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DNA Therapeutics Reports a Novel Mechanism of Action and a,Biomarker of its Lead Molecule Dbait That Open an Innovative,Pathway-Targeting in Molecular Cancer Therapy

zation at chromatin level.

It is known that the kinases ATM/ATR and DNA-PK are involved in the general cellular response to DNA damage by signaling proteins that control cell cycle, apoptosis and DNA repair. In particular, ATM and DNA-PK both phosphorylate the H2AX, an isoform of the histone H2A, after exposure to ionizing radiation. The appearance of the phosphorylated H2AX, called gamma-H2AX, in cell nuclei is an indicator of DSB repair foci.

Our data provides evidence that Dbait specifically activates DNA-PK but not ATM kinase activity, as the phosphorylation of H2AX is in a DNA-PK dependent manner. Dbait-induced H2AX phosphorylation persists more than 6 days as compared to 1 day for irradiation. Cell imaging shows that the global distribution of gamma-H2AX in the nucleus of the cells transfected by Dbait was radically different of the gamma-H2AX foci organization observed when H2AX phosphorylation was triggered by irradiation alone. This indicates that DSB repair system is dramatically disorganized in Dbait transfected cells, and points out a new mechanism of action.

In addition, the amount of gamma-H2AX is proportional to that of Dbait in cultured cells and in the cells from tumor samples. gamma-H2AX appears to be a specific biomarker of Dbait in cells. It can be easily used to monitor the bioactivity of Dbait molecules in further preclinical and clinical development.

M. Dutreix who conducted this study said "this additional mechanism of antitumoral action of Dbait molecules confirms that the use of Dbait which baits, hijacks the enzyme complexes that repair DSBs and disorganizes DSB repair system, is a novel and attractive pathway-targeting approach in molecular cancer therapy".

J.S. Sun, CEO of DNA Therapeutics comments "The elucidation of Dbait's mechanisms of action and the identification of gamma-H2AX as a specific biomarker of Dbait activity are important milestones of DNA Therapeutics' development". He adds "DNA Therape
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