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Future Procedure and Potential Applications

April 12, 2025

While the results of this research have not yet been ascertained, there are some potential applications, and a lot of future procedure.

Future Directions

Future procedure would include waiting for mostly successful IL-22 knock-outs. A significant amount of successful knock-out would have about a (???) fluorescence. After achieving a batch of mostly modified cells, one would put them through the cell sorter to separate out the green cells. The cell sorter would separate the cells and not kill them, unlike the flow cytometer. Afterwards, the modified cells would be placed in well plates and grown into cell lines; eventually, the cell lines would be homogenous, and all the 4T1 cells would be modified. The cells would then be reinserted into balb/c mice into the mammary glands to imitate human breast cancer. The tumors would be allowed to grow and metastasize. The population of MDSCs around the tumor and in the bloodstream would be studied and analyzed. Simultaneously, the spread of the modified cells would be tracked, which was the purpose of the TdTomato addition. Periodically, the mouse cells would be removed and the knock-out would be re-verified. If the population of MDSCs went down after the IL-22 knockout, the hypothesis that cancer cells use IL-22 to communicate with MDSCs would be proven. Significant data would have to be gathered.

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Future Applications

If IL-22 did reduce the concentration of MDSCs, there would be many potential applications. The most relevant would be in cancer treatment, specifically immunotherapy. Immunotherapy is designed to improve upon and strengthen the existing immune system: one of the main setbacks in developing effective immunotherapy is the difficulty with MDSCs suppressing it. Potentially, a drug limiting IL-22 production could be tested and produced. It would greatly improve the efficacy of immunotherapy and other cancer treatments. It would not be a treatment in and of itself but has the potential to support and amplify the effects of treatment. On the other hand, using CRISPR as a cancer treatment would be unreasonable. It was effective for study the effect of the IL-22 gene, but it would be impossible, not to mention unethical to modify human cells and reinsert them to knock-out IL-22.

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