Regression of high-grade cervical intraepithelial neoplasia with TG4001 targeted immunotherapy.

OBJECTIVE: We sought to evaluate the safety and efficacy of TG4001 in patients with human papillomavirus (HPV) 16-related cervical intraepithelial neoplasia (CIN) 2/3 at 6 and 12 months. STUDY DESIGN: In all, 21 patients with HPV 16-related CIN 2/3 received 3 weekly subcutaneous injections of TG4001. Regression of the CIN 2/3 lesion and the clearance of HPV 16 infection were monitored by cytology, colposcopy, and HPV DNA/messenger RNA (mRNA) detection. A clinical response was defined by no CIN 2/3 found on conization, or no conization performed because not suspected at cytology or colposcopy. RESULTS: Ten patients (48%) were evaluated as clinical responders at month 6. Nine patients experienced an improvement of their HPV 16 infection, by mRNA ± DNA eradication. HPV 16 mRNA clearance was associated with CIN 2/3 cytologic and colposcopic regression in 7 of 10 patients. At month 12, 7 of 8 patients without conization reported neither suspicion of CIN 2/3 relapse nor HPV 16 infection. The remaining patient was lost to follow-up. CONCLUSION: These promising data warrant further development of TG4001 in CIN 2/3 treatment.

Adenoviral-mediated skeletal muscle transcriptional targeting using chimeric tissue-specific promoters.

BACKGROUND: Adenoviral vectors are promising tools to achieve skeletal muscle gene transfer for the treatment of peripheral ischemia. However, the use of ubiquitous viral promoters represents a major safety issue that could limit their use. Cellular regulatory sequences that allow strong and tissue-specific expression could circumvent this problem. MATERIAL/METHODS: Adenoviral vectors encoding the firefly luciferase under the control of the human skeletal a-actin promoter, alone or combined with the b-enolase or creatine kinase enhancer, were studied in vitro in murine C2C12 cells and in vivo in C57BL/6 mice. The expression of the reporter gene was measured in cell lysates and animal tissue homogenates. Adenoviral distribution was evaluated by PCR on DNA extracted from liver, spleen, heart and lungs. RESULTS: Skeletal a-actin promoter-based expression cassettes follow the regulation of the endogenous skeletal a-actin gene in vitro as luciferase expression strongly increases with myoblast differentiation into myotubes. The addition of the cellular b enolase or the creatine kinase enhancer improves the specificity of the skeletal a-actin promoter in vitro as well as in vivo. When adenoviral vectors are locally injected into skeletal muscles, the chimeric promoters drive a relatively strong gene expression, ranging from 16 to 28% of the Rous sarcoma virus promoter-related expression. CONCLUSIONS: Chimeric regulatory sequences based on the skeletal a-actin promoter are highly specific and allow transgene expression in vivo at high levels. These results indicate that expression cassettes designed for the treatment of peripheral ischemia by gene therapy can efficiently target gene expression to skeletal muscle.

Stimulation of angiogenesis by Cyr61 gene: a new therapeutic candidate.

Cyr61 is a secreted, cysteine-rich heparin-binding protein that is associated with extracellular matrix and cell surface, and has been demonstrated to be proangiogenic in vitro. In the present study we evaluated the angiogenic effect of human Cyr61 in an adenoviral context in the rabbit ischemic hindlimb model. For this purpose, three randomized groups of New Zealand White rabbits received intramuscular injections of 5 x 10(8) infectious units of an adenovirus carrying either the Cyr61 gene (Ad-Cyr61), the vascular endothelial growth factor gene (Ad-VEGF(165)) used as the angiogenic gene of reference, or no transgene (Ad-Null), 10 days after femoral artery excision in one limb. Perfusion of the ischemic limb was evaluated before adenoviral treatment (day 10) and 30 days postinjection (day 40). Angiographic, hemodynamic, and histologic parameters indicated that animals in the Ad-Cyr61 group had significantly better perfusion than in the Ad-Null group. Interestingly, this improvement exceeded that achieved with Ad-VEGF(165). In conclusion, Cyr61 gene transfer appears potent in stimulating limb revascularization, thereby promoting great improvement in tissue perfusion in the ischemic limb. These findings indicate that Cyr61 could be a promising therapeutic candidate for treating severe peripheral ischemic diseases.