Generating morphogen gradients to engineer human integumentary organoids

生成形态发生素梯度来设计人类外皮类器官

基本信息

批准号：
10434938
负责人：
Hasan Erbil Abaci
金额：
$ 24.3万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10434938
关键词：
3-Dimensional Affect Afferent Neurons Alopecia Anatomy Animal Model BMP4 Biomedical Engineering Brain Cell Communication Cell Line Cells Chemicals Complex Cyst Dermal Development Disease Disease model Embryo Engineering Epidermis Event Exons Exposure to Generations Genetic Diseases Genetic Skin Diseases Germ Goals Growth Factor Hair Hair follicle structure Human Human Engineering Hydrogels In Vitro Individual Kidney Lamin Type A Liver Mendelian disorder Mesenchymal Messenger RNA Microfluidic Microchips Microfluidics Modeling Molecular Target Morphogenesis Morphology Mutation Neural Crest Neural Crest Cell Organ Organ Model Organogenesis Organoids Patients Pattern Pharmaceutical Preparations Pharmacotherapy Phenotype Physiological Pigmentation physiologic function Pregnancy Premature aging syndrome Progeria Proteins Reporter Research Reverse engineering Role Side Skin Skin Abnormalities Suspension Culture Syndrome System Testing Time Tissues Toxic effect Tretinoin Work appendage base cell type design disease phenotype drug development efficacy evaluation gastrulation induced pluripotent stem cell inhibitor innovation melanocyte morphogens natural Blastocyst Implantation precision drugs rare genetic disorder reconstruction self organization skin disorder skin morphogenesis skin organogenesis success tongue papilla tool transcriptome sequencing treatment strategy

项目摘要

Project Summary Recent success in generating human organoids from embryonic bodies (EBs) of induced pluripotent stem cells (iPSCs) offers a new tool to understand human organ morphogenesis and genetic diseases. However, the current organoid generating approaches, where cells are symmetrically exposed to differentiation factors/morphogens in culture, only allow for the generation of partial components of a tissue, thereby do not support the spatially-controlled generation of multicomponent tissues. The current challenge in organoid research is to achieve a physiologically-relevant organization of cells, tissue components, and anatomical features. In this project, we postulate that generating defined asymmetrical chemical gradients in EBs will lead to controlled differentiation of iPSCs into multicomponent organoids comprising anatomical features. This approach will overcome the current limitations of symmetrical culture conditions. The skin represents a great model organ to test this hypothesis, because (i) the skin morphogenesis strongly relies on interactions of cells from multiple lineages; and (ii) generation of hair follicles and pigmentation can be used as functional read-outs to assess the robustness of this bioengineering approach, which can later be adapted for other organoid systems. In Specific Aim 1, we will generate precise cross-gradients of differentiation factors at the single EB level using microfluidics. We postulate that this will induce simultaneous generation of epidermal cells and neural crest cell- derived melanocytes and dermal papilla cells. Our success criteria will be the recapitulation of the early events of skin morphogenesis, such as the formation of skin appendages and pigmentation of the epidermis. In Specific Aim 2, we will extend this approach to model a genetic disease. We will focus on Hutchinson-Gilford progeria syndrome (HGPS), which is a rare monogenic premature aging disease with distinct skin abnormalities including sclerotic skin, dyspigmentation, and alopecia. We will first induce skin morphogenesis using patient iPSCs to develop a skin disease phenotype. Subsequently, we will use our microphysiological skin model to identify early developmental abnormalities in progeria skin, which are largely unknown for humans, and further evaluate the efficacy and toxicity of three drugs with different molecular targets on the reversal of HGPS skin phenotype in a human relevant-context. This innovative approach represents a critical step towards engineering fully-developed integumentary organoids and will have an immediate and overwhelming impact on our understanding of human skin morphogenesis and developmental skin diseases.

项目概要最近成功从诱导多能干的胚胎体（EB）生成人类类器官细胞（iPSC）提供了一种了解人体器官形态发生和遗传疾病的新工具。然而，目前的类器官生成方法，其中细胞对称地进行分化培养物中的因子/形态发生素仅允许产生组织的部分成分，因此不支持多成分组织的空间控制生成。当前类器官面临的挑战研究的目的是实现细胞、组织成分和解剖学的生理相关组织特征。在这个项目中，我们假设在 EB 中生成定义的不对称化学梯度将导致控制 iPSC 分化为包含解剖特征的多组分类器官。这方法将克服目前对称培养条件的局限性。皮肤代表着伟大模型器官来检验这一假设，因为（i）皮肤形态发生强烈依赖于细胞的相互作用来自多个血统； (ii) 毛囊的生成和色素沉着可用作功能读数评估这种生物工程方法的稳健性，该方法随后可以适用于其他类器官系统。在具体目标 1 中，我们将使用以下方法在单个 EB 水平上生成分化因子的精确交叉梯度：微流体学。我们假设这将诱导表皮细胞和神经嵴细胞的同时生成- 衍生的黑素细胞和真皮乳头细胞。我们的成功标准将是早期事件的重演皮肤形态发生的过程，例如皮肤附属器的形成和表皮色素沉着。具体来说目标 2，我们将扩展这种方法来模拟遗传疾病。我们将重点关注哈钦森-吉尔福德早衰症综合征（HGPS），这是一种罕见的单基因早衰疾病，具有明显的皮肤异常，包括皮肤硬化、色素沉着和脱发。我们将首先使用患者 iPSC 诱导皮肤形态发生形成皮肤病表型。随后，我们将使用我们的微生理皮肤模型来识别早期早衰症皮肤的发育异常，这对人类来说在很大程度上是未知的，并进一步评估具有不同分子靶标的三种药物对逆转 HGPS 皮肤表型的功效和毒性人类相关背景。这种创新方法代表了工程全面开发的关键一步外皮类器官将对我们对人类的理解产生直接和压倒性的影响皮肤形态发生和发育性皮肤病。