Magnetic Flow Sorter for Pancreatic Islet Isolation

描述（由申请人提供）：SHOT, Inc.与明尼苏达大学（UofM），印第安纳大学医学院（IUSoM）和IKOtech LLC合作，提出了一项磁分离技术的研究，该技术可提高用于移植的胰岛的效力和产量。胰岛移植是一种在临床试验中成功逆转1型糖尿病患者胰岛素依赖的治疗方法。然而，将胰岛移植推向更广大公众的最重要问题之一是，很难从单个供体胰腺中分离出足够数量的健康、有活力的胰岛，以实现成功的临床移植。目前的分离和纯化技术使胰岛原有质量损失30-50%，导致细胞凋亡和/或丧失生存能力。这种可存活胰岛的损失几乎总是导致需要在一次移植中使用两个胰腺（“埃德蒙顿方案”）。提出的创新是一种革命性的装置，能够磁隔离胰岛，目标是成功的单器官胰岛移植。本研究的最终目标是提高在分离过程中恢复的健康、有活力、分离和纯化的胰岛的质量，从而产生理想的适合移植到I型糖尿病患者的纯化胰岛。这种新的净化系统基于四极磁流分选（QMS）技术，该技术已获得俄亥俄州立大学和克利夫兰诊所基金会IKOtech的许可。现有的QMS硬件能够分离细胞大小的颗粒（6-40米直径）；然而，处理较大的小岛（150-350米）需要对QMS硬件进行重大修改。在一期研究中，SHOT成功开发了QMS处理完整胰岛所需的关键技术。新的质量管理体系将解决与人类和异种胰岛制剂相关的一些众所周知的问题。对于这个拟议的II期项目，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，研究和确定可用的磁性试剂，其粒度和磁化率在三个猪分离试验中被优化用于分离胰岛，并满足适当的生物相容性（即细胞毒性和免疫原性）要求。(3)与UofM胰岛分离团队合作，在三个标准化的猪胰岛分离中测试QMS系统和试剂，验证该技术的性能。胰岛分离质量将通过胰岛计数、耗氧量（OCR）和每毫克DNA的耗氧量（OCR/DNA）测量、蛋白质组学和基因组分子分析以及糖尿病裸鼠生物测定来评估。II期工作旨在证明QMS技术用于分离胰岛的合理安全性和有效性，并将产生临床前数据，以支持所需测试的II期竞争继续拨款，最终导致临床试验器械豁免（IDE）批准QMS进行临床试验。

DESCRIPTION (provided by applicant): 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 5m diameter); however, processing the larger islets (150-350 5m) 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.