Collaborative Research: EAGER: 3D Bioprinted Organoids for Studying the Mechanism of Cerebrovascular Aging

合作研究：EAGER：用于研究脑血管衰老机制的 3D 生物打印类器官

• 批准号: 2317758
• 负责人: Michal Masternak
• 金额: $ 13.5万
• 依托单位: The University of Central Florida Board of Trustees
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2317758&HistoricalAwards=false
• 关键词: Collaborative; Research; EAGER; 3D; Bioprinted

Cognitive health is associated with the maintenance of a well-functioning cerebrovascular system throughout life. However, age-dependent changes in morphology/structure and function in different cells in the cerebrovascular system contribute to the development of different aging-associated diseases. One of these changes during aging is the induction of a complex cellular stress response termed cellular senescence. Senescent cells are viable cells that undergo alterations in metabolic activity and can compromise tissue repair and regeneration processes, thereby contributing toward aging. The use of novel, highly efficient, and reproducible 3D bioprinted organoids for cerebrovascular models will enable production of specific organoids with precisely controlled proportions and different types of senescent cells. This approach can mimic natural aging or different neurodegenerative diseases by decreasing the time burden without moving the research onto lengthy, more complex, and expensive in vivo experiments. Finally, the project will provide excellent educational training opportunities in multi-disciplinary subjects for students at all levels, especially by involving underrepresented minority participants in STEM and the development of interesting and effective outreach activities.The goal of this research is to develop 3D cerebrovascular organoids composed of human-induced pluripotent stem cells (iPSC) derived neurons, astrocytes, and cerebromicrovascular endothelial cells to study the impact of senescent cells on differential expression and exosomal secretion of senescence-associated miRNAs. Cellular senescence is recognized as one of the major mechanisms of tissue deterioration and the cause of many age-associated diseases. The innovative approach of using a microfluidic-based 3D cell culture model will enable the study of the aging process by producing a 3D organoid resembling a young organ, a pre-aging phenotype, or an old organoid, which will be achieved by controlled input of different percentages of senescent cells during the 3D organoid bioprinting process. In addition, an organoid model with senescent cells overexpressing senescence-associated miRNAs will be studied for cellular delivery of miRNA mimics to regulate the expression of senescence-associated secretory phenotype (SASPs) miRNA in healthy organoids. There is little data on the regulatory mechanisms of miRNAs during the process of cellular senescence and the role of miRNAs in cerebrovascular responses to senescence inducers such as radiation exposure. Moreover, in vitro investigation of such mechanisms and associated pathways has been extraordinarily challenging, and most studies have been limited to two-dimensional (2D) culture systems failing to capture complex vascular tissue functionalities. Importantly, the successful completion of this high-risk novel research to study the aging process will enable the establishment of highly precise models allowing manipulations and studies of different age-related diseases and will also provide a base for developing similar studies related to senescence in various organ-specific studies.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.

认知健康与一生中保持良好的脑血管系统功能有关。然而，脑血管系统中不同细胞的形态/结构和功能的年龄依赖性变化有助于不同年龄相关疾病的发展。衰老过程中的这些变化之一是诱导复杂的细胞应激反应，称为细胞衰老。衰老细胞是有活力的细胞，经历代谢活动的改变，可以损害组织修复和再生过程，从而导致衰老。新型、高效、可复制的生物3D打印类器官用于脑血管模型，将使生产具有精确控制比例和不同类型衰老细胞的特定类器官成为可能。这种方法可以通过减少时间负担来模拟自然衰老或不同的神经退行性疾病，而无需将研究转移到冗长，更复杂和昂贵的体内实验。最后，该项目将为各级学生提供优秀的多学科教育培训机会，特别是让代表性不足的少数族裔参与STEM，并开展有趣而有效的外展活动。本研究的目的是开发由人诱导多能干细胞（iPSC）衍生的神经元、星形胶质细胞和脑血管内皮细胞组成的三维脑血管类器官，研究衰老细胞对衰老相关miRNAs的差异表达和外泌体分泌的影响。细胞衰老被认为是组织退化的主要机制之一，也是许多与年龄相关的疾病的原因。使用基于微流体的3D细胞培养模型的创新方法将通过产生类似于年轻器官，预衰老表型或老年类器官的3D类器官来研究衰老过程，这将通过在3D类器官生物打印过程中控制不同百分比的衰老细胞输入来实现。此外，我们还将研究一个衰老细胞过度表达衰老相关miRNA的类器官模型，通过细胞递送miRNA模拟物来调节健康类器官中衰老相关分泌表型（sasp） miRNA的表达。关于miRNAs在细胞衰老过程中的调控机制，以及miRNAs在脑血管对衰老诱导剂（如辐射暴露）的反应中的作用，目前的数据很少。此外，这种机制和相关途径的体外研究非常具有挑战性，大多数研究仅限于二维（2D）培养系统，无法捕获复杂的维管组织功能。重要的是，这项研究衰老过程的高风险新研究的成功完成将使高精度模型的建立成为可能，从而可以操纵和研究不同的年龄相关疾病，也将为在各种器官特异性研究中开展与衰老相关的类似研究提供基础。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。