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 红细胞在有支持性巨噬细胞存在的情况下在体内成熟；这个细胞细胞 室被称为红细胞岛，这个实体在 50 多年前首次被描述。红细胞 与巨噬细胞的相互作用对于最佳营养获取、生存、增殖和生长至关重要 红细胞的分化。 我们提出的实验提出了一个令人兴奋的想法，即红细胞 Krüppel 样因子（EKLF； KLF1) 不仅在红细胞中建立正确的基因表达模式方面发挥着内在作用 在红细胞岛内，但它还通过外在机制影响这一过程 基于其在岛巨噬细胞内的令人惊讶的表达以及在巨噬细胞内的早期发育 红骨髓祖细胞（EMP）。我们的研究建立在初级或最低限度的观察基础上 在体内测定和 EKLF 救援系统的帮助下操纵细胞。 我们发现EKLF在卵黄囊的EMP中由E8.5表达。的实验 目标 1 将研究 EKLF 在稍后指导这些细胞的功能潜力中的作用 胚胎和成人阶段。 我们发现巨噬细胞限制的 EKLF 消融会从外部影响红细胞。 目标 2 的实验将检验与铁相关的红细胞效应假说，该假说可能 对于压力恢复很重要。 我们发现胎儿肝岛巨噬细胞表现出独特的基因表达特征 受到 EKLF 的广泛监管。 Aim 3 的实验将评估 EKLF 在岛屿中的作用 巨噬细胞身份的研究将扩展到人类细胞的分析。 了解这些基本机制最终将有助于理解以下问题： 出现在循环中有高水平有核红细胞的贫血中，例如 先天性红细胞生成障碍性贫血 (CDA) IV 型，以及培养系统的设计使得 有效扩增和去核人类细胞源以供临床使用。

项目成果

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

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.

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