Coordinate regulation of erythroid and macrophage lineages in development by EKLF/KLF1

EKLF/KLF1 对发育中红细胞和巨噬细胞谱系的协调调节

• 批准号: 10553699
• 负责人: JAMES J BIEKER
• 金额: $ 48.68万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553699
• 关键词: Ablation; Address; Adult; Affect; Anemia; Biological Assay; Blood Cells; Blood Island; Cell Communication; Cell Compartmentation; Cell Line; Cells; Circulation; Clinical; Congenital dyserythropoietic anemia; Data; Data Set; Dependence; Development; Embryo; Erythro; Erythroblasts; Erythrocytes; Erythroid; Erythroid Cells; Erythropoiesis; Event; Exhibits; Fetal Liver; Gene Expression Profile; Generations; Genes; Genetic Transcription; Human; In Vitro; Iron; Island; Link; LoxP-flanked allele; Macrophage; Molecular; Monitor; Morbidity - disease rate; Morphology; Mus; Mutation; Myelogenous; Nutrient; Patients; Pattern; Phenotype; Play; Process; Production; Proliferating; Property; Protocols documentation; Publishing; Reagent; Recovery; Regulation; Research; Role; Source; Stress; Supporting Cell; Surrogate Markers; System; Testing; Time; Tissues; Yolk Sac; Zinc Fingers; design; erythroid Kruppel-like factor; experimental study; in vivo; iron metabolism; novel; postnatal; progenitor; reconstitution; single cell analysis; transcription factor

SUMMARY Erythroid cells mature in vivo in the presence of a supportive macrophage; this cell-cell compartment is known as the erythroblastic island, an entity first described over 50 years ago. Red cell interactions with the macrophage are critical for optimal nutrient access, survival, proliferation, and differentiation of the erythron. Our proposed experiments address the exciting idea that Erythroid Krüppel-like Factor (EKLF; KLF1) not only plays an intrinsic role in establishing the proper gene expression patterns in the red cell within the erythroblastic island, but that it additionally affects this process by an extrinsic mechanism based on its surprising expression within the island macrophage and early in development within the erythro-myeloid progenitor (EMP). Our studies build on observations made in primary or minimally manipulated cells, aided by in vivo assays and EKLF rescue systems. We have found that EKLF is expressed in the EMP in the yolk sac by E8.5. The experiments of Aim 1 will investigate the role of EKLF in directing the functional potential of these cells at later stages in the embryo and in the adult. We have found that macrophage-restricted ablation of EKLF extrinsically affects the red cell. The experiments of Aim 2 will examine an iron-related hypothesis for the red cell effect that may be important for stress recovery. We have found that fetal liver island macrophage display a unique gene expression signature that is extensively regulated by EKLF. The experiments of Aim 3 will assess EKLF's role in island macrophage identity, studies that will be extended to analysis of human cells. Understanding these basic mechanisms will ultimately aid in understanding the problems that arise in anemias that present with high levels of nucleated red blood cells in circulation, such as congenital dyserythropoietic anemia (CDA) type IV, and in the design of culture systems that enable efficient expansion and enucleation of human cell sources for clinical use.

概括 在辅助巨噬细胞存在下，体内成熟的红细胞细胞；这个细胞细胞 车厢被称为红细胞岛，这是一个50年前最初描述的实体。红细胞 与巨噬细胞的相互作用对于最佳养分获取，生存，增殖和 erythron的区分。 我们提出的实验探讨了类似Krüppel样因子（EKLF）的令人兴奋的想法。 KLF1）不仅在建立红细胞中适当的基因表达模式中起着固有的作用 在红细胞岛内，但它还通过外在机制影响了这一过程 基于它在岛上巨噬细胞中的惊喜表达以及在 红细胞叶状体祖细胞（EMP）。我们的研究以初级或最少的观察结果为基础 受到体内测定和EKLF救援系统的帮助，操纵细胞。 我们发现EKLF在蛋黄囊中的EMP中以E8.5表示。实验 AIM 1将研究EKLF在引导这些细胞的功能潜力的作用 胚胎和成人的阶段。 我们发现，巨噬细胞限制的EKLF外在影响红细胞。 AIM 2的实验将检查与铁相关的假设，以实现红细胞效应 对于恢复压力很重要。 我们发现胎儿肝岛巨噬细胞显示独特的基因表达特征 这是由EKLF广泛调节的。 AIM 3的实验将评估EKLF在岛上的作用 巨噬细胞身份，将扩展到人类细胞分析的研究。 了解这些基本机制将最终有助于理解 在贫血中出现，循环中存在高水平的核化红细胞 先天性孕育性贫血（CDA）IV类 人类细胞来源的有效扩展和枚举临床使用。

项目成果

Isolation of Healthy F4/80+ Macrophages from Embryonic day E13.5 Mouse Fetal Liver Using Magnetic Nanoparticles for Single Cell Sequencing.

• DOI: 10.21769/bioprotoc.4243
• 发表时间: 2021-12
• 期刊: Bio-protocol
• 影响因子: 0.8
• 作者: Kaustav Mukherjee;J. Bieker

Modelling the erythroblastic island niche of dyserythropoietic anaemia type IV patients using induced pluripotent stem cells.

• DOI: 10.3389/fcell.2023.1148013
• 发表时间: 2023
• 期刊: FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
• 影响因子: 5.5
• 作者: May, Alisha;Ventura, Telma;Fidanza, Antonella;Volmer, Helena;Taylor, Helen;Romano, Nicola;D'Souza, Sunita L.;Bieker, James J.;Forrester, Lesley M.
• 通讯作者: Forrester, Lesley M.

Transcriptional Control of Gene Expression and the Heterogeneous Cellular Identity of Erythroblastic Island Macrophages.

• DOI: 10.3389/fgene.2021.756028
• 发表时间: 2021
• 期刊: Frontiers in genetics
• 影响因子: 3.7
• 作者: Mukherjee K;Bieker JJ
• 通讯作者: Bieker JJ