Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models.
Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models.
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DOI：
10.3390/cancers13061429
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发表时间：
2021-03-20
期刊：
影响因子：
5.2
通讯作者：
Sant S
中科院分区：
文献类型：
--
作者：
Ardila DC;Aggarwal V;Singh M;Chattopadhyay A;Chaparala S;Sant S
本研究的目的是探讨两种微环境因素，即肿瘤内生性缺氧和分泌组在诱导集体迁移中的作用。我们利用三维（3D）离散大小的微肿瘤模型，概括了导管原位癌（DCIS）向浸润性导管癌（IDC）过渡的特征。肿瘤内禀缺氧诱导大缺氧微肿瘤的定向迁移，而大微肿瘤分泌组诱导非缺氧微肿瘤的径向迁移。这突出了癌细胞在响应不同微环境刺激时出现的表型异质性和可塑性。为了揭示这两种不同迁移模式的机制，我们使用非迁移、非缺氧的微肿瘤作为对照，对缺氧和分泌组诱导的迁移表型进行了差异基因表达分析。我们提出了与肿瘤内生性缺氧、缺氧诱导的上皮-间质转化（EMT）以及缺氧诱导的定向迁移和分泌组诱导的径向迁移相关的独特基因标记集。集体细胞迁移是导管原位癌（DCIS）向浸润性导管癌（IDC）过渡的关键特征，但引发集体迁移的微环境因素和潜在机制尚不清楚。在这里，我们研究了两个微环境因素，肿瘤内生性缺氧和肿瘤分泌因子（分泌组），作为集体迁移的触发因素，使用三维（3D）离散大小的微肿瘤模型，概括了DCIS-IDC过渡的特征。有趣的是，这两个因素在同一乳腺癌细胞系模型T47D产生的三维微肿瘤中诱导了两种不同的集体迁移模式：定向和径向迁移。在没有外界刺激的情况下，大的（600µm） T47D微肿瘤表现出肿瘤内禀缺氧和定向迁移，而小的（150µm）非缺氧微肿瘤只有在暴露于大微肿瘤分泌组时才表现出径向迁移。为了研究缺氧和分泌组诱导的定向和径向迁移模式的机制，我们对缺氧和分泌组诱导的迁移微肿瘤与非缺氧、非迁移的小微肿瘤进行了差异基因表达分析。我们提出了与肿瘤内生性缺氧、缺氧诱导的上皮-间质转化（EMT）以及缺氧诱导的定向迁移和分泌组诱导的径向迁移相关的独特基因标记集。基因集富集分析（GSEA）和蛋白-蛋白相互作用（PPI）网络分析揭示了缺氧、EMT和迁移基因特征之间的富集和潜在相互作用，这是缺氧诱导的定向迁移的主要原因。相比之下，缺氧和EMT在分泌组诱导的径向迁移中不富集，这表明完全的EMT可能不需要径向迁移。生存分析从肿瘤-内在缺氧基因标记（CXCR4, FOXO3， LDH, NDRG1），缺氧诱导的EMT基因标记（EFEMP2， MGP）和定向迁移基因标记（MAP3K3, PI3K3R3）中确定了tcga -乳腺浸润性癌数据集中与低生存率和不良预后相关的独特基因。NOS3在缺氧和迁移基因标记之间是共同的。从分泌组诱导的径向迁移的生存分析中发现，ATM、KCNMA1（缺氧基因特征）和KLF4、IFITM1、EFNA1、TGFBR1（迁移基因特征）与低生存率相关。综上所述，我们独特的三维微环境控制培养物通过不同的集体迁移模式响应不同的微环境因素、肿瘤内生性缺氧和分泌组，其基因表达分析突出了上皮癌细胞的表型异质性和可塑性。
The objective of this study was to investigate the role of two microenvironmental factors, namely, tumor-intrinsic hypoxia and secretome in inducing collective migration. We utilized three-dimensional (3D) discrete-sized microtumor models, which recapitulate hallmarks of transition of ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC). Tumor-intrinsic hypoxia induced directional migration in large hypoxic microtumors while secretome from large microtumors induced radial migration in non-hypoxic microtumors. This highlights the emergence phenotypic heterogeneity and plasticity in cancer cells in response to different microenvironmental stimuli. To unravel mechanisms underlying these two distinct modes of migration, we performed differential gene expression analysis of hypoxia- and secretome-induced migratory phenotypes using non-migratory, non-hypoxic microtumors as controls. We proposed unique gene signature sets related to tumor-intrinsic hypoxia, hypoxia-induced epithelial-mesenchymal transition (EMT), as well as hypoxia-induced directional migration and secretome-induced radial migration. Collective cell migration is a key feature of transition of ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) among many other cancers, yet the microenvironmental factors and underlying mechanisms that trigger collective migration remain poorly understood. Here, we investigated two microenvironmental factors, tumor-intrinsic hypoxia and tumor-secreted factors (secretome), as triggers of collective migration using three-dimensional (3D) discrete-sized microtumor models that recapitulate hallmarks of DCIS-IDC transition. Interestingly, the two factors induced two distinct modes of collective migration: directional and radial migration in the 3D microtumors generated from the same breast cancer cell line model, T47D. Without external stimulus, large (600 µm) T47D microtumors exhibited tumor-intrinsic hypoxia and directional migration, while small (150 µm), non-hypoxic microtumors exhibited radial migration only when exposed to the secretome of large microtumors. To investigate the mechanisms underlying hypoxia- and secretome-induced directional vs. radial migration modes, we performed differential gene expression analysis of hypoxia- and secretome-induced migratory microtumors compared with non-hypoxic, non-migratory small microtumors as controls. We propose unique gene signature sets related to tumor-intrinsic hypoxia, hypoxia-induced epithelial-mesenchymal transition (EMT), as well as hypoxia-induced directional migration and secretome-induced radial migration. Gene Set Enrichment Analysis (GSEA) and protein-protein interaction (PPI) network analysis revealed enrichment and potential interaction between hypoxia, EMT, and migration gene signatures for the hypoxia-induced directional migration. In contrast, hypoxia and EMT were not enriched in the secretome-induced radial migration, suggesting that complete EMT may not be required for radial migration. Survival analysis identified unique genes associated with low survival rate and poor prognosis in TCGA-breast invasive carcinoma dataset from our tumor-intrinsic hypoxia gene signature (CXCR4, FOXO3, LDH, NDRG1), hypoxia-induced EMT gene signature (EFEMP2, MGP), and directional migration gene signature (MAP3K3, PI3K3R3). NOS3 was common between hypoxia and migration gene signature. Survival analysis from secretome-induced radial migration identified ATM, KCNMA1 (hypoxia gene signature), and KLF4, IFITM1, EFNA1, TGFBR1 (migration gene signature) to be associated with poor survival rate. In conclusion, our unique 3D cultures with controlled microenvironments respond to different microenvironmental factors, tumor-intrinsic hypoxia, and secretome by adopting distinct collective migration modes and their gene expression analysis highlights the phenotypic heterogeneity and plasticity of epithelial cancer cells.
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影响因子：
21.3
作者：
Ilina O;Gritsenko PG;Syga S;Lippoldt J;La Porta CAM;Chepizhko O;Grosser S;Vullings M;Bakker GJ;Starruß J;Bult P;Zapperi S;Käs JA;Deutsch A;Friedl P
通讯作者：
Friedl P
影响因子：
64.5
作者：
Cheung KJ;Gabrielson E;Werb Z;Ewald AJ
通讯作者：
Ewald AJ
DOI：
10.1039/c9an01358j
发表时间：
2019-12-02
期刊：
The Analyst
影响因子：
--
作者：
Chen YC ;Sahoo S ;Brien R ;Jung S ;Humphries B ;Lee W ;Cheng YH ;Zhang Z ;Luker KE ;Wicha MS ;Luker GD ;Yoon E
通讯作者：
Yoon E
影响因子：
4
作者：
Dillekas, Hanna;Rogers, Michael S.;Straume, Oddhjorn
通讯作者：
Straume, Oddhjorn
影响因子：
4.5
作者：
Chen JL;Lucas JE;Schroeder T;Mori S;Wu J;Nevins J;Dewhirst M;West M;Chi JT
通讯作者：
Chi JT