Intestinal UDP-Glucuronosyltransferase 1A1 and Protection against Irinotecan-Induced Toxicity in a Novel UDP-Glucuronosyltransferase 1A1 Tissue-Specific Humanized Mouse Model
Intestinal UDP-Glucuronosyltransferase 1A1 and Protection against Irinotecan-Induced Toxicity in a Novel UDP-Glucuronosyltransferase 1A1 Tissue-Specific Humanized Mouse Model
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DOI：
10.1124/dmd.121.000644
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发表时间：
2021-10
影响因子：
3.9
通讯作者：
Elvira Mennillo;Xiaojing Yang;A. Weber;Y. Maruo;Mélanie Verreault;O. Barbier;Shujuan Chen;R. Tukey
中科院分区：
文献类型：
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作者：
Elvira Mennillo;Xiaojing Yang;A. Weber;Y. Maruo;Mélanie Verreault;O. Barbier;Shujuan Chen;R. Tukey
人UDP-葡萄糖醛酸基转移酶（UGT）是一个重要的药物代谢酶家族，其中UGT 1A 1靶向许多外源性物质（包括药物）的结合和解毒。在这项研究中，我们产生了在肝脏（hUGT 1A 1HEP）或肠道（hUGT 1A 1GI）中表达人UGT 1A 1基因的人源化UGT 1A 1小鼠，使实验能够检查UGT 1A 1特异性葡萄糖醛酸化的组织特异性特性。证实了UGT 1A 1的肝和肠组织特异性表达和功能。虽然肝脏被认为是解毒的主要器官，但肠道UGT 1A 1是药物清除的重要贡献者。用伊立替康（CPT-11）激发小鼠，伊立替康是一种前药，可被羧酸酯酶水解形成活性代谢产物7-乙基-10-羟基喜树碱（SN-38），并被UGT 1A 1解毒。当暴露于CPT-11时，没有肠道UGT 1A 1的人源化UGT 1A 1HEP小鼠显示出比hUGT 1A 1GI小鼠更高的致死率。当暴露于低剂量CPT-11（10 mg/kg）时，除了p53触发的凋亡反应外，hUGT 1A 1HEP小鼠还显示出更大的肠道炎症（IL-1β和IL-6）损伤。暴露于CPT-11的肠隐窝类器官的体外研究证实了体内观察到的结果，并表明CPT-11影响UGT 1A 1缺陷类器官中的干性、细胞凋亡和内质网（ER）应激。当我们用毒胡萝卜素（一种肌质/内质网Ca 2 + ATP酶抑制剂）检测类器官中ER应激的诱导时，hUGT 1A 1HEP小鼠中发生的细胞凋亡和半胱天冬酶峰在hUGT 1A 1GI类器官中被阻断。这项研究揭示了肠道UGT 1A 1在预防炎症、细胞凋亡和干细胞能力丧失方面的重要性，这些都是在用一种重要的化疗药物进行全身性激发后发生的。肝和肠UGT 1A 1在内源性和外源性化合物的代谢和解毒中起关键作用。在肝脏或肠道中表达UGT 1A 1的组织特异性人源化模型的使用证实了肠道在伊立替康解毒中的相关性。使用肠类器官的机制研究强调了UGT 1A 1在减少炎症、细胞凋亡和干性丧失方面的重要性。这些新模型为研究人UGT 1A 1代谢物质的组织特异性葡萄糖醛酸化提供了有价值的工具。
The human UDP-glucuronosyltransferases (UGTs) represent an important family of drug-metabolizing enzymes, with UGT1A1 targeting the conjugation and detoxification of many exogenous substances, including pharmaceutical drugs. In this study we generated humanized UGT1A1 mice expressing the human UGT1A1 gene in either liver (hUGT1A1HEP) or intestine (hUGT1A1GI), enabling experiments to examine tissue-specific properties of UGT1A1-specific glucuronidation. Hepatic and intestinal tissue-specific expression and function of UGT1A1 were demonstrated. Although the liver is considered a major organ for detoxification, intestinal UGT1A1 is an important contributor for drug clearance. Mice were challenged with irinotecan (CPT-11), a prodrug hydrolyzed by carboxylesterases to form the active metabolite 7-ethyl-10-hydroxycamptothecin (SN-38) and detoxified by UGT1A1. Humanized UGT1A1HEP mice that have no intestinal UGT1A1 displayed a greater lethality rate when exposed to CPT-11 than hUGT1A1GI mice. When exposed to a low dose of CPT-11 (10 mg/kg), hUGT1A1HEP mice displayed greater intestinal inflammatory (IL-1β and IL-6) insult in addition to p53-triggered apoptotic responses. In vitro studies with intestinal crypt organoids exposed to CPT-11 confirmed the results observed in vivo and indicated that CPT-11 impacts stemness, apoptosis, and endoplasmic reticulum (ER) stress in organoids deficient in UGT1A1. When we examined the induction of ER stress in organoids with thapsigargin, an inhibitor of sarco/endoplasmic reticulum Ca2+ ATPase, apoptosis and the caspase surge that occurred in hUGT1A1HEP mice were blocked in hUGT1A1GI organoids. This study reveals the importance of intestinal UGT1A1 in preventing inflammation, apoptosis, and loss of stemness capacity upon systemic challenge with an important chemotherapeutic agent. SIGNIFICANCE STATEMENT Hepatic and intestinal UGT1A1 play a key role in the metabolism and detoxification of endogenous and exogenous compounds. The use of tissue-specific humanized models expressing UGT1A1 in liver or intestine has confirmed the relevance of the intestinal tract in the detoxification of irinotecan. Mechanistic studies using intestinal organoids highlighted the importance of UGT1A1 in reducing inflammation, apoptosis, and loss of stemness. These new models provide valuable tools for studying tissue-specific glucuronidation of substances that are metabolized by human UGT1A1.