It is hypothesised that genetically engineered KM110r will target and selectively destroy all U2OS cells. Specifically, living U2OS cell counts will decrease steadily post KM11or infection, and all will be completely infected and dead by the fourth and final time period of each experimental trial. KM110r will not be toxic to hFOB cells being grown at 34°C and 37°C incubation conditions. These undifferentiated hFOB cells will continue to proliferate up to confluence at constant rates with no infection being identified. KM110r will be an effective and safe potential cancer treatment.

Under 39°C incubatory conditions, hFOB cells will begin to differentiate, as reported[1]. It is hypothesized that the point during in vitro differentiation at which the hFOB cells respond to viral infection will be identified, and the potential underlying changes will be investigated. Given that hFOB cells proliferate as if immortalized at 34°C but differentiate at 39°C, the hFOB cells will commence their response to viral infection during the 39°C incubation trial. The identification of such a point will enhance understanding of which genetic changes permit oncolytic virus infection with the intention of identifying the point along the differentiation pathway to maximize treatment efficacy.

It is hypothesized that at 34°C and 37°C infected co-cultures (MIX+) consisting of 50% hFOB and 50% U2OS will grow at steady, even rates post-infection since the death of the U2OS cells will be balanced by the proliferation of hFOB cells not responding to viral infection.  

Ultimately, it is hypothesized that:

·        KM110r will be an effective and safe oncolytic virus for cancer treatment

·        KM110r will target and selectively destroy all U2OS cells

·        U2OS cell counts will decrease steadily post KM110r infection

·        All U2OS cells will be completely infected and dead by the fourth and final time period of each experimental trial

·        KM110r will not be toxic to hFOB cells being grown at 34°C and 37°C incubation conditions. These undifferentiated hFOB cells will continue to proliferate up to confluence at constant rates with no infection being identified.

·        Under 39°C incubation conditions, as hFOB cells experience normal genetic changes, KM110r will recognize global genetic changes and the hFOB cells will respond to viral infection. The point (during in vitro differentiation) at which this occurs will be identified and the potential underlying changes will be investigated.