Characterizing the Role of Pancreatic Progenitors in Regeneration

描述胰腺祖细胞在再生中的作用

• 批准号: 10160892
• 负责人: Michael J Parsons
• 金额: $ 37.43万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10160892
• 关键词: Ablation; Adult; Affect; Architecture; Automobile Driving; B cell differentiation; Behavior; Beta Cell; Binding; Biological; Biological Models; Biology; Cell Line; Cells; Cilia; Collection; Data; Data Set; Development; Developmental Biology; Diabetes Mellitus; Disease; Endocrine; Fishes; Genes; Genetic Transcription; Genome; Genomic approach; Genomics; Goals; Grant; Haploidy; Heterozygote; Homeostasis; Human; Human Cell Line; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Lead; Maps; Mediating; Mediator of activation protein; Methods; Modeling; Molecular; Morphology; Natural regeneration; Non-Insulin-Dependent Diabetes Mellitus; Organogenesis; Pancreas; Pancreatic Centroacinar Cell; Pathway interactions; Pharmacology; Phenotype; Play; Prevalence; Process; Production; Recovery; Regenerative capacity; Regulation; Reporter; Resources; Role; Route; Source; TACSTD1 gene; Techniques; Testing; Therapeutic; Transcript; Transgenic Organisms; Vertebrates; Work; World Health Organization; Zebrafish; beta cell replacement; cell regeneration; cell type; chromatin immunoprecipitation; cost effective; deep sequencing; dosage; insight; interest; knock-down; mutant; notch protein; novel; pancreas development; progenitor; regenerative; stem cells; therapeutically effective; tool; transcription factor; transcriptome

Project Summary The World Health Organization estimates that the global prevalence of diabetes in adults is 9%. Both Type 1 and Type 2 diabetes involve the issue of reduced β-cell mass; subsequently a cure for diabetes must involve β-cell replacement. Ideally, a cure would involve inducing regeneration via β-cell neogenesis from endogenous pancreatic progenitors. For these reasons we are interested in explicating the process of β-cell neogenesis, i.e., how β cells are formed from progenitors in the pancreas. Unlike their mammalian counterparts, we have shown that zebrafish readily regenerate their β cells following cell-specific ablation. Our goal is to identify the mechanisms behind the zebrafish’s capacity for β-cell neogenesis. Such molecular pathways could then be pharmacologically exploited in humans to induce β-cell neogenesis. We have recently made two discoveries critical to understanding how zebrafish so easily recover following β- cell ablation. First, we identified the progenitor source for β-cell neogenesis—namely a cell type called the centroacinar cell (CAC); second, we discovered that diminished activity of the Sox9b transcription factor leads to significantly accelerated regeneration. From these insights we hypothesized 1) Sox9b acts cell autonomously to maintain progenitor potency in adult CACs; 2) Diminishing Sox9b activity alters the behavior of CACs or their progeny in regeneration; and 3) The identification of downstream genes of SOX9 will elucidate molecular mechanisms that regulate β-cell differentiation. By testing these hypotheses in three complementary yet independent aims, we expect to discover if the differences in regeneration between sox9b heterozygotes and wildtypes is due either to pre-existing differences in morphology or the behavior of the CACs during regeneration. Furthermore, we will use genomic approaches to identify the direct downstream transcriptional targets of Sox9 homologs because we expect these targets will be the mediators of the sox9b haploinsufficient phenotype. As part of our preliminary data we knocked down SOX9 levels in the human PANC-1 cell line, a surrogate for pancreatic progenitors, and identified affected transcript levels. We have also used chromatin immunoprecipitation and deep sequencing (ChIP-seq) to identify where SOX9 binds in the PANC-1 genome. Putting these results together has allowed us to find direct targets of SOX9 transcriptional activity in PANC-1 cells. We have identified interesting genes downstream of SOX9, such as EpCAM, and identified biological pathways that SOX9 controls, such as cilia function and Notch regulation. Greatly encouraged by our preliminary results we now aim to expand this analysis to find SOX9 targets during development and regeneration. By the end of this proposed work we expect a better understanding of SOX9 function and the discovery of potential therapeutic routes to alleviate β-cell paucity in humans.

项目摘要 世界卫生组织估计，全球成人糖尿病患病率为9%。两种类型均为1 而2型糖尿病涉及β细胞质量减少的问题；因此，糖尿病的治疗必须包括 β-细胞更换。理想情况下，治疗方法包括通过内源性β细胞新生诱导再生。 胰腺祖细胞。由于这些原因，我们有兴趣解释β细胞新生的过程， 也就是说，β细胞是如何从胰腺的祖细胞形成的。与它们的哺乳动物不同，我们有 研究表明，斑马鱼在细胞特异性消融后很容易再生β细胞。我们的目标是找出 斑马鱼β细胞新生能力背后的机制。这样的分子途径可以是 在人类中用来诱导β细胞新生的药理作用。 我们最近有了两个发现，对于理解斑马鱼是如何在β之后如此轻松地恢复至关重要的-- 细胞消融。首先，我们确定了β的祖细胞来源--细胞新生--即一种称为 第二，我们发现Sox9b转录因子活性降低导致 显著加快了再生速度。根据这些洞察力，我们假设Sox9b行为细胞 自主地维持成年CACs的祖细胞潜能；2)SOX9B活性减弱改变 CACs或其后代在再生过程中的行为；3)CACs下游基因的鉴定 SOX9将阐明调控β细胞分化的分子机制。通过测试这些 在三个互补但独立的目标中的假设，我们希望发现 Sox9b杂合子和野生型之间的再生是由于先前存在的 再生过程中CACs的形态或行为。此外，我们将使用基因组方法来 确定Sox9同源物的直接下游转录靶点，因为我们预计这些靶点将 是Sox9b单倍体不足表型的介体。 作为我们初步数据的一部分，我们下调了人类PANC-1细胞系中SOX9的水平，SOX9是人类PANC-1细胞的替代品 胰腺祖细胞，并确定受影响的转录水平。我们还使用了染色质 免疫沉淀和深度测序(CHIP-SEQ)以确定SOX9在Panc-1基因组中的结合位置。 将这些结果放在一起，我们就可以找到PANC-1中SOX9转录活性的直接靶点 细胞。我们已经确定了SOX9下游有趣的基因，如EpCAM，并确定了生物 SOX9控制的通路，如纤毛功能和Notch调节。受到我们的大力鼓舞 初步结果我们现在的目标是扩展这一分析，以在开发和开发期间找到SOX9目标 再生。 在这项拟议的工作结束时，我们希望对SOX9的功能有更好的理解，并发现 缓解人类β细胞缺乏的潜在治疗途径。

项目成果

Adoption of the Q transcriptional regulatory system for zebrafish transgenesis.

• DOI: 10.1016/j.ymeth.2013.06.012
• 发表时间: 2014-04-01
• 期刊: METHODS
• 影响因子: 4.8
• 作者: Subedi, Abhignya;Macurak, Michelle;Gee, Stephen T.;Monge, Estela;Goll, Mary G.;Potter, Christopher J.;Parsons, Michael J.;Halpern, Marnie E.
• 通讯作者: Halpern, Marnie E.

Retinoic acid plays an evolutionarily conserved and biphasic role in pancreas development.

视黄酸在胰腺发育中起进化保守和双相作用。

• DOI: 10.1016/j.ydbio.2014.07.021
• 发表时间: 2014-10-01
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Huang W;Wang G;Delaspre F;Vitery Mdel C;Beer RL;Parsons MJ
• 通讯作者: Parsons MJ

Differential in vivo tumorigenicity of diverse KRAS mutations in vertebrate pancreas: A comprehensive survey.

• DOI: 10.1038/onc.2014.223
• 发表时间: 2015-05-21
• 期刊: Oncogene
• 影响因子: 8
• 作者: Park JT;Johnson N;Liu S;Levesque M;Wang YJ;Ho H;Huso D;Maitra A;Parsons MJ;Prescott JD;Leach SD
• 通讯作者: Leach SD

Rbm24a and Rbm24b are required for normal somitogenesis.

Rbm24a 和 Rbm24b 是正常体细胞发生所必需的。

• DOI: 10.1371/journal.pone.0105460
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Maragh S;Miller RA;Bessling SL;Wang G;Hook PW;McCallion AS
• 通讯作者: McCallion AS