Engineered Biomarker cell lines

CXR Biosciences is continually developing new in vitro models for the rapid assessment of mechanisms of toxicity. With the Engineered Biomarker cell lines specific genes associated with the response to toxic insult have been linked to reporter systems. These models provide a more rapid, more informative means to assess  mechanistic toxicology in vivo.

The following Engineered Biomarker models have been generated;

• Antioxidant Response Reporter (ARE)* more
• Xenobiotic Response Reporter (XRE)** more
• Oxidative stress Reporter  (Hmox-1)** more
• AP-1 Reporter** more
• Inflammatory Response Reporter  (NFkB)** more

*- developed in collaboration with CRUK; **- developed in collaboration with Millipore Corporation.

Antioxidant Response Reporter Cell Line : Reports on the up-regulation of the antioxidant response element gene battery. An MCF7 cell line has been made that contains a luciferase reporter gene under the regulation of multiple antioxidant response element (ARE) enhancer sequences.

Uses of the ARE reporter Cell line:

• As an efficacy screen to identify compounds with chemoprotective properties.
• As a toxicity /carcinogenicity screen to identify compounds that induce oxidative stress.

Xenobiotic Response Reporter Cell Line: Heterodimerization of the aryl hydrocarbon receptor (AhR) and AhR nuclear translocator (ARNT) then binding of the complex to Xenobiotic Response Elements (XRE) of target genes induces the expression of a variety of drug metabolising enzymes such as the cytochrome P450s (CYP1A and  CYP1B), UGT1A and GSTs.

• XRE responds to a variety of environmental pollutants.

Oxidative Stress Reporter Cell Line:  A CHO cell line has been created, containing a reporter based upon a mouse Hmox1 gene promoter that regulates the expression of an epitope-tagged hCG gene.  The induction of the Hmox1 gene generally indicates an oxidative stress response in the cell. The cell line response, to inducers of oxidative stress, has been shown to be reproducible and consistent, and a robust assay has been developed. The assay has been transferred successfully to several other laboratories.

AP-1 Reporter Cell Line:  A HeLa cell line containing an epitope tagged hCG gene construct has been generated which is controlled by the AP-1 enhancer element. The nuclear transcription factor AP-1 is a dimer complex composed by DNA-binding proteins of Jun, Fos, and ATF families. AP-1 mediates cellular response to growth factors, cytokines, neurotransmitters and other intercellular signaling molecules. AP-1 activity is mediated by G-proteins, adapter proteins, MAP kinases and other elements of cellular signaling systems. AP-1 dependent genes play a pivotal role in regulation of cell proliferation, morphogenesis, apoptosis, and differentiation.

• The crucial role of AP-1 renders it an appealing target for future nuclear-directed anticancer therapeutic and chemoprevention approaches.

Inflammatory Response Reporter Cell Line:   A HeLa cell line has been generated, which contains an epitope tagged hCG reporter protein that is regulated by multiple Nuclear Factor Kappa B (NFκB) response element enhancer sequences. NFκB plays a major role in the regulation of inflammatory response, apoptosis and tumorigenesis, and is activated by a wide variety of stimuli including pro-inflammatory cytokines, free radicals, ultraviolet radiation and bacterial or viral protein. CXR has generated data from in vitro use of this cell line, and are currently using it as an assay for inflammatory response.

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