Nano-intervention to stop therapy-induced cellular senescence.

HENSCHKE, A; GRZESKOWIAK, B. ; MIELCAREK, A.; PERRIGUE, P; JASKOT, K; IVASHCHENKO, O.; SÁNCHEZ CERVIÑO, M. CELINA; COY, E; MOYA, S

Barcelona

Congreso; NanoBio&Med20 23; 2023

NanoBio&Med20 23

Cellular senescence is a phenomenon that causes genetic, epigenetic, metabolic, and structuralchanges to original cells, causing cell cycle arrest and prevents them from growing. It is naturallycaused by telomere shortening, when cells are unable to divide anymore, and is an inherent part ofour lives. In general, this mechanism is a protector from over proliferation, what makes this a tumorsuppressive mechanism [1]. Senescent induction is an effect of various factors, such as epigenetic stress, proteotoxic stress, oxidative stress, telomere damage and DNA damage [2]. Such stressful environment may lead to accumulation of those cells in tissues which are a source of inflammation or tissue disfunction and causes many age-related diseases. Chemotherapy is one of the sources to induce cellular senescence but after induction it is ineffective for those cells making them apoptosisresistant. It is caused by their growth arrest, while chemotherapeutics are mostly working on highly proliferative cells such as cancer cells. Moreover, Senescent Associated Secretory Phenotype (SASP), which is a very characteristic feature of senescent cells, containing cytokines chemokines, growth factors and proteases, plays a crucial role in tumorigenesis and inflammation [1]. In summary, it is necessary to use drugs that are working selectively, which are senolytics, causing induction of apoptosis on senescent cells (senolysis). Increase of cancersenescent cells after chemotherapy is a considerable problem, as those cells can reenter cellcycle to become cancer stem cells and leads to cancer relapse. The idea is to introduce additionaltherapy – senotherapy – to eliminate remaining senescent cells [3-5]. This study focused onpreparing the protocol of therapy-induced senescence (TIS) with doxorubicin on two cell lines– WI38 (fibroblasts) and A549 (cancer cells). To confirm the effectiveness of the method experiments with senescence markers were conducted, such as evaluation of β-galactosidase level and proliferation of the cells. Followed by investigation of its morphology on confocal microscopy to prove the size changes which were additionally corroborated by the change in complexity as measured by flow cytometry. Next, liposomes were prepared encapsulating the senolytic drug, Fisetin, and administrated to senescent cells. Liposomes were characterized CryoSEM and DLS. Cell uptake of liposomes with encapsulated Fisetin was studied by flow cytometry. Cell viability studies after administration of liposomes were conducted by IN Cell Analyzer, using Live/Dead Assay. This investigation showed that therapy-induced senescence by doxorubicin is greatly effective and senotherapy through fisetin encapsluated in liposomes is highly potential.