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Mechanisms and consequences of sickle cell disease-induced cycling in hematopoietic stem cells

镰状细胞病诱导造血干细胞循环的机制和后果

基本信息

批准号：
10464657
负责人：
Aditya Shirish Barve
金额：
$ 6.76万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10464657
关键词：
ATAC-seq Abnormal Hemoglobins Age Age-Months Aging Animal Diseases Animals Aspirate substance Autologous Autologous Transplantation Biochemical Biological Assay Bone Marrow Cancer Biology Carrying Capacities Cell Compartmentation Cell Cycle Cell Proliferation Cells Chronic Clinical Clinical Trials DNA Damage Data Analyses Data Set Environment Epigenetic Process Erythrocytes Erythropoiesis Exposure to Flow Cytometry Frequencies Functional disorder Genes Genetic Transcription Glean Goals Health Hematologic Neoplasms Hematological Disease Hematology Hematopoietic Hematopoietic Neoplasms Hematopoietic Stem Cell Mobilization Hematopoietic Stem Cell Transplantation Hematopoietic Stem Cell subsets Hematopoietic stem cells Hemolysis Human Immune Impairment In Vitro Inflammation Inherited Investigation Knowledge Laboratories Lead Learning Life Mediating Mentorship Molecular Molecular Probes Molecular Profiling Morbidity - disease rate Morphology Mus Mutation Myelogenous Organ Output Oxygen Pain Pathway interactions Patients Phenotype Pilot Projects Ploidies Positioning Attribute Postdoctoral Fellow Regimen Research Personnel Resource Development Resources Risk Safety Saint Jude Children&apos s Research Hospital Sickle Cell Sickle Cell Anemia Source Stress Testing Time Training Transgenic Organisms Translating Transplantation Work Xenograft procedure aged career development cell age cell injury chemotherapy curative treatments cytokine disorder control exhaustion fitness gene therapy hematopoietic stem cell differentiation hemoglobin B hemoglobin polymer immune activation improved in vivo insight mortality mouse model novel polymerization post-transplant premature promoter self-renewal sickling stem cell biology stem cell function systemic inflammatory response training opportunity transcriptome sequencing

项目摘要

Project Summary Sickle cell disease (SCD) is a painful debilitating life-long condition resulting from mutations in the gene encoding hemoglobin β subunit, causing abnormal hemoglobin polymerization leading to hemolysis, repeated vasooclusion, and chronic systemic inflammation resulting in substantial global morbidity and early mortality. Curative therapy for SCD relies on hematopoietic stem cell (HSC) transplantation, however the damaging effects of SCD pathophysiology on HSCs remain uncharacterized and we seek to fill this gap in current knowledge. Our preliminary studies show that HSCs display increased cycling in a transgenic SCD mouse model upon aging, and myeloid lineage biased in vitro differentiation of SCD patient derived HSCs. As a postdoctoral fellow in the McKinney-Freeman laboratory, I will investigate the functional consequences and molecular mechanisms underlying SCD mediated HSC cycling in a murine model and translate these findings to human HSC during SCD. In Aim 1, I will use colony formation assays, limiting dilution primary and secondary HSC transplantation, and serial exposure to chemotherapy to assess the detrimental impact of SCD-induced cycling on HSC frequency and function. In Aim 2, I will probe the molecular and epigenetic dysregulation underlying increased HSC cycling during SCD. HSCs isolated from SCD and control mice will be subjected to bulk RNA-seq and ATAC-seq to define transcriptional dysregulation correlated with changes in gene promoter accessibility meditating increased cycling. Finally, I will translate our findings to humans by interrogating cell cycle dysregulation in HSCs isolated from SCD patients (Aim 3). SCD patient-derived bone marrow aspirates will be profiled for frequency of phenotypic HSCs and alterations in cell cycle status by flow cytometry and ex vivo EdU incorporation. Lineage potential and hematopoietic output of SCD HSCs will be analyzed by colony formation and in vitro single HSC differentiation assays. Lastly, SCD HSC repopulating potential and self-renewal will be examined by limiting dilution primary and secondary transplantation into human cytokine expressing immune deficient mice. Together the proposed studies serve to deepen our understanding of a previously unexplored aspect of HSC biology by examining the impact of SCD. Greater understanding of SCD mediated mechanisms of HSC impairment become exceptionally important as autologous HSC transplantation following gene editing or gene therapy for SCD improve and increase in frequency. These aims draw on my prior training in cancer biology of hematologic malignancies but also provide abundant novel training opportunities in the field of HSC biology and for professional career development. The McKinney-Freeman lab and St. Jude Children’s Research Hospital are ideal environments in which to receive training in the study HSC biology, combining state-of-the-art institutional resources, career development resources, and excellent mentorship with the singular goal of advancing to an independent academic primary investigator position.

项目总结