PO.CT01.04 · 临床试验
术前低剂量azacitidine在高危早期乳腺癌中的机会窗试验
Window of opportunity trial of preoperative low dose azacitidine in high-risk early-stage breast cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：低甲基化药物如5-azacitidine（5-aza）可解除对人内源性逆转录病毒（HERV）基因的抑制，可能生成免疫原性新表位，同时逆转表观遗传性T细胞耗竭。尽管在血液系统恶性肿瘤中已被广泛研究，但5-aza作为免疫刺激剂在实体瘤中的作用，尤其是在免疫治疗难治性的luminal型和高危乳腺癌中，仍未被充分探索。我们在一项针对因高危肿瘤前来接受初次多学科根治性管理的患者的单机构机会窗临床试验中检验了这一假设。
目的：主要目的是确定低剂量5-aza治疗对高危早期乳腺癌患者肿瘤浸润淋巴细胞（TIL）的影响。相关目的包括azacitidine治疗对高危受试者免疫应答标志物的影响。
方法：UICC BRE-04（NCT04891068）是一项单机构术前"机会窗"试验，在T1b-T3期三阴性乳腺癌或伴至少一项不良特征（HER2⁺、淋巴结⁺、PR⁻或高危基因组特征）的ER⁺疾病患者（年龄18-99岁；ECOG 0-2）中，检验连续5天皮下注射低剂量5-aza 50 mg/m²的效果。保存于FFPE中的治疗前和机会窗后组织被评估用于TIL浸润变化（定量和定性）的主要结局，以及通过定量PCR测定的HERV基因表达。从时间匹配的血浆标本中测定细胞因子浓度。
结果：27例患者已完成研究治疗。在本中期报告中，来自9例患者的18份配对标本已通过多重免疫荧光进行分析。总体CD8⁺和CD4⁺ T细胞密度及亚群比例（PD-1⁻、CD45RO⁻、FOXP3⁻）未发生显著改变（所有p>0.05），然而，治疗后肿瘤显示免疫细胞与PanCK⁺肿瘤上皮细胞之间的平均距离减少了45.3%（p<0.05）。观察到瘤内和基质CD8⁺效应亚群及基质CD4⁺效应T细胞的方向性增加，而调节性T细胞未扩增，与异质但协调的免疫参与一致。qPCR HERV基因表达分析正在进行中。任何患者均未观察到严重不良事件。
结论：我们证明了在短程5-aza治疗后出现与治疗相关的免疫重塑。具体而言，效应CD8+和CD4+群体向靶肿瘤细胞更近距离迁移，尽管未观察到对总体免疫细胞数量的影响。应答具有异质性，提示其他因素可能影响5-aza在免疫启动中的有效性。整个入组队列的最终结果将在会议报告时提供。
查看英文原文 English abstract
Introduction: Hypomethylating agents such as 5-azacitadine (5-aza) can derepress human endogenous retroviral (HERV) genes and potentially generate immunogenic neoepitopes while reversing epigenetic T-cell exhaustion. Although extensively studied in hematologic malignancies, the role of 5-aza as an immune stimulant in solid tumors, particularly immunotherapy-refractory luminal and high-risk breast cancers, remains underexplored. We tested this hypothesis in a window of opportunity single-institution clinical trial in patients presenting for initial multidisciplinary curative management of high-risk tumors.
Objectives: The primary objective is to determine the effect of low dose 5-aza therapy on tumor infiltrating lymphocytes (TILs) in patients with high-risk early-stage breast cancer. Correlative objectives include the effects of azacitidine therapy in high-risk subjects on immune response markers.
Methods: UICC BRE-04 (NCT04891068) is a single-institution preoperative “window” trial testing 5 consecutive days of low-dose subcutaneous 5-aza 50 mg/m² in patients (Age 18-99; ECOG 0-2) with stage T1b-T3 triple-negative breast cancer or ER⁺ disease with at least one adverse feature (HER2⁺, node⁺, PR⁻, or high-risk genomic profile). Pre-treatment and post-window tissues preserved in FFPE were evaluated for the primary outcome of changes in TIL infiltration (quantitative and qualitative) and for HERV gene expression determined by quantitative-PCR. Cytokine concentrations were determined from time-matched plasma specimens.
Results: 27 patients have completed study treatments. At this interim report, 18 paired specimens from 9 patients have been analyzed by multiplex immunofluorescence. Overall CD8⁺ and CD4⁺ T-cell densities and subset proportions (PD-1⁻, CD45RO⁻, FOXP3⁻) were not significantly altered (all p > 0.05), however, post-treatment tumors exhibited a 45.3% reduction in the average distance between immune cells and PanCK⁺ tumor epithelial cells ( p < 0.05). Directional increases in intratumoral and stromal CD8⁺ effector subsets and stromal CD4⁺ effector T cells were observed without expansion of regulatory T cells, consistent with heterogeneous but coordinated immune engagement. qPCR HERV gene expression analysis is ongoing. No serious adverse events were observed in any patient.
Conclusion: We demonstrate treatment-associated immune remodeling following a short-course of treatment with 5-aza. Specifically, effector CD8+ and CD4+ populations migrate in closer proximity to target tumor cells even though no observed effects on overall immune cell numbers were identified. Responses were heterogenous and suggest other factors may influence the effectiveness of 5-aza in immune priming. Final results from the entire enrolled cohort will be available at the time of meeting presentation.
利益披露 Disclosure
M. Godoy Calderon, None.
A. Kodali,
Eli Lilly and Company ).
M. Patwardhan, None..
S. Sethi, None..
M. Russell, None..
E. Gauchat, None..
M. Karan, None..
Z. Chen, None.
O. Danciu,
AstraZeneca Independent Contractor, Other, Advisory board member.
Novartis Independent Contractor, ), Other, Advisory board member.
Genentech ).
Pfizer ).
Seagen ).
Sanofi ).
BioAlta ).
Biotheranostics (Hologic) Other, Advisory board member.
K. Hoskins,
Pfizer ).
Merck ).
Novartis ).
AbbVie ).
Genentech ).
Agendia Other, Nonfinancial support.
Hologic Other, Nonfinancial support.
A. Ibraheem,
Eli Lilly and Company ).
Gilead Sciences ).
Bristol Myers Squibb ).
V. Gadi,
SEngine Precision Medicine Stock, Other Business Ownership.
Novilla Stock, Other Business Ownership.
XTRemisBio Stock, Other Business Ownership.
3rdEyeBio Stock, Other Business Ownership.
Puma Biotechnology Other, Scientific advisory board member.
Phoenix Molecular Designs Other, Scientific advisory board member.
Hologic Independent Contractor.
Pfizer Independent Contractor.
Novartis Independent Contractor, ).
AstraZeneca Independent Contractor, ).
Stemline Therapeutics Independent Contractor.
Gilead Sciences Independent Contractor, ).
Daiichi Sankyo Independent Contractor.
Genentech/Roche Independent Contractor.
Bicycle Therapeutics Independent Contractor.
Agendia ).
GlaxoSmithKline ).
Illumina ).