Konferenssiposteri
Dasatinib decreases male germline stem cell colony growth in vitro, but do not induce apoptosis in germ cells with clinically relevant doses ex vivo
Tekijät: Eggert, Anna; Olotu, Opeyemi; Laasanen, Sini; Nieminen, Miisael; Jahnukainen, Kirsi; Kotaja, Noora; Mäkelä, Juho-Antti; Toppari, Jorma
Konferenssin vakiintunut nimi: EUROTOX 2025
Julkaisuvuosi: 2025
Lehti: Toxicology Letters
Vuosikerta: 411
Numero: Suppl.
Aloitussivu: S355
Lopetussivu: S356
DOI: https://doi.org/10.1016/j.toxlet.2025.07.824
Verkko-osoite: https://doi.org/10.1016/j.toxlet.2025.07.824
Tiivistelmä
In oncology, imatinib-resistance and -intolerance is a clinical challenge (Millot et al., 2011; Quintás-Cardama et al., 2009). Dasatinib is a second-generation tyrosine kinase inhibitor (TKI) and it is used for the BCR-ABL-driven diseases, such as CML and Philadelphia-chromosome-positive acute lymphoblastic leukemia (Ph+ALL) especially when imatinib-resistance or -intolerance occurs (Lindauer & Hochhaus, 2014). Dasatinib inhibits BCR-ABL, the SRC family kinases, receptor tyrosine kinases (c-KIT, PDGFR, DDR1 and 2, c-FMS, ephrin receptors), and TEC family kinases. Dasatinib-treatment is also used for children and adults at fertile age (Lindauer & Hochhaus, 2014). We have previously shown that imatinib decreases germ cell survival and germline stem cell(mGSC) proliferation in rodent testis ex vivo and in vitro (Eggert et al., 2024). Because dasatinib inhibits e.g. c-KIT and PDGFR, it poses like imatinib, a potential risk for male germ cells (Yan et al., 2000). Therefore, we investigated whether dasatinib mimics the imatinib-mediated effects on spermatogenic cells and mGSCs in a dose-dependent manner as imatinib does. We exposed adult seminiferous tubules from mice and rats (epithelial stages VIII-IX and IX-I) ex vivo as well as germline stem cells (mGSCs) from mice in vitro to dasatinib at doses of 0-10 µM. Cleaved caspase-3 (CC3) staining showed that dasatinib induced apop-tosis in dasatinib-treated cultured rat seminiferous tubules after 24 hours only at the highest dose (10 µM). The clinically relevant doses of dasatinib (0.01-1 µM) did not increase apoptosis in germ cells ex vivo. Dasatinib treatment of mGSCs decreased the growth of mGSC colonies in vitro, but not the proliferation of mGSCs ex vivo. The dasat-inib-treated mGSCs behaved slightly controversially in vitro compared to the imatinib-treated mGSCs, which prompted us to investigate whether the culture conditions in the presence of dasatinib affect the survival of mGSCs in vitro. Accordingly, we performed cell death and apoptosis analyses (Annexin-V, CC3, Trypan blue). Dasatinib doses of 0.3 and 1 µM induced apoptosis in mGSCs in vitro. Our immunofluorescence staining of dasatinib-treated mGSCs showed that the apoptotic (CC3+) mGSCs were mainly proliferating (KI67+) mGSCs, providing a plausible mechanism of dasatinib action on mGSC colony growth. Because the outcomes observed with dasatinib differed from those seen with imatinib, the findings of one TKI may not be generalizable to the effects of all TKIs.
In oncology, imatinib-resistance and -intolerance is a clinical challenge (Millot et al., 2011; Quintás-Cardama et al., 2009). Dasatinib is a second-generation tyrosine kinase inhibitor (TKI) and it is used for the BCR-ABL-driven diseases, such as CML and Philadelphia-chromosome-positive acute lymphoblastic leukemia (Ph+ALL) especially when imatinib-resistance or -intolerance occurs (Lindauer & Hochhaus, 2014). Dasatinib inhibits BCR-ABL, the SRC family kinases, receptor tyrosine kinases (c-KIT, PDGFR, DDR1 and 2, c-FMS, ephrin receptors), and TEC family kinases. Dasatinib-treatment is also used for children and adults at fertile age (Lindauer & Hochhaus, 2014). We have previously shown that imatinib decreases germ cell survival and germline stem cell(mGSC) proliferation in rodent testis ex vivo and in vitro (Eggert et al., 2024). Because dasatinib inhibits e.g. c-KIT and PDGFR, it poses like imatinib, a potential risk for male germ cells (Yan et al., 2000). Therefore, we investigated whether dasatinib mimics the imatinib-mediated effects on spermatogenic cells and mGSCs in a dose-dependent manner as imatinib does. We exposed adult seminiferous tubules from mice and rats (epithelial stages VIII-IX and IX-I) ex vivo as well as germline stem cells (mGSCs) from mice in vitro to dasatinib at doses of 0-10 µM. Cleaved caspase-3 (CC3) staining showed that dasatinib induced apop-tosis in dasatinib-treated cultured rat seminiferous tubules after 24 hours only at the highest dose (10 µM). The clinically relevant doses of dasatinib (0.01-1 µM) did not increase apoptosis in germ cells ex vivo. Dasatinib treatment of mGSCs decreased the growth of mGSC colonies in vitro, but not the proliferation of mGSCs ex vivo. The dasat-inib-treated mGSCs behaved slightly controversially in vitro compared to the imatinib-treated mGSCs, which prompted us to investigate whether the culture conditions in the presence of dasatinib affect the survival of mGSCs in vitro. Accordingly, we performed cell death and apoptosis analyses (Annexin-V, CC3, Trypan blue). Dasatinib doses of 0.3 and 1 µM induced apoptosis in mGSCs in vitro. Our immunofluorescence staining of dasatinib-treated mGSCs showed that the apoptotic (CC3+) mGSCs were mainly proliferating (KI67+) mGSCs, providing a plausible mechanism of dasatinib action on mGSC colony growth. Because the outcomes observed with dasatinib differed from those seen with imatinib, the findings of one TKI may not be generalizable to the effects of all TKIs.
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