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， 预计将上升。尽管经过数十年的研究，目前对I型糖尿病患者的护理标准 糖尿病（TID）仍然是严格的血糖监测方案， 外源性胰岛素和糖尿病饮食，这些人仍然面临着不利的副作用， 这种疾病的并发症部分是日常依从性问题的结果。胰岛移植具有巨大的 虽然有潜力，但仍然存在严重的技术限制。缺乏合适的生物材料用于细胞 封装是临床应用的关键障碍之一。迄今为止使用的细胞包封材料是 免疫原性，并导致组织囊形成和细胞移植失败。 为了应对这一挑战，在这里，我们提出了7000个候选免疫调节藻酸盐的合成- 用于细胞包封的基于水凝胶的胶囊制剂和高通量体内方法 鉴定能够长期保护移植供体并使其存活的新材料 胰岛细胞第一代筛选的774藻酸盐配方指导我们的设计， 第二代7000模拟结构。为了提高筛选的通量，我们建议 在同一植入部位植入不同材料的混合物。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，以确定 通过共包封的HuVEC细胞的单核苷酸多态性（SNP）基因型确定材料同一性。 使用84重SNP NGS小组对23名志愿者的初步结果表明， 使用有限的NGS读取进行识别。因此，典型的植入200个藻酸盐珠（1.5mm直径） 允许同时评估20种不同的植入材料，仍然有足够的独立珠 以进行统计分析。 在本研究项目的过程中，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 混合物进入促纤维化的C57 BL/6模型啮齿动物，这使我们能够同时筛选高达20倍以上的 不同的材料配方在一个单一的啮齿动物。接下来，PI将检查电极导线的能力（5-10） 在T1 D啮齿类动物模型中，本发明提供了用于保护移植的胰岛并恢复正常血糖长达100天的制剂。 最后，将在健康的非人类灵长类动物研究中进一步审查铅制剂。我们预计， 完成该项目后，将确定用于包封胰岛细胞的海藻酸铅材料， 两种不同的动物模型（C57 BL/6小鼠和猕猴），并将准备进行临床前试验 在GLP/GMP条件下，为FDA IND提交做准备。