{
  "abstract": "Background Adoptive cell therapy (ACT) has had limited success in solid tumors, and patient access is a concern. Target antigen downregulation may limit ACT success and cell trafficking remains challenging. We produced CD8αβ T cells from inducible pluripotent stem cells (iPSCs) with TCR-dependent and innate killing, which were comprehensively engineered to address solid tumor TME challenges.Methods CD8αβ iPSC-derived-T cells (iT) cells were differentiated from the QHJI iPSC line. In vitro cytotoxicity was assessed by luciferase, cellular impedance, and/or kinetic fluorescence. PC9-GPC3 tumor cells were injected intravenously (IV) or intra-splenic into NSG mice to establish lung or liver metastases (LuM, LM). iT cells were administered IV via tail vein for systemic or portal vein (PV) for regional infusion. We assessed tumor response with bioluminescence and cellular kinetics by flow cytometry.Results We produced CD8αβ iT cells expressing a TCR targeting HLA-A24-restricted GPC3 peptides, with constitutive IL-15/IL-21 secretion and immune evasion edits to prevent cellular and humoral rejection. CD8 T cell markers (CD3, TCRαβ, CD8αβ, CCR7, CD45RA, CD45RO) and innate killing markers (NKG2D, DNAM1) were expressed. iT cells killed GPC3-negative HLA-A24+ PC9 tumor cells in vitro, which was enhanced with GPC3 peptide pulsing, across E:T ratios (70% vs 60% at 0.3:1, p=0.01; 100% v 92% at 5:1, p=0.18). iT also exhibited TCR-independent killing of GPC3-negative tumor (K562) in vitro, which was diminished 36% by NKG2D and DNAM1 blockade (p<0.01). Negligible killing of normal B cells from two donors was observed (E:T, 1:25:1) despite efficient killing of PC9. IL-15/IL-21 enhanced acute killing (48h) of GPC3+SK-Hep in vitro (58% increase, p=0.01). IL-15/IL-21 armored iT exhibited serial killing in vitro over 2 weeks with GPC3+SK-Hep (E:T 2.5:1-10:1), while GPC3-negative cells were only killed at highest E:T. Non-armored iT lacked serial-killing activity. In the LuM model, armored iT showed anti-tumor activity against GPC3+PC9 (91-fold decrease v CTRL, D10 p<0.0001). iT cells via PV, directly into the liver, resulted in an 11-fold decrease in LM growth (D4 v CTRL, P<0.0001). IV infusion resulted in 4.4-fold greater detection of iPS-T in the lung, compared with PV (D0,p=0.17). When administered via PV, there was a 31-fold (p<0.01) improvement in intra-hepatic cell trafficking relative to IV infusion on DO, and 7.8-fold on D4 (p=0.014).Conclusions CD8αβ iT cells killed tumor cells through TCR-dependent and antigen-independent mechanisms. Cytokine armoring promoted serial killing and comprehensive immune evasion edits may enable enhanced persistence in an upcoming phase 1/1b trial for liver and lung cancer patients.Acknowledgements This work is partly supported by the funding provided by the Japan Agency for Medical Research and Development.",
  "authors": [
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Sara Peyrot"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Chiyomi Sasaki"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Xiao Guo"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Jay Chen"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Fabian Oceguera-Yanez"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Taiki Nampo"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Yasuyuki Miyake"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Hirokazu Koizumi"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Yui Tatsumi"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Mitsujiro Osawa"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Hiroshi Hamana"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Lauren Gauthier-Burdick"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Parvez Vora"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Yueting Zhang"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Daniel Kemp"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, Kyoto-shi, Kyoto, Japan"
      ],
      "name": "Yasumichi Hitoshi"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, San Francisco, CA, USA"
      ],
      "name": "Luis G Borges"
    },
    {
      "affiliations": [
        "Shinobi Therapeutics, East Greenwich, RI, USA",
        "Brown University, Providence, RI, USA"
      ],
      "name": "Steven C Katz"
    }
  ],
  "title": "1010 Systemic and regionally infused iPSC-derived CD8 αβ T cells mediate tumor killing via TCR- and innate receptor-dependent mechanisms enhanced by cytokine armoring",
  "uid": "10044f24-5762-57b5-90b5-cb4312875a29"
}