AUTOMATED CLOSED SYSTEM FOR DMSO REMOVAL FROM PERIPHERAL BLOOD STEM CELLS

描述（由申请人提供）：大剂量化疗后外周血造血干细胞（PBSC）移植常用于治疗各种血液恶性肿瘤。为了促进这一过程，通常收集自体PBSC，冷冻保存并保存一段时间。目前的PBSC冷冻保存需要使用摩尔浓度的冷冻保护剂添加剂二甲基亚砜（DMSO）。通常，冷冻解冻的细胞与DMSO一起移植回患者体内。然而，长期以来，DMSO回输与各种不良事件相关，从皮肤潮红、头痛、发烧、呼吸困难、腹部痉挛、恶心和腹泻到更严重的影响，如溶血、心血管症状和脑血管缺血导致神经系统事件。这些可能导致发病率增加、住院时间延长和治疗相关费用增加。目前，当试图去除DMSO时，通常需要使用离心机对细胞进行“清洗”。这些方法引入机械力和渗透应力，导致细胞堆积/结块和潜在的重大细胞损失。他们还需要额外的实验室工作人员，并且每个病人需要3到4个小时的工作，这使得许多机构的手术更加昂贵和实际困难。最后，在细胞洗涤过程中，很难将细胞保持在一个“封闭”的系统中，这可能导致污染。本提案的目标是在适当的设计控制下完成、构建和测试第一阶段开发的自动化、便携式、封闭系统扩散驱动洗涤装置，用于清除PBSC中的DMSO，以防止在临床移植期间将DMSO输注到患者体内，而不损失细胞数量或细胞功能。在这项II期研究中，该设备将在临床前和最终临床测试中进行评估，以便向FDA提交上市前申请510(k)，以便在III期销售该设备。临床前测试将依赖于体外流式细胞术和克隆生成能力，以及在NOD/SCID小鼠模型中植入。临床试验将包括在多达20名多发性骨髓瘤患者中使用，等待可接受的临床前结果。外周血干细胞移植（PBSC）用于治疗多种疾病，如淋巴瘤、白血病、多发性骨髓瘤、乳腺癌、睾丸癌、神经母细胞瘤等。这些细胞被收集起来，冷冻并储存几段时间，以确保有足够的细胞可供移植，并在冷冻这些细胞时加入一种叫做二甲基亚砜（DMSO）的化合物。这种DMSO在冷冻期间保护细胞，但在解冻后可能对细胞造成损害，并可能导致PBSC移植并发症。本研究的目标是制造一种设备，在移植前容易地从PBSC中去除DMSO，提高细胞的有效性并减少潜在的并发症。

DESCRIPTION (provided by applicant): High dose chemotherapy followed by peripheral blood hematopoietic stem cell (PBSC) transplantation is frequently used for treating various hematologic malignancies. To facilitate this process, autologous PBSC are typically collected, cryopreserved and stored for some period of time. Current PBSC cryopreservation requires the use of molar concentrations of the cryoprotectant additive dimethyl sulfoxide (DMSO). Typically, frozen-thawed cells are transplanted back into patients along with DMSO. However, reinfusion of DMSO has long been associated with various adverse events, ranging from skin flushing, headache, fever, dyspnea, abdominal cramping, nausea, and diarrhea to more sever effects such as, hemolysis, cardiovascular symptoms and cerebrovascular ischemia leading to neurological events. These can result in increased morbidity, prolonged hospitalization, and increased treatment-related costs. Currently, when removal of DMSO is attempted, the procedures typically involve cell "washing" using centrifugation. These methods introduce mechanical forces and osmotic stress causing cell packing/clumping and potential significant cell loss. They also require additional laboratory staff as well, and take between 3 and 4 hours of work per patient, which makes the procedure more expensive and practically difficult for many institutions. Finally, it is difficult to kept the cells in a "closed" system during cell washing, which may lead to contamination. It is the goal of this proposal to finalize, build under appropriate design control and test the automated, portable, closed-system diffusion-driven washing device developed in Phase I for use in clearance of DMSO from PBSC, to prevent infusion of DMSO into patients during clinical transplant without loss of cell numbers or cell functionality. In this Phase II research, the device will be evaluated in preclinical and ultimately clinical testing for 510(k) premarket application to the FDA for clearance to market the device in Phase III. Preclinical testing will rely on in vitro flow cytometry and clonogenic ability as well as engraftment in a NOD/SCID mouse model. Clinical testing will include use in up to 20 multiple myeloma patients, pending acceptable preclinical results. PUBLIC HEALTH RELEVANCE STATEMENT Peripheral blood stem cell transplant (PBSC) is used to treat many types of diseases, such as lymphoma, leukemia, multiple myeloma, breast cancer, testicular cancer, neuroblastoma and others. These cells are collected, frozen and stored over several periods to ensure enough are available for transplant, and to freeze these cells a compound called dimethyl sulfoxide (DMSO) is added. This DMSO protects the cells during freezing, but can be damaging to cells after they are thawed and can cause complications with transplantation of PBSC. The goal of this research is to produce a device to easily remove DMSO from PBSC before they are transplanted, increasing effectiveness of the cells and reducing potential complications.