大多数致癌驱动因素是细胞内蛋白，限制了其免疫治疗靶向由个体人类白细胞抗原（HLA）同种异型1呈现的突变肽（新抗原）。然而，大多数癌症具有适度的突变负担，不足以使用基于新抗原的疗法产生反应2,3。神经母细胞瘤是一种很少有突变的儿科癌症，而是由表观遗传失调的转录网络驱动的。在这里，我们表明神经母细胞瘤免疫肽组富含源自肿瘤发生必需的蛋白质的肽。我们重点关注在 HLA-A*24:02 上发现的未突变肽 QYNPIRTTF，该肽源自神经母细胞瘤依赖性基因和主转录调节因子 PHOX2B。为了靶向 QYNPIRTTF，我们使用预测的潜在交叉反应肽通过反淘选策略开发了以肽为中心的嵌合抗原受体 (PC-CAR)。我们进一步提出，当呈现相似的整体分子表面时，PC-CAR 可以识别其他 HLA 同种异型上的肽。根据我们的计算建模结果，我们表明 PHOX2B PC-CAR 还可以识别 HLA-A*23:01 呈现的 QYNPIRTTF，HLA-A*23:01 是非洲血统人群中最常见的非 A2 等位基因。最后，我们证明了在体外对表达这些 HLA 的神经母细胞瘤细胞的有效和特异性杀伤，并在小鼠中完成肿瘤消退。这些数据表明，PC-CAR 有潜力扩大免疫治疗靶点库，包括非免疫原性细胞内癌蛋白，并允许在临床环境中通过其他 HLA 同种异型进行靶向。© 2023。作者。

The majority of oncogenic drivers are intracellular proteins, constraining their immunotherapeutic targeting to mutated peptides (neoantigens) presented by individual human leukocyte antigen (HLA) allotypes1. However, most cancers have a modest mutational burden that is insufficient for generating responses using neoantigen-based therapies2,3. Neuroblastoma is a paediatric cancer that harbours few mutations and is instead driven by epigenetically deregulated transcriptional networks4. Here we show that the neuroblastoma immunopeptidome is enriched with peptides derived from proteins essential for tumorigenesis. We focused on targeting the unmutated peptide QYNPIRTTF discovered on HLA-A*24:02, which is derived from the neuroblastoma-dependency gene and master transcriptional regulator PHOX2B. To target QYNPIRTTF, we developed peptide-centric chimeric antigen receptors (PC-CARs) through a counter panning strategy using predicted potentially cross-reactive peptides. We further proposed that PC-CARs can recognize peptides on additional HLA allotypes when presenting a similar overall molecular surface. Informed by our computational modelling results, we show that PHOX2B PC-CARs also recognize QYNPIRTTF presented by HLA-A*23:01, the most common non-A2 allele in people with African ancestry. Finally, we demonstrate potent and specific killing of neuroblastoma cells expressing these HLAs in vitro and complete tumour regression in mice. These data suggest that PC-CARs have the potential to expand the pool of immunotherapeutic targets to include non-immunogenic intracellular oncoproteins and allow targeting through additional HLA allotypes in a clinical setting.© 2023. The Author(s).