Creation of an In vivo cytosensor using engineered mesangial cells. Automatic sensing of glomerular inflammation controls transgene activity.

Kitamura, Masanori; Kawachi, Hiroshi

doi:10.1172/jci119659

Cited by 37 publications

(22 citation statements)

References 44 publications

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“…1,4,21 After the cell injection, glomeruli were isolated from both kidneys by using the conventional sieving method. 22 In all, 1000 glomeruli were incubated in 100 ml medium containing 1% FBS, and after 12 h, media were collected for SEAP assay.…”

Section: Ex Vivo Sensing Of Glomerulonephritismentioning

confidence: 99%

“…1,4 In this method, mesangial cells are transferred selectively into rat glomeruli by renal artery injection. Within glomeruli, transferred cells attach and extend along the endothelium or occupy the capillary lumen.…”

Section: Sensing Of Glomerular Inflammation In Vitro and Ex Vivomentioning

confidence: 99%

“…3 By combining the ex vivo gene transfer method with SRE, we established a local biosensor that achieves automatic sensing of glomerular inflammation and subsequent expression of a marker gene, b-galactosidase. 4 As a monitoring system for glomerular inflammation, however, this system has obvious limitations. One limitation is that b-galactosidase is not a secreted protein, and tissue biopsy is inevitable to evaluate the reporter expression.…”

mentioning

confidence: 99%

“…4 However, the promoter activity of SRE to drive foreign gene expression is relatively weak in mesangial cells. In this report, we therefore utilized an alternative, inflammation-responsive element, the kB enhancer element, as a sensor for glomerulonephritis.…”

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confidence: 99%

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

Meng

Kasai

Hiramatsu

et al. 2005

Laboratory Investigation

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Using an inflammation-responsive regulatory element as a molecular sensor, we established a cell-based biosensor for continuous, noninvasive monitoring of local microscopic inflammation in vivo. Glomerular mesangial cells were stably transfected with a marker gene encoding secreted alkaline phosphatase (SEAP) under the control of the jB enhancer elements. The established cells secreted SEAP in vitro in response to proinflammatory cytokines as well as to soluble factors produced by inflamed glomeruli. To examine feasibility of using the established cells for in vivo monitoring of local microscopic inflammation, the sensor cells were transferred selectively into rat glomeruli via the renal circulation. After induction of acute glomerulonephritis, the serum level of SEAP was increased transiently in cell-transferred nephritic rats. The kinetics of serum SEAP was closely correlated with the natural course of the inflammation, and the increase in SEAP was attenuated by suppression of inflammation using an immunosuppressive drug, cyclophosphamide. Neither cell-transferred normal rats nor nephritic rats without cell transfer exhibited increase in the serum level of SEAP. When the sensor cells were transferred extrarenally, elevation of serum SEAP was not observed in nephritic rats, confirming that the locally settled sensor cells responded only to local inflammation. These results suggested that, without invasive procedures like tissue biopsies, continuous monitoring of microscopic inflammation is feasible in vivo via locally created, cell-based biosensors.

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Section: Ex Vivo Sensing Of Glomerulonephritismentioning

confidence: 99%

Section: Sensing Of Glomerular Inflammation In Vitro and Ex Vivomentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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

Meng

Kasai

Hiramatsu

et al. 2005

Laboratory Investigation

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“…pCI-␤gal introduces lacZ under control of the immediate-early enhancer/promoter of human CMV. After transfection, cells were incubated for 48 h in 0.5% FCS, stimulated with MG132 (10 -25 M) for 24 h, and subjected to 5-bromo-4-chloro-3-indolyl-␤-D-galactopyranoside (X-gal) assay (36). Activity of AP-1 was evaluated by counting X-gal-positive cells in each well.…”

Section: Reporter Assaymentioning

confidence: 99%

Unexpected Transcriptional Induction of Monocyte Chemoattractant Protein 1 by Proteasome Inhibition: Involvement of the c-Jun N-Terminal Kinase-Activator Protein 1 Pathway

Nakayama

Furusu

et al. 2001

The Journal of Immunology

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Proteasome inhibitors, the well-known inhibitors of NF-κB, are recently considered therapeutic agents for inflammation. However, the anti-inflammatory properties of these agents have not been fully evaluated. In this report we describe a novel effect of proteasome inhibitors on the expression of monocyte chemoattractant protein 1 (MCP-1) in mesangial cells. We found that proteasome inhibitor MG132 dose-dependently induced expression of MCP-1 at the transcriptional level. The stimulatory effect was similarly observed with other proteasome inhibitors (proteasome inhibitor 1 and lactacystin) and in other cell types (NRK fibroblasts). The 5′-flanking region of the MCP-1 gene contains multiple AP-1 sites. To explore the mechanisms involved, we examined the effects of proteasome inhibition on the AP-1 pathway. Northern blot analysis showed that MG132 rapidly induced the expression of c-jun, but not c-fos. Immunoblot analysis showed that MG132 prevented degradation of c-Jun protein. Kinase assay revealed that c-Jun N-terminal kinase (JNK) was rapidly activated by MG132. Consistent with these results, a reporter assay showed that AP-1 activity was up-regulated after treatment with MG132. Curcumin, a pharmacological inhibitor of the JNK-AP-1 pathway, abrogated the induction of MCP-1 by MG132. Similarly, stable transfection with a dominant-negative mutant of c-Jun attenuated both MG132-induced activation of AP-1 and expression of MCP-1. The transcriptional activation by proteasome inhibitors was observed not only in MCP-1, but also in other AP-1-dependent genes, including stromelysin and mitogen-activated protein kinase phosphatase 1. These data revealed that proteasome inhibition triggered the expression of MCP-1 and other genes via the multistep induction of the JNK-c-Jun/AP-1 pathway.

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Unexpected Suppression of α-Smooth Muscle Actin, the Activation Marker of Mesangial Cells, by pp60v-srcTyrosine Kinase

Ishikawa

Kitamura

1998

Biochemical and Biophysical Research Communications

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Creation of an In vivo cytosensor using engineered mesangial cells. Automatic sensing of glomerular inflammation controls transgene activity.

Abstract: Automatic control over exogenous gene expression in response to the activity of disease is a crucial hurdle for gene transfer-based therapies. Towards achieving this goal, we created a "cytosensor" that perceives local inflammatory states and subsequently regulates foreign gene expression.

Cited by 37 publications

References 44 publications

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

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

Unexpected Transcriptional Induction of Monocyte Chemoattractant Protein 1 by Proteasome Inhibition: Involvement of the c-Jun N-Terminal Kinase-Activator Protein 1 Pathway

Unexpected Suppression of α-Smooth Muscle Actin, the Activation Marker of Mesangial Cells, by pp60v-srcTyrosine Kinase

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