Human Stem Cell Fate Decisions Dictated by Decoupled Biophysical Cues

人类干细胞的命运决定由解耦的生物物理线索决定

• 批准号: 1917618
• 负责人: Yan Li
• 金额: $ 40万
• 依托单位: Florida State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1917618&HistoricalAwards=false
• 关键词: Human; Stem; Cell; Fate; Decisions

Pluripotent stem cells can turn into all types of cells that make up the tissues and organs in our body. The specific cell type (stem cell fate) depends on the biochemical and biophysical cues they experience. The biophysical cues depend on the surrounding scaffold's mechanical properties, such as the elastic modulus and the Poisson's ratio (which is the ratio of a material's lateral contraction to its increase in length upon stretching). This project seeks to understand how these mechanical properties determine stem cell fate by tuning both elastic modulus and Poisson's ratio to reveal the synergistic roles of biophysical and biochemical signaling on stem cell fate. This research will benefit bioengineering applications and biomanufacturing (e.g., novel microcarriers), leading to better drug screening and disease modeling for the biotechnology and pharmaceutical industries. The project will also establish an interactive learning platform to reduce gender disparity and increase the participation of minority students in engineering. Efforts will be made to stimulate African American and women students to pursue an advanced training and career by participating in the proposed research and educational activities. This project will include working with Women in Math, Science and Engineering organization, National Society of Black Engineers, Quality Education for Minorities Network, and the SciGirls program at National High Magnetic Field Laboratory to attract young girls and African American students for advanced science and engineering training and career. The goal of this project is to elucidate the interactions between biophysical cues of elastic modulus and Poisson's ratio of 3D polyurethane (PU) scaffolds and their influence on the secretion of endogenous ECMs and Yes-associated protein (YAP) localization by iPSCs during lineage-specific commitment. The project's central hypothesis is that the scaffolds with tunable elastic modulus and Poisson's ratio affect cell organization and transduce biophysical signals to modulate the profile of endogenous ECMs and YAP expression and influence lineage commitment of iPSCs. This hypothesis is based on preliminary results that demonstrated enhanced neural and vascular differentiation of pluripotent stem cells (PSCs) using 3D scaffolds with re-entrant structures - structures with angles pointing inward that expand in all three directions if stretched in one direction, i.e., structures with negative Poisson's ratios. The research plan is organized under three objectives. The FIRST Objective is to fabricate and characterize scaffold arrays with different Poisson's ratio and elastic modulus. Regular PU scaffolds with reticulate structure will be heated/softened and controlled buckling will be used to produce a spectrum of auxetic scaffolds with varying Poisson's ratio (0.3 to -0.4) at a fixed elastic modulus. Likewise, auxetic scaffolds with different elastic modulus (1-100 kPa) at a fixed Poisson's ratio will be fabricated by compressing the regular scaffold with different modulus to the same degree of buckling. The scaffolds will be fabricated according to the prediction of finite element modeling with the inputs of temperature-dependent elastic modulus and on video data. The SECOND Objective is to examine the differential effects of Poisson's ratio and elastic modulus on iPSC lineage commitment. Undifferentiated iPSCs or iPSC-derived neural progenitor cells (NPCs) will be seeded into different scaffolds and induced toward neural lineage or vascular lineage. The cells will be characterized for neuronal markers or vascular markers with expectations that auxetic scaffolds that mimic tissue elasticity will promote neural differentiation of iPSCs and that elastic modulus and Poisson's ratio have differential effects on neural lineage commitment. The THIRD Objective is to determine the influences of Poisson's ratio and elastic modulus on YAP localization and the secretion of endogenous ECMs, which modulate canonical Wnt signaling and contribute to the lineage commitment of iPSCs. YAP localization will be examined and the influence of YAP on Wnt signaling will be revealed with expectations that auxetic scaffolds induce cytoplasmic YAP localization, that nuclear YAP localization activates Wnt signaling, that cells will secrete different profiles of ECMs in response to the scaffold elasticity and influence Wnt signaling and finally, that interactions of YAP with Wnt signaling contribute to the lineage commitment of iPSCs.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

多能干细胞可以转化为构成我们身体组织和器官的所有类型的细胞。特定的细胞类型（干细胞命运）取决于它们经历的生化和生物物理线索。生物物理线索取决于周围支架的机械性能，例如弹性模量和泊松比（材料的横向收缩与其拉伸时长度增加的比率）。 该项目旨在通过调整弹性模量和泊松比来了解这些机械特性如何决定干细胞的命运，以揭示生物物理和生化信号对干细胞命运的协同作用。这项研究将有利于生物工程应用和生物制造（例如新型微载体），从而为生物技术和制药行业带来更好的药物筛选和疾病建模。该项目还将建立一个互动学习平台，以减少性别差异并增加少数族裔学生对工程的参与。将努力刺激非裔美国学生和女学生通过参与拟议的研究和教育活动来追求高级培训和职业生涯。该项目将包括与数学、科学和工程领域的女性组织、国家黑人工程师协会、少数民族优质教育网络以及国家高磁场实验室的 SciGirls 计划合作，以吸引年轻女孩和非裔美国学生学习先进的科学和工程培训和职业。该项目的目标是阐明 3D 聚氨酯 (PU) 支架的弹性模量和泊松比的生物物理线索之间的相互作用，及其对谱系特异性过程中 iPSC 分泌内源 ECM 和 Yes 相关蛋白 (YAP) 定位的影响承诺。该项目的中心假设是，具有可调弹性模量和泊松比的支架会影响细胞组织并转导生物物理信号以调节内源性 ECM 和 YAP 表达的概况，并影响 iPSC 的谱系定型。这一假设基于初步结果，该结果证明使用具有可重入结构的 3D 支架可增强多能干细胞 (PSC) 的神经和血管分化，这种结构具有向内的角度，如果沿一个方向拉伸，则可在所有三个方向上扩展，即结构具有负泊松比。该研究计划根据三个目标进行组织。第一个目标是制造和表征具有不同泊松比和弹性模量的支架阵列。具有网状结构的常规 PU 支架将被加热/软化，并使用受控屈曲来生产一系列在固定弹性模量下具有不同泊松比（0.3 至 -0.4）的拉胀支架。同样，通过将具有不同模量的常规支架压缩至相同的屈曲程度来制造在固定泊松比下具有不同弹性模量（1-100kPa）的拉胀支架。支架将根据有限元模型的预测、输入温度相关弹性模量和视频数据来制造。第二个目标是检查泊松比和弹性模量对 iPSC 谱系承诺的不同影响。未分化的 iPSC 或 iPSC 衍生的神经祖细胞 (NPC) 将被接种到不同的支架中，并诱导向神经谱系或血管谱系发展。这些细胞将被表征为神经元标记或血管标记，期望模拟组织弹性的拉胀支架将促进 iPSC 的神经分化，并且弹性模量和泊松比对神经谱系定向具有不同的影响。 第三个目标是确定泊松比和弹性模量对 YAP 定位和内源性 ECM 分泌的影响，内源性 ECM 调节典型的 Wnt 信号传导并有助于 iPSC 的谱系定型。将检查 YAP 定位，并揭示 YAP 对 Wnt 信号传导的影响，期望拉胀支架诱导细胞质 YAP 定位，核 YAP 定位激活 Wnt 信号传导，细胞将根据支架弹性和影响分泌不同的 ECM 特征Wnt 信号传导，最后，YAP 与 Wnt 信号传导的相互作用有助于 iPSC 的谱系承诺。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力评估进行评估，被认为值得支持优点和更广泛的影响审查标准。

项目成果

Feeder-free differentiation of human iPSCs into natural killer cells with cytotoxic potential against malignant brain rhabdoid tumor cells

• DOI: 10.1016/j.bioactmat.2024.02.031
• 发表时间: 2024-06-01
• 期刊: BIOACTIVE MATERIALS
• 影响因子: 18.9
• 作者: Kiran，Sonia;Xue，Yu;Sang，Qing-Xiang Amy
• 通讯作者: Sang，Qing-Xiang Amy

Influence of Microenvironment on Mesenchymal Stem Cell Therapeutic Potency: From Planar Culture to Microcarriers

• DOI: 10.3389/fbioe.2020.00640
• 发表时间: 2020-06-24
• 期刊: FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
• 影响因子: 5.7
• 作者: Tsai, Ang-Chen;Jeske, Richard;Li, Yan
• 通讯作者: Li, Yan

Extracellular vesicle biogenesis of three‐dimensional human pluripotent stem cells in a novel Vertical‐Wheel bioreactor

• DOI: 10.1002/jex2.133
• 发表时间: 2024-01
• 期刊: Journal of Extracellular Biology
• 作者: Laureana Muok;Li Sun;Colin Esmonde;Hannah Worden;Cynthia Vied;Leanne Duke;Shaoyang Ma;Olivia Z Zeng;Tristan Driscoll;Sunghoon Jung;Yan Li

Studying the Inflammatory Responses to Amyloid Beta Oligomers in Brain-Specific Pericyte and Endothelial Co-Culture From Human Stem Cells

• DOI: 10.3389/fceng.2022.927188
• 发表时间: 2022-07
• 作者: Mark Marzano;Xingchi Chen;Teal A. Russell;Angelica Medina;Zizheng Wang;Timothy Hua;Changchun Zeng;Xueju Wang;Q. Sang;Hengli Tang;Y. Yun;Yan Li

Upscaling human mesenchymal stromal cell production in a novel vertical-wheel bioreactor enhances extracellular vesicle secretion and cargo profile

• DOI: 10.1016/j.bioactmat.2022.07.004
• 发表时间: 2023-07-01
• 期刊: BIOACTIVE MATERIALS
• 影响因子: 18.9
• 作者: Jeske, Richard;Liu, Chang;Li, Yan
• 通讯作者: Li, Yan