Bortezomib Prevents Acute Doxorubicin Ovarian Insult and Follicle Demise, Improving the Fertility Window and Pup Birth Weight in Mice

Roti, Elon C. Roti; Ringelstetter, Ashley K.; Schmidt, Jenna Kropp; Abbott, David H.; Salih, Sana M.

doi:10.1371/journal.pone.0108174

Cited by 36 publications

(38 citation statements)

References 42 publications

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“…This temporal insult pattern in granulosa cells could contribute to the initial diameter increase and fibrin degradation we observed in the beginning of culture with lower concentrations of DXR. The slower timeline for granulosa cell damage is suggestive of a window to prevent acute DXR ovarian insult and follicle demise as recently reported [ 27 ]. A treatment with Bortezomib, a DXR competitive binder to the proteasome and translocation to the DNA, prior to DXR exposure attenuated DXR-induced DNA damage in all ovarian cell types, resulting in apoptosis of preantral follicles in vivo [ 27 ].…”

Section: Resultssupporting

confidence: 58%

“…The slower timeline for granulosa cell damage is suggestive of a window to prevent acute DXR ovarian insult and follicle demise as recently reported [ 27 ]. A treatment with Bortezomib, a DXR competitive binder to the proteasome and translocation to the DNA, prior to DXR exposure attenuated DXR-induced DNA damage in all ovarian cell types, resulting in apoptosis of preantral follicles in vivo [ 27 ]. These findings support our observations of continued follicle growth during culture but impaired oocyte maturation on D12.…”

Section: Resultssupporting

confidence: 58%

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Hydrogel Based 3-Dimensional (3D) System for Toxicity and High-Throughput (HTP) Analysis for Cultured Murine Ovarian Follicles

et al. 2015

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Various toxicants, drugs and their metabolites carry potential ovarian toxicity. Ovarian follicles, the functional unit of the ovary, are susceptible to this type of damage at all stages of their development. However, despite of the large scale of potential negative impacts, assays that study ovarian toxicity are limited. Exposure of cultured ovarian follicles to toxicants of interest served as an important tool for evaluation of toxic effects for decades. Mouse follicles cultured on the bottom of a culture dish continue to serve an important approach for mechanistic studies. In this paper, we demonstrated the usefulness of a hydrogel based 3-dimensional (3D) mouse ovarian follicle culture as a tool to study ovarian toxicity in a different setup. The 3D in vitro culture, based on fibrin alginate interpenetrating network (FA-IPN), preserves the architecture of the ovarian follicle and physiological structure-function relationship. We applied the novel 3D high-throughput (HTP) in vitro ovarian follicle culture system to study the ovotoxic effects of an anti-cancer drug, Doxorobucin (DXR). The fibrin component in the system is degraded by plasmin and appears as a clear circle around the encapsulated follicle. The degradation area of the follicle is strongly correlated with follicle survival and growth. To analyze fibrin degradation in a high throughput manner, we created a custom MATLAB® code that converts brightfield micrographs of follicles encapsulated in FA-IPN to binary images, followed by image analysis. We did not observe any significant difference between manually processed images to the automated MATLAB® method, thereby confirming that the automated program is suitable to measure fibrin degradation to evaluate follicle health. The cultured follicles were treated with DXR at concentrations ranging from 0.005 nM to 200 nM, corresponding to the therapeutic plasma levels of DXR in patients. Follicles treated with DXR demonstrated decreased survival rate in greater DXR concentrations. We observed partial follicle survival of 35% ± 3% (n = 80) in 0.01nM treatment and 48% ± 2% (n = 92) in 0.005nM, which we identified as the IC50 for secondary follicles. In summary, we established a 3D in vitro ovarian follicle culture system that could be used in an HTP approach to measure toxic effects on ovarian follicles.

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Section: Resultssupporting

confidence: 58%

Section: Resultssupporting

confidence: 58%

Hydrogel Based 3-Dimensional (3D) System for Toxicity and High-Throughput (HTP) Analysis for Cultured Murine Ovarian Follicles

et al. 2015

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“…n = 3 blots/quantification, error bars indicate the SE of the mean, * p< 0.05, **p<0.001, one-way ANOVA, Tukey means comparison. The control and DXR-only treatment groups adapted from [ 27 ].…”

Section: Resultsmentioning

confidence: 99%

“…All efforts were taken to decrease the number of animals used. Data from vehicle control and DXR-only treated animals in this manuscript have been reported previously in a study examining bortezomib protection against DXR toxicity [ 27 ]. The initial study design comprised eight treatment groups, including control-, bortezomib- and dexrazoxane-treated mice ( S1 Table ).…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2015

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Advances in cancer treatment utilizing multiple chemotherapies have dramatically increased cancer survivorship. Female cancer survivors treated with doxorubicin (DXR) chemotherapy often suffer from an acute impairment of ovarian function, which can persist as long-term, permanent ovarian insufficiency. Dexrazoxane (Dexra) pretreatment reduces DXR-induced insult in the heart, and protects in vitro cultured murine and non-human primate ovaries, demonstrating a drug-based shield to prevent DXR insult. The present study tested the ability of Dexra pretreatment to mitigate acute DXR chemotherapy ovarian toxicity in mice through the first 24 hours post-treatment, and improve subsequent long-term fertility throughout the reproductive lifespan. Adolescent CD-1 mice were treated with Dexra 1 hour prior to DXR treatment in a 1:1 mg or 10:1 mg Dexra:DXR ratio. During the acute injury period (2–24 hours post-injection), Dexra pretreatment at a 1:1 mg ratio decreased the extent of double strand DNA breaks, diminished γH2FAX activation, and reduced subsequent follicular cellular demise caused by DXR. In fertility and fecundity studies, dams pretreated with either Dexra:DXR dose ratio exhibited litter sizes larger than DXR-treated dams, and mice treated with a 1:1 mg Dexra:DXR ratio delivered pups with birth weights greater than DXR-treated females. While DXR significantly increased the “infertility index” (quantifying the percentage of dams failing to achieve pregnancy) through 6 gestations following treatment, Dexra pretreatment significantly reduced the infertility index following DXR treatment, improving fecundity. Low dose Dexra not only protected the ovaries, but also bestowed a considerable survival advantage following exposure to DXR chemotherapy. Mouse survivorship increased from 25% post-DXR treatment to over 80% with Dexra pretreatment. These data demonstrate that Dexra provides acute ovarian protection from DXR toxicity, improving reproductive health in a mouse model, suggesting this clinically available drug may provide ovarian protection for cancer patients.

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“…The store of resting, primordial follicles is reduced after exposure, with evidence both of a direct damaging effect of CIS on primordial follicles (Nguyen et al 2018(Nguyen et al , 2019, and of an indirect effect on primordial follicles resulting from accelerated activation of primordial follicles due to a loss of inhibitory factors previously secreted by more developed follicles that have themselves been targeted by the drugs ('burnout' (Kalich-Philosoph et al 2013)); currently, there is some controversy over the extent to which each of these different pathways is primarily responsible for the loss of primordial follicles. DOX exposure results in a loss of both primordial and growing follicles, mainly through effects on mitotically active granulosa cells (GCs), and leads to reduced ovulation rates (Perez et al 1997, Ben-Aharon et al 2010, Morgan et al 2013, Roti Roti et al 2014. DOX also damages ovarian microvasculature and induces necrosis of stroma cells (Ben-Aharon et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Single and combined effects of cisplatin and doxorubicin on the human and mouse ovary in vitro

et al. 2020

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Chemotherapy drugs are administered to patients using combination regimens, and as such the possibility of multiplicative effects between drugs need to be investigated. This study examines the individual and combined effects of the chemotherapy drugs cisplatin and doxorubicin on the human ovary. Although cisplatin and doxorubicin are known to affect female fertility, there is limited information about their direct effects on the human ovary, and none examining the possibility of combined, multiplicative effects of co-exposure to these drugs. Here, human ovarian biopsies were obtained from 14 women at the time of caesarean section, with 38 mouse ovaries also obtained from neonatal C57Bl/6J mice. Tissue was cultured for 6 days prior to analyses, with chemotherapy drugs added to culture medium on the second day of culture only. Treatment groups of a single (5 μg/mL human; 0.5 μg/mL mouse) or double (10 μg/mL human; 1.0 μg/mL mouse) dose of cisplatin, a single (1 μg/mL human; 0.05 μg/mL mouse) or double (2 μg/mL human; 0.01 μg/mL mouse) dose of doxorubicin or a combination of a single dose of both drugs together were compared to controls without drug exposure. Exposure to cisplatin or doxorubicin significantly decreased follicle health in human and mouse, supporting the suitability of the mouse as a model for the human ovary. There was also a significant reduction of mouse follicle number. Human ovarian stromal tissue exhibited increased apoptosis and decreased cell proliferation. Crucially, there was no evidence indicating the occurrence of multiplicative effects between cisplatin and doxorubicin.

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Bortezomib Prevents Acute Doxorubicin Ovarian Insult and Follicle Demise, Improving the Fertility Window and Pup Birth Weight in Mice

Cited by 36 publications

References 42 publications

Hydrogel Based 3-Dimensional (3D) System for Toxicity and High-Throughput (HTP) Analysis for Cultured Murine Ovarian Follicles

Hydrogel Based 3-Dimensional (3D) System for Toxicity and High-Throughput (HTP) Analysis for Cultured Murine Ovarian Follicles

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

Single and combined effects of cisplatin and doxorubicin on the human and mouse ovary in vitro

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