Purpose: Disruption of lipid bilayer asymmetry is a common feature observed in cancer cells and offers novel routes for therapeutic targeting. We utilized the natural immune receptor TIM-4 to interrogate for loss of plasma membrane phospholipid polarity in primary acute myelogenous leukemia (AML) samples and evaluated the anti-leukemic activity of TIM-4-L-directed T cell therapy in preclinical AML models. Methods: We performed FACs analysis on 33 primary AML bone marrow specimens and correlated TIM-4-L expression frequency and intensity with molecular disease characteristics. Using Kasumi-1 and MV-4-11 AML cell lines, we further tested the anti-leukemic effects of TIM-4-L-directed engineered T cells in vitro and in vivo. Results: 86% of untreated AML blasts displayed upregulation of cell surface TIM-4-L. These observations were agnostic to AML genetic classification, as samples with mutations in TP53, ASXL1, and RUNX1 displayed TIM-4-L upregulation similar to that seen in favorable and intermediate subtypes. TIM-4-L dysregulation was also stably present in AML cell lines. To evaluate the potential of targeting upregulated TIM-4-L with adoptive T cell therapy (ACT), we constructed TIM-4-L-directed engineered T cells, which demonstrated potent anti-leukemic effects, effectively eliminating AML cell lines with a range of endogenous TIM-4-L expression levels both in vitro and in vivo. Conclusions: These results highlight TIM-4-L as a highly prevalent target on AML across a range of genetic classifications and novel target for T cell-based therapy in AML. Further investigations into the role of TIM-4-L in AML pathogenesis and its potential as an anti-leukemic target for clinical development are warranted.

中文翻译：

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使用工程 T 细胞靶向 TIM-4-L 治疗急性髓系白血病

目的：脂质双层不对称性的破坏是癌细胞中观察到的一个常见特征，并为治疗靶向提供了新的途径。我们利用天然免疫受体 TIM-4 来检测原发性急性髓性白血病 (AML) 样本中质膜磷脂极性的丧失，并评估 TIM-4-L 定向 T 细胞疗法在临床前 AML 模型中的抗白血病活性。方法：我们对 33 例原发性 AML 骨髓标本进行 FAC 分析，并将 TIM-4-L 表达频率和强度与分子疾病特征相关联。使用 Kasumi-1 和 MV-4-11 AML 细胞系，我们进一步测试了 TIM-4-L 定向工程 T 细胞在体外和体内的抗白血病作用。结果：86% 未经处理的 AML 母细胞显示细胞表面 TIM-4-L 上调。这些观察结果与 AML 遗传分类无关，因为 TP53、ASXL1 和 RUNX1 突变的样本显示 TIM-4-L 上调，与有利亚型和中间亚型相似。TIM-4-L 失调也稳定存在于 AML 细胞系中。为了评估通过过继性 T 细胞疗法 (ACT) 靶向上调的 TIM-4-L 的潜力，我们构建了 TIM-4-L 定向工程 T 细胞，该细胞表现出有效的抗白血病作用，有效消除了一系列 AML 细胞系。体外和体内内源性 TIM-4-L 表达水平的影响。结论：这些结果强调 TIM-4-L 是一系列遗传分类中非常普遍的 AML 靶点，也是 AML 中基于 T 细胞的治疗的新靶点。有必要进一步研究 TIM-4-L 在 AML 发病机制中的作用及其作为临床开发抗白血病靶点的潜力。