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.

认知健康与终生保持良好的脑血管系统有关。然而，脑血管系统中不同细胞的形态/结构和功能的年龄相关性变化有助于不同衰老相关疾病的发展。衰老过程中的这些变化之一是诱导一种复杂的细胞应激反应，称为细胞衰老。衰老细胞是代谢活动发生变化的活细胞，可以损害组织修复和再生过程，从而促进衰老。使用新颖、高效和可重复的3D生物打印有机化合物用于脑血管模型将能够生产比例精确控制的特定有机化合物和不同类型的衰老细胞。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，而不会将研究转移到漫长、更复杂和昂贵的体内实验上。最后，该项目将为各级学生提供良好的多学科教育培训机会，特别是通过让代表不足的少数族裔参与者参与STEM和开展有趣而有效的外展活动。本研究的目标是开发由人类诱导多能干细胞(IPSC)来源的神经元、星形胶质细胞和脑微血管内皮细胞组成的3D脑血管器官，以研究衰老细胞对衰老相关miRNAs差异表达和胞外分泌的影响。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能够通过产生类似于年轻器官、衰老前表型或老年器官的3D器官来研究衰老过程，这将通过在3D有机物生物打印过程中控制输入不同百分比的衰老细胞来实现。此外，还将研究衰老细胞过度表达衰老相关miRNAs的器官模型，以期通过细胞递送miRNA模拟物来调节健康器官中衰老相关分泌表型(SASPs)miRNA的表达。关于miRNAs在细胞衰老过程中的调控机制，以及miRNAs在脑血管对辐射等衰老诱导剂的反应中所起的作用，目前还鲜有报道。此外，这种机制和相关途径的体外研究一直是非常具有挑战性的，大多数研究都局限于二维(2D)培养系统，未能捕捉到复杂的血管组织功能。重要的是，这项研究衰老过程的高风险新奇研究的成功完成，将能够建立高精度的模型，允许操纵和研究不同的年龄相关疾病，并将为在各种器官特定的研究中开发与衰老相关的类似研究提供基础。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。