APOPTOSIS INTERVENTION IN CELL AND ORGAN PRESERVATION

BioLife Technologies是冷冻医学科学公司(CMS)的全资子公司，于1998年3月31日根据特拉华州公司法成立。BioLife的主要使命是开发改进的低温(冷藏)解决方案，旨在使细胞、组织和器官在4摄氏度的温度下保持近乎暂停的状态。心脏停搏液、器官移植和选定的非监管市场是目标。本文提供的数据表明，BioLife的低温解决方案HypoThermosol(HTS)系列在保护肾脏、心脏和皮肤细胞方面比杜邦-默克公司生产的Viaspan产品更好。Viaspan目前在移植解决方案业务中占有最大的市场份额，但自10年前推出以来，它从未进行过重大修改。更重要的是，Viaspan的设计并不是基于现代分子生物学研究。BioLife启动了一项激进的计划，以确定延长低温储存期间细胞死亡的分子基础，以便了解这些事件可以导致新一代低温解决方案。这项提案中提出的DNA凝胶表明，冷储存太长时间的细胞在返回常温(37摄氏度)两天后会因细胞凋亡(程序性细胞死亡)而死亡。HTS补充了细胞凋亡抑制剂，阻止了这一过程，改善了HTS的性能。CMS专利律师建议BioLife Technologies，在下一代FITS解决方案系列中包括细胞凋亡抑制剂，需要申请专利。该项目第一阶段的具体目标是确定：(1)在低温事件之前、期间和/或之后添加凋亡抑制剂是否能最好地防止极端低温期间发生的细胞凋亡；(2)低温是否会导致人皮肤、肾脏、肝脏、肺和主动脉细胞发生凋亡；(3)细胞死亡抑制基因的激活是否可以防止低温诱导的细胞凋亡；(4)抑制被认为发生在低温保存期间(-180摄氏度)的细胞凋亡也可以增加HTS低温保存液的效力。第一阶段SBIR的成功完成将带来(1)基于可靠的分子生物学研究的改进的冷藏解决方案和(2)专利保护包，该专利保护包将阻止任何其他公司开发基于细胞凋亡抑制的类似冷藏解决方案。拟议的商业应用：BioLife Technologies计划开发用于细胞、组织和器官的低温储存的解决方案。应用可能包括工程组织、心脏停搏液和器官运输/移植。

BioLife Technologies is a wholly owned subsidiary of Cryomedical Sciences (CMS) incorporated on March 31, 1998 under the General Corporation Law of the State of Delaware. BioLife's primary mission is to develop improved hypothermic (cold storage) solutions designed to maintain cells, tissues and organs in a near state of suspended animation at 4 degrees C. Cardioplegia, organ transplant and selected non-regulated markets have been targeted. Data presented herein demonstrate that BioLife's hypothermic solutions, the HypoThermosol (HTS) series, are better at cold-protecting kidney, heart and skin cells than is ViaSpan - a product produced by DuPont-Merck. ViaSpan currently commands the largest market share in the transplant solution business, but it has not been significantly modified since its introduction 10 years ago. More importantly, the design of ViaSpan is not based on modern molecular biology investigations. BioLife has launched an aggressive program to determine the molecular basis of cell death during extended hypothermic storage so that knowledge of these events can lead to a new generation of hypothermic solutions. DNA gels presented in this proposal demonstrate that cells cold stored for too long die by apoptosis (programmed cell death) two days after they are returned to normothermic (37 degrees C) temperatures. HTS supplemented with apoptosis inhibitors blocks this process and improves HTS's performance. CMS patent lawyers have advised BioLife Technologies that the inclusion of apoptosis inhibitors in a future generation series of FITS solutions warrants a patent application. The Specific Aims of this Phase I project are to determine if: (1) apoptosis that occurs during extreme hypothermia is best prevented when apoptosis inhibitors are added before, during and/or after the hypothermic event; (2) if hypothermia-induced apoptosis occurs to human skin, kidney, liver, lung and aortic cells; (3) activation of cell death inhibiting genes prevents hypothermia-induced apoptosis; (4) inhibition of apoptosis thought to occur during cryopreservation (-180 degrees C) also increases the efficacy of HTS cryopreservative solutions. Successful completion of this Phase I SBIR will lead to (1) improved cold storage solutions based on sound molecular biology investigations and (2) a patent protection package that will preclude any other company from developing similar cold storage solutions based on apoptosis inhibition. PROPOSED COMMERCIAL APPLICATIONS: BioLife Technologies proposes to develop solutions to be used for the hypothermic storage of cells, tissues and organs. Applications may include engineered tissues, cardioplegia and organ transport/transplant.