Encapsulated Porcine Islets into Rhesus Macaques

描述(申请人提供)：异体胰岛移植作为一种治疗I型糖尿病的概念已经被过去五年在埃德蒙顿、迈阿密、明尼阿波利斯、费城和其他地点所做的开创性工作证明是可行的。然而，供移植的组织仍然存在相当大的短缺。Microlslet，Inc.的重点是通过提供免疫隔离的成年猪胰岛异种移植来满足对内分泌组织日益增长的需求。到目前为止，我们已经开发了新的冷藏和胰岛加工解决方案，并产生了改进的胰岛培养条件。在撰写本文时，我们已经处理了超过87个胰腺，我们的平均产量已经增加到每个胰腺437,000 q34,000 LEQ，纯度大约90%(&gt；90%的胰岛分离成功率)。虽然在第一阶段的AIMS中没有提出，但在本建议的第一阶段中产生的胰岛被包裹在两种类型的钙-海藻酸盐微囊中，并在两种链脲佐菌素诱导的糖尿病小鼠模型中进行了功能测试。在4个实验中，将裸露和包裹的猪胰岛移植到47只免疫缺陷、糖尿病小鼠(NOD/SCID)的腹膜腔中。每只动物接受的移植物组织量从500到104 IE不等，其中100%的动物在移植后2天内恢复到正常血糖。到目前为止，这些动物已经在移植后6个月以上保持了移植功能。在第二系列研究中，将包裹的胰岛移植到具有免疫能力的小鼠(C57BL/6)中，在没有任何免疫抑制的情况下，100%的动物在2天内恢复到正常血糖。目前已有3组链脲佐菌素诱导的糖尿病大鼠C57BL/6移植了我们的核心技术包被的胰岛，其中40%的动物血糖正常超过200天。这些数据表明，将包裹的成年猪胰岛移植到糖尿病啮齿动物模型中是有功能和生物相容性的。为了确定这种异种移植物是否有可能在人类身上发挥作用，下一步是在糖尿病的非人类灵长类动物身上进行测试，这是一个已经建立的大型动物临床前模型。因此，第二阶段的目标是展示包裹的猪胰岛在非人类灵长类动物中的长期功能。具体目的1：证实藻酸盐微囊化猪胰岛异种移植在链脲佐菌素诱导的糖尿病非人类灵长类动物中的安全性和有效性。目标I的目的是确定成年猪胰岛异种移植在糖尿病恒河猴中的安全性、有效性和命运，并将其包裹在基于藻酸盐的专利制剂中。特定目的2：胰岛分离、微囊化和移植末端制备的方法和材料的鉴定。目标2的目的是验证和鉴定良好实验室操作规范(GLP)标准所描述的方法和材料，为研究性新药/装置(IND)的应用做准备。

DESCRIPTION (provided by applicant): The concept of islet allotransplantation as a treatment for Type I Diabetes has been demonstrated as feasible by the pioneering work done in Edmonton, Miami, Minneapolis, Philadelphia and other sites during the last five years. However, there still exists a considerable shortage of tissue for transplantation. The focus of Microlslet, Inc is to meet the increased demand for endocrine tissue by providing immune-isolated adult porcine islet xenografts. To date we have developed novel cold storage and islet processing solutions and generated modified islet culturing conditions. We have processed over 87 pancreata at the time of this writing and our average yields have increased to 437,000 q34,000 lEq per pancreas with approximately 90% purity (>90% successful islet isolation rate). Though not proposed in the Phase 1 aims, islets generated during Phase I of this proposal were encapsulated in 2 types of Ca2+-alginate microcapsules and were tested for function in two streptozotocin-induced diabetic mouse models. In 4 experiments, naked and encapsulated porcine islets were transplanted into the peritoneal cavities of 47 immunodeficient, diabetic mice (NOD/SCID). Animals received varying amounts of graft tissue ranging from 500 to 104 IE per animal, with a return to normoglycemia occurring in 100% of the animals within 2 days of transplantation. To date, these animals have maintained graft function for more than 6 months post-transplantation. In the second series of studies, encapsulated islets were transplanted into immune competent mice (C57BL/6) with a return to normoglycemia achieved within 2 days in 100% of animals without any immunosuppression. Presently, 3 groups of streptozotocin-induced diabetic C57BL/6 transplanted with islets coated with our core technology, a gelled-core Ca2+-alginate bead, where 40% of the animals have been normoglycemic for over 200 days. These data demonstrate that encapsulated adult porcine islets are functional and biocompatible when transplanted intraperitoneally in a diabetic rodent model. To ascertain whether this xenograft has the potential to function in humans, the next step is to test it in diabetic nonhuman primates, an established large animal preclinical model. Therefore, the objective of Phase II is to demonstrate the long-term function of encapsulated porcine islets in non-human primates. SPECIFIC AIM 1: Demonstrate the safety and efficacy of alginate microencapsulated pig islet xenografts in streptozotocin-induced diabetic non-human primates. The goal of Aim I is to determine the safety, efficacy and fate of adult porcine islet xenografts encapsulated in a proprietary alginate-based formulation, in diabetic rhesus macaques. SPECIFIC AIM 2: Qualification of methods and materials for islet isolation, microencapsulation and transplantation for END preparation. The objective of Aim 2 is to validate and qualify the methods and materials described by Good Laboratory Practice (GLP) standards in preparation for an Investigative New Drug/Device (IND) application.