Synthesis and High-Throughput In Vivo Characterization of Alginate Encapsulation Materials for Long-Term Islet

用于长期胰岛的海藻酸盐封装材料的合成和高通量体内表征

• 批准号: 9982065
• 负责人: Omid Veiseh
• 金额: $ 77.51万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9982065
• 关键词: Address; Alginates; Allogenic; Animal Model; Bar Codes; Biocompatible Materials; Biological Assay; Blood Glucose; C57BL/6 Mouse; Caliber; Cells; Chemicals; Chemistry; Chronic Disease; Data; Detection; Diabetes Mellitus; Diabetic Diet; Diabetic mouse; Disease; Donor person; Encapsulated; Epidemic; Evaluation; Exhibits; Failure; Fibrosis; Foreign Bodies; Formulation; Gene Frequency; Generations; Genetic; Genomics; Genotype; Glucose; Graft Rejection; Graft Survival; Histology; Human; Hydrogels; Immune; Immunologic Monitoring; Immunosuppression; Implant; In Vitro; Incidence; Individual; Inflammation; Injections; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Islets of Langerhans Transplantation; Isomerism; Kinetics; Lead; Libraries; Macaca; Mediating; Methods; Modeling; Modification; Monkeys; Mus; Patients; Performance; Preclinical Testing; Preparation; Property; Regimen; Research; Research Project Grants; Rodent; Rodent Model; SNP genotyping; Single Nucleotide Polymorphism; Site; Statistical Data Interpretation; Structure; Testing; Tissues; Transplantation; Triazoles; Validation; analog; base; biomaterial compatibility; capsule; clinical application; combinatorial; density; design; genetic variant; glucose monitor; graft failure; high throughput screening; immunogenic; implant material; implantation; improved; in vitro testing; in vivo; insulin secretion; islet; next generation sequencing; nonhuman primate; novel; response; screening; side effect; standard of care; subcutaneous; theories; treatment research; volunteer

Diabetes remains a global epidemic afflicting more than 300 million people worldwide, with incidence only expected to rise. Despite decades of research the current standard of care for those suffering from type I diabetes (TID) remains a rigorous regimen of blood glucose monitoring together with daily administrations of exogenous insulin and diabetic diet, these individuals still are challenged with untoward side effects due to complications of the disease in part the result of daily compliance issues. Islet transplantation has tremendous potential, but serious technological limitations remain. The lack of suitable biomaterials used for cell encapsulation is one of the key obstacles to clinical application. Cell encapsulation materials used to date are immunogenic, and lead to tissue capsule formation and cell graft failure. To address this challenge, here we propose the synthesis of 7000 candidate immune modulatory alginate- based hydrogel capsule formulations for cell encapsulation and a high-throughput in vivo method for identification of novel materials which can enable long-term protection and viability of transplanted donor pancreatic islet cells. A first-generation screen of 774 alginate formulations guides our design of this proposed second generation 7000 analogue structures. To enable increased throughput of screening, we propose to implant mixtures of different materials in the same implantation site. To pair the material identity with the observed material function, we are developing a next-generation sequencing (NGS) assay to determine material identity via a single nucleotide polymorphism (SNP) genotype of co-encapsulated HuVEC cells. Preliminary results using a 84-plex SNP NGS panel on 23 volunteers indicate high confidence genomic identification using limited NGS reads. Thus, a typical implantation of 200 alginate beads (1.5mm diameter) allows the simultaneous evaluation of 20 different implanted materials, with still sufficient independent beads per material to allow statistical analysis. In the course of this research project, the PI will first synthesize the new analogue library compounds. Subsequently, the PI will screen these materials in vivo using a xenogeneic transplantation of islets/HuVEC mixtures into a profibrotic C57BL/6 model rodents, that enables us to simultaneously screen up to 20-fold more distinct materials formulations in a single rodent. Next, the PI will examine the ability of leads (5-10) formulations to protect transplanted islet and restore normoglycemia in a T1D rodent model for up to 100 days. Finally lead formulations will be further vetted in healthy non-human primate studies. We anticipate that upon completion of this project, lead alginate materials for encapsulating islet cells will be identified that succeed in two different animal models (C57BL/6 mouse and macaque monkeys) and will be ready for pre-clinical tests under GLP/GMP conditions in preparation for an FDA IND submission.

糖尿病仍然是一种全球流行病，困扰着全世界3亿多人，其发病率仅为 预计会上升。尽管进行了数十年的研究，但目前对I型患者的护理标准 糖尿病(TID)仍然是一种严格的血糖监测方案，同时每天给药 外源性胰岛素和糖尿病饮食，这些人仍然面临着由于以下原因而产生的不良副作用 这种疾病的并发症部分是由于日常依从性问题造成的。胰岛移植有巨大的 有潜力，但仍然存在严重的技术限制。用于细胞的合适生物材料的缺乏 胶囊化是制约其临床应用的关键因素之一。到目前为止使用的细胞封装材料有 免疫原性，并导致组织包膜形成和细胞移植失败。 为了应对这一挑战，我们建议合成7000候选免疫调节性海藻酸盐- 用于细胞包埋的水凝胶胶囊配方和体内高通量方法 能够对移植供者进行长期保护和存活的新材料的鉴定 胰岛细胞。774种海藻酸盐配方的第一代筛选指导我们设计这项提议 第二代7000个模拟结构。为了增加筛查的吞吐量，我们建议 在同一种植部位使用不同材料的种植体混合物。将物质身份与 观察到的物质功能，我们正在开发一种下一代测序(NGS)分析来确定 通过共包裹的HUVEC细胞的单核苷酸多态(SNP)基因型进行材料鉴定。 在23名志愿者身上使用84个SNP NGS小组的初步结果表明高可信的基因组 使用有限的NGS读取进行身份识别。因此，一个典型的植入200颗海藻酸盐珠(直径1.5毫米) 允许同时评估20种不同的植入材料，仍然有足够的独立珠子 每种材料，以便进行统计分析。 在这个研究项目的过程中，PI将首先合成新的模拟库化合物。 随后，PI将使用异种胰岛/人脐静脉内皮细胞移植在体内筛选这些材料 混合成促纤维化的C57BL/6啮齿动物模型，使我们能够同时筛查多达20倍的 在一只啮齿动物身上有不同的材料配方。接下来，PI将检查引线的能力(5-10) 在T1D啮齿动物模型中保护移植胰岛并恢复正常血糖长达100天的配方。 最后，铅配方将在健康的非人类灵长类动物研究中进一步审查。我们期待着那一天 该项目完成后，将确定用于包裹胰岛细胞的海藻酸铅材料 两种不同的动物模型(C57BL/6小鼠和猕猴)，并将准备进行临床前测试 在GLP/GMP条件下为FDA IND提交做准备。