Combinatorial targeting of the macropinocytotic pathway in leukemia and lymphoma cells
Combinatorial targeting of the macropinocytotic pathway in leukemia and lymphoma cells
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DOI：
10.1074/jbc.m708849200
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发表时间：
2008-04-25
影响因子：
4.8
通讯作者：
Kuniyasu, Akihiko
中科院分区：
文献类型：
--
作者：
Nishimura, Shinpei;Takahashi, Shunsuke;Kuniyasu, Akihiko
配体定向向白血病和淋巴瘤细胞递送药物有可能产生新的疾病机制见解和靶向治疗。在这里，我们着手通过组合方法来靶向巨胞饮途径。通过使用随机噬菌体展示肽库筛选急性 T 淋巴细胞白血病 Molt-4 细胞，我们分离出了展示序列 CAYHRLRRC 的噬菌体。该肽含有淋巴结归巢基序 (Cys-Ala-Tyr) 和细胞穿透基序 (Arg-Leu-Arg-Arg)。这种配体导向的噬菌体与大量白血病/淋巴瘤细胞以及患者来源的样品的结合比与非白血病对照细胞的结合要高得多。 CAYHRLRRC 噬菌体内化到 Molt-4 细胞中是能量和温度依赖性的。使用荧光素标记的肽进行的流式细胞术和使用特定抑制剂阻断内吞作用的结果表明，CAYHRLRRC 确实通过 Molt-4 和 K562 人白血病细胞中的巨胞饮作用被摄取。出乎意料的是，CAYHRLRRC 肽的细胞表面受体不是硫酸乙酰肝素蛋白聚糖，正如其他细胞穿透肽所预测的那样。为了证实这一解释，我们评估了所有白血病和淋巴瘤细胞中 CAYHRLRRC 指导的肽模拟物诱导的细胞死亡，而转录蛋白 (tat) 指导的促凋亡肽模拟物的对照反式激活子是非选择性的。总之，靶向肽 CAYHRLRRC 通过白血病和淋巴瘤细胞中的巨胞饮作用选择性内化，并具有作为配体导向抗白血病治疗的药物先导物的潜力。
Ligand-directed delivery of agents to leukemia and lymphoma cells has the potential to yield new mechanistic disease insights and targeted therapies. Here we set out to target the macropinocytotic pathway with a combinatorial approach. From the screening of acute T-lymphoblastic leukemia Molt-4 cells with a random phage-display peptide library, we isolated a phage displaying the sequence CAYHRLRRC. This peptide contains a lymph node-homing motif (Cys-Ala-Tyr) and a cell-penetrating motif (Arg-Leu-Arg-Arg). Binding of this ligand-directed phage to a large panel of leukemia/lymphoma cells and to patient-derived samples was much higher than to non-leukemia control cells. CAYHRLRRC phage internalization into Molt-4 cells is both energy- and temperature-dependent. Flow cytometry with fluorescein-labeled peptide and endocytosis blocking with specific inhibitors revealed that CAYHRLRRC is indeed taken up through macropinocytosis in Molt-4 and K562 human leukemia cells. Unexpectedly, the cell surface receptor for the CAYHRLRRC peptide is not a heparan sulfate proteoglycan as it would be predicted for other cell-penetrating peptides. Confirming this interpretation, a CAYHRLRRC-directed peptidomimetic-induced cell death in all the leukemia and lymphoma cells was evaluated, whereas a control transactivator of transcription protein (tat)-directed proapoptotic peptidomimetic was non-selective. In summary, the targeting peptide CAYHRLRRC is selectively internalized through macropinocytosis in leukemia and lymphoma cells and has potential as a drug lead for ligand-directed anti-leukemia therapies.