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From preclinical mouse models through clinical trials, IsoPlexis’ low-volume, highly multiplexed single-cell proteomics technologies with murine, nonhuman primate, and human cytokine panels reveal deep functional insights.

IsoPlexis’ single-cell proteomics technologies are applicable from preclinical R&D in mouse models through patient characterization and stratification during clinical trials as a standard tool for leading institutions worldwide.

IsoPlexis addresses the challenges of limited volumes of precious samples in small animal studies with low-volume, highly multiplexed bulk and single-cell proteomics. Murine, nonhuman primate, and human panels are available on IsoCode & CodePlex chips, enabling immune profiling for a full range of cytokines and phosphoproteins in different model systems. IsoPlexis’ technology dives deep to provide functional data previously inaccessible by driving the convergence of dynamic proteomics and single-cell biology for the first time to enable highly multiplexed analyses optimized for single-cell resolution.

Understanding immune persistence

Li et al. (Gastroenterology 2020) hypothesized that the function of CARs is affected by their epitope specificity, affinity, and functional activity. To improve antitumor activity, the study employed IsoPlexis’ functional single-cell proteomics to characterize experimental CAR-T designs for high affinity against glypican-3 (GPC3), an antigen expressed in hepatocellular carcinoma (HCC) and other solid tumors.

Preclinical tests were performed in NOD/SCID/IL-2Rgcnull (NSG) mice. Mice injected with GPC3-specific humanized YP7 (hYP7) CAR-T cells had persistent expansion of T cells and unique subsets of highly polyfunctional CAR-T cells via antigen-induced selection observed in the tumor microenvironment and spleen for up to seven weeks post administration.

The study demonstrated a positive correlation of enhanced polyfunctionality by functional single-cell proteomics with persistence of T-cell response and in vivo killing capacity. Importantly, this antigen-driven persistence and expansion of polyfunctional hYP7 CAR-T cells in the tumor microenvironment caused tumor regression in the HCC mouse model, while mice treated with the HN3 CAR-T cells showed no reduction in tumor burden and demonstrated remarkably lower polyfunctionality than the hYP7 CAR-T cells.

From preclinical mouse models to clinical trials

Adoptive cell therapy (ACT) aims to generate a robust antitumor response through naturally-occurring or genetically modified T cells. Supplemental IL-2 administration is required after T-cell adoptive transfer both in mice models and humans to support cellular expansion and function, but significant toxicities can result from high doses.

Although an attractive target, the IL-2 receptor can induce both immune stimulatory and suppressive responses. Therefore, approaches look to expand effector T cells and natural killer (NK) cells.

A study by Parisi et al. (Nature Communications 2020) sought to understand the functional drivers of T-cell persistence in response to a novel engineered IL-2 receptor agonist, PEG-IL-2, from Nektar Therapeutics. Functional single-cell proteomics was used to profile translational insights between early-stage development of combination PEG-IL-2 with ACT therapy, and to study clinical impact on various immune cell types.

The mechanism resulted in superior antitumor activity in a B16-F10 murine melanoma model as well as the promotion of unique subsets of highly polyfunctional T and NK cells in peripheral blood of melanoma patients in a Phase I clinical trial.

Compared to IL-2, the novel agonist increased the number of highly polyfunctional T cells in murine spleens and tumors, which correlated to increases in proliferation, homing, and persistence of antitumor T cells in vivo. The ability of the infiltrated T cells to secrete IFN-γ, granzymes, and perforin both in preclinical and clinical settings supports the ability of PEG-IL-2 to strongly activate a pool of cytotoxic effector T cells.

Conclusions

Data from both studies suggest that polyfunctionality and functional immune profiling obtained via IsoPlexis’ single-cell proteomics may provide a more comprehensive biomarker in solid tumor indications, accelerating insights in cancer immunology.

Download the paper summary for the Parisi et al. case now.  info.isoplexis.com/act-nature-communications-summary

 

References
Li D, et al., Persistent Polyfunctional Chimeric Antigen Receptor T Cells That Target Glypican 3 Eliminate Orthotopic Hepatocellular Carcinomas in Mice. Gastroenterology. 2020 Jun;158(8):2250-2265.e20. doi: 10.1053/j.gastro.2020.02.011. Epub 2020 Feb 12.
Parisi G, et al. Persistence of Adoptively Transferred T Cells with a Kinetically Engineered IL-2 Receptor Agonist. Nat Commun. 2020 Jan 31;11(1):660. doi: 10.1038/s41467-019-12901-3.