Targeted gene delivery for induced apoptosis of transitional cell carcinomas was carried out in vivo in mice via utilization of the murine cyclooxygenase type 2 (Cox-2) promoter (Tis10). MB49 cells, which constitutively overexpress Cox-2 like numerous other carcinomas, selectively expressed delivered genes that utilized this transcriptional control element. The products of the delivered genes were artificially inducible forms of caspases 3 and 9, which remained inactive until a chemical inducer of dimerization was later injected intraperitoneally. The genes were delivered intravesically as plasmids complexed with poly(ethylenimine). Significant improvements, in the form of reduced bladder mass, reduced tumor volume, anti-angiogenesis and inhibition of tumor growth were seen versus untreated or unactivated controls. In some instances, tumors were seen to go into complete remission. There were no apparent bystander effects associated with the treatments. This targeted gene therapy regimen could have wide applicability to numerous cancers due to constitutive overexpression of Cox-2.

Three drugs were compared for their efficacy in treating murine transitional cell carcinoma (TCC) of the bladder. Intravesical gene therapy treatments utilizing expression-targeted plasmids, where the murine cyclooxygenase-2 (Cox-2) promoter was used to drive the expression of exogenously inducible forms of caspases 3 and 9, were compared with treatment modalities employing Bacille Calmette-Guérin (BCG) and celecoxib. When administered via lavage, only the gene therapy regimen was found to be effective at restricting tumor progression following a 7-day incubation of tumor tissues. Celecoxib was also administered via the diet to allow for systemic delivery of the drug. The most efficacious celecoxib use tested yielded tumors with masses of (18.3 ± 8.4 mg) versus the gene delivery method, which yielded tumors with masses of (3.6±7.7 mg). The difference was significant (t-test, nX4, Po0.025). The results showed that the Cox-2 expression-targeted gene therapy system could efficiently bypass the bladder permeability barrier and more effectively inhibit tumor growth and development than either BCG or celecoxib treatments. Long-term data further demonstrated that the gene therapy system could effectively inhibit tumor growth and elongate life expectancy.

Concern about rising rates of obesity has prompted searches for its genetic risk determinants in genome-wide association studies (GWAS). Most genetic variants that contribute to the increased risk of a given trait are probably regulatory single nucleotide polymorphisms (SNPs). However, identifying plausible regulatory SNPs is difficult because of their varied locations relative to their target gene and linkage disequilibrium, which makes most GWAS-derived SNPs only proxies for many fewer functional SNPs. We developed a systematic approach to prioritizing GWAS-derived obesity SNPs using detailed epigenomic and transcriptomic analysis in adipose tissue vs. heterologous tissues. From 50 obesity-related GWAS and 121,064 expanded SNPs, we prioritized 47 potential causal regulatory SNPs (Tier-1 SNPs) for 14 gene loci. A detailed examination of seven of these genes revealed that four (CABLES1, PC, PEMT, and FAM13A) had Tier-1 SNPs that might regulate alternative use of transcription start sites resulting in different polypeptides being generated or different amounts of an intronic microRNA gene being expressed. HOXA11 and long noncoding RNA gene RP11-392O17.1 had Tier-1 SNPs in their 3’ or promoter region, respectively, and strong preferences for expression in subcutaneous vs. visceral adipose tissue. ZBED3-AS1 had two intragenic Tier-1 SNPs, each of which might contribute to mediating obesity risk through modulating long-distance chromatin interactions. We conclude that prioritization of regulatory SNP candidates should focus on their surrounding epigenetic features in a trait-relevant tissue. Our approach not only revealed especially credible novel regulatory SNPs, but also helped evaluate previously highlighted obesity GWAS SNPs that were candidates for transcription regulation.

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