Nutlin-3a Enhances Natural Killer Cell–Mediated Killing of Neuroblastoma by Restoring p53-Dependent Expression of Ligands for NKG2D and DNAM-1 Receptors
Nutlin-3a Enhances Natural Killer Cell–Mediated Killing of Neuroblastoma by Restoring p53-Dependent Expression of Ligands for NKG2D and DNAM-1 Receptors
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DOI：
10.1158/2326-6066.cir-20-0313
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发表时间：
2020-12
影响因子：
10.1
通讯作者：
I. Veneziani;Paola Infante;E. Ferretti;O. Melaiu;Cecilia Battistelli;V. Lucarini;M. Compagnone;C. Nicoletti;A. Castellano;S. Petrini;M. Ognibene;A. Pezzolo;L. Di Marcotullio;R. Bei;L. Moretta;V. Pistoia;D. Fruci;V. Barnaba;F. Locatelli;L. Cifaldi
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文献类型：
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作者：
I. Veneziani;Paola Infante;E. Ferretti;O. Melaiu;Cecilia Battistelli;V. Lucarini;M. Compagnone;C. Nicoletti;A. Castellano;S. Petrini;M. Ognibene;A. Pezzolo;L. Di Marcotullio;R. Bei;L. Moretta;V. Pistoia;D. Fruci;V. Barnaba;F. Locatelli;L. Cifaldi
Nutlin-3a是一种拮抗MDM2-P53相互作用的小分子，对神经母细胞瘤具有P53依赖的免疫调节作用。Nutlin-3a增加了神经母细胞瘤对NK细胞杀伤的敏感性，强调了这种化合物如何有望用于基于NK细胞的免疫治疗。在这项研究中，我们探讨了已知的无毒小分子化合物Nutlin-3a是否可以恢复神经母细胞瘤细胞上自然杀伤(NK)细胞激活受体(NK-AR)的配体，以增强NK细胞介导的杀伤。用Nutlin-3a处理神经母细胞瘤细胞系，检测NKG2D和dNaM-1 NK-ARs配体的表达以及神经母细胞瘤对NK细胞的敏感性。在移植神经母细胞瘤的NSG小鼠模型中过继转移人NK细胞。对神经母细胞瘤患者的两个数据集进行了研究，以期将P53的表达与配体的表达联系起来。用荧光素酶和染色质免疫沉淀分析P53与PVR启动子的功能结合。原代神经母细胞瘤细胞也用Nutlin-3a处理，并检测从一名高危患者获得的神经母细胞瘤球体的NK细胞毒性。我们提供的证据表明，神经母细胞瘤细胞中依赖Nutlin-3a的P53功能的挽救导致(I)NK-AR配体的表面表达增加，从而使神经母细胞瘤细胞系显著更容易受到NK细胞介导的杀伤；(Ii)携带神经母细胞瘤小鼠过继转移NK细胞后，与总体生存相关的人神经母细胞瘤肿块缩小；(Iii)原代神经母细胞瘤细胞中配体表达增加，并促进NK细胞介导的神经母细胞瘤球体的解聚。我们还发现，P53是一个直接的转录因子，调节dNaM-1识别的PVR配体的表达。我们的研究结果表明，Nutlin-3a具有免疫调节作用，有望用于神经母细胞瘤的NK细胞免疫治疗。
Nutlin-3a, a small molecule antagonizing the inhibitory MDM2–p53 interaction, has a p53-dependent immunomodulatory effect on neuroblastoma. Nutlin-3a increases neuroblastoma's susceptibility to NK-cell killing, highlighting how this compound could be prospectively employed for an NK cell–based immunotherapy. In this study, we explored whether Nutlin-3a, a well-known, nontoxic small-molecule compound antagonizing the inhibitory interaction of MDM2 with the tumor suppressor p53, may restore ligands for natural killer (NK) cell–activating receptors (NK-AR) on neuroblastoma cells to enhance the NK cell–mediated killing. Neuroblastoma cell lines were treated with Nutlin-3a, and the expression of ligands for NKG2D and DNAM-1 NK-ARs and the neuroblastoma susceptibility to NK cells were evaluated. Adoptive transfer of human NK cells in a xenograft neuroblastoma-bearing NSG murine model was assessed. Two data sets of neuroblastoma patients were explored to correlate p53 expression with ligand expression. Luciferase assays and chromatin immunoprecipitation analysis of p53 functional binding on PVR promoter were performed. Primary neuroblastoma cells were also treated with Nutlin-3a, and neuroblastoma spheroids obtained from one high-risk patient were assayed for NK-cell cytotoxicity. We provide evidence showing that the Nutlin-3a–dependent rescue of p53 function in neuroblastoma cells resulted in (i) increased surface expression of ligands for NK-ARs, thus rendering neuroblastoma cell lines significantly more susceptible to NK cell–mediated killing; (ii) shrinkage of human neuroblastoma tumor masses that correlated with overall survival upon adoptive transfer of NK cells in neuroblastoma-bearing mice; (iii) and increased expression of ligands in primary neuroblastoma cells and boosting of NK cell–mediated disaggregation of neuroblastoma spheroids. We also found that p53 was a direct transcription factor regulating the expression of PVR ligand recognized by DNAM-1. Our findings demonstrated an immunomodulatory role of Nutlin-3a, which might be prospectively used for a novel NK cell–based immunotherapy for neuroblastoma.