Understanding Pancreatic Progenitors for Diabetes Cell-Replacement Therapy

了解胰腺祖细胞用于糖尿病细胞替代疗法

• 批准号: 8350255
• 负责人: Danwei Huangfu
• 金额: $ 39.78万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8350255
• 关键词: Acinus organ component; Address; Beta Cell; Biological Assay; Biology; Cell Culture Techniques; Cell Therapy; Cell surface; Cells; Cellular biology; Characteristics; Chemicals; Complement; Development; Developmental Biology; Diabetes Mellitus; Duct (organ) structure; EGF gene; Embryo; Embryonic Development; Endocrine; Epithelial; Exocrine pancreas; FGF10 gene; Fibroblasts; Fluorescence-Activated Cell Sorting; Foundations; Genetic; Goals; Growth Factor; Human; In Vitro; Individual; Insulin; Investigation; Knowledge; Laboratories; Lead; Molecular; Multipotent Stem Cells; Mus; Outcome; Pancreas; Patients; Physiological; Pluripotent Stem Cells; Property; Protocols documentation; Replacement Therapy; Research; Role; Signal Transduction; Staging; Stem cells; Structure of beta Cell of islet; Therapeutic; Work; base; beta cell replacement; cell type; diabetic patient; global health; human embryonic stem cell; in vivo; induced pluripotent stem cell; islet; mouse development; novel; novel strategies; progenitor; research study; self-renewal; stem cell differentiation; type I and type II diabetes

DESCRIPTION (provided by applicant): Diabetes mellitus is an escalating global health problem. Both Type 1 and Type 2 diabetes lead to the gradual loss of insulin-producing beta cells. Our long-term goal is to develop beta cell replacement strategies to overcome the insulin deficiency in diabetic patients. To achieve this goal, we have developed novel, safer approaches to convert patient fibroblasts to induced pluripotent stem cells (iPSCs) using chemicals together with genetic factors. These iPSCs can be differentiated into multipotent pancreatic progenitors, the precursors to beta cells, using protocols developed for human embryonic stem cells (hESCs). However, our understanding of the basic biology of pancreatic progenitor cells is still rudimentary, which presents hurdles in the development of effective strategies to expand and further differentiate these progenitor cells for therapy. Evidence emerged from our own studies and from those of other laboratories suggests that pancreatic progenitor cells are heterogeneous, and likely consist of subpopulations with different physiological functions. Yet, molecular and functional distinctions between these subpopulations are poorly understood. It is unclear which subpopulation(s) in the developing pancreas is(are) capable of generating functional beta cells, and to which extent a specific subpopulation is capable of self-renewal. Even less is known about the physiological functions and self-renewal properties of pancreatic progenitor cells differentiated from hESCs. We aim to address these critical questions in this proposal. We hypothesize that the pancreatic progenitor cell compartment in vivo contains heterogeneous subpopulations: (i) a common multipotent pancreatic progenitor responsible for generating all three pancreatic lineages; and (ii) distinct lineage-specific progenitors responsible for generating one or two specific pancreatic lineages. Based on this hypothesis, the overall objective of this proposal is to define pancreatic progenitor subpopulations in terms of their molecular characteristics and physiological functions, as well as to identify mechanisms for self-renewal. We anticipate that our study will yield the following outcomes. Aim 1 will reveal the presence of distinct pancreatic progenitor subpopulations both during embryo development and in hESC differentiation culture. This will identify the progenitor subpopulation(s) capable of generating functional cells. Aim 2 will identify cell surface markers for enrichment of the appropriate progenitor subpopulation(s) for further ¿ differentiation into ¿ cells, and set the stage for studies on expansion of progenitor cells. Aim 3 will elucidate the mechanism of self-renewal of distinct progenitor subpopulations, which are critical for the development of novel, effective strategies to expand pancreatic progenitors for beta cell replacement therapy. Additionally, the ability to generate a large quantity of human pancreatic progenitor cells will provide a new way to study the biology of these cells to complement mouse genetics approaches. Broadly, the proposed research will lead to novel findings to fill in critical gaps in our current knowledge of human pancreatic development. PUBLIC HEALTH RELEVANCE: Generating functional pancreatic beta cells from human pluripotent stem cells offers a promising cure for diabetes, a growing global health problem resulting from the loss of beta cells. However, a major challenge in fulfilling such a promise lies in the rudimentary understanding of the distinct types of pancreatic progenitor cells, which are the precursors that generate mature, insulin-secreting beta cells during embryo development. The goal of this research is to use combined stem cell and developmental biology approaches to understand pancreatic progenitor subtypes in terms of their molecular characteristics, physiological functions, and self-renewal properties, which will form the foundation for developing rational approaches to effective beta cell replacement therapy.

描述（由申请人提供）：糖尿病是一个不断升级的全球健康问题。 1 型和 2 型糖尿病都会导致产生胰岛素的 β 细胞逐渐丧失。我们的长期目标是开发β细胞替代策略来克服糖尿病患者的胰岛素缺乏。为了实现这一目标，我们开发了新颖、更安全的方法，使用化学物质和遗传因子将患者成纤维细胞转化为诱导多能干细胞 (iPSC)。使用为人类胚胎干细胞 (hESC) 开发的方案，这些 iPSC 可以分化为多能胰腺祖细胞，即 β 细胞的前体。然而，我们对胰腺祖细胞的基本生物学的了解仍然很初级，这给开发有效策略来扩展和进一步分化这些祖细胞进行治疗带来了障碍。 我们自己的研究和其他实验室的研究得出的证据表明，胰腺祖细胞是异质的，并且可能由具有不同生理功能的亚群组成。然而，人们对这些亚群之间的分子和功能差异知之甚少。目前尚不清楚发育中的胰腺中哪些亚群能够产生功能性β细胞，以及特定亚群能够在多大程度上自我更新。对于从 hESC 分化而来的胰腺祖细胞的生理功能和自我更新特性知之甚少。我们的目标是在本提案中解决这些关键问题。 我们假设体内胰腺祖细胞区室包含异质亚群：（i）共同的多能胰腺祖细胞，负责产生所有三个胰腺谱系； (ii) 负责产生一种或两种特定胰腺谱系的不同谱系特异性祖细胞。基于这一假设，该提案的总体目标是定义胰腺祖细胞 亚群的分子特征和生理功能，以及确定自我更新的机制。我们预计我们的研究将产生以下结果。目标 1 将揭示胚胎发育和 hESC 分化培养过程中不同胰腺祖细胞亚群的存在。这将鉴定出能够产生功能细胞的祖细胞亚群。目标 2 将鉴定细胞表面标记，以富集适当的祖细胞亚群，进一步分化为细胞，并为祖细胞扩增研究奠定基础。目标 3 将阐明不同祖细胞亚群的自我更新机制，这对于开发新的、有效的策略来扩展胰腺祖细胞以用于 β 细胞替代疗法至关重要。此外，产生大量人类胰腺祖细胞的能力将为研究这些细胞的生物学提供一种新方法，以补充小鼠遗传学方法。总的来说，拟议的研究将带来新的发现来填补关键问题 我们目前对人类胰腺发育的了解存在差距。 公共健康相关性：从人类多能干细胞生成功能性胰腺β细胞为糖尿病提供了一种有希望的治疗方法，糖尿病是由于β细胞丧失而导致的日益严重的全球健康问题。然而，实现这一承诺的一大挑战在于 对不同类型的胰腺祖细胞的初步了解，这些细胞是胚胎发育过程中产生成熟、分泌胰岛素的β细胞的前体。这项研究的目标是利用干细胞和发育生物学相结合的方法来了解胰腺祖细胞亚型的分子特征、生理功能和自我更新特性，这将为开发有效的β细胞替代疗法的合理方法奠定基础。