Identification of Novel Senescent Markers in Small Extracellular Vesicles

Abstract

Senescent cells exhibit several typical features, including the senescence-associated secretory phenotype (SASP), promoting the secretion of various inflammatory proteins and small extracellular vesicles (EVs). SASP factors cause chronic inflammation, leading to age-related diseases. Recently, therapeutic strategies targeting senescent cells, known as senolytics, have gained attention; however, noninvasive methods to detect senescent cells in living organisms have not been established. Therefore, the goal of this study was to identify novel senescent markers using small EVs (sEVs). sEVs were isolated from young and senescent fibroblasts using three different methods, including size-exclusion chromatography, affinity column for phosphatidylserine, and immunoprecipitation using antibodies against tetraspanin proteins, followed by mass spectrometry. Principal component analysis revealed that the protein composition of sEVs released from senescent cells was significantly different from that of young cells. Importantly, we identified ATP6V0D1 and RTN4 as novel markers that are frequently upregulated in sEVs from senescent and progeria cells derived from patients with Werner syndrome. Furthermore, these two proteins were significantly enriched in sEVs from the serum of aged mice. This study supports the potential use of senescent markers from sEVs to detect the presence of senescent cells in vivo.

Senescent Cells: A Therapeutic Target in Cardiovascular Diseases

“…In addition to other SASP factors, senescent cells can release small extracellular vesicles (EVs), including exosomes. Molecules in EVs obtained from the blood could be novel gerodiagnostic markers [76,77]. The microRNA (miRNA) expression and secretion profile of senescent cells has recently come into the spotlight [78,79].…”

Renal dysfunction, malignant neoplasms, atherosclerotic cardiovascular diseases, and sarcopenia as key outcomes observed in a three-year follow-up study using the Werner Syndrome Registry

“…It has been suggested that the inclusion of the WRN gene in genetic analyses for early-onset diabetes, lipodystrophy, or dyslipidemia may offer an opportunity to diagnose patients with Werner syndrome long before the presentation of the full spectrum of symptoms and complications, enabling earlier interventions, including malignant neoplasm screenings and prevention of diseases listed in the clinical criteria [ 58 ]. Recently, novel senescent markers, ATP6V0D1 and RTN4, were shown to be increased in cells derived from patients with Werner syndrome, raising a possibility that they may serve as markers of disease progression [ 59 ]. Similarly, ribonuclease H2 subunit A (RNaseH2A) was shown to be downregulated in Werner syndrome cells as well as selected cancer cells [ 60 ].…”

Unraveling the Multifaceted Roles of Extracellular Vesicles: Insights into Biology, Pharmacology, and Pharmaceutical Applications for Drug Delivery

“…EVs are involved in diverse biological processes, such as cell motility [ 254 , 255 , 256 ], differentiation [ 257 , 258 , 259 ], proliferation [ 260 , 261 ], apoptosis [ 262 , 263 ], reprogramming [ 264 , 265 , 266 ], and immunity [ 267 , 268 ], through regulation of intercellular communication. Understanding the regulatory mechanisms of EVs in these biological processes and in intercellular communication is crucial for their clinical potential [ 269 , 270 ] and applications [ 271 , 272 , 273 , 274 , 275 , 276 , 277 , 278 , 279 ].…”