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Biomarker system for studying muscle, stem cells, and cancer in vivo

Koichi Nishijo^*, Tohru Hosoyama^*, Christopher R. R. Bjornson, Beverly S. Schaffer^*, Suresh I. Prajapati^*, Ali N. Bahadur^*, Mark S. Hansen, Mary C. Blandford, Amanda T. McCleish^*, Brian P. Rubin, Jonathan A. Epstein^||, Thomas A. Rando, Mario R. Capecchi and Charles Keller^*,1

^* Greehey Children’s Cancer Research Institute and Departments of Cellular and Structural Biology and Pediatrics, University of Texas Health Science Center, San Antonio, Texas, USA;

Department of Neurology and Neurological Sciences, Stanford University, Palo Alto, California, USA;

Department of Human Genetics, University of Utah, Salt Lake City, Utah, USA;

Departments of Anatomic Pathology and Molecular Genetics, Taussig Cancer Center and Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA; and

^|| Department of Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA

¹ Correspondence: 8403 Floyd Curl Dr., MC-7784, San Antonio, TX 78229-3900, USA. E-mail: kellerc2{at}uthscsa.edu

Bioluminescent reporter genes are sensitive in situ tools for following disease progression in preclinical models, albeit they are subject to scattering and absorption in deep tissues. We have generated a bicistronic Cre/LoxP reporter mouse line that pairs the expression of firefly luciferase with quantifiable expression of a human placental alkaline phosphatase that is secreted into the serum (SeAP). With the use of this dual-modality bioreporter with a novel, inducible Pax7-CreER line for tracking muscle satellite cells, we demonstrate the longitudinal kinetics of muscle stem cell turnover, accounting for a doubling of the signal from satellite cell and progeny every 3.93 wk in the transition from adolescence to early adulthood. We also show that this dual-modality bioreporter can be incorporated in preclinical cancer models, whereby SeAP activity is reflective of tumor burden. Thus, this dual bioreporter permits both spatial localization and accurate quantification of biological processes in vivo even when the tissue of interest is deep within the animal.—Nishijo, K., Hosoyama, T., Bjornson, C. R. R., Schaffer, B. S., Prajapati, S. I., Bahadur, A. N., Hansen, M. S., Blandford, M. C., McCleish, A. T., Rubin, B. P., Epstein, J. A., Rando, T. A., Capecchi, M. R., Keller, C. Biomarker system for studying muscle, stem cells, and cancer in vivo.

Key Words: optical imaging • reporter gene • satellite cell