Microfluidic Impedance Red Cell Assay (MIRCA) for Emerging Pharmacologic and Gene based Therapies for Sickle Cell Disease

项目摘要 获得性或遗传性疾病可改变红细胞（RBC），导致严重的临床并发症 影响脉管系统和器官的健康遗传性红细胞疾病、肾衰竭、糖尿病和血库 储存损伤都可以产生坚硬的、不可变形的RBC。红细胞僵硬是特别有问题的镰状 细胞疾病（SCD），一种使人衰弱的遗传性血液疾病。美国估计有10万人， 全世界还有数百万人患有SCD。该领域已经经历了新SCD疗法的爆炸， 旨在针对过去五年中RBC中的特定个体异常。许多基于基因的 旨在治愈SCD的疗法正处于临床试验中。SCD疗法的这一显著转变产生了对 评估RBC健康和质量的诊断和分析工具。我们需要红细胞功能的生物标志物， 作为临床试验的终点，协助最佳药物选择和个体化监测， 并确定基因治疗是否使红细胞正常化。红细胞的许多方面必须 评估，但一个关键特征是可变形性。缺乏足够灵敏的坚固、廉价的设备， 捕获少量变形性差的红细胞，这些红细胞能够继续溶血，损伤血管， 导致器官损伤和过早死亡至关重要的是，我们要审查治疗方法是否有能力使其正常化。 所有的红细胞。在这里，我们建议开发和翻译微流控阻抗红细胞 功能性测量红细胞变形性的测定（MIRCA），报告为仿生化合物的闭塞指数（OI）。 芯片上的微毛细管网络阵列。我们提出了一种创新的方法和一种新颖的合作， 工程师和血液学专业的医生科学家。我们将通过以下步骤实现这一目标： 1.我们将实现红细胞OI的MIRCA测量的仪器化和分析验证。解析 将在PI实验室（Dr. Umut Gurkan，CWRU）和Co-I血液学实验室进行验证 实验室（Vivien Sheehan博士，埃默里），以证明普遍性，并允许利益相关者的反馈和设计 改性 2.我们将通过评估OI和临床并发症之间的关系， SCD中疾病严重程度的传统实验室测量。 3.我们将通过证明其预后价值，将优化的MIRCA的使用扩展到临床空间 在经历临床并发症和过渡期的SCD患者的纵向评估中， 疗法 利用我们独特的生物医学工程师和血液学训练的医生科学家之间的合作， 我们将在临床试验和病人护理中优化、验证并实现MIRCA技术的临床应用 在美国和世界范围内患有SCD的个人。

PROJECT SUMMARY Acquired or inherited diseases can alter the red blood cell (RBC), causing severe clinical complications affecting the vasculature and organ health. Inherited red cell disorders, renal failure, diabetes, and blood bank storage lesions can all produce stiff, non-deformable RBCs. Red cell stiffness is particularly problematic in sickle cell disease (SCD), a debilitating inherited blood disorder. An estimated 100,000 individuals in the US, and millions more world-wide, have SCD. The field has experienced an explosion of novel SCD therapeutics, designed to target specific individual abnormalities in the RBC in the last five years. Numerous gene-based therapies intending to cure SCD are in clinical trials. This marked shift in SCD therapeutics produced a need for diagnostics and analytical tools that assess RBC health and quality. We need biomarkers of red cell function to serve as endpoints in clinical trials, assist in optimal drug selection and personalized monitoring for the individual, and to determine if gene-based therapy has normalized the red cell. Many aspects of the red cell must be assessed, but a key feature is deformability. There is a lack of robust, inexpensive devices sensitive enough to capture small populations of poorly deformable red cells able to continue to hemolyze, damage the blood vessels, and cause organ damage and early mortality. It is essential that we vet therapies for their ability to normalize the entire population of red cells. Here we propose to develop and translate the Microfluidic Impedance Red Cell Assay (MIRCA) to functionally measure red cell deformability, reported as Occlusion Index (OI) of a biomimetic microcapillary network array on a chip. We propose an innovative approach and a novel collaboration between engineers and a hematology-trained physician scientist. We will do so with the following steps: 1. We will achieve instrumentation and analytical validation of MIRCA measurement of red cell OI. Analytic validation will be performed both in PI’s laboratory (Dr. Umut Gurkan, CWRU) and in Co-I’s hematology laboratory (Dr. Vivien Sheehan, Emory) to prove generalizability and permit stakeholder feedback and design modification. 2. We will clinically validate MIRCA by assessing the association between OI and clinical complications and traditional laboratory measures of disease severity in SCD. 3. We will expand the use of the optimized MIRCA to the clinical space by demonstrating its prognostic value in longitudinal assessments of patients with SCD experiencing clinical complications and transitions in therapy. Using our unique collaboration between biomedical engineers and a hematology trained physician-scientist, we will optimize, validate, and achieve clinical adoption of MIRCA technology in clinical trials and patient care for individuals living with SCD in the US and world-wide.