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SA-β-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs

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Senolytic-Resistant Senescent Cells Have a Distinct SASP Profile and Functional Impact: The Path to Developing

Utkarsh Tripathi^1,2, Masayoshi Suda^3,4, Vagisha Kulshreshtha⁵

¹Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota, USA.

|December 30, 2025

Summary

Senolytics eliminate senescent cells (SCs), but some SCs resist treatment. Resistant SCs have altered inflammatory profiles and cause less physical dysfunction, suggesting senescent cell plasticity.

Keywords:

cellular senescence senescent cell subtypes senolytics senosensitizers

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Published on: July 12, 2022

Area of Science:

Cellular senescence
Aging research
Immunology

Background:

Senescent cells (SCs) contribute to aging and disease.
Senolytics target SCs but leave a resistant population.
Senescent cell antiapoptotic pathways (SCAPs) protect SCs from death.

Purpose of the Study:

Investigate the characteristics of senolytic-resistant SCs.
Determine the functional differences between senolytic-sensitive and -resistant SCs.
Explore the plasticity between senolytic-sensitive and -resistant SC states.

Main Methods:

Utilized senolytics (Dasatinib, Quercetin) and JAK/STAT inhibitor (Ruxolitinib).
Analyzed senescent human preadipocytes and HUVECs.
Assessed SC markers (p16INK4a, p21CIP1, SAβgal, γH2AX) and senescence-associated secretory phenotype (SASP) factors.
Transplanted SCs into mice to evaluate physical dysfunction.

Main Results:

A subset of SCs (30%-70%) exhibited resistance to senolytics.
Senolytic-resistant SCs had altered SASP with reduced pro-inflammatory factors but increased growth/fibrotic factors.
Resistant SCs released less mitochondrial DNA and expressed more GPNMB.
Transplantation of resistant SCs caused less physical dysfunction in mice.

Conclusions:

Senolytic-resistant SCs possess distinct functional and molecular profiles.
Senescent cell plasticity allows interconversion between sensitive and resistant states.
Targeting senolytic-resistant SCs may offer new therapeutic avenues for aging-related diseases.