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Stem cells are promising therapeutic delivery vehicles; however pre‐clinical and clinical applications of stem cell‐based therapy would benefit significantly from the ability to simultaneously determine therapeutic efficacy and pharmacokinetics of therapies delivered by engineered stem cells. In this study, we engineered and screened numerous fusion variants that contained therapeutic (TRAIL) and diagnostic (luciferase) domains designed to allow simultaneous investigation of multiple events in stem cell‐based therapy  in vivo . When various stem cell lines were engineered with the optimized molecule, SRL O L 2 TR, diagnostic imaging showed marked differences in the levels and duration of secretion between stem cell lines, while the therapeutic activity of the molecule showed the different secretion levels translated to significant variability in tumor cell killing.  In vivo , simultaneous diagnostic and therapeutic monitoring revealed that stem cell‐based delivery significantly improved pharmacokinetics and anti‐tumor effectiveness of the therapy compared to intravenous or intratumoral delivery. As treatment for highly malignant brain tumor xenografts, tracking SRL O L 2 TR showed stable stem cell‐mediated delivery significantly regressed peripheral and intracranial tumors. Together, the integrated diagnostic and therapeutic properties of SRL O L 2 TR answer critical questions necessary for successful utilization of stem cells as novel therapeutic vehicles. S TEM  C ELLS :2010;28:832–841

A Novel Molecule Integrating Therapeutic and Diagnostic Activities Reveals Multiple Aspects of Stem Cell‐based Therapy