Bone Marrow Niche dysfunction in sickle cell disease

镰状细胞病的骨髓生态位功能障碍

基本信息

批准号：
9927706
负责人：
Nadia Carlesso
金额：
$ 34.6万
依托单位：
BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-15 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9927706
关键词：
Abnormal Hemoglobins Acute Acute Pain Affect African American Age Allogenic American Anemia Animal Model Aspirate substance Autologous Biological Assay Biology Birth Blood flow Bone Marrow Bone Marrow Diseases Bone Marrow Transplantation Bone marrow biopsy Bone remodeling CD34 gene Cell physiology Cells Cessation of life Chronic Cities Clinical Trials Coagulation Process Complex Development Disease Disease-Free Survival Dissection Endothelial Cells Endothelium Engineering Engraftment Enrollment Erythrocytes Event Evolution Exposure to Fetal Hemoglobin Flow Cytometry Frequencies Functional disorder Gene-Modified Genes Globin Hematological Disease Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hemoglobin Hemolysis Hispanics Human Immune Immunohistochemistry Impairment In Vitro Incidence Individual Infection Inflammation Inflammatory Inherited Injury Institution Intervention Kidney Failure Lead Life Expectancy Mesenchymal Mus Mutation Myelogenous NF-kappa B Natural regeneration Organ Organ failure Outcome Oxidative Stress Pain Pain management Pathway interactions Patients Phase I Clinical Trials Prevention Process Production Property Pulmonary Hypertension Quality of life Recurrence Reperfusion Injury Resolution Retinal Diseases Risk Safety Sampling Serum Siblings Sickle Cell Sickle Cell Anemia Signal Transduction Specimen Stress Stroke Supportive care TLR4 gene Testing Therapeutic Therapeutic Intervention Time Translations Transplant Recipients Transplantation Up-Regulation acute chest syndrome aged base chronic pain comorbidity curative treatments cytokine disease phenotype experience fitness gene correction gene therapy graft failure hematopoietic cell transplantation hematopoietic stem cell self-renewal improved in vivo insight nano-string novel peripheral blood polymerization premature preservation programs reconstitution sickle vasoocclusion stem cells success transcriptome sequencing

项目摘要

SUMMARY/ABSTRACT Background. Sickle cell disease (SCD) is caused by a single mutation of the -globin gene and is the most common form of inherited blood disorder. In the US, approximately 100,000 of Americans are affected. Patients with SCD suffer from repeated red cell sickling and vaso-occlusion episodes, which, since early age, lead to acute and chronic pain, stroke, anemia, infections, organ failure and premature death. Vaso-occlusion episodes cause ischemia-reperfusion injury, activation of endothelial cells, immune cells and the coagulation cascade. These processes lead to a state of chronic inflammation in SCD, associated with increased pro- inflammatory cytokines, TLR4 activation and oxidative stress in all organs, including the bone marrow (BM). To date, little is known about how inflammation impacts the BM microenvironment and HSC functions in SCD patients. Notably, few and limited studies have been conducted on human samples. Preliminary results. Our previous studies using different animal models have shown that TLR4 activation and upregulation of miR155 and NF-kB signaling during inflammation results in the remodeling of the BM niche, myeloid lineage skewing and loss of hematopoietic stem cell self-renewal. Preliminary results that we obtained in SCD patient samples enrolled in a HSC transplantation (HSCT) clinical trial at City of Hope demonstrated a decrease in frequency and number of CD34+ progenitor cells, decreased ability to generate colonies and increased miR155 and NF-kB signaling in SCD BM-derived mesenchymal cells (MSC). Hypothesis. We hypothesize that SCD-induced chronic inflammation remodels the BM niche and impairs HSC function in SCD patients, and that these changes impact the engraftment of donor HSC during transplant. Aims and Strategy. This study will be conducted on BM specimens of SCD patients (n=21) prior HSCT and at day +15, +30 and +1yr after HSCT. Characterization of the cellular components and inflammatory pathways will be carried on BM biopsies and aspirates by using complementary approaches such as multiparametric flow cytometry analysis, immunohistochemistry and nanostring assays; CD34+ cells will be isolated at baseline, used for RNA-seq and tested for colony-forming assay and ability to engraft NSG mice in vivo; BM and peripheral blood serum will be probed for inflammatory cytokines; MSC will be BM derived in vitro and analyzed characterization and persistence of the inflammatory signature. Results will be correlated with HSCT outcomes. Aged matched healthy donors will be used as controls. Relevance. This study represents a unique opportunity to better understand the biology of SCD in human specimens. Important, the dissection of the mechanisms involved in BM niche dysfunction and inflammatory state in severe SCD should offer avenues for therapeutic intervention to improve engraftment and hematopoietic reconstitution in patients with SCD.

摘要/摘要背景。镰状细胞疾病（SCD）是由珠蛋白基因的单个突变引起的，是最多的遗传性血液疾病的常见形式。在美国，大约有10万美国人受到影响。 SCD患者患有反复的红细胞疾病和血管封闭发作，从小就开始导致急性和慢性疼痛，中风，贫血，感染，器官衰竭和过早死亡。血管封闭发作会导致缺血 - 重新灌注损伤，内皮细胞的激活，免疫细胞和凝结级联。这些过程导致SCD中的慢性炎症状态包括骨髓（BM）在内的所有器官中的炎性细胞因子，TLR4激活和氧化应激。迄今患者。值得注意的是，对人类样本进行的研究很少和有限的研究。初步结果。我们先前使用不同动物模型的研究表明，TLR4激活和更新炎症期间的miR155和NF-kb信号传导导致BM生阵（Myeloid）谱系的重塑倾斜和造血干细胞自我更新的损失。我们在SCD患者中获得的初步结果在希望之城参加HSC移植（HSCT）临床试验的样品表明有所减少 CD34+祖细胞的频率和数量，产生菌落的能力降低并增加 SCD BM衍生的间充质细胞（MSC）中的miR155和NF-KB信号传导。假设。我们假设 SCD诱导的慢性炎症会重塑BM的利基市场，并损害SCD患者的HSC功能，这些变化会影响移植过程中供体HSC的植入。目的和策略。这研究将对HSCT前SCD患者的BM标本（n = 21）进行，在+15，+30和 +1年 HSCT之后。细胞成分和炎症途径的表征将在BM上携带活检和抽吸方法通过使用完整的方法，例如多参数流式细胞仪分析，免疫组织化学和纳米弦测定； CD34+细胞将在基线时分离，用于RNA-seq 并测试了菌落形成测定和在体内植入NSG小鼠的能力； BM和外周血系列将探测炎性细胞因子； MSC将是BM在体外衍生和分析的表征和炎症签名的持久性。结果将与HSCT结果相关。老年匹配健康的捐助者将被用作对照。关联。这项研究代表了更好的机会了解人类标本中SCD的生物学。重要的是，解剖涉及的机制严重SCD中的BM利基功能障碍和炎症状态应为治疗干预提供途径改善SCD患者的植入和造血重建。