Clinical Microfluidic Assessment of Red Blood Cell Adhesion, Deformability, Cellular Hemoglobin Distribution, Cellular Density, and Blood Rheology for Curative Therapies in Sickle Cell Disease

项目摘要 在本提案中，我们建立在我们相关的已发表工作1 -14和正在进行的研究活动的基础上， 一个活跃的NHLBI R 01（R 01 HL 133574，PI：Gurkan，Little），标题为：“标准化监测细胞 粘附改善镰状细胞病临床护理”，一项有效的FDA R 01（R 01 FD 005341，PI： Manwani），标题为：“Gamunex（静脉注射丙种球蛋白）治疗镰状细胞急性疼痛的2期试验”， 和一个活跃的NSF奖（1552782，PI Gurkan）：“职业：红细胞粘附的生物力学 变形能力”。 我们已经开发了红细胞（RBC）和白色血细胞（WBC）生物标志物测定法， 镰状细胞病（SCD）1-14的表型分析，得到NHLBI R 01（R 01 HL 133574）和 FDA R 01（R 01 FD 005341）奖，如上所述。在这个项目中，我们建立在我们的专业知识， SCD生物标志物测定开发，以独特地解决OTA中确定的以下高优先级领域- 19-007治愈镰状细胞倡议： (1)开发并验证定量测定细胞分布和半定量测定的测定方法 胎儿、成人和/或镰状血红蛋白的细胞表达。 (2)开发和验证用于评估红细胞流变学的分析方法，包括微流体和成像 流式细胞仪镰状化测定。 在这个项目中，我们提出了先进的分析和临床验证的集成临床微流体 测定以评估RBC粘附、变形性、细胞血红蛋白分布、细胞密度（如 每个单个细胞的质量与体积比）和全血流变学作为候选生物标志物， 评估SCD患者对治愈性治疗的反应。

PROJECT SUMMARY In this proposal we build on our relevant published work1-14, and ongoing research activities supported by an active NHLBI R01 (R01HL133574, PI: Gurkan, Little), titled: “Standardized Monitoring of Cellular Adhesion to Improve Clinical Care in Sickle Cell Disease”, an active FDA R01 (R01FD005341, PI: Manwani), titled: “Phase 2 Trial of Gamunex (Intravenous Gammaglobulin) for Sickle Cell Acute Pain”, and an active NSF Award (1552782, PI Gurkan): “CAREER: Biomechanics of Red Blood Cell Adhesion and Deformability”. We have developed red blood cell (RBC) and white blood cell (WBC) biomarker assays for more robust phenotypic analyses in sickle cell disease (SCD)1-14, supported by an NHLBI R01 (R01HL133574) and an FDA R01 (R01FD005341) award, as described above. In this project we build on our expertise in novel SCD biomarker assay development to uniquely address the following high priority areas identified in OTA- 19-007 for Cure Sickle Cell Initiative: (1) Develop and validate assays to quantitatively determine cellular distribution and semi-quantitative cellular expression of fetal, adult and/or sickle hemoglobin. (2) Develop and validate assays to assess red blood cell rheology, including microfluidics and imaging flow cytometry sickling assays. In this project we propose advanced analytical and clinical validation of integrated clinical microfluidic assays to assess RBC adhesion, deformability, cellular hemoglobin distribution, cellular density (as mass to volume ratio for each individual cell), and whole blood rheology as candidate biomarkers to assess response to curative therapies in SCD.