Snapshots of nascent RNA reveal cell- and stimulus-specific responses to acute kidney injury.
Snapshots of nascent RNA reveal cell- and stimulus-specific responses to acute kidney injury.
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DOI：
10.1172/jci.insight.146374
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发表时间：
2022-03-22
期刊：
影响因子：
8
通讯作者：
Barasch J
中科院分区：
文献类型：
--
作者：
Shen TH;Stauber J;Xu K;Jacunski A;Paragas N;Callahan M;Banlengchit R;Levitman AD;Desanti De Oliveira B;Beenken A;Grau MS;Mathieu E;Zhang Q;Li Y;Gopal T;Askanase N;Arumugam S;Mohan S;Good PI;Stevens JS;Lin F;Sia SK;Lin CS;D'Agati V;Kiryluk K;Tatonetti NP;Barasch J
目前检测肾小管细胞急性损伤的策略依赖于血清肌酐水平的变化。然而，血清肌酐（sCr）是功能和病理过程的标志，不能充分测定肾小管损伤。此外，sCr可能需要数天才能达到诊断阈值，而管状细胞在几分钟或几小时内就能对损伤和修复程序做出反应。为了检测对临床相关刺激的急性反应，我们创造了表达rosa26 - floxd -stop尿嘧啶磷酸核糖基转移酶（Uprt）的小鼠，并在选定的时间点接种4-硫脲嘧啶（4-TU）来标记新生RNA。从完整肾脏中分离出由cred驱动的4- tu标记RNA，结果表明，体积减少和缺血在收集管和插层细胞中诱导了不同的遗传程序。甚至与谱系相关的细胞类型也表达了不同的基因来响应这两种应激源。TU标记也证明了响应的瞬态性质。由于我们将Uprt放置在普遍活跃的Rosa26位点上，因此可以在体内标记来自许多细胞类型的新生rna，并在各种条件下询问它们的作用。简而言之，4-TU标记识别刺激特异性、细胞特异性和时间依赖性的急性反应，否则很难用其他技术检测到，当sCr是肾脏损伤的唯一指标时，这些急性反应完全被掩盖了。
The current strategy to detect acute injury of kidney tubular cells relies on changes in serum levels of creatinine. Yet serum creatinine (sCr) is a marker of both functional and pathological processes and does not adequately assay tubular injury. In addition, sCr may require days to reach diagnostic thresholds, yet tubular cells respond with programs of damage and repair within minutes or hours. To detect acute responses to clinically relevant stimuli, we created mice expressing Rosa26-floxed-stop uracil phosphoribosyltransferase (Uprt) and inoculated 4-thiouracil (4-TU) to tag nascent RNA at selected time points. Cre-driven 4-TU–tagged RNA was isolated from intact kidneys and demonstrated that volume depletion and ischemia induced different genetic programs in collecting ducts and intercalated cells. Even lineage-related cell types expressed different genes in response to the 2 stressors. TU tagging also demonstrated the transient nature of the responses. Because we placed Uprt in the ubiquitously active Rosa26 locus, nascent RNAs from many cell types can be tagged in vivo and their roles interrogated under various conditions. In short, 4-TU labeling identifies stimulus-specific, cell-specific, and time-dependent acute responses that are otherwise difficult to detect with other technologies and are entirely obscured when sCr is the sole metric of kidney damage.