Leveraging 3D bioprinted organoid constructs to pattern and model human brain development

利用 3D 生物打印类器官结构来模拟人类大脑发育

• 批准号: 10184225
• 负责人: Vahid Serpooshan
• 金额: $ 68.01万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10184225
• 关键词: 3-Dimensional; Achievement; Address; Architecture; Bathing; Behavior; Benchmarking; Biocompatible Materials; Biological; Biological Assay; Biological Models; Biomechanics; Biomedical Engineering; Brain; Cell Differentiation process; Cell Survival; Cells; Complex; Cues; Custom; Development; Diffusion; Distress; Dorsal; Electrophysiology (science); Elements; Emotional; Encapsulated; Environment; Extracellular Matrix; Financial Hardship; Generations; Geometry; Glycoproteins; Glycosaminoglycans; Goals; Health; Histologic; Histology; Human; Image; In Vitro; Individual; Kinetics; Laboratories; Leadership; Light; Measurement; Measures; Methods; Modeling; Molecular; Nanotechnology; Nature; Nervous System Physiology; Neurodevelopmental Disorder; Neurons; Neurosciences; Organoids; Output; Pattern; Physiological; Physiology; Property; Proteins; Proteoglycan; Protocols documentation; Public Health; Reproducibility; Research; Resolution; Robotics; Role; SHH gene; Scheme; Shapes; Signal Transduction; Signaling Molecule; System; Technology; Testing; Tissue Engineering; Tissues; Work; bioprinting; brain parenchyma; brain tissue; cell behavior; cost; cranium; design; extracellular; human model; human stem cells; hydrogel scaffold; in vitro Model; induced pluripotent stem cell; innovation; matrigel; morphogens; nano; nanoparticle; nervous system development; nervous system disorder; neural circuit; neural model; neurodevelopment; novel; photoactivation; physical property; prevent; relating to nervous system; scaffold; single-cell RNA sequencing; skills; small molecule; smoothened signaling pathway; spatiotemporal; stem cell fate; stem cell model; stem cells; success; synergism; tool

Project Summary In addition to significant human distress, neurological disorders cost the U.S. economy more than $1.5 trillion per year—8.8 percent of the gross domestic product. This physical, emotional and financial burden underscores the potential benefit from developing innovative platforms to study brain development, physiology, and associated diorders. The advent of human induced pluripotent stem cell (hiPSC)-derived 3D cortical organoid cultures has shown great promise as a model system, yet there remain a number of technical limitations that have stymied their ability to recapitulate critical non-cell autonomous aspects of human brain development. These components include extrinsic influences of the extracellular matrix (ECM), skull, and axial morphogen gradients that together help shape and diversify regions of the developing brain. Currently, standard organoid protocols involve embedding organoids in Matrigel droplets or in suspended bath culture, which prevents reproducible user-control of the extracellular environment. This limits the ability to create morphogen gradients that topographically polarize stem cells, or to ask how molecular and physical properties of the ECM outside the brain parenchyma guide neurodevelopment. To address these challenges, we propose developing 3D bioprinted cortical organoid constructs that recapitulate key microenvironmental cues of native brain tissue. This project builds upon our recent technological achievements, enabling embedded bioprinting of custom tissue constructs at high spatial resolution (20 µm) specifically designed for long-term organoid culture. Our goal is to bioprint cortical brain organoids into 3D scaffolds with customizable molecular and physiomechanical compositions. The synergy of brain organoid and bioprinting technologies provides a nearly unlimited potential to manipulate extrinsic developmental cues of a complex multicellular human model system. We will pursue two integrated Specific Aims using the multi-PI leadership mechanism to combine complementary research skills and expertise in the Sloan (neurodevelopment) and Serpooshan (tissue engineering) laboratories. In Aim 1, we will decouple the molecular composition and physical stiffness paramaters of the ECM, and ask how these factors influence neural development, differentiation, maturation and architecture. In Aim 2, we will use three separate approaches to generate a stable morphogen gradient within bioprinted constructs, which we will use to induce intra-organoid polarization of both dorsal (pallial) and ventral (subpallial) regional identities. Together, these approaches offer a novel platform for 3D stem cell modeling that could be applied broadly to numerous systems and usher a new generation of neurodevelopmental modeling.

项目摘要 除了巨大的人类困扰外，神经系统疾病还使美国经济损失超过1.5万亿美元 每年为国内生产总值的8.8％。这种身体，情感和经济燃烧强调了 开发创新平台来研究大脑发育，生理学和 相关的第三次。人类诱导多能干细胞（HIPSC）衍生的3D皮质器官的冒险 文化作为模型系统表现出了巨大的希望，但仍有许多技术限制 阻碍了他们概括人脑发育的关键非细胞自主方面的能力。 这些成分包括细胞外基质（ECM），头骨和轴向形态的外部影响 共同塑造和多样化大脑区域的渐变。 目前，标准的器官协议涉及将类器官嵌入矩阵液滴或悬挂浴中 培养，可防止细胞外环境的可再现用户控制。这限制了创建的能力 形态上极化干细胞或询问分子和物理特性的形态梯度 大脑实质之外的ECM指导神经发育。为了应对这些挑战，我们建议 开发3D生物打印的皮质器官构建体，概括了天然的关键微环境线索 脑组织。该项目以我们最近的技术成就为基础 高空间分辨率（20 µM）的定制组织构建体专为长期器官培养而设计。 我们的目标是将皮质脑器官生物构成分子和可定制分子和的3D支架 物理成分。脑器官和生物打印技术的协同作用提供了附近 操纵复杂的多细胞人类模型系统外部发育提示的无限潜力。 我们将使用多PI领导力机制来实现两个集成的特定目标 Sloan（神经发育）和Serpooshan（组织工程）的研究技能和专业知识 实验室。在AIM 1中，我们将使分子组成和物理刚度参数分解 ECM，并询问这些因素如何影响神经元发展，分化，成熟和结构。在 AIM 2，我们将使用三种单独的方法在生物报中生成稳定的形态梯度 构造，我们将用来诱导背侧和腹侧的甲内极化 （下）区域身份。这些方法一起为3D干细胞建模提供了一个新颖的平台 可以广泛地应用于众多系统，并引入新一代的神经发育建模。