{
  "abstract": "Background CAR T cells targeting CLDN18.2 have shown promising efficacy and safety in clinical trials for gastric cancer, but have struggled to achieve complete, durable responses due to challenges posed by the solid tumor microenvironment (TME). The therapeutic benefits of CLDN18.2 CAR Ts are further restricted by the difficulties of ex-vivo manufacturing, resulting in low patient accessibility and high treatment costs. Here, we developed a dual-armored CAR design, WAY-102, that addresses critical challenges in the gastric TME and has dramatically superior efficacy compared to multiple benchmark CAR T designs. We have further developed this asset for off-the-shelf delivery with a lentivirus formulation to manufacture armored CAR T cells in vivo.Methods We investigated the efficacy and safety of ex vivo manufactured WAY-102 CAR T cells relative to multiple benchmark CAR T designs in cell-line derived xenograft (CDX) models. Specifically, we used AsPC-1-CLDN18.2+ as a pancreatic cancer CDX, and GSU cells as a gastric cancer CDX with endogenous CLDN18.2 expression. We measured CAR T cell expansion by flow cytometry, and assessed CAR T tumor infiltration and cytotoxicity through spatial analysis of FFPE tumor sections. To manufacture WAY-102 CAR T cells in vivo, we used a lentivirus engineered to specifically target T cells, and injected this virus into tumor-bearing PBMC-humanized mice.Results Using our in vivo spatial pooled screening platform to optimize clinically-relevant features of CAR T performance, we developed WAY-102, a dual-armored CLDN18.2 CAR. We found that ex vivo manufactured WAY-102 cells had greater anti-tumor efficacy at lower doses than benchmark designs, including unarmored-, TGFβRIIDN-armored, and IL15-armored CLDN18.2 CAR T cells. WAY-102 CAR T cells resulted in complete tumor clearance in gastric and pancreatic cancer CDX models, for both subcutaneous and intraperitoneal tumors. Complete tumor clearance occurred with as few as 100-200k CAR T cells and resulted in mouse survival without measurable tumors for >100 days after CAR T cell injection. Mechanistically, we show that the armors in WAY-102 increase T cell infiltration-capacity, cytotoxicity, and expansion. To develop an off-the-shelf treatment with WAY-102, we used an engineered lentivirus for in vivo CAR T manufacturing, which specifically transduced WAY-102 into T cells in PBMC-humanized mice and resulted in complete CLDN18.2+ tumor clearance.Conclusions WAY-102 CAR T cells show promising anti-tumor efficacy against multiple in vivo CLDN18.2+ solid tumor models, with successful in vivo manufacturing of WAY-102 paving the way for an off-the-shelf therapy for CLDN18.2+ cancers.Ethics Approval All animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) of Mispro Biotech Services, under the approved protocols 2023-WPT-01 (amendments A1-A20) and 2024-WPT-02.",
  "authors": [
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Cassandra Kontur"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Ashley Chui"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Reeti Sanghrajka"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Liang He"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Gundula Povysil"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Megha Sah"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Annie Dyatel"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Stephen Pu"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Ujas Patel"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "Xinchen Wang"
    },
    {
      "affiliations": [
        "Waypoint Bio, Inc., New York, NY, USA"
      ],
      "name": "David Phizicky"
    }
  ],
  "title": "240 WAY-102: an in vivo lentiviral dual-armored CLDN18.2 CAR T engineered for long-term durability & cytotoxicity in the tumor microenvironment",
  "uid": "2536015a-164b-56e8-a57a-3a498b95d6c1"
}