The Transformative Power of Advanced Flow Cytometry in Cell and Gene Therapy

Cell and gene therapy is one of the most promising areas of biomedical research with numerous breakthrough treatments targeting cancer and genetic disorders. Despite rapid growth, companies face high costs and technical difficulties when it comes to phenotyping and profiling cell products. In this post, we will focus on the role of flow cytometry in chimeric antigen receptor (CAR) T cell development and how advanced systems can transform workflows and help bring novel therapies to bedside.

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What are cell and gene therapies?

Cell and gene therapies are used to treat, and in some cases cure, diseases by addressing the underlying genes or proteins. While there is overlap between the fields, they are different. 

Cell therapies involve living cells that are either derived from the patient (autologous), or a donor (allogenic). For example, stem cell therapies are used to treat diseases of the bone marrow and blood. Gene therapies introduce genetic code into cells in order to restore the function of a defective or missing gene. Glybera was the first gene therapy approved for use in Europe and USA for the treatment of a rare genetic disorder called lipoprotein lipase deficiency.
   

What is CAR-T cell therapy?

CAR-T cells are a hot area of biomedical research at the intersection of cell and gene therapy. CAR-T cells are gene-modified cell therapies that are designed to interact with a specific surface protein on the tumor cell, triggering cell death. This targeted approach to cancer treatment is preferred to traditional chemotherapies and has worked remarkably well in treating blood cancers like leukemia and lymphoma. 

   
Development of CAR-T cells

The CAR-T cell development process starts with isolated T cells from a patient or healthy donor. The cells are then reprogrammed to express the CAR construct, which recognizes the target antigen on the tumor cell. CAR-T cells are then expanded before being administered back to the patient. Like any cell and gene therapy product, CAR-T cells require phenotypic and functional characterization at every stage to ensure safety and efficacy.  
   

Role of flow cytometry 

Flow cytometry is used in every stage of CAR-T cell development. Early on, flow cytometry can be used to identify specific populations of the donor T cells that are associated with a better prognosis for CAR-T cell therapy. The cells are then continuously characterized for quality, safety, and function until transfer to patient, and through follow-up monitoring. Below are all the ways flow cytometry analysis is used in CAR-T cell development and manufacturing:

• Identify and quantify cell composition in the donor sample 
• Assess cell viability and functionality
• Evaluate CAR expression on the therapeutic cells
• Estimate effector cell activation and cytotoxic killing
• Quantify tumor expression of antigen and immunosuppressive molecules
• Monitor cell persistence and therapeutic effect in the patient

   
Limitations of traditional flow cytometry

Phenotyping, purifying, and expanding immune cells from patients is both expensive and technically difficult. Traditional flow cytometers used to analyze cell and secreted proteins are low throughput and can’t support pharma-level data generation. There is also a need for connecting insights from both immune cell health, phenotype and proliferation to cytokine secretion profiles, from the same sample. Perhaps the most common pain points to traditional flow are the disjointed data analysis workflows that slow down progress.  
   

Advanced flow cytometry simplifies workflows

To overcome these issues, many scientists have turned to advanced flow cytometry. Simply put, advanced flow cytometry systems give you throughput, speed and more data per assay well. With the iQue^® Advanced Flow Cytometry Platform, for example, you can get immunophenotyping data with measurements of cell health, and bead-based cytokine secretion, in a single experiment; this saves sample and avoids the need for multiple assays. Advanced flow technologies also integrate powerful software for instant access to real-time data, better visualization tools and dynamic gating capabilities, so it’s easy to interpret complex datasets.  
   

Conclusion

Advanced flow cytometry adds tremendous value at multiple stages in the development and expansion of cell and gene therapy products, like CAR-T cells. By combining multiplexed analysis in a single well, modern labs working on the next breakthrough immunotherapy can simultaneously learn about immune cell phenotype and function in a single well.

Read more about the iQue^® Advanced Flow Cytometry Platform and compatible kits for T-cell activation, T-cell killing, T-cell exhaustion and T-cell memory.