1991.196. [DOI] [PubMed] [Google Scholar]
  3. Bosch R. J., Woolf A. S., Fine L. G. Gene transfer into the mammalian kidney: direct retrovirus-transduction of regenerating tubular epithelial cells. Exp Nephrol. 1993 Jan-Feb;1(1):49–54. [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Davidson B. L., Allen E. D., Kozarsky K. F., Wilson J. M., Roessler B. J. A model system for in vivo gene transfer into the central nervous system using an adenoviral vector. Nat Genet. 1993 Mar;3(3):219–223. doi: 10.1038/ng0393-219. [DOI] [PubMed] [Google Scholar]
  7. Floege J., Eng E., Young B. A., Alpers C. E., Barrett T. B., Bowen-Pope D. F., Johnson R. J. Infusion of platelet-derived growth factor or basic fibroblast growth factor induces selective glomerular mesangial cell proliferation and matrix accumulation in rats. J Clin Invest. 1993 Dec;92(6):2952–2962. doi: 10.1172/JCI116918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ginsberg J. M., Chang B. S., Matarese R. A., Garella S. Use of single voided urine samples to estimate quantitative proteinuria. N Engl J Med. 1983 Dec 22;309(25):1543–1546. doi: 10.1056/NEJM198312223092503. [DOI] [PubMed] [Google Scholar]
  9. Isaka Y., Fujiwara Y., Ueda N., Kaneda Y., Kamada T., Imai E. Glomerulosclerosis induced by in vivo transfection of transforming growth factor-beta or platelet-derived growth factor gene into the rat kidney. J Clin Invest. 1993 Dec;92(6):2597–2601. doi: 10.1172/JCI116874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kawachi H., Iwanaga T., Toyabe S., Oite T., Shimizu F. Mesangial sclerotic change with persistent proteinuria in rats after two consecutive injections of monoclonal antibody 1-22-3. Clin Exp Immunol. 1992 Oct;90(1):129–134. doi: 10.1111/j.1365-2249.1992.tb05844.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kawachi H., Orikasa M., Matsui K., Iwanaga T., Toyabe S., Oite T., Shimizu F. Epitope-specific induction of mesangial lesions with proteinuria by a MoAb against mesangial cell surface antigen. Clin Exp Immunol. 1992 Jun;88(3):399–404. doi: 10.1111/j.1365-2249.1992.tb06461.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kitamura M., Mitarai T., Maruyama N., Nagasawa R., Yoshida H., Sakai O. Mesangial cell behavior in a three-dimensional extracellular matrix. Kidney Int. 1991 Oct;40(4):653–661. doi: 10.1038/ki.1991.257. [DOI] [PubMed] [Google Scholar]
  13. Kitamura M., Shirasawa T., Maruyama N. Gene transfer of metalloproteinase transin induces aberrant behavior of cultured mesangial cells. Kidney Int. 1994 Jun;45(6):1580–1586. doi: 10.1038/ki.1994.208. [DOI] [PubMed] [Google Scholar]
  14. Lin H., Parmacek M. S., Morle G., Bolling S., Leiden J. M. Expression of recombinant genes in myocardium in vivo after direct injection of DNA. Circulation. 1990 Dec;82(6):2217–2221. doi: 10.1161/01.cir.82.6.2217. [DOI] [PubMed] [Google Scholar]
  15. Morgenstern J. P., Land H. Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res. 1990 Jun 25;18(12):3587–3596. doi: 10.1093/nar/18.12.3587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Orikasa M., Matsui K., Oite T., Shimizu F. Massive proteinuria induced in rats by a single intravenous injection of a monoclonal antibody. J Immunol. 1988 Aug 1;141(3):807–814. [PubMed] [Google Scholar]
  17. Palmer T. D., Rosman G. J., Osborne W. R., Miller A. D. Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1330–1334. doi: 10.1073/pnas.88.4.1330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Price J., Turner D., Cepko C. Lineage analysis in the vertebrate nervous system by retrovirus-mediated gene transfer. Proc Natl Acad Sci U S A. 1987 Jan;84(1):156–160. doi: 10.1073/pnas.84.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Remuzzi A., Brenner B. M., Pata V., Tebaldi G., Mariano R., Belloro A., Remuzzi G. Three-dimensional reconstructed glomerular capillary network: blood flow distribution and local filtration. Am J Physiol. 1992 Sep;263(3 Pt 2):F562–F572. doi: 10.1152/ajprenal.1992.263.3.F562. [DOI] [PubMed] [Google Scholar]
  20. Rizzino A. Soft agar growth assays for transforming growth factors and mitogenic peptides. Methods Enzymol. 1987;146:341–352. doi: 10.1016/s0076-6879(87)46035-7. [DOI] [PubMed] [Google Scholar]
  21. Scharfmann R., Axelrod J. H., Verma I. M. Long-term in vivo expression of retrovirus-mediated gene transfer in mouse fibroblast implants. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4626–4630. doi: 10.1073/pnas.88.11.4626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sedor J. R. Cytokines and growth factors in renal injury. Semin Nephrol. 1992 Sep;12(5):428–440. [PubMed] [Google Scholar]
  23. Shah S. V. Oxidant mechanisms in glomerulonephritis. Semin Nephrol. 1991 May;11(3):320–326. [PubMed] [Google Scholar]
  24. Tomita N., Higaki J., Morishita R., Kato K., Mikami H., Kaneda Y., Ogihara T. Direct in vivo gene introduction into rat kidney. Biochem Biophys Res Commun. 1992 Jul 15;186(1):129–134. doi: 10.1016/s0006-291x(05)80784-3. [DOI] [PubMed] [Google Scholar]

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