{
  "abstract": "Background FDA-approved companion diagnostic biomarkers for immune checkpoint inhibitors (ICIs) have limited accuracy in predicting response. Cytokine profiling of live tumor samples has shown promise in improving accuracy for predicting response to PD-1 blockade. 1 However, this approach has been limited to tumor resections, given the need for large amounts of tissue, and cryopreserved samples, which can lead to a reduction in specific immune cell populations. Here we present a cytokine profiling platform for live transported core needle biopsies (CNBs). Using data collected on the platform, we provide external validation of a weighted classifier published by Voabil et al.1 for predicting response to PD-1 blockade. Additionally, we train and validate a novel classifier using >250 tumor specimens profiled on the platform with several different ICI regimens.Methods Tumor specimens were obtained from patients being considered for standard of care ICI therapy participating in one of three prospective multi-center observational clinical trials ( NCT05478538, NCT05520099 and NCT06349642) or from a prospective biobank study. Live pretreatment biopsies were transported overnight and processed within 24 hours of collection. Biopsies were cut into live tumor fragments, encapsulated in hydrogel, and treated using a strategy where control (IgG) and ICI treatments were performed sequentially on the same tissue in a single well. ICI treatment (αPD-1, αPD-L1 or αPD-1+αCTLA-4) was aligned with the anticipated clinical regimen for individual patients. Changes in cytokine concentrations were assessed longitudinally using multiplexed assays encompassing >45 cytokines. Differences in cytokine production rates were calculated between the ICI and control treatment phases. Unsupervised clustering was performed to identify clusters of cytokine responsive and nonresponsive patient samples.Results Using area under the receiver operating characteristic curve analysis, we built a weighted classifier using the most discriminative cytokines between the two clusters. We then validated its performance, along with the Voabil et al. classifier, on an independent set of >15 tumor specimens collected from patients who received ICI therapy and were assessed for radiological response.Conclusions Here we show that our classifier, built on a large multicenter dataset, can distinguish between clinical responder and non-responder patients on ICI. Furthermore, we externally validate the Voabil classifier and show that the limited tissue available from a single live transported CNB is sufficient to support functional ex vivo cytokine profiling to characterize response to ICI treatment. This approach has great potential for guiding ICI treatment decisions in the clinic.Reference Voabil P, de Bruijn M, Roelofsen LM, Hendriks SH, Brokamp S, van den Braber M, Broeks A, Sanders J, Herzig P, Zippelius A, Blank CU, Hartemink KJ, Monkhorst K, Haanen JBAG, Schumacher TN, Thommen DS. An ex vivo tumor fragment platform to dissect response to PD-1 blockade in cancer. Nat Med. 2021 Jul;27(7):1250-1261. doi: 10.1038/s41591-021-01398-3. Epub 2021 Jul 8. PMID: 34239134.",
  "authors": [
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Nicholas Dana"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Hilary R Hernan"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Pichet Adstamongkonkul"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Sean Caenepeel"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Catarina Costa"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Laura CF Hrycyniak"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Nathan Marhefke"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Payton McDonnell"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Amreen Nasreen"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "TS Ramasubramanian"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Katherine Rexroad"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Jordyn C Richardson"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Sidney Schneider"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Mikaela Schultz"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Christina Scribano"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Aishwarya Sunil"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Rebecca Tarlazzi"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Veronica Wang"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Ellen Wargowski"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Jackie Derrick"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Steven Fuhrman"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Ann Leonard"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Chetan Sood"
    },
    {
      "affiliations": [
        "Elephas Biosciences, Madison, WI, USA"
      ],
      "name": "Hinco J Gierman"
    },
    {
      "affiliations": [
        "Mayo Clinic, Jacksonville, FL, USA"
      ],
      "name": "Hani M Babiker"
    },
    {
      "affiliations": [
        "Orlando Health, Orlando, FL, USA"
      ],
      "name": "Barbara (BJ) Broome"
    },
    {
      "affiliations": [
        "Frederick Health, Fredrick, MD, USA"
      ],
      "name": "Heather Chalfin"
    },
    {
      "affiliations": [
        "EmpiriQA LLC, Long Grove, IL, USA"
      ],
      "name": "Tarun Chandra"
    },
    {
      "affiliations": [
        "Meritus Health, Hagerstown, MD, USA"
      ],
      "name": "Victoria Giffi"
    },
    {
      "affiliations": [
        "Baylor Scott and White Health, Dallas, TX, USA"
      ],
      "name": "Alan Gowan"
    },
    {
      "affiliations": [
        "CARTI, Little Rock, AR, USA"
      ],
      "name": "David Hays"
    },
    {
      "affiliations": [
        "UNC School of Medicine, Chapel Hill, NC, USA"
      ],
      "name": "Nima Kokabi"
    },
    {
      "affiliations": [
        "Mayo Clinic, Jacksonville, FL, USA"
      ],
      "name": "Yanyan Lou"
    },
    {
      "affiliations": [
        "Mayo Clinic, Jacksonville, FL, USA"
      ],
      "name": "Umair Majeed"
    },
    {
      "affiliations": [
        "University of Louisville School of Medicine, Louisville, KY, USA"
      ],
      "name": "Robert Martin"
    },
    {
      "affiliations": [
        "UW Health University Hospital, Madison, WI, USA"
      ],
      "name": "Giuseppe Toia"
    },
    {
      "affiliations": [
        "New York Cancer and Blood Specialists, New York, NY, USA"
      ],
      "name": "Richard Zuniga"
    },
    {
      "affiliations": [
        "Mayo Clinic, Jacksonville, FL, USA"
      ],
      "name": "Roxana S Dronca"
    },
    {
      "affiliations": [
        "Orlando Health, Orlando, FL, USA"
      ],
      "name": "Pooja Advani"
    },
    {
      "affiliations": [
        "Mayo Clinic, Jacksonville, FL, USA"
      ],
      "name": "Debabrata Mukhopadhyay"
    },
    {
      "affiliations": [
        "Mayo Clinic, Jacksonville, FL, USA"
      ],
      "name": "Ruqin Chen"
    }
  ],
  "title": "54 Development and characterization of cytokine-based classifiers to predict response to immunotherapy using an ex vivo live tumor fragment platform",
  "uid": "4a9ed00c-2b0f-57f5-a660-abfcecb63ba8"
}