Endothelialized microfluidic assays for emerging therapies in sickle cell disease

用于镰状细胞病新兴疗法的内皮微流体检测

• 批准号: 10476681
• 负责人: Chiara Federici
• 金额: $ 85.21万
• 依托单位: BIOCHIP LABS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-05 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10476681
• 关键词: Abnormal Red Blood Cell; Adhesions; Adhesives; American; Anemia; Antisickling Agents; Biological Assay; Biological Markers; Biomimetics; Blood; Blood Cells; Blood Transfusion; Blood Vessels; Cell Adhesion; Clinical; Clinical Research; Complex; Deoxygenated Sickle Hemoglobin; Development; Drug Screening; Drug Targeting; Economic Burden; Endothelial Cells; Endothelium; Erythrocytes; Exhibits; Experimental Designs; Functional disorder; Goals; Health Expenditures; Hematological Disease; Hemoglobin; Hemoglobin concentration result; Hypoxia; Impairment; In Vitro; Individual; Inflammation; Inherited; Ischemia; Lactate Dehydrogenase; Lead; Letters; Measurement; Mediating; Membrane; Methodology; Microfluidics; Modernization; Monitor; Monoclonal Antibodies; Morbidity - disease rate; Organ; P-Selectin; Pain; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Physiological; Polymers; Priapism; Procedures; Property; Quality Control; Quality of life; Reproducibility of Results; Reticulocyte count; Sampling; Sickle Cell; Sickle Cell Anemia; Sickle Hemoglobin; Site; Small Business Technology Transfer Research; Standardization; Stroke; Technology; Testing; Therapeutic; Thrombophilia; Translating; United States Food and Drug Administration; Validation; Variant; Vascular Diseases; Whole Blood; acute chest syndrome; base; chromatin immunoprecipitation; comorbidity; design; drug candidate; drug discovery; hydroxyurea; in vitro Assay; mortality; novel; novel therapeutics; patient population; patient response; personalized medicine; polymerization; pre-clinical; pre-clinical assessment; precision drugs; prevent; response; screening; sickling; social; targeted agent; targeted treatment; tool; vaso-occlusive crisis

PROJECT SUMMARY Sickle Cell Disease (SCD) afflicts 100,000 Americans and millions of people worldwide, and is characterized by anemia, painful vaso-occlusive crises, ischemia, inflammation, significant morbidity, and early mortality. In SCD, deoxygenated sickle hemoglobin (deoxy-HbS) polymerizes, deforms red blood cells (RBCs), changes membrane properties, and triggers inflammation, thrombophilia, and vasculopathy. Traditional treatment approaches include routine blood transfusions and hydroxyurea therapy while both options exhibit inherent challenges and limitations. Recent efforts in developing new treatment approaches in SCD focused on targeted therapies, aiming to prevent hypoxia-mediated polymerization of sickle hemoglobin and the abnormal adhesive interactions between blood and endothelial cells. Accordingly, the Food and Drug Administration (FDA) has recently approved two new treatments for SCD: ADAKVEO (Crizanlizumab, Novartis) and Oxbryta (Voxelotor, Global Blood Therapeutics) to prevent abnormal RBC adhesion and sickling. In addition to those, many other drug candidates targeting a specific adhesion pathway are currently in the development stage. Because SCD pathophysiology is extremely complex and heterogenous, it is crucial to predict how/if a patient will benefit from a specific targeted therapy. An anti-adhesive therapy may not benefit a certain patient population or may need to be supplemented with additional therapeutic approaches (i.e., anti-sickling drugs). Further, patient-specific responses to such therapies may be impacted by or associated with clinical variables (e.g., lactate dehydrogenase level, reticulocyte count, hemoglobin levels, etc.). Being able to predict a patient's response to a certain targeted therapy, or combination of multiple, using biomimetic in vitro tools will substantially help the entire SCD patient population. Therefore, there is an urgent need for novel biomimetic in vitro assays that can serve as a drug screening platform, both in understanding the effect of emerging therapies on a single patient level and in screening drug candidates for potential mitigation of SCD pathophysiology. Our objective in this STTR Phase I/II proposal is to develop a standardized and validated endothelialized microfluidic cellular adhesion assay to predict in vitro patient-specific responses to a targeted therapy by probing blood cell adhesion to activated endothelial cells. In Phase I, we propose to standardize and validate the culture and activation of endothelial cells as well as blood cell adhesion within the Endothelium-on- a-chip under physiologically relevant flow conditions. Phase II aims and milestones focus on activities related to determining baseline cellular adhesion levels, establishing the testing methodologies for proof-of-concept, and clinically validating the Endothelium-on-a-chip. Our goal is to demonstrate the utility of the Endothelium-on- a-chip in predicting patient-specific responses to single and combined targeted therapies in SCD.

项目摘要 镰状细胞病（SCD）折磨着10万美国人和全世界数百万人，其特征是： 贫血、疼痛性血管闭塞危象、缺血、炎症、显著发病率和早期死亡率。在SCD中， 脱氧镰状血红蛋白（脱氧-HbS）聚合，使红细胞（RBC）变形， 膜特性，并触发炎症、血栓形成倾向和血管病变。传统治疗 方法包括常规输血和羟基脲治疗，而这两种选择都表现出固有的 挑战和限制。最近在SCD中开发新治疗方法的努力集中在靶向治疗上。 治疗，旨在防止缺氧介导的镰状血红蛋白聚合和异常粘附 血液和内皮细胞之间的相互作用。因此，美国食品和药物管理局（FDA） 最近批准了两种新的SCD治疗方法：ADAKVEO（Crizanlizumab，Novartis）和Oxbryta（Voxelotor， Global Blood Therapeutics），以防止异常RBC粘附和镰状化。除此之外，许多其他 靶向特定粘附途径的候选药物目前处于开发阶段。因为SCD 病理生理学是极其复杂和异质的，预测患者如何/是否将受益于 一种特定的靶向治疗抗粘连治疗可能不会使某些患者群体受益，或者可能需要 以补充额外的治疗方法（即，抗镰状化药物）。此外，患者特异性 对这种治疗的反应可能受到临床变量的影响或与临床变量相关（例如，乳酸 脱氢酶水平、网织红细胞计数、血红蛋白水平等）。能够预测病人对 使用仿生体外工具的某种靶向治疗或多种组合将实质上帮助 整个SCD患者人群。因此，迫切需要新的仿生体外测定，其可以 作为一个药物筛选平台，既可以了解新兴疗法对单个患者的影响， 水平和筛选候选药物的潜在缓解SCD的病理生理。 我们在STTR I/II期提案中的目标是开发一种标准化和经验证的内皮化 预测体外患者对靶向治疗的特异性反应的微流控细胞粘附测定 通过探测血细胞与激活的内皮细胞的粘附。在第一阶段，我们建议标准化和 验证内皮细胞的培养和活化以及内皮细胞上的血细胞粘附， 在生理相关流动条件下的A芯片。第二阶段的目标和里程碑侧重于与下列活动有关的活动： 确定基线细胞粘附水平，建立概念验证的测试方法，以及 临床验证内皮细胞芯片我们的目标是证明内皮细胞的效用， a芯片预测SCD患者对单一和联合靶向治疗的特异性反应。