Noninvasive imaging of protein–protein interactions in living animals

Luker, Gary D.; Sharma, Vijay; Pica, Christina M.; Dahlheimer, Julie L.; Li, Wei; Ochesky, Joseph; Ryan, Christine E.; Piwnica‐Worms, Helen; Piwnica‐Worms, David

doi:10.1073/pnas.092022399

Cited by 155 publications

(62 citation statements)

References 27 publications

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“…The same genetically encoded reporter system was then used to validate the lead compound from the cell culture screen (PU-H71) for its ability to disrupt Hsp90a/p23 and Hsp90h/ p23 interactions in living mice. In addition to SRL-PFAC, split FL protein fragment-assisted complementation assays (SFL-PFAC) have also been used to monitor other protein-protein interactions in living mice (40)(41)(42). The smaller fragment sizes for the split RL strategy (23 kDa for NRL and 13 kDa for CRL) relative to split FL fragments (f32 kDa) may minimize steric hindrance for Hsp90a/ p23 and Hsp90h/p23 interactions in the split reporters.…”

Section: Discussionmentioning

confidence: 99%

Molecular Imaging of the Efficacy of Heat Shock Protein 90 Inhibitors in Living Subjects

et al. 2008

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Heat shock protein 90A (Hsp90A)/p23 and Hsp90B/p23 interactions are crucial for proper folding of proteins involved in cancer and neurodegenerative diseases. Small molecule Hsp90 inhibitors block Hsp90A/p23 and Hsp90B/p23 interactions in part by preventing ATP binding to Hsp90. The importance of isoform-selective Hsp90A/p23 and Hsp90B/p23 interactions in determining the sensitivity to Hsp90 was examined using 293T human kidney cancer cells stably expressing split Renilla luciferase (RL) reporters. Interactions between Hsp90A/p23 and Hsp90B/p23 in the split RL reporters led to complementation of RL activity, which was determined by bioluminescence imaging of intact cells in cell culture and living mice using a cooled charge-coupled device camera. The three geldanamycin-based and seven purinescaffold Hsp90 inhibitors led to different levels of inhibition of complemented RL activities (10-70%). However, there was no isoform selectivity to both classes of Hsp90 inhibitors in cell culture conditions. The most potent Hsp90 inhibitor, PU-H71, however, led to a 60% and 30% decrease in RL activity (14 hr) in 293T xenografts expressing Hsp90A/p23 and Hsp90B/p23 split reporters respectively, relative to carrier control-treated mice. Molecular imaging of isoform-specific Hsp90A/p23 and Hsp90B/p23 interactions and efficacy of different classes of Hsp90 inhibitors in living subjects have been achieved with a novel genetically encoded reporter gene strategy that should help in accelerating development of potent and isoformselective Hsp90 inhibitors. [Cancer Res 2008;68(1):216-26]

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Section: Discussionmentioning

confidence: 99%

Molecular Imaging of the Efficacy of Heat Shock Protein 90 Inhibitors in Living Subjects

et al. 2008

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“…5 Accordingly, several experimental studies have shown that PET can be used to quantify gene expression by measuring the accumulation of radiolabeled tracers which are trapped only in tissues expressing reporter proteins such as the Herpes simplex virus type-1 thymidine kinase (HSV1-TK) in vivo. 6,7 Several mutant HSV1-TK enzymes (mHSV1-TK) have been developed to further improve the sensitivity of PET imaging to detect and quantify the cellular accumulation of radioactive tracers. 7,8 To date, the validity of this strategy to quantify mHSV1-tk reporter gene expression has been demonstrated in the liver, 9 in tumor xenografts in rodents, 10 and in myocardium; 11 recently, we have confirmed these results in rat lungs.…”

Section: Introductionmentioning

confidence: 99%

Imaging the spatial distribution of transgene expression in the lungs with positron emission tomography

Richard

2003

Gene Ther

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“…MicroPET images were corrected for decay, but not attenuation or scatter, and stacked regions-of-interest (ROI) of relevant tissues and organs were analyzed with AnalyzePC 6.0 software. Data for accumulation of 18 F-FDG on microPET images were expressed as standard uptake values (SUV), representing counts per gram of tissue divided by injected dose of radioactivity per gram of animal weight (55,56). Data are presented as mean values Ϯ SEM.…”

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confidence: 99%

Loss of the Par-1b/MARK2 polarity kinase leads to increased metabolic rate, decreased adiposity, and insulin hypersensitivity in vivo

Hurov

Huang

White

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Obesity is a major factor central to the development of insulin resistance and type 2 diabetes. The identification and characterization of genes involved in regulation of adiposity, insulin sensitivity, and glucose uptake are key to the design and development of new drug therapies for this disease. In this study, we show that the polarity kinase Par-1b/MARK2 is required for regulating glucose metabolism in vivo. Mice null for Par-1b were lean, insulin hypersensitive, resistant to high-fat diet-induced weight gain, and hypermetabolic. 18 F-FDG microPET and hyperinsulinemic-euglycemic clamp analyses demonstrated increased glucose uptake into white and brown adipose tissue, but not into skeletal muscle of Par-1b null mice relative to wild-type controls. Taken together, these data indicate that Par-1b is a regulator of glucose metabolism and adiposity in the whole animal and may be a valuable drug target for the treatment of both type 2 diabetes and obesity.glucose ͉ obesity ͉ EMK

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Noninvasive imaging of protein–protein interactions in living animals

Cited by 155 publications

References 27 publications

Molecular Imaging of the Efficacy of Heat Shock Protein 90 Inhibitors in Living Subjects

Molecular Imaging of the Efficacy of Heat Shock Protein 90 Inhibitors in Living Subjects

Imaging the spatial distribution of transgene expression in the lungs with positron emission tomography

Loss of the Par-1b/MARK2 polarity kinase leads to increased metabolic rate, decreased adiposity, and insulin hypersensitivity in vivo

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