Magnetic Flow Sorter for Pancreatic Islet Isolation

项目摘要/摘要：Sort，Inc.，与印第安纳州明尼苏达大学(UOFM)合作 大学医学院(IUSoM)和IKOtech LLC提出了一项导致磁选的研究 提高胰岛移植的效力和产量的技术。小岛 移植是一种成功逆转1型糖尿病患者胰岛素依赖的治疗方法 在临床试验中。然而，将胰岛移植引入美国，最重要的问题之一 更普遍的是，从单个捐赠者的胰腺中分离出临界数量的健康、有活力的胰岛是很困难的 实现了成功的临床移植。目前的分离和纯化技术损失了30%-50%的 原胰岛肿块并导致细胞凋亡和/或丧失活力。这种可存活的小岛的丧失几乎 总是需要在一次移植中使用两个胰腺(“埃德蒙顿方案”)。这个 拟议的创新是一种革命性的设备，能够通过磁力隔离胰岛 成功的单器官胰岛移植的目标。这项研究的最终目标是增加 在分离过程中恢复的健康的、活的、分离的和纯化的胰岛的质量 胰岛非常适合移植给I型糖尿病患者。这种新的净化系统是基于 四极磁流分选(QMS)技术，已获得来自俄亥俄州的IKOtech许可 州立大学和克利夫兰临床基金会。现有的QMS硬件能够分离细胞大小的 颗粒(直径6-40？m)；然而，处理较大的胰岛(150-350？m)需要显著 对质量管理体系硬件的修改。在第一阶段的研究中，肖特成功地开发出了 质量管理体系加工完整胰岛所需的技术。新的质量管理体系将解决一些良好的- 与人类和异种胰岛制剂相关的已知问题。对于这一拟议的第二阶段项目， Shot将(1)完成能够分离胰岛的QMS系统的设计和制造，(2) 与外部同事合作，研究并确定可用的颗粒大小的磁性试剂 以及在三个猪隔离试验中为隔离胰岛而优化的磁化率，并且满足 适当的生物兼容性(即细胞毒性和免疫原性)要求，以及(3)验证性能 三种标准化猪胰腺QMS系统和试剂联合检测的技术研究 与密歇根大学的小岛隔离团队合作进行小岛隔离。胰岛分离质量将通过以下方式进行评估 胰岛计数、耗氧率(OCR)和每毫克DNA的OCR(OCR/DNA)测量，蛋白质组 以及基因组分子图谱和糖尿病裸鼠生物检测。第二阶段的努力旨在证明 QMS技术用于分离胰岛的合理安全性和有效性，并将导致 支持所需测试的第二阶段竞争延续拨款的临床前数据，最终导致 批准用于临床试验的QMS的研究设备豁免(IDE)。项目简介：用于胰岛隔离的磁流分选器项目是相关的 作为胰岛移植的使能技术，这是一种 目前正在进行临床试验的1型糖尿病有希望的微创治疗方法。 目前，全国估计有80多万人患有1型糖尿病，有60,000例或更多的病例 每年都会确诊。严格的器官采购限制目前限制了 可以通过胰岛移植治疗的患者(这是由于需要更多的 每一次移植一个器官)，所以拟议技术的成功将使这一数字翻一番 可以通过从身体供体组织移植的胰岛治愈的患者 随着猪的异种移植作为一种 进一步的使能技术。

Project Summary/Abstract: SHOT, Inc., in collaboration with the University of Minnesota (UofM), Indiana University School of Medicine (IUSoM) and IKOtech LLC, proposes research leading to a magnetic separation technology that provides an improvement in potency and yield of pancreatic islets for transplantation. Islet transplantation is a method of treatment that has successfully reversed insulin dependence in Type 1 diabetics in clinical trials. However, one of the most significant issues with bringing pancreatic islet transplantation to the greater public is the difficulty of isolating a critical mass of healthy, viable islets from a single donor pancreas to achieve a successful clinical transplant. Current isolation and purification techniques lose 30-50% of the original islet mass and result in cellular apoptosis and/or loss of viability. This loss of viable islets almost always results in the need to utilize two pancreases for a single transplant (the "Edmonton Protocol"). The proposed innovation is a revolutionary device that is capable of magnetically isolating pancreatic islets with the goal of successful single-organ islet transplantations. The ultimate goal of this research is to increase the quality of healthy, viable, isolated, and purified islets recovered during the isolation process yielding purified islets ideally suited for transplantation into Type I diabetic patients. This new purification system is based on Quadrupole Magnetic flow Sorting (QMS) technology, which has been licensed by IKOtech from The Ohio State University and Cleveland Clinic Foundation. Available QMS hardware is capable of separating cell-sized particles (6-40 ¿m diameter); however, processing the larger islets (150-350 ¿m) requires significant modifications to the QMS hardware. During Phase I research SHOT successfully developed the key technologies required for the processing of intact islets by QMS. The new QMS will solve some of the well- known problems associated with human and xenogeneic islet preparations. For this proposed Phase II project, SHOT will (1) complete the design and manufacture of a QMS system capable of isolating pancreatic islets, (2) in conjunction with external colleagues, research and identify available magnetic reagents with a particle size and magnetic susceptibility that is optimized for isolating islets in three porcine isolation trials and that meets appropriate biocompatibility (i.e. cytotoxicity and immunogenicity) requirements, and (3) verify the performance of the technology by testing the QMS system and reagents together in three standardized porcine pancreatic islet isolations in collaboration with the islet isolation team at UofM. Islet isolation quality will be assessed via islet count, Oxygen Consumption Rate (OCR) and OCR per mg of DNA (OCR/DNA) measurements, proteomic and genomic molecular profiling, and diabetic nude mouse bioassays. The Phase II effort is designed to prove the reasonable safety and efficacy of the QMS technology for isolating pancreatic islets and will result in preclinical data in support of a Phase II Competing Continuation grant for required testing, eventually leading to an Investigational Device Exemption (IDE) approving QMS for clinical trials. Project Narrative: The project ¿Magnetic Flow Sorter for Pancreatic Islet Isolations¿ is relevant to public health as an enabling technology for pancreatic islet transplantation, which is a promising and minimally invasive cure for Type 1 Diabetes currently undergoing clinical trials. Type 1 Diabetes currently afflicts an estimated 800,000+ nationally with 60,000 or more cases diagnosed each year. Severe organ procurement limitations currently restrict the number of patients that can be treated via islet transplantation (which is compounded by the need for more than one organ per transplant), so success with the proposed technology will double the number of patients that can be cured with islets transplanted from cadaver donor tissue and will potentially facilitate unlimited treatment with the advent of porcine xenotransplantation as a further enabling technology.