Bone Marrow Niche dysfunction in sickle cell disease

镰状细胞病的骨髓生态位功能障碍

• 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
• 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

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。镰状细胞病（SCD）是由 -珠蛋白基因的单一突变引起的，是最常见的疾病。 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镰状化和血管闭塞发作，从很小的时候起， 导致急性和慢性疼痛、中风、贫血、感染、器官衰竭和过早死亡。血管闭塞 发作引起缺血再灌注损伤，激活内皮细胞、免疫细胞和凝血 级联。这些过程导致 SCD 的慢性炎症状态，与促炎症反应增加相关。 包括骨髓 (BM) 在内的所有器官中的炎症细胞因子、TLR4 激活和氧化应激。 迄今为止，我们对炎症如何影响 SCD 中的 BM 微环境和 HSC 功能知之甚少 患者。值得注意的是，对人类样本进行的研究很少且有限。初步结果。 我们之前使用不同动物模型的研究表明，TLR4 的激活和上调 炎症期间的 miR155 和 NF-kB 信号传导导致 BM 生态位、骨髓谱系的重塑 造血干细胞自我更新的偏差和丧失。我们在 SCD 患者中获得的初步结果 参加 City of Hope 的 HSC 移植 (HSCT) 临床试验的样本显示， CD34+ 祖细胞的频率和数量，产生集落的能力下降并增加 SCD BM 来源的间充质细胞 (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 SCD 患者的 BM 生态位并损害 HSC 功能， 这些变化会影响移植过程中供体 HSC 的植入。目标和战略。这 研究将对 SCD 患者 (n=21) 之前 HSCT 以及第 +15、+30 和 +1 年的 BM 样本进行 造血干细胞移植后。细胞成分和炎症途径的表征将在 BM 上进行 使用多参数流式细胞术分析等补充方法进行活检和抽吸， 免疫组织化学和纳米线测定； CD34+细胞将在基线时分离，用于RNA-seq 并测试了集落形成试验和 NSG 小鼠体内移植能力； BM和外周血清 将检测炎症细胞因子； MSC 将在体外衍生自 BM，并进行表征和分析 炎症特征的持续存在。结果将与 HSCT 的结果相关。年龄匹配 健康的捐赠者将用作对照。关联。这项研究提供了一个独特的机会，可以更好地 了解人类标本中 SCD 的生物学。重要的是，对涉及的机制的剖析 严重 SCD 中的 BM 生态位功能障碍和炎症状态应为治疗干预提供途径 改善 SCD 患者的移植和造血重建。