PO.CH02.01 · 化学

BRAF-V600E突变型转移性结直肠癌FFPE组织的蛋白质组学分析揭示WEE1表达可能参与治疗耐药

Proteomic analysis of BRAF-V600E mutant metastatic colorectal cancer FFPE tissues reveals potential involvement of WEE1 expression in therapeutic resistance

编号 7694 展板 18 时间 4/22 09:00–12:00 区域 Section 39 主讲 Shotaro Yamaguchi

分会场 Proteomics: Biomarker Discovery and Signaling Networks

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作者与单位 Authors & Affiliations

Shotaro Yamaguchi¹, Satoshi Muraoka², Yosui Nojima³, Toshiharu Hirose¹, Hidekazu Hirano¹, Natsuko Okita¹, Atsuo Takashima¹, Jun Adachi², Hirokazu Shoji¹, Ken Kato⁴

¹Gastrointestinal Medical Oncology, National Cancer Center Hospital, Tokyo, Japan,²Laboratory of Proteomics for Drug Discovery, Center for Drug Design Research, National Institute of Biomedical Innovation, Health and Nutrition, Osaka, Japan,³Center for Mathematical Modeling and Data Science, The University of Osaka, Osaka, Japan,⁴Head and Neck, Esophageal Medical Oncology / Department of Gastrointestinal Medical Oncology, National Cancer Center Hospital, Tokyo, Japan

摘要 Abstract

中文摘要

背景与目的：约10%的转移性结直肠癌（mCRC）携带BRAF-V600E突变。尽管已针对该亚型开发出靶向治疗方案，如encorafenib联合cetuximab并可选择性联合binimetinib（BEACON方案），但临床疗效仍然较差，克服治疗耐药是一项关键的未满足需求。为阐明这种耐药的分子机制，我们利用取自BRAF-V600E突变型mCRC患者的福尔马林固定石蜡包埋（FFPE）组织进行了蛋白质组学分析。本研究旨在鉴定可能与BRAF靶向治疗耐药相关的蛋白，并探索该人群的治疗靶点。方法：在接受任何全身治疗之前，从BRAF-V600E突变型mCRC患者采集FFPE肿瘤样本。将切片（厚度5 µm）贴于PEN玻片上，对宏观切割的肿瘤区域（每份样本12–130 mm²）进行蛋白提取处理。采用单管固相增强样本制备（SP3）方案纯化并消化蛋白，随后在Orbitrap Exploris 480上进行质谱分析。此外，我们回顾性分析了2020年12月至2024年4月期间在我院接受BEACON方案作为mCRC二线治疗患者的临床数据。采用log-rank检验评估蛋白定量表达与临床结局（包括无进展生存期[PFS]）之间的相关性。结果：共纳入15例患者，其中12例样本成功实现蛋白定量。在这些样本中，对9,078种蛋白进行了定量分析，并考察了蛋白表达与PFS之间的相关性。接受BEACON方案作为二线治疗患者的中位PFS（mPFS）为6.9个月（95% CI，2.6–11.0）。在定量蛋白中，13种蛋白与PFS呈负相关（相关系数[r] < -0.8），其中WEE1显示出最强的相关性（r = -0.94）。相比之下，在一线治疗期间未观察到WEE1表达与PFS之间存在显著相关性（r = -0.48）。在二线治疗中按WEE1表达水平对患者进行分层时，高WEE1组的PFS显著短于低WEE1组（mPFS：3.6个月 vs 10.8个月[HR，21.88，95% CI，3.84–124.6]；p = 0.0005）。结论：我们的蛋白质组学分析将WEE1表达确定为接受BEACON方案治疗的BRAF-V600E突变型mCRC患者临床结局不佳的潜在生物标志物。这些发现提示存在一种情境依赖性耐药机制，可作为有前景的治疗靶点，值得在临床前和临床环境中进一步研究。

查看英文原文 English abstract

Background and Aims: Approximately 10% of metastatic colorectal cancers (mCRC) harbor the BRAF -V600E mutation. Although targeted therapies such as encorafenib plus cetuximab with or without binimetinib (BEACON regimen) have been developed for this subtype, clinical outcomes remain poor, and overcoming therapeutic resistance represents a critical unmet need. To elucidate the molecular mechanisms underlying this resistance, we performed a proteomic analysis using formalin-fixed paraffin-embedded (FFPE) tissues obtained from patients with BRAF -V600E mutant mCRC. The aim of this study was to identify proteins potentially associated with resistance to BRAF -targeted therapy and to explore therapeutic targets for this population. Methods: FFPE tumor samples were collected from patients with BRAF -V600E mutant mCRC prior to any systemic treatment. Sections (5 µm thick) were mounted on PEN glass slides, and macro-dissected tumor areas (12-130 mm 2 per sample) were processed for protein extraction. Proteins were purified and digested using the Single-pot, solid-phase-enhanced sample preparation (SP3) protocol, followed by mass spectrometric analysis on an Orbitrap Exploris 480. Furthermore, we retrospectively analyzed clinical data from patients who received the BEACON regimen as second-line treatment for mCRC at our hospital between December 2020 and April 2024. Correlations between quantitative protein expression and clinical outcomes, including progression-free survival (PFS), were assessed using the log-rank test. Results: A total of 15 patients were included, and protein quantification was successfully achieved in samples of 12 patients. Across these samples, 9,078 proteins were quantitatively profiled, and correlations between protein expression and PFS were examined. The median PFS (mPFS) for patients who received the BEACON regimen as second-line therapy was 6.9 months (95% CI, 2.6-11.0). Among the quantified proteins, 13 proteins showed a negative correlation with PFS (correlation coefficient [r] < -0.8), with WEE1 demonstrating the strongest correlation (r = -0.94). In contrast, no significant correlation was observed between WEE1 expression and PFS during first-line treatment (r = -0.48). When patients were stratified by WEE1 expression levels for the second-line setting, the high-WEE1 group exhibited significantly shorter PFS than the low-WEE1 group (mPFS: 3.6 months vs 10.8 months [HR, 21.88, 95% CI, 3.84-124.6]; p = 0.0005). Conclusion: Our proteomic analysis identified WEE1 expression as a potential biomarker associated with poor clinical outcomes in patients with BRAF -V600E mutant mCRC treated with the BEACON regimen. These findings suggest a context-dependent resistance mechanism as a promising therapeutic target warranting further investigation in preclinical and clinical settings.

利益披露 Disclosure

S. Yamaguchi, None.. S. Muraoka, None.. Y. Nojima, None. T. Hirose, Bristol Myers Squibb ). ONO Pharmaceutical, CO. LTD. ). MSD ). TAIHO Pharmaceutical, CO. LTD. ). Astellas Pharma Inc. ). H. Hirano, Bristol Myers Squibb ), Other, Honoraria. Ono Pharmaceutical, CO. LTD Other, Honoraria. Novartis ), Other, Honoraria. Daiichi-Sankyo, CO. LTD ), Other, Honoraria. Taiho Pharmaceutical, CO. LTD ), Other, Honoraria. PPD ). Boehringer Ingelheim ). ALX Oncology ). BeiGene ). Amgen ). Seagen ). N. Okita, None. A. Takashima, MSD ). AstraZeneca plc ). Amgen ). Eisai, CO. LTD ). Bristol Myers Squibb ). Seagen Inc ). Ono Pharmaceutical, CO. LTD ). Eli Lilly Other, Honoraria. Taiho Pharmaceutical, CO. LTD Other, Honoraria. Chugai Pharmaceutical, CO. LTD Other, Honoraria. Takeda Pharmaceutical, CO.LTD Other, Honoraria. Merck Serono Other, Honoraria. J. Adachi, Proteobiologics Employment, Stock. ONO Pharmaceutical, CO. LTD ). Boelinger Ingelheim ). Mitsubishi Tanabe Pharma ). Takeda Pharma ). Stemrim ). Kirin Holdings ). AMED JP19ck0106465h0001 ). JP23ak0101203h0001 ). JSPS KAKENHI 20K17069, 20H03544 ). H. Shoji, MSD ). Astellas Pharma Inc ). AstraZeneca plc ). AbbVie Inc ). Taiho Pharmaceutical CO, LTD ). Daiichi Sankyo CO, LTD ). Ono Pharmaceutical CO, LTD ). Elevation Oncology Inc ). Chugai Pharmaceutical, CO, LTD ). Metagen Therapeutics Inc ). K. Kato, Bristol Myers Squibb ), Other, Honoraria. MSD ). BeiGene ). Roche ). AstraZeneca plc ). Bayer ). Ono Pharmaceutical, CO. LTD ), Other, Honoraria. Taiho pharmaceutical, CO. LTD Other, Honoraria. Chugai Pharmaceutical, CO. LTD ). Shionogi Inc ).

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