Transfusion-driven hyperhemolysis in sickle cell disease

镰状细胞病中输血引起的高溶血症

• 批准号: 10690278
• 负责人: Xiuli An
• 金额: $ 80.4万
• 依托单位: NEW YORK BLOOD CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-13 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10690278
• 关键词: Acute; Address; Adult; Anemia; Antibodies; Autologous; Binding; Biological Assay; Biological Markers; Blocking Antibodies; Blood Circulation; Bone Marrow; Bone Marrow Suppression; CFU-E; Cell Culture System; Cell Line; Cells; Cessation of life; Complication; Data; Defect; Development; Diagnosis; EPOR gene; Eating; Erythroblasts; Erythrocyte Transfusion; Erythrocytes; Erythroid; Erythroid Cells; Erythrophagocytosis; Erythropoiesis; Fc Receptor; Growth; Heme; Hemoglobin; Hemoglobinopathies; Hemolysis; Hemolytic Anemia; Human; Immune; Impairment; In Vitro; Inflammatory; Interferon Type I; Interferons; Iron Chelation; Knock-out; Knockout Mice; Life; Ligands; Mediating; Modeling; Molecular; Mus; Pathologic; Pathway interactions; Patients; Phagocytes; Phagocytosis; Phosphorylation; Plasma; Prevention; Production; Proteins; Reaction; Recovery; Reticulocytes; Role; STAT1 gene; STAT2 gene; SYK gene; Sampling; Savings; Severities; Sickle Cell; Sickle Cell Anemia; Signal Transduction; Signaling Molecule; Testing; Therapeutic; Therapeutic Effect; Thrombospondin 1; Thrombospondins; Time; Transfusion; Up-Regulation; base; cell age; cell type; crosslink; effective therapy; experience; in vivo; macrophage; monocyte; mouse model; new therapeutic target; patient population; premature; prevent; receptor; risk stratification; sickling; targeted treatment; therapeutic evaluation; uptake

Red blood cell (RBC) transfusions remain a cornerstone treatment in the management of sickle cell disease (SCD). However, patients may experience delayed hemolytic transfusion reaction (DHTR) which in this patient population has an unpredictable progression from mild to life-threatening severe reactions where both transfused and patient’s own RBCs are destroyed along with reticulocytopenia at the time of hemolytic crisis, exacerbating the anemia. The mechanisms underlying severe DHTR progression are poorly understood, posing challenges for prevention and effective treatments for this transfusion complication which is disproportionately encountered in patients with SCD. We recently found that acute hemolysis induces type I interferon (IFN-I) production in innate immune cells, leading to increased differentiation and activation of monocyte-derived macrophages (MoMΦ) in SCD, and exacerbating destruction of antibody (Ab)-coated transfused RBCs. Our preliminary data showed that Ab-sensitized RBC destruction alone also led to IFN-I production but with even higher levels if destruction occurred under hemolytic conditions, which interestingly also induced bystander hemolysis of sickle RBCs, mimicking hyperhemolysis reaction in SCD. We also found that hemolysis-induced IFN-I impairs erythropoiesis along with inhibition of EPO/EPOR signaling. Based on these data, we hypothesize that Fc receptor crosslinking in a hemolytic backdrop of SCD leads to increase in IFN-I levels, causing heightened IFN-I signaling in phagocytes and erythroid cells which trigger increased RBC destruction and further suppression of RBC production, respectively, leading to severe DHTR. In aim 1, we will focus on identifying mechanisms of bystander hemolysis by examining the role of key phagocytosis activation molecules, specifically SCD associated eat me signals including thrombospondin (TSP-1) and its ligands which are upregulated in hyperhemolytic models. We will compare the role of Ab-mediated erythrophagocytosis versus Ab-independent RBC engulfment in triggering bystander hemolysis and interrogate the relative contribution of inhibiting FcR/SYK phosphorylation and heme activation pathways in autologous sickle RBC destruction. We will also examine the potential of IFN-I as a biomarker of DHTR severity by examining IFN-I/STAT1 driven changes in monocytes in SCD patient samples, comparing patients experiencing severe and mild DHTR and after recovery to steady state. For aim 2, we will define the mechanisms by which IFN-I suppresses erythropoiesis using primary erythroid cell culture system and targeted deletion of key downstream pathways in human erythroblast cell lines and mouse models. We will also test the therapeutic effects of inhibiting IFN-I production/IFN-I signaling or/and increasing EPO/EPOR signaling on reversing impaired BM erythropoiesis in SCD mice and on human erythropoiesis in vitro and in cultures treated with SCD patient plasma. We believe that our proposed studies to examine the basis for progression to DHTR severity may help stratify risk and aid in development of novel targeted therapies to reverse or prevent hyperhemolysis, a devastating complication of an otherwise life-saving treatment in SCD.

红细胞（RBC）输血仍然是镰状细胞疾病治疗的基石治疗 （SCD）。但是，患者可能会遭受延迟的溶血翻译反应（DHTR） 人口从轻度到威胁生命的严重反应的进展不可预测 患者自己的RBC在溶血危机时与网状细胞减少症一起破坏，加剧 贫血。严重DHTR进展的基础机制知之甚少，提出了挑战 用于预防和有效治疗这种输血并发症，这是不成比例的 在SCD患者中。我们最近发现，急性溶血会诱导I型干扰素（IFN-I）的先天产生 免疫细胞，导致单核细胞衍生的巨噬细胞（MOMφ）在中的分化增加和激活 SCD和加剧抗体（AB）涂层的输血RBC的破坏。我们的初步数据表明 仅对AB敏感的RBC破坏也导致IFN-I产生 发生在溶血条件下，有趣的是，这也引起了镰状RBC的旁观者溶血， 模仿SCD中的过度分解反应。我们还发现溶血引起的IFN-I损害了红细胞生成 以及抑制EPO/EPOR信号传导。基于这些数据，我们假设FC受体交联 在SCD的溶血背景下，IFN-I水平的增加，导致IFN-I信号提高 吞噬细胞和红细胞细胞会触发RBC破坏并进一步抑制RBC 生产分别导致严重的DHTR。在AIM 1中，我们将专注于识别旁观者的机制 通过检查关键吞噬作用激活分子的作用，特别是SCD相关的食用我 信号在内，包括血小板传播（TSP-1）及其配体，这些信号在超血模型中被上调。我们 将比较AB介导的红细胞增多症与非AB依赖性RBC在触发中的作用 旁观者溶血和询问抑制FCR/SYK磷酸化和血红素的相对贡献 自体镰刀rbc破坏中的激活途径。我们还将研究IFN-I作为一个的潜力 DHTR严重程度的生物标志物通过检查IFN-I/STAT1驱动的SCD患者样本中单核细胞的变化， 比较患有严重和轻度DHTR的患者以及恢复到稳态后。对于目标2，我们将 定义IFN-I使用原发性红细胞培养系统和 人类红细胞细胞系和小鼠模型中关键下游途径的靶向删除。我们也会 测试抑制IFN-I产生/IFN-I信号传导或/和增加EPO/EPOR信号传导的治疗效应 逆转SCD小鼠中的BM红细胞生成障碍以及在体外和培养物中的人类红细胞生成 用SCD患者血浆治疗。我们认为，我们的拟议研究是为了研究发展的基础 DHTR的严重程度可能有助于分层风险并有助于开发新颖的靶向疗法以逆转或防止 过度分解，这是SCD中原本挽救生命治疗的毁灭性并发症。