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

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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.