PO.MCB02.01 · 分子与细胞生物学
解析中性粒细胞异质性以增强TRAIL（肿瘤坏死因子相关凋亡诱导配体）在三阴性乳腺癌中的治疗疗效
Deciphering neutrophil heterogeneity to enhance therapeutic efficacy of TRAIL (Tumor necrosis factor-related apoptosis-inducing ligand) in triple negative breast cancers
该海报暂无可下载的资料
AACR 官方页面
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
TRAIL可在包括乳腺癌模型在内的多种临床前癌症模型中诱导凋亡，并作为一种潜在的癌症治疗靶点被广泛研究。然而，TRAIL激动剂的临床疗效有限，提示存在未知的调节机制导致其在患者中缺乏活性。我们前期的工作表明，在三阴性乳腺癌(TNBC)中，TRAIL通过NFKB2依赖性通路诱导多种细胞因子(CXCL1、CXCL2、CXCL3、CXCL8、CXCL11、IL6)，促进免疫抑制性中性粒细胞的募集，提示中性粒细胞介导的肿瘤免疫微环境调节可调控TRAIL在TNBC中的活性。因此，本研究旨在：1)表征由TRAIL以及经TRAIL处理的TNBC所产生因子诱导的中性粒细胞异质性；2)识别可将中性粒细胞重编程为抗肿瘤表型、从而增强TRAIL疗效的可靶向通路。为此，将从一名健康供者分离的中性粒细胞在4种不同条件下孵育：无血清培养基(SFM)、添加TRAIL的SFM(SFM-T)、来自TNBC的条件培养基(CM)，或来自经TRAIL处理的TNBC的条件培养基(T-CM)。对4种条件下的中性粒细胞(43,420个细胞)进行单细胞RNA测序(scRNA-seq)显示存在10个转录上不同的细胞簇。这些发现通过对来自5名不同健康个体的混合中性粒细胞(35,500个细胞)的scRNA-seq得到进一步验证。t-SNE分析识别出类似的转录上不同的状态，涵盖炎症、抗原呈递和核糖体生物合成程序，展示出显著的中性粒细胞异质性。基于文献和功能相关性，我们聚焦于在两个实验中均识别出的、在中性粒细胞经T-CM处理后增加的2个关键细胞簇。其一富集抗原呈递基因和免疫抑制标志物，如CCL4、CD274、IL1A和IL1B；另一个富集核糖体生物合成相关基因，包括DDX21、UTP18、LAGE3和NPM1。这些细胞簇还表现出IFN-γ/α反应性基因的低表达，进一步提示T-CM的免疫抑制作用。相比之下，SFM-T处理产生了一个具有调控中性粒细胞特异性功能标志物（如NETosis、脱颗粒）的细胞群。目前正通过基因表达分析和表面染色方法确认scRNA-seq的发现。总体而言，我们的研究阐明了经TRAIL处理的TNBC细胞上清液与中性粒细胞可塑性的潜在机制作用。这些发现提示，靶向固有免疫系统可能调节TRAIL对TNBC肿瘤的作用，从而增强治疗效果。
查看英文原文 English abstract
TRAIL induces apoptosis in many preclinical cancer models including breast cancer models and has been extensively studied as a potential cancer therapeutic target. However, the clinical efficacy of TRAIL agonists is limited, suggesting that there are unknown modulatory mechanisms responsible for the lack of activity in patients. Our prior work demonstrated that TRAIL induces various cytokines (CXCL1, CXCL2, CXCL3, CXCL8, CXCL11, IL6) via NFKB2-dependent pathway in triple negative breast cancers (TNBC), promoting the recruitment of immunosuppressive neutrophils, suggesting that neutrophil-mediated regulation of the tumor immune microenvironment modulates TRAIL activity in TNBC. Thus, this study aims 1) to characterize the neutrophil heterogeneity induced by TRAIL as well as derived factors from TRAIL treated TNBC, and 2) to identify targetable pathways that could reprogram neutrophils towards an anti-tumor phenotype, thus enhancing TRAIL efficacy. To address this, neutrophils isolated from a healthy donor were incubated with 4 different conditions: serum-free media (SFM), SFM supplemented with TRAIL (SFM-T), conditioned media from TNBC (CM), or conditioned media from TRAIL-treated TNBC (T-CM). Single cell RNA sequencing (scRNA-seq) of neutrophils (43,420 cells) across the 4 conditions indicated that there are 10 transcriptionally distinct clusters. These findings were further validated with scRNA-seq of pooled neutrophils from 5 different healthy individuals (35,500 cells). t -SNE analysis identified similar transcriptionally distinct states encompassing inflammation, antigen presentation, and ribosome biogenesis programs, demonstrating remarkable neutrophil heterogeneity. Based on literature and functional relevance, we focused on 2 key clusters that were increased upon treatment of the neutrophils with T-CM identified in both experiments. One is enriched for antigen-presentation genes and immunosuppressive markers such as CCL4 , CD274 , IL1A , and IL1B , and a second enriched for ribosome biogenesis-related genes including DDX21 , UTP18 , LAGE3 , and NPM1 . These clusters also exhibited low expression of IFN-gamma/alpha-responsive genes, further suggesting the immunosuppressive role of T-CM. In contrast, SFM-T treatment resulted in a population with markers regulating neutrophil-specific function (e.g. NETosis, degranulation). The Sc-RNA-seq findings are currently being confirmed by gene expression analysis and surface staining methods. Collectively, our study delineates a potential mechanistic role of supernatants from TRAIL treated TNBC cells and neutrophil plasticity. These findings imply that targeting the innate immune system may modulate the effects of TRAIL on TNBC tumors enhancing the therapeutic outcomes.
利益披露 Disclosure
M. Kundu Sil, None..
D. Kumar, None..
W. Wu, None..
L. Ma, None..
S. Lipkowitz, None.