PO.CL01.13 · 临床研究
三阴性乳腺癌中与新辅助治疗反应相关的空间转录组特征
Spatial transcriptomic features associated with response to neoadjuvant therapy in triple-negative breast cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：KEYNOTE-522 试验通过显著改善病理完全缓解（pCR）和无事件生存期，确立了帕博利珠单抗联合新辅助化疗作为早期三阴性乳腺癌（TNBC）的标准疗法。然而，预测反应的生物标志物以及残留疾病背后的生物学机制——尤其是仅接受化疗与化疗加免疫治疗的残余癌负荷 2 级（RCB2）患者之间观察到的不同结局——仍未明确界定。我们假设诊断时的空间细胞结构及其随治疗的纵向重塑决定了 TNBC 的治疗反应和耐药。
方法：我们对来自接受含或不含帕博利珠单抗新辅助化疗患者的 131 份诊断活检、161 份切除标本和 38 处转移病灶进行了单细胞分辨率空间转录组刻画，其中包括 50 对配对的治疗前/后样本用于纵向分析。一个 60 标志物的空间蛋白质组面板提供了跨模态验证。在 pCR 与非 pCR 组、RCB 分级（0-3）以及每个治疗组内的纵向转变之间，评估了空间特征，包括细胞类型丰度、共定位、肿瘤-免疫界面结构和生态位组织。
结果：在 6 个病理学家标注的组织学区域中，我们解析出 17 种主要细胞类型、72 种基因定义的亚型和 96 个空间群落。早期分析凸显了对比鲜明的 CD8⁺ T 细胞空间状态：肿瘤内 CD8⁺ 浸润是应答者的特征，而基质局限的 CD8⁺ 定位连同 CD33⁺ 髓系程序则标志着无应答和 T 细胞排斥。在成纤维细胞密度、基质结构以及髓系-淋巴系邻近关系方面观察到额外的队列特异性变异。通过整合诊断和纵向空间特征，我们正在开发一种可解释的空间风险评分，以区分治疗反应和 RCB 分级。
结论：本研究是迄今为止 TNBC 新辅助治疗中规模最大的整合空间转录组和蛋白质组分析之一。新出现的模式提示，细胞毒性 T 细胞进入肿瘤巢标志着有效的抗 PD-1 应答，而成纤维细胞驱动的基质和髓系介导的排斥则是耐药的基础。这些见解为设计克服新辅助免疫治疗中基质和髓系屏障的联合疗法提供了路线图。
查看英文原文 English abstract
Background: The KEYNOTE-522 trial established pembrolizumab plus neoadjuvant chemotherapy as a standard for early-stage triple-negative breast cancer (TNBC) by significantly improving pathologic complete response (pCR) and event-free survival. Yet biomarkers predicting response and the biology underlying residual disease-particularly the divergent outcomes observed among patients with residual cancer burden class 2 (RCB2) from chemotherapy only and chemotherapy plus immunotherapy-remain poorly defined. We hypothesized that spatial cellular architectures at diagnosis, and their longitudinal remodeling with therapy, govern TNBC treatment response and resistance.
Methods: We performed single-cell resolution spatial transcriptomic profiling on 131 diagnostic biopsies, 161 resection specimens, and 38 metastatic lesions from patients treated with neoadjuvant chemotherapy with or without pembrolizumab, including 50 paired pre/post-treatment samples for longitudinal analysis. A 60-marker spatial proteomic panel provided cross-modality validation. Spatial features, including cell-type abundance, co-localization, tumor-immune interface structure, and niche organization, were evaluated across pCR versus non-pCR groups, RCB classes (0-3), and longitudinal transitions within each therapeutic arm.
Results: Across 6 pathologist-annotated histologic regions, we resolved 17 major cell types, 72 gene-defined subtypes, and 96 spatial communities. Early analyses highlight contrasting CD8⁺ T-cell spatial states: intra-tumoral CD8⁺ infiltration characterizes responders, whereas stromal-restricted CD8⁺ localization, together with CD33⁺ myeloid programs, marks non-response and T-cell exclusion. Additional cohort-specific variation was observed in fibroblast density, stromal architecture, and myeloid-lymphoid proximities. Integrating diagnostic and longitudinal spatial features, we are developing an interpretable spatial risk score to distinguish treatment response and RCB class.
Conclusions: This study represents one of the largest integrated spatial transcriptomic and proteomic analyses of TNBC neoadjuvant therapy to date. Emerging patterns suggest that cytotoxic T-cell access to tumor nests marks effective anti-PD-1 response, whereas fibroblast-driven stroma and myeloid-mediated exclusion underlie resistance. These insights provide a roadmap for designing combination therapies to overcome stromal and myeloid barriers to neoadjuvant immunotherapy.
利益披露 Disclosure
S. Liao, None..
T. Noel, None..
J. Lownik, None..
P. Nedumaran, None..
Y. Yang, None..
R. Mebane, None..
A. Shah, None..
A. Martinez, None..
S. Knott, None.