Developing second generation SCID pig models: filling the gaps to improve translation of therapeutics in regenerative medicine

项目摘要/摘要 开发再生医学疗法和最大限度地实现安全移植，迫切需要大型动物模型 提供给临床，因为FDA建议提供关于小动物和大动物模型的数据，以便批准这种 治疗。使用R24资金(OD019813)，我们开发了生物遏制设施和方案来饲养Artemis (DCLRE1C)突变的SCID猪模型(ART)以及在IL2RG中也发生突变的改进模型(ART-IL2RG)。SCID的使用 临床前研究中的猪模型需要能够安全地将健康的SCID猪运送到美国 没有能力自己饲养这种猪的感兴趣的科学家。以下两个目标解决了这一问题 SCID猪研究领域的现有不足： 目的1.创造在HEPA过滤隔离器内安全运输SCID猪的方法 给美国和加拿大研究人员的预告片 目标2.通过建立淋浴设施和购买专用设备来改善SCID猪的生物安全和健康 消毒设备。 由于在保持生物遏制的同时运输SCID猪还没有既定的方法，目标1的工作将集中在 由定制拖车和内部隔离器组成的运输系统的开发将使这一点变得至关重要 下一步有可能。可移动的内部隔离器既是安全饲养SCID猪所必需的，也是短时间内需要的。 从设施生物遏制气泡到灭菌拖车的长途运输。 此外，这一修订计划扩大了可以使用SCID猪的客户类型，作为包含SCID猪的隔离器 可以在外部站点维护，以进行短期研究。在目标2中，我们继续开发有效的 用于SCID养猪的协议，并已确定隔离器是最初提议的和 经过验证的气泡程序。通过增加淋浴装置，SCID猪的生物安全性将显著提高 设施和购买专用过氧化氢气化器，其大小足以对这些新舱位进行消毒 设备。我们建议使用A&R资金翻新现有生物围堵气泡附近的未使用空间，以 提供这种淋浴设施，并将SCID猪舍的生物安全水平提高到行业标准。这个 灭菌设备，由能够处理现有气泡的专用过氧化氢汽化器组成 由于现有设备不足，目标1中提议的新运输方法是非常必要的。这个 拟议的补充目标不会扩大现有父母赠款的范围，而是直接响应呼吁 用于A&R补充剂。

Project Summary/Abstract Large animal models are critically needed to develop regenerative medicine therapeutics and maximize safe translation to the clinic, as the FDA recommends data be provided on both small and large animal models for approval of such therapies. Using R24 funding (OD019813), we developed biocontainment facilities and protocols to raise an Artemis (DCLRE1C) mutant SCID pig model (ART) as well as an improved model also mutant in IL2RG (ART-IL2RG). The use of SCID pig models in preclinical research necessitates the ability to safely ship healthy SCID pigs across the United States to interested scientists who do not have the ability to raise such pigs themselves. The following two Aims address this existing deficit in the SCID pig research field: Aim 1. Create methods to safely transport SCID pigs within a HEPA filtered isolator transported in a HEPA filtered trailer to researchers in the US and Canada Aim 2. Improve the biosecurity and health of SCID pigs by creating a shower-in facility and purchase dedicated sterilization equipment. As there is no established method to ship SCID pigs while maintaining biocontainment, work in Aim 1 will focus on development of a transportation system consisting of a customized trailer and internal isolators will make this critical next step possible. The transportable internal isolators are needed both to safely house the SCID pigs as well as short- distance transportation from facility biocontainment bubbles to the sterilized trailer for long-distance transport. Further, this revised plan expands the type of clientele who can use the SCID pig, as the isolators containing the SCID pig can be maintained at an external site for short-term research studies. In Aim 2, we continue to develop effective protocols for SCID pig rearing and have identified that isolators are a useful adjunct to the originally proposed and validated bubble procedures. Biosecurity of the SCID pigs will be dramatically improved by the addition of a shower-in facility and purchase of dedicated hydrogen peroxide vaporizer adequately sized for sterilizing these new spaces and equipment. We propose to use A&R funds to renovate unused space adjacent to the existing biocontainment bubbles to provide this shower-in facility and raise the level of biosecurity for SCID pig housing to the industry standard. The sterilization equipment, consisting of a dedicated hydrogen peroxide vaporizer capable of handling the existing bubbles and the proposed new transportation approach in Aim 1, is critically needed as existing equipment is inadequate. The proposed Supplemental Aims do not expand the scope of the existing parent grant and are directly responsive to the call for A&R supplements.