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，仅发病率 预计会上升。尽管经过数十年的研究，目前对 I 型患者的护理标准 糖尿病（TID）仍然需要严格的血糖监测以及每日服用药物 外源性胰岛素和糖尿病饮食，这些人仍然面临着不良副作用的挑战 该疾病的并发症部分是由于日常依从性问题造成的。胰岛移植具有巨大的 潜力，但仍然存在严重的技术限制。缺乏合适的细胞生物材料 封装是临床应用的主要障碍之一。迄今为止使用的电池封装材料有 免疫原性，并导致组织被膜形成和细胞移植失败。 为了应对这一挑战，我们在这里提出合成 7000 种候选免疫调节藻酸盐—— 基于水凝胶胶囊制剂的细胞封装和高通量体内方法 鉴定能够长期保护移植供体并提高其生存能力的新型材料 胰岛细胞。 774 种海藻酸盐配方的第一代筛选指导了我们这一提议的设计 第二代7000模拟结构。为了提高筛选通量，我们建议 在同一植入部位植入不同材料的混合物。将材料特性与 观察材料功能，我们正在开发下一代测序 (NGS) 测定法来确定 通过共封装 HuVEC 细胞的单核苷酸多态性 (SNP) 基因型确定材料身份。 对 23 名志愿者使用 84 重 SNP NGS 检测组得出的初步结果表明基因组可信度高 使用有限的 NGS 读取进行识别。因此，通常植入 200 个藻酸盐珠（直径 1.5 毫米） 允许同时评估 20 种不同的植入材料，并且仍然有足够的独立珠子 每个材料允许统计分析。 在这个研究项目的过程中，PI将首先合成新的类似物库化合物。 随后，PI 将使用胰岛/HuVEC 异种移植在体内筛选这些材料 将混合物注入促纤维化 C57BL/6 模型啮齿动物中，这使我们能够同时筛选多达 20 倍的啮齿动物 单一啮齿动物体内的不同材料配方。接下来，PI将考察潜在客户的能力（5-10） 在 T1D 啮齿动物模型中保护移植胰岛并恢复正常血糖的制剂长达 100 天。 最后，先导制剂将在健康的非人类灵长类动物研究中进一步审查。我们预计，届时 该项目完成后，将确定用于封装胰岛细胞的海藻酸铅材料能够成功 两种不同的动物模型（C57BL/6 小鼠和猕猴），将为临床前测试做好准备 符合 GLP/GMP 条件，准备 FDA IND 提交。