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IsoPlexis’ platform has been a critical tool for the development and evaluation of several CAR-T products and other immune therapies. Heterogeneity among CAR-T products makes functional immune profiling a key component of evaluating product quality. Many researchers have utilized this unique technology for product manufacturing applications, two cases of which are discussed here.
Single-Cell Proteomics Accelerates Therapeutic Development of CAR-T Products
IsoPlexis provides the crucial functional insights researchers need to accelerate the development of their therapeutics and optimize manufacturing. Here, we’d like to highlight research by Srivastava et al., which used IsoPlexis’ technology to assess the efficacy of two methods of CAR-T cell product manufacturing. CAR-T cell products were produced using two different methods. The function of each product was characterized using IsoPlexis’ unique functional proteomics. Method 1 (M1) concluded with a wash step at day 5, while Method 2 (M2) concluded at day 3. IsoPlexis’ functional single-cell data revealed that the CD4+ and CD8+ CAR-T cell products produced with M2 showed a consistently more robust polyfunctional response compared to those produced with M1.1
The single-cell polyfunctional data (see figure) clearly demonstrates that M2 significantly improved the overall polyfunctional responses of bispecific CD19/CD22 CAR-T cell products compared to the previously used M1. Moreover, the data revealed that CAR-T cell products manufactured by M2 secreted many more cytokines and at higher intensities than with M1. The enhanced polyfunctionality was mainly driven by effector and stimulatory cytokines. Chemoattractive (in CD4+) and regulatory (in CD8+) cytokine secretions were also enhanced in the M2 group. M2 improved the quality of CAR-T cell products in multiple dimensions.1 Through the use of IsoPlexis’ platform, researchers concluded that M2 was the superior method for optimization of CAR-T cell product quality. With IsoPlexis’ unique single-cell proteomics and polyfunctionality metric, researchers can evaluate the quality of pre-infusion CAR-T cell products and provide predictive metrics for the efficacy and persistence of novel CAR-T products.
IsoPlexis Accelerates Development of Engineered Cell Therapies by Confirming Function
The applications of IsoPlexis’ platform extend beyond just CAR-T therapies, as demonstrated by a study published in Cell Stem Cell in which IsoPlexis characterized the polyfunctionality and anti-cancer function of CRISPR-edited NK cells. To improve in vivo persistence, researchers Zhu, et al. derived NK cells from induced pluripotent stem cells (iPSCs) and used CRISPR to delete CISH, a negative regulator of IL-15 signaling in NK cells.2 Researchers then used IsoPlexis’ single-cell proteomics to evaluate the function of these CRISPR-edited cells and identify whether they were functioning as intended. The CRISPR-engineered NK cells demonstrated increased persistence in vivo and polyfunctionality compared to control NK cells in a mouse leukemia xenograft model. These polyfunctional CRISPR- edited NK cells, detected by IsoPlexis’ platform, showed enhanced anti-tumor function.2 Using IsoPlexis’ unique functional proteomics, researchers were able to characterize the functional drivers in a novel therapy.2
Whether developing novel CAR-T cell products or gene-edited cell therapies, IsoPlexis’ single-cell proteomics provides the functional immune profiling needed to evaluate a product’s function and identify highly functional cell subsets driving response. The automated IsoLight and IsoSpark proteomic hubs and IsoSpeak software can accelerate optimization of therapy products across all stages of development.
1. Srivastava S, et al. Abbreviated T-cell activation on the automated CliniMACS Prodigy device enhances bispecific CD19/22 Chimeric Antigen Receptor T-Cell viability and fold expansion, reducing total culture duration. ASH Annual Meeting 2018. Blood 2018; 132: 4551
2. Zhu H, et al. Metabolic reprogramming via deletion of CISH in human iPSC-derived NK cells promotes in vivo persistence and enhances anti-tumor activity. Cell Stem Cell 2020; 27: 1–14.
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