PO.TB09.03 · 肿瘤生物学
用于神经母细胞瘤细胞状态可视化的新型报告工具
Novel reporter tool for visualization of neuroblastoma cell state
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
超过50%被诊断为神经母细胞瘤（NBL）的患者会因治疗耐药性肿瘤而复发。NBL复发的5年生存率不足10%，因此迫切需要理解这些肿瘤是如何复发的。NBL肿瘤具有异质性，由源自增殖性交感神经母细胞的两大主要细胞群体组成：间充质型（MES）和肾上腺素能型（ADRN）。核心假说是MES细胞群体能够逃避标准治疗，并在NBL中重建肿瘤复发。为更好地理解肿瘤复发过程中ADRN和MES细胞状态的动态，我们设计了一种报告工具来可视化细胞状态。利用一段驱动ADRN或MES细胞状态的超级增强子小区域，置于最小启动子和绿色荧光蛋白（GFP）上游，这些报告工具在成熟的细胞系中显示出前景。我们构建了四种报告工具，两种特异于ADRN细胞，两种特异于MES细胞。使用流式细胞术、蛋白质印迹分析和qPCR对这些报告工具进行验证，结果显示MES报告工具稳健且特异，而ADRN报告工具在两种细胞状态下均组成性激活。此外，这些报告工具在患者来源的细胞系中进行了测试。初步数据显示，MES报告工具在这些细胞系中可能同样有效。通过这些研究，我们预期能够确定对化疗药物耐药的细胞群体，并识别驱动肿瘤复发的细胞状态。目前迫切需要理解复发性NBL肿瘤的细胞构成和特征。这些研究旨在确定肿瘤如何重新增殖，并可能带来治疗复发性NBL肿瘤的新方法。
St. Jude - 机密
查看英文原文 English abstract
Over fifty percent of patients diagnosed with neuroblastoma (NBL) will relapse with therapy resistant tumors. The 5-year survival rate of NBL relapse is less than 10%, leaving a desperate need to understand how these tumors recur. NBL tumors are heterogeneous and made up of two major cell populations derived from proliferating sympathoblasts: mesenchymal (MES) and adrenergic (ADRN). The central hypothesis is that the MES cell population evades standard of care treatments and re-establishes tumor recurrence in NBL. To better understand the dynamic of ADRN and MES cell states in tumor recurrence, a reporter was designed to visualize cell states. Using a small region of super-enhancers driving ADRN or MES cell states upstream of a minimal promoter and green fluorescent protein (GFP), these reporters show promise in well-established cell lines. Four reporters were created, two specific to ADRN cells and two specific to MES cells. Validation of these reporters using flow cytometry, western blot analysis, and qPCR showed that MES reporters were robust and specific, while ADRN were constitutively active in both cell sates. Additionally, these reporters were tested in patient-derived cell lines. Preliminary data shows that the MES reporters may also function in these lines. From these studies, we anticipate determining the cell population resistant to chemotherapeutics and identify the cell state that drives tumor recurrence. There is a drastic need for understanding the cellular makeup and features of relapsed NBL tumors. These studies are designed to determine how the tumors repopulate and may lead to a novel way to treat relapsed NBL tumors.
St. Jude - Confidential
利益披露 Disclosure
S. Turk, None..
M. Allensworth, None..
J. McEvoy, None..
M. Lupo, None..
J. Norrie, None.