1. Membrane-anchored  -glucuronidase Steve Roffler Institute of Biomedical Sciences Academia Sinica Taipei, Taiwan

2. MEMzyme Transgene Transgene Expression ß-glucuronidase glucuronide Glucuronide probe or drug B7 TM glucuronide Potential advantage: Signal amplification due to enzymatic hydrolysis of glucuronide probes or drugs Memzyme activation of imaging or therapeutic agents

3. ßG in lysosomes – inaccessible to glucuronides Low serum ßG – little systemic activation Low toxicity Fast renal clearance – rapid imaging Endogenous - low immunogenicity Many possible glucuronide probes and drugs Potential advantages of a ßG reporter

4. Fluorescent probe can be activated and accumulate at cells that express enzyme In vitro imaging of gene expression red = enzyme expression green = activated probe accumulation

5. Sites of Ad transgene expression could be imaged Imaging of adenovirus memzyme expression Kodak IS2000MM

6. In vitro activation of HAMG HAMG can be activated at EJ/mßG cells O HO OH COOH O N Cl HAMG Glucuronide prodrug N Cl HO pHAM Active drug

7. HAMG prodrug toxicity to mice HAMG is well tolerated by mice

8. Prodrug therapy of EJ tumors HAMG displayed selective activity against EJ/mßG tumors 01020304050 0 1000 2000 EJ PBS EJ HAMG EJ/mßG PBS EJ/mßG HAMG Days Tumor volume (mm 3 ) (150 mg/kg HAMG on days 15 and 19)

9. Summary Surface enzyme allow both imaging and therapy Immunogenicity is low but selectivity is high May possess advantages for cancer gene therapy

10. Dr. Kuang-Wen Liao Shey-Cherng Tzo Tang-Bi Liu Surface scFv T cell activation Surface expression Shih-en Chang Wen-Jun Chung Bing-Mae Chen Dr. Yu-Ling Leu Chia-Nan College of Pharmacy and Science Dr. Ji-Wang Chern National Taiwan University Dr. Tian-Lu Cheng Kaohsiung Medical University Chin-Chuan Chen Dr. Hsin-Ell Wang National Yang Ming University Yi-Hsuan Chiang Pei-Yu Wu Chien-I Su http://www.ibms.sinica.edu.tw/~sroff/sr_refs.html