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.

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 j3-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 f8-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 13-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. (J. Clin. Invest. 1994. 94:497-505.)

Priming of Glomerular Mesangial Cells by Activated Macrophages Causes Blunted Responses to Proinflammatory Stimuli

“…Acute anti-Thy 1 glomerulonephritis was induced in rats (Sprague-Dawley rats; 250 -300 g body weight) by a mAb 1-22-3 (22), as described previously (23). In this experimental model, accumulation of macrophages in glomeruli is observed within 24 h (11).…”

“…In this experimental model, accumulation of macrophages in glomeruli is observed within 24 h (11). Twenty-four hours after the Ab injection, SM/NF B-SEAP5 cells (1 ϫ 10 6 cells) were trypsinized and injected into the nephritic rat kidneys, as described previously (23). In this experimental setting, 90% of glomeruli are transferred with the reporter mesangial cells (24).…”

Continuous, noninvasive monitoring of local microscopic inflammation using a genetically engineered cell-based biosensor

“…Mesangial cells (SM43) established from isolated glomeruli of a male Sprague-Dawley rat 1 were maintained in medium containing 5% fetal bovine serum (FBS). Medium containing 1% FBS was generally used for studies.…”

“…1 This method utilized the glomerular mesangial cell as a vector for gene delivery. That is, mesangial cells were cultured from isolated glomeruli, stably transfected with foreign genes in vitro and transferred back into glomeruli via the renal circulation.…”

β-Galactosidase marker genes to tag and track human hematopoietic cells

“…LacZ was transferred to the cotton rat or to nonhuman primate airway epithelium using adenoviral vectors in preparation for a clinical trial of cystic fibrosis. 34 -36 The gene encoding ␤-gal was also transferred to neuronal [37][38][39][40] and retinal cells, [41][42][43] keratinocytes, 44,45 bladder 46 and gallbladder cells, 47 mesangial and tubular cells, 48,49 endothelial and other vascular cells, 50 -54 skeletal and heart muscle cells, [55][56][57][58][59] hepatocytes, 60 -63 salivary gland cells, 64 and even thyroid follicular cells 65 using different, mostly adenoviral vectors in different species. In these experiments, limited side effects were observed and were believed to be related to the vector itself, 34,51,57,65,66 rather than to the LacZ gene or its product.…”