A Functional Bioluminescent Zebrafish Screen for Enhancing Hematopoietic Cell Homing.
A Functional Bioluminescent Zebrafish Screen for Enhancing Hematopoietic Cell Homing.
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DOI：
10.1016/j.stemcr.2016.12.004
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发表时间：
2017-01-10
影响因子：
5.9
通讯作者：
Lund TC
中科院分区：
文献类型：
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作者：
Astuti Y;Kramer AC;Blake AL;Blazar BR;Tolar J;Taisto ME;Lund TC
为了发现在过继转移后调节造血细胞归巢的小分子，我们创建了在泛素启动子（ubi：luc）下游表达萤火虫荧光素酶的转基因斑马鱼以充当造血供体。生物发光成像（BLI）用于检测和跟踪早在移植后1天就归巢到骨髓的ubi：luc造血细胞。BLI能够检测前列腺素E2对供体造血细胞早期归巢/植入的生物学效应。该系统用于小分子的功能性筛选以增强归巢/植入。我们发现了一种植物甾醇，麦角甾醇，可以增加斑马鱼和小鼠的造血细胞归巢。此外，麦角固醇增加CXCR 4表达，并促进体外Lin−SCA-1+KIT+细胞的扩增。我们已经证明了体内BLI的实用性，以非侵入性监测供体造血细胞活性在成年斑马鱼作为一个功能性屏幕介质的细胞归巢。生物发光成像技术（BLI）可用于造血细胞归巢增强剂的筛选利用生物发光成像技术，发现麦角甾醇可增强造血细胞归巢，并在体外影响造血祖细胞的迁移、生长和存活，加速过继转移后的细胞归巢，可加速造血移植后的恢复。隆德及其同事开发了一种新的斑马鱼模型，利用生物发光成像跟踪造血细胞归巢，从而允许小分子文库的功能筛选方法。
To discover small molecules that modulate hematopoietic cell homing after adoptive transfer, we created a transgenic zebrafish expressing firefly luciferase downstream of the ubiquitin promoter (ubi:luc) to serve as a hematopoietic donor. Bioluminescence imaging (BLI) was used to detect and follow ubi:luc hematopoietic cells that homed to the marrow as early as 1 day post-transplant. BLI was able to detect the biological effect of prostaglandin E2 on early homing/engraftment of donor hematopoietic cells. This system was utilized in a functional screen of small molecules to enhance homing/engraftment. We discovered a phytosterol, ergosterol, that could increase hematopoietic cell homing in zebrafish and mice. In addition, ergosterol increased CXCR4 expression and promoted expansion of Lin−SCA-1+KIT+ cells in vitro. We have demonstrated the utility of in vivo BLI to non-invasively monitor donor hematopoietic cell activity in adult zebrafish as a functional screen for mediators of cellular homing. Bioluminescent imaging (BLI) can track engrafting hematopoietic cells BLI can be used for screening of enhancers of hematopoietic cell homing Using BLI, ergosterol was found to increase hematopoietic cell homing Ergosterol affects hematopoietic progenitor migration, growth, and viability in vitro Accelerating cell homing after adoptive transfer may expediate recovery after hematopoietic transplant. Lund and colleagues developed a novel zebrafish model utilizing bioluminescent imaging to track hematopoietic cell homing, thus allowing for a functional screening approach of small molecule libraries.