SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

Abstract

SUMMARY Cellular senescence irreversibly arrests cell proliferation, accompanied by a multi-component senescence-associated secretory phenotype (SASP) that participates in several age-related diseases. Using stable isotope labeling with amino acids (SILACs) and cultured cells, we identify 343 SASP proteins that senescent human fibroblasts secrete at 2-fold or higher levels compared with quiescent cell counterparts. Bioinformatic analysis reveals that 44 of these proteins participate in hemostasis, a process not previously linked with cellular senescence. We validated the expression of some of these SASP factors in cultured cells and in vivo. Mice treated with the chemotherapeutic agent doxorubicin, which induces widespread cellular senescence in vivo, show increased blood clotting. Conversely, selective removal of senescent cells using transgenic p163MR mice showed that clearing senescent cells attenuates the increased clotting caused by doxorubicin. Our study provides an in-depth, unbiased analysis of the SASP and unveils a function for cellular senescence in hemostasis.

A proteomic atlas of senescence-associated secretomes for aging biomarker development

“…Data from several laboratories, including our own, strongly support the idea that senescent cells and the SASP drive multiple age-related phenotypes and pathologies, including atherosclerosis [10], osteoarthritis [11], cancer metastasis, cardiac dysfunction [12,13], myeloid skewing [14,15], kidney dysfunction [16], and overall decrements in health span [17]. Recently, senescent cells were shown to secrete bioactive factors into the blood that alter hemostasis and drive blood clotting [18]. SASP factors therefore hold potential as plasma biomarkers for aging and age-related diseases that are marked by the presence of senescent cells.…”

“…Thus, plasma biomarkers of aging are highly enriched with sSASP factors. Complement and coagulation cascade proteins [18], particularly protease inhibitors such as SERPINs, were also noted as prominent plasma biomarkers of aging [48]. These proteins and their pathway networks were robustly altered in the sSASPs of cells induced to senesce by all the tested stressors (Figs 2B and 7A), in addition to other top biomarker candidates: MMP1, STC1, and GDF15 ( Fig 7B, 7C and 7D).…”

“…By virtue of this proteomic analysis of secreted proteins, SASP profiles are also ideal candidates for plasma-based biomarkers, which are enriched among secreted proteins [73]. Senescent cells were recently shown to modulate hemostasis and clotting phenotypes in plasma in vivo [18], demonstrating that senescent cells secrete bioactive factors into circulation in vivo. The use of SASP factors as biomarker candidates is further supported by our analysis, which has indicated that core SASP factors are enriched among plasma biomarkers of aging in humans.…”

The aged hematopoietic system promotes hippocampal‐dependent cognitive decline

“…In these studies, CyPA plasma levels were also found to be elevated with aging (Ohtsuki et al, 2017; Ramachandran et al, 2014; Satoh et al, 2013). While little is known about the role of CyPA in aging, recent proteomic analysis using mass spectrometry has identified CyPA as part of the senescence‐associated secretory phenotype (SASP) (Wiley et al, 2019). Ultimately, our data identify the aged hematopoietic system, and downstream circulating immune factors, as potential therapeutic targets to restore cognitive function in the elderly.…”

Regulation of the mitochondrial permeability transition pore and its effects on aging

“…In relation to this, NF-κB was shown to downregulate ANT1 and the activity of mPTP [ 152 , 153 ]. In a hypothetic scenario, NF-κB could therefore promote senescent cell survival, by lowering mitochondrial potential in a functional and ANT1-dependent manner, allowing to send eventual inflammatory and repairing signals, as platelets where found to be activated by senescent cells [ 154 , 155 ]. However, this situation revolves around a delicate equilibrium, as excessive stress and damage could cause an increase in the levels of inflammation, further downregulating ANT1 .…”