Stable isotope tracing to assess tumor metabolism in vivo.
Stable isotope tracing to assess tumor metabolism in vivo.
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DOI：
10.1038/s41596-021-00605-2
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发表时间：
2021-11
期刊：
影响因子：
14.8
通讯作者：
DeBerardinis, Ralph J.
中科院分区：
文献类型：
--
作者：
Faubert, Brandon;Tasdogan, Alpaslan;Morrison, Sean J.;Mathews, Thomas P.;DeBerardinis, Ralph J.
癌细胞经历不同的代谢适应以满足由失调的生长和增殖施加的能量需求。评估完整肿瘤中的代谢允许研究者观察在培养模型中不能完全捕获的许多癌细胞内在和外在因素的组合代谢效应。我们已经开发了使用稳定的同位素标记的营养素的方法（例如，[13C]葡萄糖）来探测小鼠和人类体内完整肿瘤内的代谢活性。在这些方法中，在手术切除肿瘤和邻近的非恶性组织之前，通过静脉导管将标记的营养素引入循环。在输注过程中，这些组织内的代谢将同位素标记从输注营养素提供的途径转移到代谢中间体中。从手术标本中提取代谢物并分析其同位素标记模式提供了有关组织中代谢的信息。我们提供了有关这项技术的详细信息，从引入标记示踪剂通过数据分析和解释，包括简化的方法来量化从组织样本中提取的信息代谢物中的同位素标记。我们专注于输注[13 C]葡萄糖和质谱的应用，以评估同位素标记的中间体从中央代谢途径，包括糖酵解，三羧酸循环和非必需氨基酸的合成。我们概述了将这些方法应用于接受实体瘤手术切除的人类受试者的实际考虑。我们还讨论了该方法的多功能性，并考虑替代方法的相对优势和局限性，引入示踪剂，收获组织和分析数据。
Cancer cells undergo diverse metabolic adaptations to meet the energetic demands imposed by dysregulated growth and proliferation. Assessing metabolism in intact tumors allows the investigator to observe the combined metabolic effects of numerous cancer cell-intrinsic and -extrinsic factors that cannot be fully captured in culture models. We have developed methods to use stable isotope-labeled nutrients (e.g., [13C]glucose) to probe metabolic activity within intact tumors in vivo, in mice and humans. In these methods, the labeled nutrient is introduced to the circulation through an intravenous catheter prior to surgical resection of the tumor and adjacent nonmalignant tissue. Metabolism within these tissues during the infusion transfers the isotope label into metabolic intermediates from pathways supplied by the infused nutrient. Extracting metabolites from surgical specimens and analyzing their isotope labeling patterns provides information about metabolism in the tissue. We provide detailed information about this technique, from introduction of the labeled tracer through data analysis and interpretation, including streamlined approaches to quantify isotope labeling in informative metabolites extracted from tissue samples. We focus on infusions with [13C]glucose and the application of mass spectrometry to assess isotope labeling in intermediates from central metabolic pathways, including glycolysis, the tricarboxylic acid cycle and nonessential amino acid synthesis. We outline practical considerations to apply these methods to human subjects undergoing surgical resections of solid tumors. We also discuss the method’s versatility and consider the relative advantages and limitations of alternative approaches to introduce the tracer, harvest the tissue and analyze the data.
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影响因子：
29
作者：
Marin-Valencia I;Yang C;Mashimo T;Cho S;Baek H;Yang XL;Rajagopalan KN;Maddie M;Vemireddy V;Zhao Z;Cai L;Good L;Tu BP;Hatanpaa KJ;Mickey BE;Matés JM;Pascual JM;Maher EA;Malloy CR;Deberardinis RJ;Bachoo RM
通讯作者：
Bachoo RM
影响因子：
3.3
作者：
Romijn, JA;Coyle, EF;Wolfe, RR
通讯作者：
Wolfe, RR
影响因子：
50.3
作者：
Momcilovic M;Bailey ST;Lee JT;Fishbein MC;Braas D;Go J;Graeber TG;Parlati F;Demo S;Li R;Walser TC;Gricowski M;Shuman R;Ibarra J;Fridman D;Phelps ME;Badran K;St John M;Bernthal NM;Federman N;Yanagawa J;Dubinett SM;Sadeghi S;Christofk HR;Shackelford DB
通讯作者：
Shackelford DB
影响因子：
64.5
作者：
Faubert B;Li KY;Cai L;Hensley CT;Kim J;Zacharias LG;Yang C;Do QN;Doucette S;Burguete D;Li H;Huet G;Yuan Q;Wigal T;Butt Y;Ni M;Torrealba J;Oliver D;Lenkinski RE;Malloy CR;Wachsmann JW;Young JD;Kernstine K;DeBerardinis RJ
通讯作者：
DeBerardinis RJ
代谢组学和同位素跟踪。
影响因子：
64.5
作者：
Jang C;Chen L;Rabinowitz JD
通讯作者：
Rabinowitz JD