Improving Preclinical Discovery of CRISPR Engineered Immune Cell Therapies

There is an urgent need to characterize the potency and efficacy of CRISPR-Cas9-modified inducible pluripotent stem cell-derived natural killer cells for preclinical cancer immunotherapy research. IsoPlexis' single-cell proteomics system addresses this challenge by connecting each immune cell to cytokine secretion and thereby correlating them to in vivo outcome across a range of disease areas.

Mar 6, 2019
IsoPlexis

Natural killer (NK) cells, known for their ability to kill tumor cells, are promising agents for cell-based cancer immunotherapies. CRISPR-Cas9 gene editing can be used to effectively modify the genetic makeup of inducible pluripotent stem cell (iPSC)-derived NK cells towards this end, but there is an urgent need to characterize their potency and efficacy as a preclinical cancer immunotherapy. IsoPlexis' single-cell proteomics system addresses this challenge by connecting each immune cell to the many cytokines it secretes, revealing correlations to in vivo outcome across a range of disease areas.

The Polyfunctional Strength Index (PSI) delivers correlative potency data

The IsoPlexis system identifies which cells are polyfunctional (i.e., those powerful cells that secrete multiple cytokines) and quantitates the cytokine concentrations within each cell. PSI combines these two single-cell metrics to effectively identify highly potent immunotherapies.

PSI reveals cell potency of gene-edited, iPSC-derived NK cells

To study the role of cytokine-inducible SH2-containing protein (CIS) in human NK cell anti-tumor activity, IsoPlexis’s systems analyzed the potency of iPSC-derived NK cells with the CISH gene deleted using CRISPR-Cas9 technology. A 10X higher PSI of the gene-edited NK cells was found relative to control wildtype NK cells, driven by increased antitumor cytokine production (e.g., Granzyme B, IFN-γ, MIP-1α, Perforin, TNF-α). Researcher Dr. Dan Kaufman, PI of this study, said, “Our studies have used CRISPR-Cas9 gene editing in human induced pluripotent stem cells to produce natural killer cells with improved antitumor activity. Analysis of these cells using the IsoPlexis system was very valuable to characterize the improved polyfunctional cytokine response that plays a key role in improved activity of these iPSC-derived NK cells.”

Increase in PSI of gene-edited NK cells correlates with improved in vivo response of the NK cell therapy

To test CISH-/- human NK cells in vivo, immunodeficient NSG mice were inoculated with the human leukemia cell line, MOLM-13, and split into three groups: untreated mice, mice treated with control wildtype NK cells, and mice treated with the CISH-knockout NK cells. The study revealed substantial tumor growth in untreated mice and wildtype NK treated mice, while tumor growth was nearly absent in mice receiving the CISH-KO NK treatment. This strongly suggests both a correlation between in vitro PSI and in vivo mouse antitumor activity, as well as therapeutic efficacy.

Conclusion

IsoPlexis’ single-cell system improves engineered immune-cell therapy R&D, by providing metrics such as PSI, indicative of improved potency of gene-edited cells, and potentially correlating to in vivo efficacy of engineered cell therapies.

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This article is brought to you by IsoPlexis. IsoPlexis is dedicated to accelerating the fight against cancer and a range of our toughest diseases by producing the world’s most precise, award-winning detection systems. We succeed by merging top engineering with top researchers and clinicians to generate clear solutions to complex patient response. https://isoplexis.com/