Enhancement of Conditional Reprogramming Technology for Production of Functional Human Beta-Cells

增强条件重编程技术以生产功能性人类β细胞

• 批准号: 9306378
• 负责人: Brian Andrew Pollok
• 金额: $ 22.5万
• 依托单位: PROPAGENIX, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9306378
• 关键词: Alpha Cell; Animal Model; Animals; Applications Grants; B cell differentiation; Beta Cell; Biological Assay; Biomass; Blood Glucose; C-Peptide; Cell Aggregation; Cell Culture Techniques; Cell Lineage; Cell Maturation; Cell Proliferation; Cell Survival; Cell Transplantation; Cell Transplants; Cell physiology; Cell surface; Cell-Matrix Junction; Cells; Complex; Conditioned Culture Media; Cornea; Cyclic GMP; Data; Diabetic mouse; Differentiation Antigens; Disease; Endocrine; Epithelial; Epithelial Cells; Equation; Extracellular Matrix; Formulation; Gene Expression; Gene Expression Profiling; Genetic; Glucose; Growth Factor; Harvest; Human; Implant; In Vitro; Insulin; Islets of Langerhans; Kidney; Kinetics; Label; Mammary gland; Measures; Methods; Modification; Monitor; Mus; N-Cadherin; Nuclear; Nude Mice; Organ Donor; Outcome; Pathology; Pathway interactions; Patients; Phase; Phenotype; Phosphoproteins; Population; Positioning Attribute; Preparation; Process; Production; Prostate; Protocols documentation; Public Health; Quantitative Reverse Transcriptase PCR; Replacement Therapy; Reporter; Reproducibility; Research; Rotation; Sampling; Serum; Small Business Innovation Research Grant; Staining method; Stains; Stem cells; Structure of beta Cell of islet; System; Technology; Testing; Therapeutic; Tissue Expansion; Tissues; Transplantation; Work; beta cell replacement; capsule; cell growth; cell type; cost; density; diabetic patient; differentiated B cell; efficacy study; efficacy testing; embryonic stem cell; experimental study; human embryonic stem cell; in vivo; induced pluripotent stem cell; islet; keratinocyte; mouse model; phenotypic biomarker; polypeptide C; preclinical development; preclinical efficacy; programs; response; safety study; safety testing; screening; small molecule; success; tool; tumor; type I diabetic

Project Abstract Insufficient supply of donor pancreatic islets to meet the demand for transplants severely limits therapeutic options for Type 1 diabetic (T1D) patients in whom insulin therapy is insufficient to control their disease. And while there has been much recent success with the directed differentiation of human embryonic stem cells (hu-ESCs) and human induced pluripotent stem cells (hu-iPSCs) into pancreatic beta cells, the process is still quite imperfect and inefficient. To this end, we have shown that Conditional Reprogrammed (CR) cell culture technology can support the significant expansion of human islet-derived β-cells to at least one million-fold amount. Just as importantly, our preliminary data indicates that CR-propagated human β-cells can readily develop mature differentiation markers and functional phenotype in vitro. To develop this promising technology into a practical option for human β-cell replacement therapy, we seek to enhance the proliferative capacity of CR-grown human β-cells through the use of a focused screening approach that we have used with success with other CR-responsive cell types. The optimization of the post-CR expansion differentiation step is also a component of our proposal, using reproduced findings from other labs on how cell surface interactions can help drive human β-cell maturation and functionality. The ultimate test of using CR technology to expand human β-cells will be to use these cells in an animal model of T1D and to assess their ability to modulate the disease pathology; a return to euglycemia and production of human C- peptide in the transplant-receiving mice will be the positive hallmark of success. Successful completion of these specific research aims will position us to propose a Phase II SBIR application focused on the preclinical development of CR technology into GLP efficacy and safety testing and cGMP-compliant cell manufacturing.

项目摘要 供体胰岛供应不足，以满足移植的需求严重限制了 胰岛素治疗不足以治疗1型糖尿病（T1 D）患者的治疗选择 控制他们的疾病。虽然最近有很多成功的定向 人胚胎干细胞（hu-ESCs）和人诱导多能干细胞的分化 尽管将人诱导多能干细胞（hu-iPSC）转化为胰腺β细胞的过程仍然非常不完善且效率低下。到 为此，我们已经证明条件重编程（CR）细胞培养技术可以 支持人胰岛来源的β细胞显著扩增至至少一百万倍 金额同样重要的是，我们的初步数据表明，CR增殖的人类β细胞 可以在体外容易地形成成熟的分化标记和功能表型。 将这项有前途的技术发展成为人类β细胞替代的实用选择 治疗中，我们寻求增强CR生长的人β细胞的增殖能力，通过 使用我们在其他CR应答患者中成功使用的集中筛选方法 细胞类型。CR后扩增分化步骤的优化也是一个组成部分 我们的建议，使用复制的结果从其他实验室的细胞表面相互作用如何可以 有助于促进人类β细胞成熟和功能。使用CR技术的最终考验， 扩增人β细胞的方法是在T1 D动物模型中使用这些细胞，并评估其 调节疾病病理学的能力;恢复正常和产生人类C- 在接受移植的小鼠中的肽将是成功的积极标志。成功 这些具体研究目标的完成将使我们能够提出第二阶段SBIR 应用重点是CR技术临床前开发转化为GLP疗效， 安全测试和符合cGMP的细胞制造。