Spatially Resolved Single-Cell Assessment of Pancreatic Cancer Expression Subtypes Reveals Co-expressor Phenotypes and Extensive Intratumoral Heterogeneity.
Spatially Resolved Single-Cell Assessment of Pancreatic Cancer Expression Subtypes Reveals Co-expressor Phenotypes and Extensive Intratumoral Heterogeneity.
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DOI：
10.1158/0008-5472.can-22-3050
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发表时间：
2023-02-03
期刊：
影响因子：
11.2
通讯作者：
Wolpin, Brian M.
中科院分区：
文献类型：
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作者：
Williams, Hannah L.;Costa, Andressa Dias;Zhang, Jinming;Raghavan, Srivatsan;Winter, Peter S.;Kapner, Kevin S.;Ginebaugh, Scott P.;Vayrynen, Sara A.;Vayrynen, Juha P.;Yuan, Chen;Navia, Andrew W.;Wang, Junning;Yang, Annan;Bosse, Timothy L.;Kalekar, Radha L.;Lowder, Kristen E.;Lau, Mai Chan;Elganainy, Dalia;Morales-Oyarvide, Vicente;Rubinson, Douglas A.;Singh, Harshabad;Perez, Kimberly;Cleary, James M.;Clancy, Thomas E.;Wang, Jiping;Mancias, Joseph D.;Brais, Lauren K.;Hill, Emma R.;Kozak, Margaret M.;Linehan, David C.;Dunne, Richard F.;Chang, Daniel T.;Koong, Albert C.;Hezel, Aram F.;Hahn, William C.;Shalek, Alex K.;Aguirre, Andrew J.;Nowak, Jonathan A.;Wolpin, Brian M.
通过大量 RNA 测量，胰腺导管腺癌 (PDAC) 被分为经典转录亚型和基底样转录亚型。然而，最近的工作发现转录亚型的复杂性比最初使用批量 RNA 表达谱分析所认识到的要复杂得多。为了更深入地了解 PDAC 亚型，我们开发了多重免疫荧光 (mIF) 流程，可量化六种 PDAC 亚型标记物（CLDN18.2、TFF1、GATA6、KRT17、KRT5 和 S100A2）的蛋白质表达，并允许对切除标本和空心针活检中的胰腺肿瘤进行空间分辨、单细胞研究。原发性肿瘤和转移性肿瘤都表现出惊人的肿瘤内亚型异质性，这种异质性与患者的预后相关，存在于个体腺体的范围内，并且在患者来源的类器官培养物中显着减少。共表达经典和基础标记物的肿瘤细胞存在于 >90% 的肿瘤中，存在于基础-经典极化连续体上，并且在包含更多基础和经典细胞群混合的肿瘤中富集。肿瘤腺体内的细胞-细胞邻近分析进一步表明，共表达细胞可能代表表达亚型极之间的中间状态。通过这一临床适用的 mIF 流程确定的广泛的肿瘤内异质性可以为 PDAC 患者的预后和治疗选择提供信息。
Pancreatic ductal adenocarcinoma (PDAC) has been classified into classical and basal-like transcriptional subtypes by bulk RNA measurements. However, recent work has uncovered greater complexity to transcriptional subtypes than was initially appreciated using bulk RNA expression profiling. To provide a deeper understanding of PDAC subtypes, we developed a multiplex immunofluorescence (mIF) pipeline that quantifies protein expression of six PDAC subtype markers (CLDN18.2, TFF1, GATA6, KRT17, KRT5, and S100A2) and permits spatially resolved, single-cell interrogation of pancreatic tumors from resection specimens and core needle biopsies. Both primary and metastatic tumors displayed striking intra-tumoral subtype heterogeneity that was associated with patient outcomes, existed at the scale of individual glands, and was significantly reduced in patient-derived organoid cultures. Tumor cells co-expressing classical and basal markers were present in >90% of tumors, existed on a basal-classical polarization continuum, and were enriched in tumors containing a greater admixture of basal and classical cell populations. Cell-cell neighbor analyses within tumor glands further suggested that co-expressor cells may represent an intermediate state between expression subtype poles. The extensive intra-tumoral heterogeneity identified through this clinically applicable mIF pipeline may inform prognosis and treatment selection for patients with PDAC.