Improving granulocyte transfusion in neutropenia-related infections

改善中性粒细胞减少相关感染的粒细胞输注

基本信息

批准号：
10682602
负责人：
Hongbo R Luo
金额：
$ 66.47万
依托单位：
BOSTON CHILDREN'S 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：
2022-08-15 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10682602
关键词：
Affect Aging Animal Experiments Animal Model Animals Antibiotics Antifungal Agents Antioxidants Apoptosis Apoptotic Aspergillus fumigatus Bacteria Bacterial Infections Binding Blood Component Removal Blood Transfusion CASP3 gene CSF3 gene Caspase Caspase Inhibitor Cell Death Cell Differentiation process Cell Membrane Permeability Cell physiology Cells Cessation of life Chemotaxis Clinical Clinical Trials Collection Colony-Stimulating Factors Combined Modality Therapy Complement Cytoplasmic Granules Disadvantaged Disease Effectiveness Elastases Escherichia coli FDA approved Fracture Goals Granulocyte Colony-Stimulating Factor Half-Life Heat-Shock Proteins 70 Host Defense Human Impairment Infection Inflammatory Response Invaded Investigation Knowledge Leukocyte Elastase Life Longevity Lytic Mediating Mediator Membrane Mission Modeling Molecular Molecular Profiling Morbidity - disease rate Mus Mycoses Nature Neutropenia Outcome Oxidants Oxidative Stress Pathway interactions Patients Peptide Hydrolases Persons Phagocytosis Pharmaceutical Preparations Play Pneumonia Procedures Process Production Research Resolution Role Serine Serine Protease Signal Transduction Testing Therapeutic Transfusion Translational Research Transplant Recipients Transplantation Treatment Factor United States National Institutes of Health aged bactericide clinically relevant design experimental study granulocyte hematopoietic cell transplantation improved in vivo inhibitor insight mortality mouse model neutrophil novel novel strategies novel therapeutic intervention pathogen pharmacologic phosphatidylinositol 3,4,5-triphosphate pneumonia model preservation programs response screening single-cell RNA sequencing therapeutic target transfusion medicine

项目摘要

Project Summary Granulocyte transfusion (aka. neutrophil transfusion or GTX) has been utilized as a therapeutic approach for the treatment of life-threatening bacterial and fungal infections in severe neutropenic patients, including hematopoietic cell transplantation (HCT) recipients. Clinical outcomes from GTX are disadvantaged by the short shelf life and compromised function of donor neutrophils. Spontaneous neutrophil death is heterogeneous and mediated by multiple pathways. Leveraging mechanistic knowledge and pharmacological screening, we identified a combined treatment, caspases–lysosomal membrane permeabilization–oxidant– necroptosis inhibition plus granulocyte colony-stimulating factor (CLON-G), which alters neutrophil fate by simultaneously targeting multiple cell death pathways. CLON-G treatment significantly prolongs neutrophil ex vivo survival, increasing their half-life from less than 1 day to greater than 5 days. Compared to freshly isolated neutrophils, the stored CLON-G-treated neutrophils display normal functions, including chemotaxis, ROS production, phagocytosis, and bacteria killing. These results led us to hypothesize that CLON-G treatment can significantly increase the ex vivo shelf life of granulocyte transfusion products without impairing the function of the neutrophils in the GTX recipients. Due to the prolonged intracellular retention of the drugs, CLON-G treatment may even improve the survival and increase the half-life of the transfused neutrophils in vivo and therefore further improve the efficacy of GTX. In this study, we will first test this hypothesis in a clinically relevant murine model of granulocyte transfusion in neutropenia-related pneumonia. We will determine whether the stored CLON-G-treated neutrophils can function normally and survive even longer in vivo compared to freshly isolated neutrophils (Aim 1A). Additionally, we will investigate the survival and function of transfused human neutrophils in vivo in neutrophil-depleted NSG mice (Aim 1B). Next, we will examine whether transfusion with stored CLON-G-treated neutrophils can ultimately enhance the inflammatory response and bactericidal capability of the recipients as effectively as, or even more effectively than, transfusion with untreated fresh neutrophils (Aim 2). Last, to identify novel CLON-G-inducible factors and pathways that play critical role in regulating neutrophil death and function, we will perform single-cell RNA sequencing (scRNA-seq) to reveal the molecular signature of CLON-G reprogrammed neutrophils (Aim 3) . Together, experiments proposed in the three aims will assist us to better understand the molecular and cellular mechanism leading to neutrophil death and to design novel clinical procedures for the long-term storage and application of neutrophils in transfusion medicine. This study also provides a new strategy for treatment of neutropenia-related pneumonia. The ability to utilize GTX through better understanding of granulocyte function and survival could have a major impact on patients with no other options. This will be an important and necessary complementation to the current antibiotic and G-CSF therapies.

项目概要粒细胞输注（又名中性粒细胞输注或 GTX）已被用作以下疾病的治疗方法：治疗严重中性粒细胞减少症患者危及生命的细菌和真菌感染，包括 GTX 的造血细胞移植 (HCT) 接受者的临床结果受到不利影响。供体中性粒细胞的保质期短且功能受损。异质性并由多种途径介导。筛选后，我们确定了一种联合治疗方法，半胱天冬酶-溶酶体膜透化-氧化剂- 坏死性凋亡抑制加上粒细胞集落刺激因子（CLON-G），通过改变中性粒细胞的命运同时靶向多种细胞死亡途径的 CLON-G 治疗可显着延长中性粒细胞的寿命。与新鲜分离的相比，体内存活率从不到 1 天增加到超过 5 天。中性粒细胞，储存的 CLON-G 处理的中性粒细胞显示正常功能，包括趋化性、ROS 这些结果使我们认为 CLON-G 治疗可以。显着延长粒细胞输注产品的离体保质期，且不损害其功能 GTX 受体中的中性粒细胞由于药物在细胞内滞留时间较长，CLON-G 受到影响。治疗甚至可以提高输注中性粒细胞的存活率并延长体内的半衰期因此，进一步提高GTX的功效在本研究中，我们将首先在临床上检验这一假设。我们将确定中性粒细胞减少症相关肺炎的粒细胞输注的相关小鼠模型。储存的经过CLON-G处理的中性粒细胞在体内能否正常发挥功能并存活更长时间与新鲜分离的中性粒细胞相比（目标 1A）此外，我们将研究其存活和功能。在中性粒细胞耗尽的 NSG 小鼠体内输注人中性粒细胞（目标 1B）。输注储存的 CLON-G 处理的中性粒细胞是否最终会增强炎症接受者的反应和杀菌能力与以下效果一样有效，甚至更有效：输注未经处理的新鲜中性粒细胞（目标 2），以新的方式鉴定 CLON-G 诱导因子和在调节中性粒细胞死亡和功能中发挥关键作用的途径，我们将进行单细胞 RNA 测序 (scRNA-seq) 揭示 CLON-G 重编程中性粒细胞的分子特征（目标 3）。总之，这三个目标中提出的实验将帮助我们更好地理解分子和导致中性粒细胞死亡的细胞机制并设计新的长期临床程序该研究也为中性粒细胞在输血医学中的储存和应用提供了新的策略。通过更好地了解中性粒细胞减少症相关肺炎的治疗能力。粒细胞功能和生存可能对没有其他选择的患者产生重大影响。对当前抗生素和 G-CSF 疗法的重要且必要的补充。