Magnetic Flow Sorter for Pancreatic Islet Isolation

项目概要/摘要：SHOT，Inc.，与印第安纳州的明尼苏达大学（UofM）合作 大学医学院（IUSoM）和IKOtech LLC提出了一项磁分离研究 提供用于移植的胰岛的效力和产量的改进的技术。胰岛 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 在临床试验阶段然而，将胰岛移植引入临床的最重要的问题之一是， 更大的公众是，从单个供体胰腺中分离出临界质量的健康、有活力的胰岛， 实现成功的临床移植。目前的分离和纯化技术损失了30-50%的 原始胰岛质量并导致细胞凋亡和/或活力丧失。这种活性胰岛的丧失几乎 总是导致需要利用两个胰腺进行单次移植（“埃德蒙顿方案”）。的 所提出的创新是一种革命性的装置，该装置能够将胰岛与 单器官胰岛移植成功的目标。这项研究的最终目标是增加 在分离过程中回收的健康的、有活力的、分离的和纯化的胰岛的质量 胰岛非常适合移植到I型糖尿病患者体内。这种新的净化系统是基于 四极磁流分选（QMS）技术，已获得IKOtech从俄亥俄州的许可 州立大学和克利夫兰诊所基金会。可用的QMS硬件能够分离细胞大小的 颗粒（直径6-40 º m）;然而，处理较大的胰岛（150-350 º m）需要大量的 对QMS硬件的修改。在第一阶段的研究中，肖特成功地开发了关键的 QMS处理完整胰岛所需的技术。新的QMS将解决一些问题- 与人和异种胰岛制备相关的已知问题。对于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） 与外部同事一起，研究和鉴定具有颗粒大小 和磁化率，其在三个猪分离试验中被优化用于分离胰岛，并且满足 适当的生物相容性（即细胞毒性和免疫原性）要求，以及（3）验证性能 通过在三个标准化猪胰腺中测试QMS系统和试剂， 与密歇根大学的胰岛分离团队合作进行胰岛分离。胰岛分离质量将通过 胰岛计数、耗氧率（OCR）和每毫克DNA的OCR（OCR/DNA）测量、蛋白质组学 和基因组分子谱分析以及糖尿病裸鼠生物测定。第二阶段的工作旨在证明 QMS技术分离胰岛的合理安全性和有效性，并将导致 临床前数据，以支持第二阶段竞争继续补助金所需的测试，最终导致 批准临床试验质量管理体系的试验用器械豁免（IDE）。项目叙述：项目<$用于胰岛分离的磁流分选仪<$是相关的 作为胰岛移植的一项使能技术， 目前正在进行临床试验的1型糖尿病有希望的微创治疗。 1型糖尿病目前在全国范围内约有800，000多人患有6万或更多病例 每年诊断。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 一次移植一个器官），因此，这项技术的成功将使移植数量增加一倍。 从尸体供体组织移植的胰岛可以治愈的患者， 随着猪异种移植的出现， 进一步推动技术。

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.