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354 Pulling the lever on all fronts: ICT01, a g9d2 T cell-activating monoclonal antibody, in combination with Venetoclax and 5-Azacytidine as a novel combination therapy for AML
  1. Elisabeth Wieduwild1,
  2. Anne-Charlotte Le Floch2,
  3. Céline Garulli1,
  4. Sarah Bourass1,
  5. Caroline Imbert2,
  6. Paul Frohna1,
  7. Norbert Vey3,
  8. Daniel Olive2,
  9. Aude de Gassart1 and
  10. Loui Madakamutil1
  1. 1Imcheck Therapeutics SAS, Marseille, FL, France
  2. 2Inserm, Marseille, France
  3. 3Institut Paoli-Calmettes, Marseille, France


Background Combination of Venetoclax, a BCL-2 inhibitor, and 5-Azacytidine, a hypomethylating agent, is the standard of care for patients with newly diagnosed acute myeloid leukemia (AML) who are not candidates for intensive chemotherapy.1 In addition to their anti-leukemic activity, 5-Azacytidine was shown to improve cancer cell recognition by immune effector cells through induction of stress ligand expression,2,3 and Venetoclax was described to enhance T-cell and NK-cell–mediated cytotoxicity against AML blasts [3,4]. Among attractive mediators of anti-tumor immunity, g9d2 T-cells are emerging as effector cells that harbor strong cytolytic and pro-inflammatory activities and are associated with good prognosis in cancer patients.5,6 ICT01 is a first-in-class anti-BTN3A mAb activating g9d2 T-cells and is being evaluated in a Phase 1/2a clinical study in solid tumors and hematologic malignancies (NCT04243499).7,8

Here, we explore the potential of combining ICT01 with Venetoclax and 5-Azacytidine to increase the anti-leukemic activity of g9d2 T-cells.

Methods Flow cytometry was used to assess the effects of Venetoclax and 5-Azacytidine on ICT01-mediated g9d2 T-cell activation after in vitro culture of healthy donor (HD) or patient derived PBMCs. AML cell lines’ and primary AML blasts’ sensitivity to ICT01-activated g9d2 T-cell killing was monitored after 4 or 24 hours of target:effector co-culture. Targets and/or effectors were pre-treated with Venetoclax, 5-Azacytidine or the combination. Expression of specific stress ligands upon treatment with Venetoclax, 5-Azacytidine or the combination was analyzed by flow cytometry.

Results First, we show that activation of g9d2 T-cells by ICT01 is unaltered in HD-PBMCs treated in vitro with Venetoclax or 5-Azacytidine, in naïve AML patients’ PBMCs or in PBMCs from AML patients under respective treatment. In addition, ICT01-mediated activation of g9d2 T-cells improved resistance to Venetoclax-induced cell death, which is otherwise pronounced in resting g9d2 T-cells. Next, we identified several stress ligands to be upregulated on AML cell lines and primary AML blasts upon treatment with Venetoclax, 5-Azacytidine or the combination. Consequently, pre-treatment of AML (cell lines and primary) with Venetoclax, 5-Azacytidine or the combination improved g9d2 T-cell killing capacities, which confirms previous findings on NK- and T-cells and adds further mechanistic insights. Finally, activation of g9d2 T-cells with ICT01 in combination with Venetoclax and 5-Azacytidine treatment further increased apoptosis of AML cells.

Conclusions These results demonstrate the ability of Venetoclax and 5-Azacytidine to potentiate ICT01-mediated killing of AML cells by γ9δ2 T-cells and support clinical evaluation of this combination as a novel therapeutic approach for AML patients.


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