Posted: July 9th, 2022

Flow Cytometry Methods and Applications

Flow Cytometry Methods and Applications
Please read the article “Multi Targeted CAR-T Cell therapies for B-Cell Malignancies” and discuss the follow topics in your own words:

1. How do those therapies achieve multiple targets?

2. What are the benefits of having multiple targets?

Flow Cytometry Methods and Applications
1. How do those therapies achieve multiple targets?
Multiple targets in the CERT-T therapies involve novel strategies to overcome the resistance that is considered to be a limitation in the current single targeted CERT-T therapies. The multi-targeted CERT-T cell therapy is used to combat the antigen loss problem following CERT-T cell therapy by targeting more than one antigen receptor at the same time. In the multi-targeted CERT-T cells, therapies archive multiple targets using three mechanisms. The first mechanism applied by therapies is the coadministration mechanism, which involves the generation of two or more cell populations that express different chimeric antigen receptors (CARs) and infusing them sequentially or together (Shah, Maatman, Hari, & Johnson, 2019). The second approach used by therapies to achieve multiple targets is the bicistronic mechanism. The bicistronic approach involves encoding two different chimeric antigen receptors on the same cell using a bicistronic vector. The third mechanism for achieving multiple targets by therapies is through the cotransduction. The mechanism involves T cells being simultaneously engineered with two different chimeric antigen receptors resulting in the generation of three CAR-T subsets that are comprised of dual and single CAR-expressing cells. The last mechanism of achieving multiple targets is by the use of the tandem approach, which involves the use of a single vector such as bi-specific and tandem CARs to encode two CARs on the same chimeric protein.
2. What are the benefits of having multiple targets?
The benefit of using more than one CAR or multiple targets is helping in the prevention of antigen escape and providing a better anti-tumor efficiency. The other benefit provided by studies regarding the use of multiple targets is the, specifically, the tandem CAR help in preventing the spontaneous development of the CD19-negative tumor cell variants. The multiple targets approach was also found to improve the killing of some cancer cells by binding the CD20 peptide (Shah et al., 2019). The multiple targets therapy has also been associated with the ability to overcome resistance mechanisms associated with single targeting, which involves selective pressure and tumor cell clones development that can evade CD19 CAR-T therapy. In general, multiple targets are considered beneficial in improving clinical outcomes.
References
Shah, N. N., Maatman, T., Hari, P., & Johnson, B. (2019). Multi Targeted CAR-T Cell Therapies for B-Cell Malignancies. Frontiers in oncology, 9, 146. https://doi.org/10.3389/fonc.2019.00146

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