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

项目总结 获得性或遗传性疾病会改变红细胞，导致严重的临床并发症。 影响血管系统和器官健康。遗传性红细胞疾病、肾功能衰竭、糖尿病和血库 储存损伤都能产生僵硬、不可变形的红细胞。红血球僵硬在镰刀中尤其成问题 细胞病(SCD)，一种使人衰弱的遗传性血液疾病。据估计，美国有10万人， 全世界还有数百万人患有SCD。该领域经历了新的SCD疗法的爆炸性增长， 旨在针对过去五年中红细胞的特定个体异常。众多基于基因的 打算治愈SCD的疗法正在进行临床试验。SCD疗法的这种显著转变产生了对 评估红细胞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记物来 作为临床试验的终点，协助对个体进行最佳药物选择和个性化监测， 并确定基于基因的治疗是否使红细胞正常化。红细胞的许多方面都必须是 经过评估，但一个关键特征是可变形性。目前缺乏足够灵敏的坚固、廉价的设备 捕获少量易变形的红细胞，这些红细胞能够继续溶血，破坏血管， 并导致器官损伤和早期死亡。至关重要的是，我们必须审查治疗方法，看它们是否有能力使 所有的红血球。在这里，我们建议开发和翻译微流阻抗红细胞 用于功能性测量红细胞变形性的试验(MIRCA)，报告为仿生生物的遮挡指数(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的合作方式 工程师和一位受过血液学训练的内科科学家。我们将通过以下步骤来实现这一点： 1.我们将实现红细胞OI的Mirca测量的仪器和分析验证。分析 验证将在Pi的实验室(CWRU的Umut Gurkan博士)和Co-I的血液学实验室进行 实验室(Vivien Sheehan博士，Emory)，以证明通用性并允许利益相关者反馈和设计 修改。 2.我们将通过评估OI与临床并发症和 SCD疾病严重程度的传统实验室测量方法。 3.我们将通过展示其预后价值，将优化的MIRCA的使用扩展到临床空间 在对经历临床并发症和过渡的SCD患者的纵向评估中 心理治疗。 利用我们在生物医学工程师和受过血液学训练的内科科学家之间的独特合作， 我们将在临床试验和患者护理中优化、验证和实现Mirca技术的临床采用 适用于美国和世界各地患有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.