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Return to: College of Biological Sciences: Medical School: U of M Home |
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Areas of Research Strength: Membranes and receptors Cell interactions Gene expression Gene Therapy Apoptosis back to top |
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Research Techniques: Light, confocal and electron microscopy Immunostaining and morphometric analysis FACS analysis Northern, western and Southern blotting Isolation of nuclei, polysomes and mitochondria Run-on transcription, in vitro RNA decay and RNase protection Electrophoretic mobility gel shifts Microarrrays Tissue culture both primary and cell lines Reporter gene assays - luciferase and CAT PCR, RT-PCR, quantitative PCR and RT-PCR, real-time PCR Apoptosis detection: TUNEL, annexin, mitochondrial and caspase assays Plasmid and transgene construction using the Sleeping Beauty transposon system Nonviral transgene and oligonucleotide delivery for gene augmentation and repair Biochemical assays for gene activity - specific to model system being studied Protein production and purification back to top |
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Research Interests: The Steer laboratory is involved in two major areas of research. In the first, the lab has over the last six years developed a novel gene therapy that involves the precise repair of genetic defects in cells. The lab has concentrated on the genetic repair of a variety of diseases, including hemophilia, sickle cell disease, Crigler-Najjar syndrome type I, ornithine transcarbamylase deficiency, ß-thalassemia, von Willebrand’s disease and certain neurodegenerative disorders like Huntington’s disease. In fact, Steer has successfully treated several accurate animal models for these human disorders. The technology of gene repair is remarkable for its broad application. The ability to now correct a genetic sequence in combination with the knowledge from the human genome project creates a remarkable vista of potential therapeutic clinical studies. The correction of a precise genetic defect allows the gene to be endogenously regulated without the potential problems associated with viral vectors. It may provide us with the first cures to diseases such as sickle cell and hemophilia. The Steer lab has successfully elucidated the basic science and will now apply the technology as therapy for human disease. In addition, the lab has also developed a more traditional type of gene therapy for diseases that are not candidates for genetic repair. However, while it is just as powerful as any traditional approach, it does not require potentially harmful viral vectors. The future of gene therapy is non-viral and that is the focus of our research as it applies to human therapy. In the second area research, Steer has discovered that ursodeoxycholic acid, an endogenous hydrophilic bile acid in humans, is a potent antiapoptotic agent. Several animal models have been for diseases that are relatively accurate to their human counterparts. Specifically, ursodeoxycholic acid as a therapeutic agent to treat models of Huntington’s disease, head trauma, acute stroke, as well as Parkinson’s disease. One common characteristic shared by these disorders as well as others is the role that apoptosis plays in disease progression. Bile acid has been determined as a potent antiapoptotic agent, significantly improves neurologic status in these models. In the basic science studies, the lab has delineated the molecular mechanism by which ursodeoxycholic acid acts to preserve cell survival and cell function. As a therapeutic agent, ursodeoxycholic acid is unique in that it is a natural bile acid with no toxicity, crosses the blood-brain barrier, and can be delivered easily to patients. There are, in fact, many disease states that could potentially benefit, including myocardial infarction, autoimmune diseases, and the many acute and chronic neurodegenerative disorders for which there is little available treatment. back to top |
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Selected Publications: Park CW, Park J, Kren BT, and Steer CJ: Sleeping Beauty transposition in mouse genome is associated with changes in DNA methylation at the site of insertion. Genomics 88:204-213, 2006. Solá S, Amaral JD, Castro RE, Ramalho RM, Borralho PM, Kren BT, Tanaka H, Steer CJ, and Rodrigues CMP: Nuclear translocation of UDCA by the glucocorticoid receptor is required to reduce TGF-beta1 induced apoptosis in rat hepatocytes. Hepatology 42:925-934, 2005. Park CW, Kren BT, Largaespada DA, and Steer CJ: DNA methylation of Sleeping Beauty with transposition into the mouse genome. Genes Cells 10:763-776, 2005. Yin W, Kren BT, and Steer CJ: Site-specific base changes in the coding or promoter region of the human beta- and gamma-globin genes by single-stranded oligonucleotides. Biochem J 390:253-261, 2005. Chen ZJ, Kren BT, Wong PY-P, Low WC, and Steer CJ: Sleeping Beauty-mediated down-regulation of huntingtin expression by RNA interference. Biochem Biophys Res Comm 329:646-652, 2005. Solá S, Castro RE, Kren BT, Steer CJ, and Rodrigues CMP: Modulation of nuclear steroid receptors by ursodeoxycholic acid inhibits TGF-b1-induced E2F-1/p53-mediated apoptosis of rat hepatocytes. Biochemistry 43:8429-8438, 2004. Ramalho RM, Ribeiro PS, Solá S, Castro RE, Steer CJ, and Rodrigues CMP: Inhibition of the E2F-1/p53/Bax pathway by tauroursodeoxycholic acid in amyloid b-peptide-induced apoptosis of PC12 cells. J Neurochem 90:567-575, 2004. Castro RE, Solá S, Ramalho RM, Steer CJ, and Rodrigues CMP: The bile acid tauroursodeoxycholic acid modulates phosphorylation and translocation of Bad via phosphatidylinositol 3-kinase in glutamate-induced apoptosis of rat cortical neurons. J Pharm Exp Ther 311:845-852, 2004. Park CW, Chen Z, Kren BT, and Steer CJ: Double-stranded siRNA targeted to the huntingtin gene does not induce DNA methylation. Biochem Biophys Res Comm 323:275-280, 2004. Fan G, Ma X, Wong P-Y, Rodrigues CMP, and Steer CJ: p53 dephosphorylation and p21Cip1/Waf1 translocation correlate with caspase 3 activation in TGF-b1?induced apoptosis of HuH-7 cells. Apoptosis 9:213-223, 2004. Kren BT, Gosh SS, Linehan CL, Chowdhury NR, Hackett PB, Chowdhury JR, and Steer CJ: Hepatocyte-targeted delivery of Sleeping Beauty mediates efficient gene transfer in vivo. Gene Ther Mol Biol 7:229-238, 2003. Solá S, Ma X, Castro RE, Kren BT, Steer CJ, and Rodrigues CMP: Ursodeoxycholic acid modulates E2F-1 and p53 expression through a caspase-independent mechanism in TGF-b1?induced apoptosis of rat hepatocytes. J Biol Chem 278:48831-48838, 2003. Rodrigues CMP, Solá S, Nan Z, Castro RE, Ribeiro PS, Low WC, and Steer CJ: Tauroursodeoxycholic acid reduces apoptosis and protects against neurologic injury after acute hemorrhagic stroke in rats. Proc Natl Acad Sci USA 100:6087-6092, 2003. Kren BT, Wong PY, and Steer CJ: Short, single-stranded oligonucleotides mediate targeted nucleotide conversion using extracts from isolated liver mitochondria. DNA Repair 2:531-546, 2003. Rodrigues CMP, Solá S, Sharpe JC, Moura JJG, and Steer CJ: Tauroursodeoxycholic acid prevents Bax-induced membrane perturbation and cytochrome c release in isolated mitochondria. Biochemistry 42:3070-3080, 2003. To view these and other publications visit http://www.ncbi.nlm.nih.gov/PubMed search menu should say PubMed type Steer CJ in the avaliable line back to top |
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