Potential clinical application of antioncogene ribozymes for human lung cancer

Non-small-cell lung cancer frequently contains oncogenetic defects (mutations in ras, retinoblastoma, and p53 genes) that contribute to disease pathophysiology. Recent studies and clinical trials have focused on gene therapy approaches that either replace the function of defective tumor-suppressor genes such as p53 or inactivate mutant oncogenes such as ras. Ribozymes are RNA molecules with highly specific intrinsic enzymatic activity against target RNA sequences, which can discriminate mutant sequences that differ by a single base from their wild-type counterparts. Following binding to the RNA substrate by base-pair complementation, the ribozyme cleaves the target RNA irreversibly, then releases itself for new rounds of subsequent cleavage, resulting in significantly improved target:effector stoichiometry as compared with antisense oligonucleotides of the same specificity. Transcript-specific ribozymes have been used extensively for experimental oncogene inactivation. Ribozymes are effective for targeting mutant ras, p53, or the multidrug-resistant gene product for lung cancer cells in vitro. However, their in vivo effect is not well defined against this malignancy. We recently characterized the antitumor properties of an anti-K-ras ribozyme specific for the K-ras codon 12 mutation (GGT-->GTT). When delivered as a transgene by an adenoviral vector (ADV), the K-ras ribozyme (KRbz) suppressed growth of lung tumor xenografts expressing the relevant mutation, whereas the corresponding antisense sequence lacking catalytic activity did not. Multiple intratumoral (3-5) injections of KRbz-ADV were effective in producing complete tumor regressions of preexisting tumor xenografts. Clinical trials are under consideration to examine the applicability of this anti-K-ras ribozyme for treatment of non-small-cell lung cancers expressing the relevant mutation.