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.

糖尿病仍然是全球流行病，全球超过3亿人，仅发病率 预计会上升。尽管进行了数十年的研究，但目前对患有I型患者的人的护理标准 糖尿病（TID）仍然是严格的血糖监测方案，每天管理 外源性胰岛素和糖尿病饮食，这些人仍然受到挑战，因为 该疾病的并发症部分是每日合规性问题的结果。胰岛移植具有巨大的 潜力，但仍然存在严重的技术局限性。缺乏用于细胞的合适的生物材料 封装是临床应用的关键障碍之一。迄今为止使用的单元包封装材料是 免疫原性，并导致组织囊的形成和细胞移植衰竭。 为了应对这一挑战，我们在这里提出合成7000个候选免疫调节藻酸盐 - 用于细胞封装的基于水凝胶胶囊制剂和用于体内的高通量方法 识别可以实现移植供体的长期保护和生存能力的新型材料 胰岛细胞。 774藻酸盐配方的第一代屏幕指导我们对此提出的设计 第二代7000个模拟结构。为了增加筛查的吞吐量，我们建议 在同一植入部位的不同材料的植入物混合物。将物质身份与 观察到的材料功能，我们正在开发下一代测序（NGS）测定法以确定 通过共同封装的HUVEC细胞的单核苷酸多态性（SNP）基因型的材料认同。 在23名志愿者上使用84 plex snp ngs面板的初步结果表明基因组高置信度 使用有限的NGS读取的识别。因此，典型的200藻酸珠（直径1.5mm）的植入 允许同时评估20种不同的植入材料，并具有足够的独立珠 根据材料允许统计分析。 在该研究项目的过程中，PI将首先合成新的模拟库化合物。 随后，PI将使用胰岛/HUVEC的异构移植在体内筛选这些材料 混合物成纤维化的C57BL/6型啮齿动物，使我们能够同时筛选多达20倍 单一啮齿动物中的不同材料制剂。接下来，PI将检查铅的能力（5-10） 在T1D啮齿动物模型中保护移植的胰岛和恢复正常血糖的制剂长达100天。 最终，在健康的非人类灵长类研究中将进一步审查铅配方。我们预料到这一点 完成该项目的完成，将确定用于封装胰岛细胞的铅藻酸盐材料 两种不同的动物模型（C57BL/6小鼠和猕猴），并准备好进行临床前测试 在GLP/GMP条件下，准备FDA IND提交。