Genetic screens reveal CCDC115 as a modulator of erythroid iron and heme trafficking.
Genetic screens reveal CCDC115 as a modulator of erythroid iron and heme trafficking.
复制标题
基因筛选显示 CCDC115 是红细胞铁和血红素运输的调节剂。
DOI：
10.1002/ajh.25899
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发表时间：
2020
影响因子：
12.8
通讯作者：
Vulpe,ChristopherD
中科院分区：
文献类型：
--
作者：
Sobh,Amin;Loguinov,Alex;Zhou,Jie;Jenkitkasemwong,Supak;Zeidan,Rola;ElAhmadie,Nader;Tagmount,Abderrahmane;Knutson,Mitchell;Fraenkel,PaulaG;Vulpe,ChristopherD
转铁蛋白结合铁（TBI）是一种生理循环铁形式，由细胞通过转铁蛋白受体（TfR 1）通过内吞作用获得。在红系细胞中，大多数获得的铁被并入线粒体中的血红素中。血红素的细胞运输对于红细胞生成和许多其他基本生物过程是必不可少的。为了更好地理解影响红细胞生成的生理和病理过程，需要全面阐明控制和调节细胞铁获得和血红素运输的分子途径。在这里，我们报告了人类红细胞中的第一个全基因组聚集的规则间隔短回文重复序列（CRISPR）筛选，以确定铁和血红素摄取的决定因素，以及血红素介导的红细胞分化。我们确定了几个候选调制器TBI收购包括TfR 1，表明我们的方法有效地揭示了球员机械相关的过程。有趣的是，内吞途径的成分也被揭示为转铁蛋白收购的潜在决定因素。我们破译了空泡型H+ − ATP酶（V-ATP酶）组装因子卷曲螺旋结构域（CCDC 115）在TBI摄取中的作用，并在CCDC 115缺陷的K562细胞中验证了这一作用。我们在氯化血红素处理的细胞中的筛选揭示了导致细胞适应血红素的扰动，包括那些对应于运输机制和增强红细胞分化的转录因子的扰动。途径分析表明，内吞和囊泡酸化是血红素运输到红系前体细胞的关键过程。此外，我们提供的证据表明，我们确定为TBI摄取所需的CCDC 115也参与细胞血红素分布。这项工作证明了一个以前未得到重视的共同的交叉运输转铁蛋白，铁和血红素的红细胞内吞途径。
Transferrin‐bound iron (TBI), the physiological circulating iron form, is acquired by cells through the transferrin receptor (TfR1) by endocytosis. In erythroid cells, most of the acquired iron is incorporated into heme in the mitochondria. Cellular trafficking of heme is indispensable for erythropoiesis and many other essential biological processes. Comprehensive elucidation of molecular pathways governing and regulating cellular iron acquisition and heme trafficking is required to better understand physiological and pathological processes affecting erythropoiesis. Here, we report the first genome‐wide clustered regularly interspaced short palindromic repeats (CRISPR) screens in human erythroid cells to identify determinants of iron and heme uptake, as well as heme‐mediated erythroid differentiation. We identified several candidate modulators of TBI acquisition including TfR1, indicating that our approach effectively revealed players mechanistically relevant to the process. Interestingly, components of the endocytic pathway were also revealed as potential determinants of transferrin acquisition. We deciphered a role for the vacuolar‐type H+ − ATPase (V‐ ATPase) assembly factor coiled‐coil domain containing 115 (CCDC115) in TBI uptake and validated this role in CCDC115 deficient K562 cells. Our screen in hemin‐treated cells revealed perturbations leading to cellular adaptation to heme, including those corresponding to trafficking mechanisms and transcription factors potentiating erythroid differentiation. Pathway analysis indicated that endocytosis and vesicle acidification are key processes for heme trafficking in erythroid precursors. Furthermore, we provided evidence that CCDC115, which we identified as required for TBI uptake, is also involved in cellular heme distribution. This work demonstrates a previously unappreciated common intersection in trafficking of transferrin iron and heme in the endocytic pathway of erythroid cells.