Pluripotent Stem Cell-derived Liver Organoids as a Patient-based New Model for Deliverable Assessment of GH Insensitivity Syndrome

多能干细胞衍生的肝脏类器官作为基于患者的新模型，用于 GH 不敏感综合征的可交付评估

• 批准号: 9897571
• 负责人: Vivian Hwa
• 金额: $ 19.88万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9897571
• 关键词: 3-Dimensional; Affect; Afibrinogenemia; Animals; Biochemistry; Biological Models; Caring; Cell Line; Cell surface; Cells; Child; Complement; Cues; Defect; Dependence; Development; Developmental Process; Disease model; Dominant-Negative Mutation; Evaluation; Failure; Foundations; Future; Gene Expression; Gene Expression Regulation; Genes; Growth; Growth Hormone Receptor; Growth and Development function; Height; Hepatic; Hepatocyte; Hormonal; Hormone Responsive; Human; Human Development; IGF1 gene; Immunodeficient Mouse; Individual; Insulin-Like Growth Factor I; International; Investigation; JAK2 gene; Janus kinase 2; Knockout Mice; Laron Syndrome; Lead; Liver; Liver Stem Cell; Mediating; Modeling; Molecular Profiling; Mutation; Organ; Organoids; Pathway interactions; Patients; Phase; Pituitary Gland; Pluripotent Stem Cells; Process; Production; Regulation; Reporting; Research; STAT5B gene; Signal Transduction; Somatomedins; Somatotropin; Somatropin; System; Therapeutic; Time; Transplantation; base; falls; improved; in utero; in vivo; induced pluripotent stem cell; insight; insulin regulation; novel; novel marker; personalized therapeutic; precision medicine; receptor; response; therapeutic evaluation

PROJECT SUMMARY Circulating insulin-like growth factor (IGF)-I, critical for all phases of human development, growth, and maturation, is produced predominantly by the liver, with post-natal hepatic production of IGF-I highly dependent on pituitary-secreted growth hormone (GH). The importance of the hepatic human GH-IGF-I axis for normal human growth is highlighted by the multiple genetic defects downstream of GH, in children who present with severe growth failure (height -3 to -10 standard deviation below the mean), GH insensitivity (GHI) and IGF-I deficiency (IGFD). The identification of rare mutations of STAT5B (signal transducer and activator of transcription 5B) in children with GHI and IGFD, together with the more than 80 reported GHR (GH receptor) mutations in affected children, firmly established the critical importance of the GHR-STAT5B pathway for GH- induced regulation of IGF-I production. However, a disturbing >60% of GHI, idiopathic short statured children, do not carry GHR or STAT5B defects, yet are IGF-I deficient, indicating still limited understanding of the hepatic GHR-STAT5B-IGF-I growth axis and that current model systems are clearly insufficient. We propose establishing a new patient-based model for deliverable assessment of GHI and IGFD, based on emerging human induced-pluripotent stem cells (iPSC)-derived organoids (“mini-organs”), which are promising “living replica” models for disease investigation and precision medicine. We hypothesize that human hepatic response to GH cues can be recapitulated in patient iPSC-derived liver organoids. This human- based model offers unique opportunities to probe human hepatic GHR-STAT5B dependent and independent mechanisms of gene regulation in patient-specific organoids and to follow responsiveness of the developing liver organoid to hormonal cues, paralleling in utero and post-natal developmental processes. The Specific Aims will leverage our unique panel of characterized, patient primary cells to assess molecular signatures, sensitivity, and functionality of iPSC-derived organoids to GH cues, towards establishing the first viable human based models for mechanistic investigation of the human hepatic GH-IGF-I growth axis. The Specific Aims will (1) assess the developmental expression of the GHR-STAT5B-IGF-I axis in human iPSC-derived liver organoid model; and (2) delineate hepatic STAT5B-dependent and independent GH-regulated gene expression in human iPSC-derived liver organoids. At the conclusion of these studies we will have established and demonstrated the utility of a patient-specific, iPSC-derived liver organoid system for evaluating functional disruptions by genetic defects leading to GHI and IGFD. New mechanistic insights gained include improved understanding of human, hepatic GHR-STAT5B mediated regulation of IGF-I expression, with potential for identifying new markers of human GH responsiveness. In the long term, these novel patient-based models provide a platform for future personalized therapeutic testing, towards improving patient management and care of affected children, and for future evaluation of liver responsiveness to other hormonal cues.

项目摘要 循环胰岛素样生长因子（IGF）-i，对于人类发展，生长和 成熟，主要由肝脏产生，并在产后肝后产生IGF-I高度产生 取决于垂体分泌的生长马（GH）。肝脏人类GH-igf-i轴的重要性 在GH的下游的多种遗传缺陷中强调了正常的人类生长 严重的生长失败（高度-3至-10的标准偏差低于平均值），GH不敏感（GHI）和 IGF-I缺乏症（IGFD）。 STAT5B的罕见突变（信号传感器和激活因子的罕见突变）的鉴定 转录5b）在患有GHI和IGFD的儿童中，以及80多个报道的GHR（GH受体） 受影响儿童的突变首先确立了GHR-STAT5B途径对GH-的至关重要性 诱导的IGF-1产生调节。然而，吉是60％的特发性简短说明儿童的令人不安的人， 不要携带GHR或STAT5B缺陷，但IGF-I缺陷，表明对 肝GHR-STAT5B-IGF-I生长轴和当前模型系统显然不足。 我们建议建立一个新的基于患者的模型，用于可交付的GHI和IGFD， 基于新兴的人类诱导的干细胞（IPSC）衍生的类器官（“ Mini-Organs”），这是 有望进行疾病调查和精确医学的“生活复制品”模型。我们假设人类 可以在患者IPSC衍生的肝癌中概括对GH提示的肝素反应。这个人 - 基于的模型提供了独特的机会来探测人类肝炎GHR-STAT5B依赖和独立的机会 患者特异性器官中基因调节的机制，并遵循发展的反应性 肝脏器官到激素线索，在子宫和产后发育过程中并行。具体 AIMS将利用我们独特的特征性患者原代细胞来评估分子特征， IPSC衍生的类器官对GH提示的敏感性和功能，以建立第一个可行的人 基于人肝GH-IGF-1生长轴机械投资的模型。具体目标将 （1）评估人IPSC衍生的肝脏器官中GHR-STAT5B-IGF-I轴的发育表达 模型; （2）描绘肝Static Static5b依赖性和独立的GH调节基因表达 人IPSC衍生的肝癌。在这些研究结束时，我们将建立并 证明了患者特异性，IPSC衍生的肝脏器官的实用性用于评估功能 导致GHI和IGFD的遗传缺陷的破坏。获得的新机械见解包括改进 了解人类的Hepatitic GHR-STAT5B介导的IGF-I表达调节，潜力 识别人类GH响应能力的新标记。从长远来看，这些新型基于患者的模型 提供一个未来个性化治疗测试的平台，以改善患者管理和护理 受影响的儿童，以及将来评估对其他马索线索的肝反应能力。

项目成果

Rare CNVs provide novel insights into the molecular basis of GH and IGF-1 insensitivity.

• DOI: 10.1530/eje-20-0474
• 发表时间: 2020-12
• 期刊: European journal of endocrinology
• 影响因子: 5.8
• 作者: Cottrell E;Cabrera CP;Ishida M;Chatterjee S;Greening J;Wright N;Bossowski A;Dunkel L;Deeb A;Basiri IA;Rose SJ;Mason A;Bint S;Ahn JW;Hwa V;Metherell LA;Moore GE;Storr HL
• 通讯作者: Storr HL

Human growth disorders associated with impaired GH action: Defects in STAT5B and JAK2.

• DOI: 10.1016/j.mce.2020.111063
• 发表时间: 2021-01-01
• 期刊: Molecular and cellular endocrinology
• 影响因子: 4.1
• 作者: Hwa V