Angelos Mathew G, Ruh Paige N, Webber Beau R, Blum Robert H, Ryan Caitlin D, Bendzick Laura, Shim Seonhui, Yingst Ashley M, Tufa Dejene M, Verneris Michael R, Kaufman Dan S
Division of Hematology, Oncology, and Transplantation, Department of Medicine.
Stem Cell Institute.
Blood. 2017 Jun 29;129(26):3428-3439. doi: 10.1182/blood-2016-07-730440. Epub 2017 May 22.
The aryl hydrocarbon receptor (AHR) plays an important physiological role in hematopoiesis. AHR is highly expressed in hematopoietic stem and progenitor cells (HSPCs) and inhibition of AHR results in a marked expansion of human umbilical cord blood-derived HSPCs following cytokine stimulation. It is unknown whether AHR also contributes earlier in human hematopoietic development. To model hematopoiesis, human embryonic stem cells (hESCs) were allowed to differentiate in defined conditions in the presence of the AHR antagonist StemReginin-1 (SR-1) or the AHR agonist 2,3,7,8-tetrachlorodibenzo-dioxin (TCDD). We demonstrate a significant increase in CD34CD31 hematoendothelial cells in SR-1-treated hESCs, as well as a twofold expansion of CD34CD45 hematopoietic progenitor cells. Hematopoietic progenitor cells were also significantly increased by SR-1 as quantified by standard hematopoietic colony-forming assays. Using a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-engineered hESC--tdTomato reporter cell line with deletion, we further demonstrate a marked enhancement of hematopoietic differentiation relative to wild-type hESCs. We also evaluated whether AHR antagonism could promote innate lymphoid cell differentiation from hESCs. SR-1 increased conventional natural killer (cNK) cell differentiation, whereas TCDD treatment blocked cNK development and supported group 3 innate lymphoid cell (ILC3) differentiation. Collectively, these results demonstrate that AHR regulates early human hematolymphoid cell development and may be targeted to enhance production of specific cell populations derived from human pluripotent stem cells.
芳烃受体（AHR）在造血过程中发挥着重要的生理作用。AHR在造血干细胞和祖细胞（HSPCs）中高度表达，抑制AHR会导致细胞因子刺激后人脐带血来源的HSPCs显著扩增。目前尚不清楚AHR是否也在人类造血发育的早期阶段发挥作用。为了模拟造血过程，在存在AHR拮抗剂StemReginin-1（SR-1）或AHR激动剂2,3,7,8-四氯二苯并二恶英（TCDD）的情况下，使人类胚胎干细胞（hESCs）在特定条件下分化。我们证明，在经SR-1处理的hESCs中，CD34CD31血液内皮细胞显著增加，同时CD34CD45造血祖细胞扩增了两倍。通过标准造血集落形成试验定量分析，SR-1处理也使造血祖细胞显著增加。使用具有缺失的成簇规律间隔短回文重复序列（CRISPR）/CRISPR相关蛋白9（Cas9）工程化的hESC-tdTomato报告细胞系，我们进一步证明相对于野生型hESCs，造血分化显著增强。我们还评估了AHR拮抗作用是否能促进hESCs向先天性淋巴细胞分化。SR-1增加了传统自然杀伤（cNK）细胞的分化，而TCDD处理则阻断了cNK细胞的发育并支持3型先天性淋巴细胞（ILC3）的分化。总的来说，这些结果表明AHR调节人类早期血液淋巴细胞的发育，并且可能成为增强源自人类多能干细胞的特定细胞群体产生的靶点。
