Dissection of Hematopoietic Stem and Progenitor Cell Differentiation during Embryogenesis

胚胎发生过程中造血干细胞和祖细胞分化的剖析

• 批准号: 10607210
• 负责人: Anastasia Nizhnik
• 金额: $ 4.77万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-16 至 2026-04-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607210
• 关键词: Address; Adult; Affect; Agreement; Biological Assay; Blood; Blood Cells; CRISPR/Cas technology; Candidate Disease Gene; Cell Differentiation process; Cells; Clinical; Development; Dissection; Embryo; Embryonic Development; Endothelial Cells; Endothelium; Environment; Erythroid; Excision; Experimental Genetics; Gene Mutation; Generations; Genes; Genetic; Genetic Screening; Genomic approach; Goals; Growth; Hematologic Neoplasms; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Impairment; In Vitro; Inherited; Kinetics; Knowledge; Label; Lymphoid; Lymphoid Cell; Malignant Neoplasms; Measures; Methods; Molecular; Mus; Mutagenesis; Myelogenous; Myeloid Progenitor Cells; Natural Killer Cells; Organism; Output; Patients; Pluripotent Stem Cells; Population; Process; Property; RUNX1 gene; Regenerative capacity; Replacement Therapy; Source; System; T-Lymphocyte; Therapeutic; Tissues; Transplantation; Umbilical Cord Blood; Zebrafish; cell behavior; cell type; genetic approach; granulocyte; hematopoietic differentiation; hematopoietic stem cell differentiation; hematopoietic stem cell formation; hemogenic endothelium; improved; in vivo; induced pluripotent stem cell; loss of function; molecular marker; mutant; novel; progenitor; regenerative; regenerative cell; reverse genetics; self-renewal; single-cell RNA sequencing; spatiotemporal; stem cell therapy; stem cells; tool; transcription factor; vertebrate embryos; zebrafish development

PROJECT SUMMARY Hematopoietic Stem Cells (HSCs) are clinically valuable because they display regenerative properties of self- renewal and the ability to produce an entire hematopoietic system through multilineage differentiation. In other words, HSC can make all mature blood cell types such as myeloid, erythroid, and lymphoid and they sustain those blood types long-term in the organism. These regenerative properties make them an idea therapeutic tool for treating hematological malignancies by transplantation from a healthy donor, however a shortage of donors has prompted the effort to generate HSCs from alternative sources such as patient derived iPSCs. Knowledge about molecular mechanisms of HSC formation is needed to aid those efforts. HSCs are formed in early embryonic development alongside other HSC-independent Progenitor Cells (HPCs). One major difference between them is that HPCs are more limited in their differentiation repertoire compared to HSCs. We aim to study the differences between HSCs and HPCs to further advance our understanding of the specific molecular drivers of HSC regenerative properties. Understanding how HSCs acquire the ability to differentiate and self- renew in development can aid the efforts for generating them in vitro from alternative sources such as patient derived tissues or cord blood sources. Current molecular markers are insufficient for distinguishing HSCs from all other HPC subtypes. Moreover, most experimental systems require the removal of these cells from their native environment to study their functionality using in vitro culture assays or transplantation, which will measure the potential of a cell but may not reflect its actual endogenous function. Genetic experiments in mice and zebrafish showed that HSC and HPC fate can be molecularly uncoupled, implying that unique drivers of HSC formation exist. What genes are responsible for the formation of the rare population of HSCs independently from HPCs? To address this question, we established a genetic lineage tracing approach that measures HSC and HPC differentiation without removing them from their native environment. Using this assay, we found that HSCs show a significant delay in their ability to differentiate to myeloid and lymphoid lineages relative to HPCs, and this finding agrees with others in the field. The assay allows us to assess HSC differentiation in vivo, uncoupled from HPCs. We propose to use this functional tool in combination with other molecular and genetic assays to address the following aims: (1) Determine the differentiation potential of HSCs and HPCs in vivo and (2) Perform a reverse genetic screen for regulators of HSC differentiation. Our findings can aid the efforts to generate HSCs from alternative sources for clinical use.

项目总结