Gene therapy using tissue-specific replication competent HSV.

Abstract

Tissue- or cell-specific targeting of vectors is critical to the success of gene therapy. I describe a novel approach to viral-mediated gene therapy, where viral replication and associated cytotoxicity are limited to a specific cell-type by the regulated expression of an essential immediate-early viral gene product. This is illustrated with two herpes simplex virus type 1 vectors (G92A and d12.CALP) whose growth are restricted to albumin- or calponin-expressing cells, respectively. G92A was constructed by inserting an albumin enhancer/promoter--ICP4 transgene into the thymidine kinase gene of mutant herpes simplex virus type 1 d120, deleted for both copies of the ICP4 gene. This vector also contains the Escherichia coli lacZ gene under control of the thymidine kinase promoter, a viral early promoter, to permit easy detection of infected cells containing replicating vector. In the adult, albumin is expressed uniquely in the liver and in hepatocellular carcinoma and is transcriptionally regulated. G92A efficiently replicated in vitro in two human hepatoma cell lines expressing albumin, but not in three human non-hepatoma, albumin-non-expressing tumor cell lines, while all cell lines were equally susceptible to a tissue non-specific HSV recombinant, hrR3. In vivo, G92A replicated well in subcutaneous xenografts of human hepatoma cells (Hep3B) in athymic mice, but not in non-hepatoma subcutaneous tumors (PC3 and HeLa), whereas, hrR3 replicated well in both tumor types. Intratumoral inoculation of G92A inhibited the growth of established subcutaneous hepatoma tumors in nude mice, but not prostate tumors. D12CALP also revealed the cell-specific replication to leiomyosarcoma in which calponin expression was augmented. Using hrR3, we demonstrated inhibition of re-stenosis of rat carotid arteries caused by balloon injury. The antiproliferative effects of this virus was marked in the proliferating smooth muscle cells, however, there still remained the fear for the injury of the endothelial cells. Confining a productive, cytotoxic viral infection to a specific cell-type should be useful for tumor therapy and the ablation of specific cell-types for the generation of animal models of disease. Further experiments using d12CALP will be focused on the arteriosclerosis due to balloon angioplasty or organ transplantation.