Despite ample documentation of age-related changes in physiologic parameters, mechanisms governing the process of aging of the lungs have not been well studied. Among the identified age-associated changes is the modification of the extracellular matrix (remodeling). These changes result in alteration of the aged lungs response to injury and repair. As importantly, the number of senescent cells in the lungs increases with age. However, the mechanisms by which these cells provoke aging of the lungs are uncertain. All senescent cells are in growth arrest but continue to secrete large amounts of proteins that affect their microenvironment. Seemingly contradictory data indicate that, on the one hand, senescent fibroblasts are beneficial in wound healing but, on the other hand, limiting the number of senescent alveolar epithelial cells is beneficial in pulmonary fibrosis. Thus, there is a critical need to characterize the senescence profile of the various cells present in the lungs (mainly endothelial, epithelial, and fibroblast cells) to determine their relative contribution to he aging process. Despite the detection of senescent cells in aged lungs, their role in physiological remodeling associated with aging is unknown and their secretory phenotype remains undefined. Studying senescence in vivo has been technically challenging; however, we have recently obtained the p16-3MR transgenic system that empowers in vivo identification of p16ink4A+ cells (senescent cells) and their subsequent isolation by flow cytometry as well as their conditional deletion via the administration of gancyclovir. We propose to use this mouse model to more definitely define the role of senescent cells in lung aging; we will validate our data in nonhuman primate aging lungs, a translational model to human aging. We hypothesize that each senescent cell type (alveolar epithelial, endothelial and fibroblasts) has a characteristic secretory associated senescence profile to indicate their relative contribution to aging.
Two specific aims are proposed to address this hypothesis: 1) To identify the senescence-associated secretory profile of each different senescent cell type (alveolar epithelial and endothelial cells and fibroblasts) to determine their contribution to lung tissue remodeling. Senescent and non-senescent cells will be isolated from non-lethally irradiated p16-3MR mouse lungs by flow cytometry and their transcriptional profiling will be determined. The expression of differentially expressed identified proteins will be validated in young and aged nonhuman primate lungs; and 2) To determine the contribution of senescent cells in the age-associated remodeling of the lungs. Using the p16-3MR transgenic mouse, we will determine whether the presence of senescent cells affects the remodeling process in the lungs by deleting p16ink4A+ cells after non-lethal dose irradiation. The data gathered with this project will characterize the secretory profile of various senescent cell types that have an important functional role in remodeling in the lungs but also in other organs such as the brain or the kidney (senescent endothelial cells and fibroblasts) and provide new information as to mechanism.

Public Health Relevance

The rise in life expectancy worldwide has been accompanied by an increased incidence of age-related diseases, presenting an enormous burden on healthcare services and society. Age-related pulmonary diseases continue to be a health problem associated with high disability and mortality. Accumulating evidence points to aged lung tissue been predisposed to age-related diseases in association with the accumulation of senescent cells. A clear mechanistic link between known causes of age-related structural tissue changes and dysfunctional senescent cells has not been established. This work proposes to determine whether senescence cells contribute to the age- associated structural changes. We will determine whether the presence of senescence cells is necessary to provoke the age-associated structural changes. Furthermore, we will characterize the secretome of three main cell types (endothelial and alveolar epithelial cells, and fibroblasts) and determine their relativ contribution to the structural changes

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AG052744-01
Application #
9086628
Study Section
Lung Injury, Repair, and Remodeling Study Section (LIRR)
Program Officer
Macchiarini, Francesca
Project Start
2016-06-01
Project End
2018-03-31
Budget Start
2016-06-01
Budget End
2017-03-31
Support Year
1
Fiscal Year
2016
Total Cost
$198,125
Indirect Cost
$73,125
Name
University of California San Francisco
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94118