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: 10380006
• 负责人: Vahid Serpooshan
• 金额: $ 65.05万
• 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/10380006
• 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; Tissue constructs; Tissues; Work; bioink; 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.

项目概要 除了严重的人类痛苦之外，神经系统疾病还给美国经济造成了超过 1.5 万亿美元的损失 每年——国内生产总值的8.8%。这种身体、情感和经济负担强调 开发创新平台来研究大脑发育、生理学和 相关疾病。人类诱导多能干细胞 (hiPSC) 衍生的 3D 皮质类器官的出现 作为一种模型系统，文化已显示出巨大的前景，但仍存在许多技术限制 阻碍了他们概括人类大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)、颅骨和轴向形态发生素的外在影响 梯度共同帮助塑造发育中的大脑区域并使其多样化。 目前，标准类器官方案涉及将类器官嵌入基质胶液滴或悬浮浴中 文化，这阻止了用户对细胞外环境的可重复控制。这限制了创建的能力 形状极化干细胞的形态发生素梯度，或询问分子和物理特性如何 脑实质外的 ECM 引导神经发育。为了应对这些挑战，我们建议 开发 3D 生物打印的皮质类器官结构，重现天然的关键微环境线索 脑组织。该项目建立在我们最近的技术成果的基础上，实现了嵌入式生物打印 专为长期类器官培养而设计的高空间分辨率 (20 µm) 定制组织结构。 我们的目标是将皮质脑类器官生物打印成具有可定制分子和结构的 3D 支架 物理力学成分。脑类器官和生物打印技术的协同作用提供了近乎 操纵复杂多细胞人类模型系统的外在发育线索的无限潜力。 我们将利用多 PI 领导机制，将互补性结合起来，追求两个综合的具体目标 斯隆（神经发育）和塞尔普山（组织工程）的研究技能和专业知识 实验室。在目标 1 中，我们将解耦分子组成和物理刚度参数 ECM，并询问这些因素如何影响神经发育、分化、成熟和结构。在 目标 2，我们将使用三种不同的方法在生物打印中生成稳定的形态发生素梯度 结构，我们将用它来诱导背侧（大脑皮层）和腹侧的类器官内极化 （子皮层）区域身份。这些方法共同为 3D 干细胞建模提供了一个新颖的平台， 可以广泛应用于众多系统，并引领新一代神经发育模型。