Confirming Function of Gene Edits in Allogeneic “Off-the-Shelf” Cell Therapies
CAR-T therapies are currently among the most popular immunotherapies, with the top active industry sponsors holding a combined total of 32 CAR-T cell therapies in development and about 70 ongoing trials. Additionally, many companies are developing allogeneic therapies. While autologous therapies created specifically for one patient have been effective, challenges remain. For example, not all patients can provide quality source material for autologous therapies. Allogeneic therapies are created using T cells from healthy donors, providing cells to be expanded for product development. However, the possibility exists for graft versus host disease (GVHD) mediated by immune response within the patient.
Allogeneic therapies are gaining in popularity due to their potential for mass production and use as “off the shelf” therapies. To create these therapies, many companies are employing various gene-editing techniques, although studies have shown that the correlation between genomic and transcriptomic sequencing and direct cytokine secretion remains weak, highlighting the importance of the functional characterization of the engineered cells. Given that function is key in cell therapies for determining outcome in the patient post-treatment, the ability to measure the direct function of individual immune cells is critical. IsoPlexis’ ability to identify functional heterogeneity within cell samples provides immense benefits, including aiding in overcoming the challenges associated with scaling. There is no guarantee that cellular performance will be uniform even in healthy donors, and IsoPlexis’ platform can funtionally characterize these products. The number of CAR-T therapies in development is sure to grow as more companies develop breakthrough therapies. An example of one such company is Precision Biosciences, which recently IPO’d in March of 2019. The Scientist recently spoke to Vladimir Senyukov, the Director of BioAnalytical Development at Precision Biosciences, about his work with allogeneic gene-edited CAR-T cells. Read on for highlights from this interview.
Functionally Profiling Gene-Edited and Engineered Cells for Quality and Potency
Aside from toxicity and rejection, engineered and gene-edited cells could function differently than intended and expected after initial modification. Directly measuring single-cell functionality can address this. Senyukov spoke about his current project, which “involves using IsoPlexis to profile the cytokine signatures of polyfunctional cells that we might have in a cell population.” The IsoPlexis platform can detect, within a phenotypically identical sample, functionally heterogeneous and potent cells, which are missed by flow cytometry and bulk ELISA. By using IsoPlexis’ single-cell cytokine detection system, which can analyze the true function of each cell, researchers can better understand how their potential immunotherapies will affect patients.
“To test CAR-T cells and figure out if they function properly you usually test them against their targets,” explained Senyukov. IsoPlexis’ single-cell functional proteomics is also important here, he continues. “When optimizing, or when changing manufacturing processes to make better cells, it can be used to monitor cellular changes. It is especially valuable in screening for polyfunctionality, which is key, because based on the literature, polyfunctionality is one of the most important factors driving clinical efficiency.”
Polyfunctional cells and cell subsets (cells secreting two or more cytokines simultaneously) have correlated to response in multiple studies. With a growing number of studies in allogeneic therapies, IsoPlexis’ platform can potentially help improve these types of therapies by detecting the functionally heterogeneous and potent cells that other technologies miss, meaning more accurate functional information leading to more targeted and effective therapies.