A Single-Cell Proteomic instrument for Predictive Product Quality Check in Autologous CAR-T Immunotherapies

While CAR-T therapies have advanced in the clinic, there are still two specific issues using these breakthrough therapies for blood cancers: Durable responses in patients are only at 30-50% and Cytokine Release Syndrome occurs in 60-90% of treated patients. These limitations, for a drug that costs as much as $500,000, may cost an additional $250,000+ for treating adverse events leading to caution for many clinical centers to administer these therapies. IsoPlexis is developing its technology to predict responders, non-responders, and toxicity in patients, directly from the CAR T cell therapy product prior to entry in the patients. This would allow clinical centers to provide improved product release criteria improving CAR T therapy production and potentially increasing patient response while reducing expensive side effects. IsoPlexis recently published in the journal Blood in July of 2018 showing that across a National Cancer Institute Trial of 20 NHL patients, only the IsoCode Single-Cell Chip predicted responders and non-responders (p = 0.0119), where existing product evaluation technologies like flow cytometry and bulk ELISA did not. The IsoCode’s ability to detect 40+ secreted proteins per cell identified unique polyfunctional cells in responding patients (in a metric termed PSI). This published work with Kite Pharma and the NCI also demonstrated the ability to predict certain types of toxicities in combination with in vivo metrics, including grade 3+ neurotoxicity (p = 0.0007) and cytokine release syndrome (p = 0.0085). In its NCI phase II CAR-T SBIR, IsoPlexis validated this CAR-T IsoCode Chip, and built and tested the fully automated workflow for the IsoCode Chip on the IsoLight instrument. The IsoLight is a sample-to-answer device on which users can process 8 samples simultaneously. Images are taken overnight through a 3 laser-based optics system, and back- end ELISA steps are run in an automated fashion. The CAR-T polyfunctional results are directly available in the IsoSpeak software suite. The instrument is now working and analyzing CAR-T samples. IsoPlexis’ IsoCode Chip, for use in CAR-T, was recognized as the #1 Innovation of 2017 by the Scientist Magazine, as well as Fierce Life Sciences. The IsoLight instrument was recognized for ease of use by the leading Global Red Dot Design Award, and IsoSpeak was recognized as Top 10 Innovation by Pharma Tech Outlook. The Phase IIB goal advances the IsoLight RUO system and clinically validates the IsoCode and IsoLight across centers delivering an IsoLight Dx to provide the autologous T cell therapy market predictive therapeutic product evaluation. Aim 1: Develop automation for manufacturing consumable chip to meet larger production needs. Aim 2: Development of enterprise software modules, including CFR Part 211 compliance to enable GMP pharmaceutical environment for CAR-T product release. Aim 3: Develop automated IsoLight production and calibration processes verifying with four instruments over 6 months to meet IsoLight scalability requirements in CAR-T release. Aim 4: Achieve IsoLight clinical validation according to best-in-class ROC, and statistical parameters: One CAR-T trial in NHL (Moffitt CC), a CAR-T trial in ALL (Memorial Sloan Kettering CC), and a CAR-T trial in DLBCL (Stanford CC).Adoptive transfer of autologous T cells engineered to express chimeric antigen receptors (CARs) has emerged as a promising immunotherapy for patients with hematologic malignancies, such as CD19 CARs in leukemias and lymphomas. However, challenges remain in terms of manufacturing consistency and the functional profile of the CAR-T cell product, as cytokines secreted by these cells, post-infusion, result in not only therapeutic efficacy but also life-threatening immunotoxicity. We have developed a single-cell, highly-multiplexed, barcoding device that measures all safety and efficacy functional cytokines secreted, to meet the need of cancer patients by providing full spectrum potency and toxicity profiling of CAR-T cell therapies before infusion as part of the clinical quality control process.