Recombinant Newcastle Disease Virus Encoding IL-12 and/or IL-2 as Potential Candidate for Hepatoma Carcinoma Therapy

Abstract

Interleukins as immunomodulators are promising therapeutic agents for cancer therapy. Previous studies showed that there was an improved antitumor immunity in tumor-bearing mice using recombinant Newcastle disease virus carrying for interleukin-2. Interleukin-12 is a promising antitumor cytokine too. So we investigated and compared the antitumor effect of genetically engineered Newcastle disease virus strains expressing both interleukin-12 and/or interleukin-2 (rClone30-interleukin-2, rClone30-interleukin-12, and rClone30-interleukin-12-interleukin-2). In vitro studies showed that rClone30s could efficiently infect tumor cells and express interleukin-12 and/or interleukin-2. 3-(4,5-Dimethylthiazol-2-y)-2,5-diphenyl-tetrazolium bromide results showed rClone30s possessed strong cytotoxic activities against multiple tumor cell lines (U251, HepG2, A549, and Hela). Animal studies showed that rClone30-interleukin-12-interleukin-2 was more effective in inhibition of murine hepatoma carcinoma tumors, with the mean tumor volume (day 14) of 141.70 mm 3 comparing 165.67 mm 3 of rClone30-interleukin-12 group, 210.47 mm 3 of rClone30-interleukin-2 group, 574.70 mm 3 of rClone30 group, and 1206.83 mm 3 of phosphate-buffered saline group. Moreover, the rClone30-interleukin-12-interleukin-2 treated mice secreted more interferon g (333.518 pg/mL) and its downstream cytokine interferon-g induced protein 10 (16.006 pg/mL) in tumor than the rClone30-interleukin-12 group (interferon g: 257.548 pg/mL; interferon-g induced protein 10: 13.601 pg/mL), rClone30-interleukin2 group (interferon g: 124.601 pg/mL; interferon-g induced protein 10: 9.779 pg/mL), or rClone30 group (interferon g: 48.630 pg/mL; interferon-g induced protein 10:1.650 pg/mL). For the survival study, rClone30-interleukin12-interleukin2 increased the survival rate (12 of 16) of the tumor-bearing mice versus 11 of 16 in rClone30-interleukin-12 group, 10 of 16 in rClone30-interleukin-2 group, 7 of 16 in Clone30 group, and 0/16 in phosphate-buffered saline group, respectively. To determine whether the mice treated with recombinant virus developed protective immune response, the mice were rechallenged with the same tumor cells. The results showed that viral-treated mice were significantly protected from rechallenge. These results suggest that expressing both interleukin-2 and/or interleukin-12 could be ideal approaches to enhance the antitumor ability of Newcastle disease virus, and rClone30-interleukin-12-interleukin-2 is slightly superior over rClone30-interleukin-12 and rClone30-interleukin-2 alone.

Oncolytic Virus Encoding a Master Pro-Inflammatory Cytokine Interleukin 12 in Cancer Immunotherapy

“…Several other OVs encoding IL-12 have been developed ( Table 2), including adenoviruses [18,42,[46][47][48][49][133][134][135][136][137][138], measles virus [20,50], maraba virus [51], Newcastle disease virus [52], Semliki forest virus [53][54][55][56][139][140][141], vesicular stomatitis virus [57,142], and Sindbis virus [58]. In these above-mentioned studies, IL-12 is expressed either alone [18,20,51,57,[137][138][139] or co-expressed alongside with GM-CSF [134][135][136], pericellular matrix proteoglycan decorin [47], tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) [133], IL-2 proinflammatory cytokine [52], IFN-γ inducing factor IL-18 [46], T cell co-stimulatory ligand 4-1BBL [49], CD8 + co-receptor for CD28 and CTLA-4 [48], or suicide genes [42]. The anti-tumor efficacy of these engineered OVs are...…”

“…In these above-mentioned studies, IL-12 is expressed either alone [18,20,51,57,[137][138][139] or co-expressed alongside with GM-CSF [134][135][136], pericellular matrix proteoglycan decorin [47], tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) [133], IL-2 proinflammatory cytokine [52], IFN-γ inducing factor IL-18 [46], T cell co-stimulatory ligand 4-1BBL [49], CD8 + co-receptor for CD28 and CTLA-4 [48], or suicide genes [42]. The anti-tumor efficacy of these engineered OVs are tested in various mouse or hamster pre-clinical cancer models and produce superior anti-tumor immunity either alone [18,20,42,[46][47][48]51,52,57,133,135,137] or in combination with other immunotherapeutic agents such as dendritic cell (DC) vaccine [49,134,136], ICI anti-PD-1 and anti-PD-L1 [139], or cytokine-induced killer cells [138].…”

“…Similar to the MeVac FmIL-12 virus, IL12-expressing oncolytic maraba virus (designated MG1-IL12-ICV) treatment also leads to complete eradication of peritoneal carcinomatosis, which is associated with significant recruitment of NK cells in the tumor microenvironment [51]. The lentogenic Newcastle disease virus Clone30 strain was generated to express IL-12 (designated rClone30-IL-12) that displays improved survival and reduced tumor volume [52]. In this case, co-expression of two cytokines (IL-12 and IL-2) produced synergistic effects on treatment, with the rClone30-IL-12-IL-2 virus inducing the greatest release of IFN-γ and IP-10 [52].…”

“…The lentogenic Newcastle disease virus Clone30 strain was generated to express IL-12 (designated rClone30-IL-12) that displays improved survival and reduced tumor volume [52]. In this case, co-expression of two cytokines (IL-12 and IL-2) produced synergistic effects on treatment, with the rClone30-IL-12-IL-2 virus inducing the greatest release of IFN-γ and IP-10 [52].…”

Reverse Genetics of Newcastle Disease Virus

“…NDV reverse genetics has made possible the development of a valuable recombinant vaccine system, enabling expression of its own mutated proteins or foreign proteins, thus opening opportunities to investigate its applications as recombinant vaccines, as a multivalent vaccine candidate for poultry, and as a vaccine vector for other animal species and humans [14,[17][18][19][20][21][22][23][24][25][26][27][28][29][30]. NDVs modified by reverse genetics have also become valuable candidates for anticancer therapy in humans [31][32][33][34][35].…”