PO.CL01.23 · 临床研究
炎性乳腺癌中CCR7的表达和空间分布:用于治疗靶向的基线表征
CCR7 expression and spatial distribution in inflammatory breast cancer: A baseline characterization for therapeutic targeting
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摘要 Abstract
中文摘要
目的:炎性乳腺癌(IBC)是一种罕见、高度侵袭性的亚型,以快速增殖、广泛的血管/淋巴管生成和早期转移为特征。CCR7是一种参与免疫运输的趋化因子受体,通过CCL19/CCL21促进肿瘤向淋巴管迁移。本研究对IBC中的CCR7进行了整合的基因组、蛋白、空间和功能评估,以评价其治疗潜力。
方法:临床分析使用了来自HR+ HER2-和TNBC IBC的基因组数据,并与亚型匹配的非IBC进行比较。通过免疫印迹评估IBC和非IBC细胞系中的CCR7表达;通过ELISA测量肿瘤和基质细胞中的配体分泌(CCL19/CCL21)。使用亚细胞分级分离和免疫荧光(IF)评估CCR7定位,并测试了一种CCR7阻断抗体。对代表不同CCR7基因组和蛋白状态的BCM PDX模型进行了多重IF。功能研究包括CCR7拮抗剂Navarixin、CCL19/21诱导的迁移实验以及CCR7 CRISPR敲除克隆的构建。
结果:CCR7 CNAs在IBC和非IBC中均出现,但与mRNA或蛋白水平不相关。CCR7蛋白广泛表达,在A3250中膜表达最强,而在IBC-3、SUM-190和SUM-149中水平不一。阻断抗体仅在A3250中降低了膜CCR7,表明存在亚型特异性的可及性。IBC细胞系不分泌CCL19/CCL21,支持其依赖基质配体,而基质配体因共培养而异。在六个BCM PDX中,CCR7持续表达,而配体水平呈异质性;MBI-117显示高CCL19/CCL21。空间分析显示CCR7⁺肿瘤细胞沿Podoplanin⁺淋巴管排列,伴有CCL21⁺基质细胞,且CD163⁺巨噬细胞富集于CCR7⁺瘤栓周围。Navarixin在96小时时降低了IBC3和SUM-190的增殖,而CCL19/21增强了A3250和SUM-149的迁移。已构建CCR7-KO SUM-149克隆,功能分析正在进行中。
结论:IBC中的CCR7表达独立于拷贝数,并由一个富含配体的微环境所支持,该微环境使CCR7⁺肿瘤细胞沿淋巴管定位。整合的基因组、空间和功能数据支持CCR7作为IBC的治疗靶点,相关研究正在进行中。
查看英文原文 English abstract
Purpose: Inflammatory breast cancer (IBC) is a rare, highly aggressive subtype marked by rapid proliferation, extensive angio-/lymphangiogenesis, and early metastasis. CCR7, a chemokine receptor involved in immune trafficking, promotes tumor migration toward lymphatics via CCL19/CCL21. This study provides an integrated genomic, protein, spatial, and functional evaluation of CCR7 in IBC to assess its therapeutic potential.
Methods: Clinical analysis used genomic data from HR+ HER2- and TNBC IBC compared with subtype-matched non-IBC. CCR7 expression was assessed in IBC and non-IBC cell lines by immunoblotting; ligand secretion (CCL19/CCL21) in tumor and stromal cells was measured by ELISA. CCR7 localization was evaluated using subcellular fractionation and IF, along with testing of a CCR7-blocking antibody. Multiplex IF was performed on BCM PDX models representing distinct CCR7 genomic and protein states. Functional studies included the CCR7 antagonist Navarixin, CCL19/21-induced migration assays, and generation of CCR7 CRISPR knockout clones.
Results: CCR7 CNAs occurred in both IBC and non-IBC but did not correlate with mRNA or protein levels. CCR7 protein was broadly expressed, with strongest membranous expression in A3250 and variable levels in IBC-3, SUM-190, and SUM-149. The blocking antibody reduced membranous CCR7 only in A3250, indicating subtype-specific accessibility. IBC cell lines did not secrete CCL19/CCL21, supporting reliance on stromal ligands, which varied by coculture. Across six BCM PDXs, CCR7 was consistently expressed, whereas ligand levels were heterogeneous; MBI-117 showed high CCL19/CCL21. Spatial profiling revealed CCR7⁺ tumor cells aligned along Podoplanin⁺ lymphatics with CCL21⁺ stromal cells and CD163⁺ macrophages enriched around CCR7⁺ emboli. Navarixin reduced proliferation in IBC3 and SUM-190 by 96h, and CCL19/21 enhanced migration in A3250 and SUM-149. CCR7-KO SUM-149 clones were generated, with functional analyses ongoing.
Conclusion: CCR7 expression in IBC is independent of copy number and supported by a ligand-rich microenvironment that positions CCR7⁺ tumor cells along lymphatics. Integrated genomic, spatial, and functional data support CCR7 as a therapeutic target in IBC, with studies ongoing.
利益披露 Disclosure
S. Shivhare, None..
C. Sanchez, None..
R. Larson, None..
L. Dobrolecki, None..
M. Lewis, None..
J. Chen, None..
S. Lin, None.
A. Evdokimova,
BostonGene Corporation Employment.
D. Goncharova,
BostonGene Corporation Employment.
A. Alexander, None..
A. Nasrazadani, None..
R. Layman, None..
B. Lim, None..
V. Valero, None..
A. Lucci, None..
W. A. Woodward, None.