Approach

The nature of ischemic damage and reperfusion injury obligates us to design a treatment method which is effective and safe, being able to ameliorate hypoxia by eliminating the blockage of bloodstream and preventing the ROS production as well as its related pathways such as xanthine degradation in order to maintain cell health once reperfusion has been conducted.

As a result, we have designed our approach in four levels which will enable us to manipulate the entire ischemia process. To juxtapose our project design:

1. Our cells must sense hypoxic conditions immediately in order to respond in the most efficient way.

To do this, we need to present and empower the physiological hypoxia sensing systems in normal human cells. Our plan is to combine plasmid transcription accumulators and hypoxia respond elements to achieve maximum production response in gene and protein levels.

2. The clot causing hypoxia must be eliminated under strict regulations to ensure cell survival.

To carry out a successful clot solution, we need to use the most potent and common drug popular against clotting and embolism in clinic. This is to remove the clot in a short time before the destructive effect of hypoxia on cells does not increase in time. The production of anticoagulant will be regulated by hypoxia sensing modules which allow treatment to be performed only in case of severe hypoxia.

3. Cells must be prepared for a possible oxygen burst once the clot is removed.

ROS are produced by hypoxic cells as well as cells which are exposed to high oxygen concentrations. Because of this, ROS inhibiting enzymes and proteins must be present as soon as possible in the occurrence of hypoxia. This will prevent cells from being further damaged in the presence of high oxygen presence while the clot is being removed. We also aim to inhibit the further production of ROS in hypoxic cells.

4. The power of regulating system must be compensated after normalizing the oxygen level.

Clot dissolving and ROS inhibiting systems will be under strict control using hypoxia inducible modules. These modules are expected to be shut after the clot is removed and when oxygen is present in the media. Therefore, we need to restore this regulation to keep enough production of preventive enzymes in order to end possible oxidative stress. We are aiming to do this by sensing ROS particles in the transcription level and ensuring that they no longer exist within the body environment.