Reengineering Tumor Microenvironment with Sequential Interleukin Delivery

Abstract

Some cytokines can reengineer anti-tumor immunity to modify the tumor micro-environment. Interleukin-27 (IL-27) can partially reduce tumor growth in several animal models, including prostate cancer. We hypothesized that addition of IL-18, which can induce the proliferation of several immune effector cells through inducing IFNγ could synergize with IL-27 to enhance tumor growth control. We describe our findings on the effects of IL-27 gene delivery on prostate cancer cells and how sequential therapy with IL-18 enhanced the efficacy of IL-27. The combination of IL-27 followed by IL-18 (27→18) successfully reduced cancer cell viability, with significant effects in cell culture and in an immunocompetent mouse model. We also examined a novel chimeric cytokine, comprising an IL-27 targeted at the C-terminus with a short peptide, LSLITRL (27pepL). This novel cytokine targets a receptor upregulated in tumor cells (IL-6Rα) via the pepL ligand. Interestingly, when we compared the 27→18 combination with the single 27pepL therapy, we observed a similar efficacy for both. This efficacy was further enhanced when 27pepL was sequenced with IL-18 (27pepL→18). The observed reduction in tumor growth and significantly enriched canonical pathways and upstream regulators, as well as specific immune effector signatures (as determined by bioinformatics analyses in the tumor microenvironment) supported the therapeutic design, whereby IL-27 or 27pepL can be more effective when delivered with IL-18. This cytokine sequencing approach allows flexible incorporation of both gene delivery and recombinant cytokines as tools to augment IL-27’s bioactivity and reengineer efficacy against prostate tumors and may prove applicable in other therapeutic settings.

Integrative Characterization of the Role of IL27 In Melanoma Using Bioinformatics Analysis

“…As expected, upregulated genes in response to IL27 treatment were positively co-expressed with IL27 in both human prostate cancer (GSE32448; FDR = 0.014, NES = 1.47; Figure 4D) and human melanoma cancer (TCGA-SKCM; FDR = 0.000, NES = 3.33; Figure 4E), further supporting IL27 as a driver gene. Moreover, tumor volume for the control group was observed to be significantly larger than that in IL27 overexpressing group in a previously reported study (23). Altogether, these combined analyses suggest that IL27 acts as a driver gene and has an anti-tumor effect.…”

“…Although we have observed that IL27 was linked to enhanced immune response as described above, it remained unclear whether IL27 was a driver of the immune response or simply a passenger. To further validate the effects of IL27 on the biological function of tumors in vivo , we searched and found an ideal dataset (GSE178142) ( 23 ), including RNA-seq data from control mice (GSM5380810 and GSM5380811) and IL27 overexpressing mice treated intramuscularly with plasmids containing IL27 (GSM5380806 and GSM5380807). We downloaded RNA-seq data for control mice and experimental mice, and converted the ensemble ID to gene symbol.…”

“…RNA-seq profiles, including RNA-seq data from control mice and IL27 overexpressing mice treated intramuscularly with plasmids containing IL27 , were obtained from GSE178142 ( 23 ), which was utilized to investigate the effect of overexpressed IL27 on the biological behavior of tumors in vivo .…”

A Second-Generation Nanoluc-IL27 Fusion Cytokine for Targeted-Gene-Therapy Applications

“…We have also shown a role of IL-27 in impacting bone-cell differentiation and proliferation [ 6 ]. We previously examined a first-generation IL-27 targeted at the C-terminus with a short ‘peptide L’ (pepL, LSLITRL), which binds the interleukin 6 receptor α (IL-6Rα) that is upregulated in tumor cells [ 7 ] in order to reduce prostate tumor growth (IL-27pepL or 27pepL) [ 4 , 8 , 9 ].…”

“…IL-27 can also serve as an effective bone-normalization agent due to its impact on pro-osteogenic-gene changes in both osteoblasts and osteoclasts [ 6 ]. Additionally, IL-27 exhibits immunomodulatory activity capable of promoting the accumulation of tumor-clearing effector cells at the site of prostate-cancer bone metastases [ 4 , 9 ].…”