Evaluation of a Recombinant Newcastle Disease Virus Expressing Human IL12 against Human Breast Cancer
Zahiah Mohamed Amin1, Muhamad Alhapis Che Ani2, Sheau Wei Tan1
1Institute of Bioscience, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia.
Abstract:
The Newcastle disease virus (NDV) strain AF2240 is an avian avulavirus that has been demonstrated to possess oncolytic activity against cancer cells. However, to illicit a greater anti-cancer immune response, it is believed that the incorporation of immunostimulatory genes such as IL12 into a recombinant NDV backbone will enhance its oncolytic effect. In this study, a newly developed recombinant NDV that expresses IL12 (rAF-IL12) was tested for its safety, stability and cytotoxicity. The stability of rAF-IL12 was maintained when passaged in specific pathogen free (SPF) chicken eggs from passage 1 to passage 10; with an HA titer of 29. Based on the results obtained from the MTT cytotoxic assay, rAF-IL12 was determined to be safe as it only induced cytotoxic effects against normal chicken cell lines and human breast cancer cells while sparing normal cells. Significant tumor growth inhibition (52%) was observed in the rAF-IL12-treated mice. The in vivo safety profile of rAF-IL12 was confirmed through histological observation and viral load titer assay. The concentration and presence of the expressed IL12 was quantified and verified via ELISA assay. In summary, rAF-IL12 was proven to be safe, selectively replicating in chicken and cancer cells and was able to maintain its stability throughout several passages; thus enhancing its potential as an anti-breast cancer vaccine.
Insights
Newcastle disease virus (NDV) engineered to express IL12 (rAF-IL12) shows promise as an oncolytic agent. This recombinant NDV is safe, stable, and effectively inhibits tumor growth, enhancing its potential as an anti-breast cancer vaccine.
Area of Science:
- Oncolytic virotherapy
- Immunotherapy
- Avian medicine
Background:
- Newcastle disease virus (NDV) strain AF2240 exhibits oncolytic properties.
- Enhancing the anti-cancer immune response may improve NDV's oncolytic efficacy.
- Incorporating immunostimulatory genes like IL12 into NDV is a potential strategy.
Purpose of the Study:
- To develop and evaluate a recombinant NDV expressing IL12 (rAF-IL12).
- To assess the safety, stability, and cytotoxicity of rAF-IL12.
- To determine the therapeutic potential of rAF-IL12 against breast cancer.
Main Methods:
- rAF-IL12 was passaged in specific pathogen-free (SPF) chicken eggs.
- Cytotoxicity was assessed using MTT assays on cell lines.
- In vivo efficacy was evaluated in mice, with safety confirmed by histology and viral load assays.
- IL12 expression was quantified using ELISA.
Main Results:
- rAF-IL12 remained stable through 10 passages with an HA titer of 2^9.
- rAF-IL12 demonstrated selective cytotoxicity against cancer cells while sparing normal cells.
- Significant tumor growth inhibition (52%) was observed in rAF-IL12 treated mice.
- In vivo safety was confirmed, and IL12 expression was verified.
Conclusions:
- rAF-IL12 is a safe and stable oncolytic virus.
- The recombinant virus selectively replicates in cancer cells.
- rAF-IL12 demonstrates significant anti-tumor activity and holds potential as an anti-breast cancer vaccine.
