Cancer gene therapy using a survivin mutant adenovirus
Mehdi Mesri, … , Richard W. Kim, Dario C. Altieri
Mehdi Mesri, … , Richard W. Kim, Dario C. Altieri
Published October 1, 2001
Citation Information: J Clin Invest. 2001;108(7):981-990. https://doi.org/10.1172/JCI12983.
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Article

Cancer gene therapy using a survivin mutant adenovirus

Abstract

We have constructed a replication-deficient adenovirus encoding a nonphosphorylatable Thr34→Ala mutant of the apoptosis inhibitor survivin (pAd-T34A) to target tumor cell viability in vitro and in vivo. Infection with pAd-T34A caused spontaneous apoptosis in cell lines of breast, cervical, prostate, lung, and colorectal cancer. In contrast, pAd-T34A did not affect cell viability of proliferating normal human cells, including fibroblasts, endothelium, or smooth muscle cells. Infection of tumor cells with pAd-T34A resulted in cytochrome c release from mitochondria, cleavage of approximately 46-kDa upstream caspase-9, processing of caspase-3 to the active subunits of approximately 17 and 19 kDa, and increased caspase-3 catalytic activity. When compared with chemotherapeutic regimens, pAd-T34A was as effective as taxol and considerably more effective than adriamycin in induction of tumor cell apoptosis and enhanced taxol-induced cell death. In three xenograft breast cancer models in immunodeficient mice, pAd-T34A suppressed de novo tumor formation, inhibited by approximately 40% the growth of established tumors, and reduced intraperitoneal tumor dissemination. Tumors injected with pAd-T34A exhibited loss of proliferating cells and massive apoptosis by in situ internucleosomal DNA fragmentation. These data suggest that adenoviral targeting of the survivin pathway may provide a novel approach for selective cancer gene therapy.

Authors

Mehdi Mesri, Nathan R. Wall, Jia Li, Richard W. Kim, Dario C. Altieri

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Effect of intratumor injection of pAd-T34A on tumor growth, proliferatio...
Effect of intratumor injection of pAd-T34A on tumor growth, proliferation, and apoptosis. (a) Expression of injected pAd vectors in vivo. Uninfected MCF-7 cells (1.5 × 106) were injected into the flank of female CB17 SCID/beige mice, and tumors were allowed to grow to approximately 100–150 mm3 volume (7–10 mm diameter). Tumors were injected with pAd-GFP or pAd-T34A in three sites (5 × 108 GFU/site) and harvested after 48 hours (pAd-T34A) or 96 hours (pAd-GFP). Cryostat tumor sections were analyzed for GFP expression by fluorescence microscopy. In, needle injection track. ×200. (b) Kinetics of tumor growth. The experimental conditions are as in a. Tumor growth was measured in three dimensions at the indicated time intervals after intratumor injection of pAd-GFP or pAd-T34A. Data are the mean ± SEM of the various animals per group indicated in parenthesis. *P < 0.05, **P < 0.005. (c) Histology. Tumor sections from one animal injected with pAd-GFP or two representative animals injected with pAd-T34A were analyzed after 48 hours by H&E staining (H&E), cell proliferation by Ki-67 labeling (Ki-67), or in situ apoptosis by TUNEL labeling (TUNEL). In, putative needle injection tracks; W, epidermal wound corresponding to the putative entry site of injection. ×100. Insets show loss of Ki-67–positive cells and strong TUNEL labeling along the needle injection tracks of pAd-T34A–treated tumors. Insets, ×200.