Investigating the impact of donor and environmental age on iPSC derived endothelial progenitors

PROJECT SUMMARY/ABSTRACT Induced pluripotent stem cells (iPSCs) offer immense promise for autologous therapies by using the recipient’s own cells to engineer new functional tissue. Many of the targeted diseases for iPSC treatment research, such as cardiovascular diseases, are disproportionally found in aging individuals. While iPSCs have been generated from donors of all ages, even centenarians, the influence of donor age on iPSC functionality remains a critically understudied aspect. Moreover, the role of environmental age on differentiated iPSC functionality is not well understood. This research aims to comprehensively address this gap, particularly focusing on the effects of age- related changes in both the donor and environment of iPSC-derived endothelial progenitors (iPSC-EPs). My preliminary data indicates that iPSC-EPs from mature donors (above the age of 30) exhibit a diminished capacity for self-assembling into microvasculature. Building on this observation, I propose a two-fold investigation (Fig. 1). Firstly, I will characterize differences between iPSC-EPs sourced from neonatal, mature, and aged donors, exploring differentiation yield, methylation patterns, gene expression levels, and vascular cell biopotency. This analysis will provide a holistic understanding of how donor age persists through iPSC differentiation into vascular lineages. Secondly, I will investigate the influence of age-related changes in the microenvironment on iPSC-EP functionality by employing innovative 3D hydrogel environments with tunable stiffness and variable composition properties (Fig 1.). With age, there is a general increase in tissue stiffness, but also a change in extracellular protein composition. By decoupling the effects of stiffness from ECM composition, I expect to characterize the specific contributions of these factors and their role in determining microvascular plexus structure, gene expression, and protein expression profiles. Through the examination of iPSC lines sourced from neonatal, mature, and aged donors, matched by sex and somatic cell origin, this research aims to provide crucial insights into the potential limitations of using iPSCs from older individuals for tissue-engineered vascular applications. I hypothesize that age-related differences in gene expression will result in limited microvasculature formation in iPSC-EPs from aged donors, but exposure to a young microenvironment may offer a promising avenue for rejuvenation. This study holds significance for advancing the field of iPSC research and informing the development of effective strategies for personalized vascular therapies, especially as it relates to targeted aged populations.