Dexrazoxane Ameliorates Doxorubicin-Induced Injury in Mouse Ovarian Cells1

Abstract

Doxorubicin (DXR) is a frontline chemotherapy agent implicated in unintended ovarian failure in female cancer survivors. The fertility preservation techniques currently available for cancer patients are often time and cost prohibitive and do not necessarily preserve endocrine function. There are no drug-based ovary protection therapies clinically available. This study provides the first investigation using dexrazoxane (Dexra) to limit DXR insult in ovarian tissue. In KK-15 granulosa cells, a 3-h DXR treatment increased double-strand (ds) DNA breaks 40%-50%, as quantified by the neutral comet assay, and dose-dependent cytotoxicity. Dexra exhibited low toxicity in KK-15 cells, inducing no DNA damage and less than 20% cell loss. Cotreating KK-15 cells with Dexra prevented acute DXR-induced dsDNA damage. Similarly, Dexra attenuated the DXR-induced 40%-65% increase in dsDNA breaks in primary murine granulosa cells and cells from in vitro cultured murine ovaries. DXR can cause DNA damage either through a topoisomerase II-mediated pathway, based on DXR intercalation into DNA, or through oxidative stress. Cotreating KK-15 cells with 2 μM Dexra was sufficient to prevent DXR-induced, but not H(2)O(2)-induced, DNA damage. These data indicated the protective effects are likely due to Dexra's inhibition of topoisomerase II catalytic activity. This putative protective agent attenuated downstream cellular responses to DXR, preventing H2AFX activation in KK-15 cells and increasing viability as demonstrated by increasing the DXR lethal dose in KK-15 cells 5- to 8-fold (LD(20)) and primary murine granulosa cells 1.5- to 2-fold (LD(50)). These data demonstrate Dexra protects ovarian cells from DXR insult and suggest that it is a promising tool to limit DXR ovarian toxicity in vivo.

Dexrazoxane Diminishes Doxorubicin-Induced Acute Ovarian Damage and Preserves Ovarian Function and Fecundity in Mice

“…The 20 mg/kg Dexra dose represents a 1:1 Dexra/DXR mg ratio, providing a significant dose reduction from that used in cardioprotection to limit potential side effects of Dexra. The chosen Dexra dose was based on our previous in vitro study demonstrating a 2 μM Dexra dose, 100-folds lower than that used in in vitro cardiac protection studies, preserved granulosa cell viability against DXR [ 23 ]. Animals were euthanized with CO 2 followed by cervical dislocation and ovaries removed surgically 0, 2, 4, 10, 12 or 24 h after the second injection.…”

“…In a proof-of-concept study using immortalized KK-15 mouse granulosa cells, we demonstrated that Dexra protects granulosa cells from DXR-induced DNA damage and cytotoxicity in vitro in a manner consistent with inhibiting TOPII DNA cleavage, rather than oxidative stress [ 23 ]. Furthermore, Dexra treatment of in vitro- cultured mouse and marmoset ovaries prior to DXR administration reduced the number of dsDNA breaks, and rescued primary granulosa cell viability [ 23 , 24 ]. The data demonstrated the potential for Dexra to be an effective ovoprotective agent against DXR toxicity by blocking the initial insult as well as subsequent cellular demise.…”

Modulatory Effects of Dexrazoxane Against Genotoxicity and Lipid Peroxidation Induced by Idarubicin in HepG2 Cells

“…Anti-genotoxic effects of dexrazoxane against doxorubicin in mouse ovarian cells were also reported (40). Their results revealed the ability of dexrazoxane to inhibit the topoisomerase II catalytic activity, to reduce double-strand DNA breaks and, thus, to prevent genotoxicity.…”

Activation of TGF‐β1/Smad3 signaling pathway inhibits the development of ovarian follicle in polycystic ovary syndrome by promoting apoptosis of granulosa cells

“…The largest follicles were punctured to obtain oocytes and granulosa cells (Cheng et al, ). Theca cells cells were isolated by 30‐min treatment with 0.25% collagenase at 37°C followed by cell dispersion using a 23‐Gauge needle (Roti Roti & Salih, ). Bone marrow mesenchymal cells were flushed out from the tibial and femoral bones of donor adult female rats using a syringe and 20‐Gauge needle (Wei, Fraser, Lu, Hu, & Yu, ).…”