Using genetically encoded fluorescent biosensors to interrogate ovarian cancer metabolism.
Using genetically encoded fluorescent biosensors to interrogate ovarian cancer metabolism.
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DOI：
10.1186/s13048-022-01046-5
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发表时间：
2022-10-20
影响因子：
4
通讯作者：
中科院分区：
文献类型：
--
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上皮性卵巢癌（OC）是最致命的妇科恶性肿瘤，患者具有显着的转移负担，特别是向富含脂肪的网膜微环境转移。最近的证据强调了代谢适应在实现这种转移中的重要性，导致人们对开发用于研究 OC 代谢的工具库产生了浓厚的兴趣。在这项研究中，我们展示了基因编码荧光生物传感器研究 OC 的能力，重点是更好地再现体内肿瘤微环境的 3D 类器官模型。将编码代谢生物传感器 HyPer、iNap、Peredox 和 Perceval 的质粒转染到 15 个卵巢癌细胞系中，分别检测氧化应激、NADPH/NADP+、NADH/NAD+ 和 ATP/ADP。荧光读数用于测定对网膜条件培养基 (OCM) 和 100μM 卡铂治疗的动态代谢反应。表达 HyPer 的 SKOV3 细胞通过活体网膜窗成像为 2D 单层、3D 类器官和体内转移瘤。我们进一步从患有卡铂耐药或卡铂敏感肿瘤的 III/IV 期 OC 患者（总共 n = 8）收集的腹水中建立了类器官。这些源自患者的类器官 (PDO) 被设计为表达 HyPer，并在 100μM 卡铂治疗期间进行氧化应激的代谢读数。使用代谢传感器测量，暴露于 OCM 或卡铂会导致 15 种 OC 细胞系发生异质代谢变化。通过转移 SKOV3-HyPer 细胞的活体成像测量体内网膜转移的氧化应激，SKOV3-HyPer 类器官比 2D 单层细胞更能准确地再现。最后，表达HyPer的PDO的卡铂治疗在来自卡铂耐药患者的类器官中诱导比来自卡铂敏感患者的类器官更高的氧化应激。我们的研究表明，生物传感器提供了一种研究 OC 临床前模型动态代谢变化的有用方法，包括 3D 类器官和活体成像。随着 OC 3D 模型的不断发展，生物传感器库可能会成为探索 OC 中具有临床重要性的代谢变化的有价值的工具。在线版本包含可在 10.1186/s13048-022-01046-5 获取的补充材料。
Epithelial ovarian cancer (OC) is the most lethal gynecological malignancy and patients present with significant metastatic burden, particularly to the adipose-rich microenvironment of the omentum. Recent evidence has highlighted the importance of metabolic adaptations in enabling this metastasis, leading to significant interest in evolving the arsenal of tools used to study OC metabolism. In this study, we demonstrate the capability of genetically encoded fluorescent biosensors to study OC, with a focus on 3D organoid models that better recapitulate in vivo tumor microenvironments. Plasmids encoding the metabolic biosensors HyPer, iNap, Peredox, and Perceval were transfected into 15 ovarian cancer cell lines to assay oxidative stress, NADPH/NADP+, NADH/NAD+, and ATP/ADP, respectively. Fluorescence readings were used to assay dynamic metabolic responses to omental conditioned media (OCM) and 100 μM carboplatin treatment. SKOV3 cells expressing HyPer were imaged as 2D monolayers, 3D organoids, and as in vivo metastases via an intravital omental window. We further established organoids from ascites collected from Stage III/IV OC patients with carboplatin-resistant or carboplatin-sensitive tumors (n = 8 total). These patient-derived organoids (PDOs) were engineered to express HyPer, and metabolic readings of oxidative stress were performed during treatment with 100 μM carboplatin. Exposure to OCM or carboplatin induced heterogenous metabolic changes in 15 OC cell lines, as measured using metabolic sensors. Oxidative stress of in vivo omental metastases, measured via intravital imaging of metastasizing SKOV3-HyPer cells, was more closely recapitulated by SKOV3-HyPer organoids than by 2D monolayers. Finally, carboplatin treatment of HyPer-expressing PDOs induced higher oxidative stress in organoids derived from carboplatin-resistant patients than from those derived from carboplatin-sensitive patients. Our study showed that biosensors provide a useful method of studying dynamic metabolic changes in preclinical models of OC, including 3D organoids and intravital imaging. As 3D models of OC continue to evolve, the repertoire of biosensors will likely serve as valuable tools to probe the metabolic changes of clinical importance in OC. The online version contains supplementary material available at 10.1186/s13048-022-01046-5.
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影响因子：
16.6
作者：
Nasu Y;Murphy-Royal C;Wen Y;Haidey JN;Molina RS;Aggarwal A;Zhang S;Kamijo Y;Paquet ME;Podgorski K;Drobizhev M;Bains JS;Lemieux MJ;Gordon GR;Campbell RE
通讯作者：
Campbell RE
影响因子：
11.2
作者：
通讯作者：
--
影响因子：
82.9
作者：
通讯作者：
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基因编码荧光传感器揭示 NADPH 代谢的动态调节
影响因子：
48
作者：
Tao R;Zhao Y;Chu H;Wang A;Zhu J;Chen X;Zou Y;Shi M;Liu R;Su N;Du J;Zhou HM;Zhu L;Qian X;Liu H;Loscalzo J;Yang Y
通讯作者：
Yang Y
DOI：
10.1016/j.medj.2021.08.005
发表时间：
2021-09-10
期刊：
Med (New York, N.Y.)
影响因子：
--
作者：
Bose S;Clevers H;Shen X
通讯作者：
Shen X