Nanoscale Nuclear Patterning to Detect Nuclear Envelope Remodeling During Skin Cell Aging

A significant hallmark of aging is the deterioration of the nucleus in cells, where aged and senescent cells are commonly observed to have nuclei with abnormal morphology and architecture, such as enlarged size, wrinkled and invaginated nuclear membrane, loose heterochromatin, etc. A better understanding of how aging-related genetic and epigenetic altercations is triggered by or leads to the physically mis-organized nuclear morphology is important for the discovery and evaluation of anti-aging therapeutics. However, a reliable and quantitative detection of the abnormal nuclear morphology is technically challenging. This is due to the fact that most of the abnormal features are randomly distributed across the entire nuclear space and appear in tens to hundreds of nanometers in dimension, beyond the detection limit of conventional microscopic imaging. To overcome these difficulties, we propose to introduce nanofabrication-enabled nuclear patterning to deliver quantifiable readouts of the abnormal nucleus in aged and senescent cells. Nanopillar-enabled nuclear patterning has been recently developed by the Zhao lab of the team to differentiate cancer cells with high and low malignancy. Here in this proposal, we aim to study whether or how the nuclear deformation patterns on nanopillar arrays can differentiate young and aged nucleus. Uniquely, we will focus on the aging of human fibroblasts and keratinocytes as the skin is an accessible, well-characterized and optimal model system to translate potential in vitro findings towards clinical application. Using the inducible premature aging and senescence cell models developed by the Dreesen lab of the team, we will compare different nanostructure designs and nuclear morphology markers to build assays affording quantification of nuclear alterations in aging.The success of this proposal will establish nanostructure-based nuclear patterning as a new strategy to detect cellular aging and open a new angle to develop anti-aging therapeutics.