Functional screen identifies regulators of murine hematopoietic stem cell repopulation.
Functional screen identifies regulators of murine hematopoietic stem cell repopulation.
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功能屏幕确定了鼠造血干细胞再生的调节剂。
DOI：
10.1084/jem.20150806
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发表时间：
2016-03-07
期刊：
影响因子：
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通讯作者：
McKinney-Freeman S
中科院分区：
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作者：
Holmfeldt P;Ganuza M;Marathe H;He B;Hall T;Kang G;Moen J;Pardieck J;Saulsberry AC;Cico A;Gaut L;McGoldrick D;Finkelstein D;Tan K;McKinney-Freeman S
霍姆菲尔德等人。进行基于移植的筛选以确定 HSPC 植入的调节因子，并报告 Foxa3 对于移植后最佳 HSC 功能至关重要。了解造血干细胞和祖细胞 (HSPC) 植入的分子调控对于改善移植结果至关重要。为了发现 HSPC 再增殖的新调节因子，我们在分离和转导后 24 小时内将 shRNA 转导的 HSPC 移植到超过 1,300 只小鼠中，以重点检测调节再增殖的基因。我们鉴定了 17 个 HSPC 重新增殖的调节因子：Arhgef5、Armcx1、Cadps2、Crispld1、Emcn、Foxa3、Fstl1、Glis2、Gprasp2、Gpr56、Myct1、Nbea、P2ry14、Smarca2、Sox4、Stat4 和 Zfp521。这些基因的敲除都会导致功能丧失，但 Armcx1 和 Gprasp2 除外，它们的丧失增强了造血干细胞 (HSC) 的重新增殖。调节囊泡运输、细胞表面受体周转和细胞外基质成分分泌的多个基因的发现表明，HSC 和微环境之间存在活跃的交互作用，并且 HSC 可能主动调节微环境以促进植入。我们验证了 Foxa3 是 HSC 重新增殖活动所必需的，因为 Foxa3−/− HSC 无法有效地重新增殖被消融的宿主，这首次表明 Foxa 基因作为 HSPC 的调节因子。我们进一步表明 Foxa3 可能调节 HSC 对血液应激的反应。这里发现的每个基因都为了解调节 HSPC 稳定植入消融宿主的新过程提供了一个窗口。
Holmfeldt et al. perform a transplant-based screen to identify regulators of HSPC engraftment and report that Foxa3 is critical for optimal HSC function after transplant. Understanding the molecular regulation of hematopoietic stem and progenitor cell (HSPC) engraftment is paramount to improving transplant outcomes. To discover novel regulators of HSPC repopulation, we transplanted >1,300 mice with shRNA-transduced HSPCs within 24 h of isolation and transduction to focus on detecting genes regulating repopulation. We identified 17 regulators of HSPC repopulation: Arhgef5, Armcx1, Cadps2, Crispld1, Emcn, Foxa3, Fstl1, Glis2, Gprasp2, Gpr56, Myct1, Nbea, P2ry14, Smarca2, Sox4, Stat4, and Zfp521. Knockdown of each of these genes yielded a loss of function, except in the cases of Armcx1 and Gprasp2, whose loss enhanced hematopoietic stem cell (HSC) repopulation. The discovery of multiple genes regulating vesicular trafficking, cell surface receptor turnover, and secretion of extracellular matrix components suggests active cross talk between HSCs and the niche and that HSCs may actively condition the niche to promote engraftment. We validated that Foxa3 is required for HSC repopulating activity, as Foxa3−/− HSC fails to repopulate ablated hosts efficiently, implicating for the first time Foxa genes as regulators of HSPCs. We further show that Foxa3 likely regulates the HSC response to hematologic stress. Each gene discovered here offers a window into the novel processes that regulate stable HSPC engraftment into an ablated host.