Tumor-Derived Oxidative Stress Triggers Ovarian Follicle Loss in Breast Cancer

Abstract

Breast cancer is a common indication for ovarian cryopreservation. However, whether the grafting ovarian tissue meets functional requirements, as well as the need for additional interventions, remains unclear. The current study demonstrates abnormal serum hormones in breast cancer in humans and breast cancer cell line–derived tumor-bearing mice, and for the first time shows tumor-induced loss of primordial and growing follicles, and the number of follicles being lost to either growth or atresia. A gene signature of tumor-bearing mice demonstrates the disturbed regulatory network of steroidogenesis, which links to mitochondria dysfunction in oocytes and granulosa cells via the phosphatidylinositol 3-kinase signaling pathway. Notably, increased reactive oxygen species were identified in serum and ovarian tissues in tumor-bearing mice. Furthermore, supplementation with vitamin C promoted follicular quiescence, repairing tumor-induced follicle loss via inactivation of the phosphatidylinositol 3-kinase–Akt–mammalian target of rapamycin pathway, indicating the potential of antioxidants as a fertility therapy to achieve higher numbers of healthy follicles ready for ovarian cryopreservation. Breast cancer is a common indication for ovarian cryopreservation. However, whether the grafting ovarian tissue meets functional requirements, as well as the need for additional interventions, remains unclear. The current study demonstrates abnormal serum hormones in breast cancer in humans and breast cancer cell line–derived tumor-bearing mice, and for the first time shows tumor-induced loss of primordial and growing follicles, and the number of follicles being lost to either growth or atresia. A gene signature of tumor-bearing mice demonstrates the disturbed regulatory network of steroidogenesis, which links to mitochondria dysfunction in oocytes and granulosa cells via the phosphatidylinositol 3-kinase signaling pathway. Notably, increased reactive oxygen species were identified in serum and ovarian tissues in tumor-bearing mice. Furthermore, supplementation with vitamin C promoted follicular quiescence, repairing tumor-induced follicle loss via inactivation of the phosphatidylinositol 3-kinase–Akt–mammalian target of rapamycin pathway, indicating the potential of antioxidants as a fertility therapy to achieve higher numbers of healthy follicles ready for ovarian cryopreservation. Breast cancer is the most common malignant tumor and the leading cause of cancer deaths among women, with an estimated 2.3 million new cases and 685,000 deaths worldwide in 2021.1Sung H. Ferlay J. Siegel R.L. Laversanne M. Soerjomataram I. Jemal A. Bray F. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries.CA Cancer J Clin. 2021; 71: 209-249Crossref PubMed Scopus (37538) Google Scholar The overall breast cancer onset in China is at a younger age than in many Western countries.2Fan L. Strasser-Weippl K. Li J.J. St Louis J. Finkelstein D.M. Yu K.D. Chen W.Q. Shao Z.M. Goss P.E. Breast cancer in China.Lancet Oncol. 2014; 15: e279-e289Abstract Full Text Full Text PDF PubMed Scopus (1206) Google Scholar Approximately 13.6% of all women diagnosed annually with breast cancers in China were aged <40 years, compared with 4.8% in the United States.3Co M. Kwong A. Young onset breast cancer in Southern China - a 5-year clinico-pathological study from a multi-centre database.Cancer Treat Res Commun. 2020; 24: 100182Crossref PubMed Scopus (5) Google Scholar Breast cancer in young women is a crucial health concern, especially for those who have not completed their family planning, and evidence supports that childbearing is not contraindicated.4Hickey M. Peate M. Saunders C.M. Friedlander M. Breast cancer in young women and its impact on reproductive function.Hum Reprod Update. 2009; 15: 323-339Crossref PubMed Scopus (126) Google Scholar Thus, topics of reproductive options and fertility outcomes are becoming more important as life expectancy and quality of life after breast cancer are increasing.5Gonçalves V. Sehovic I. Quinn G. Childbearing attitudes and decisions of young breast cancer survivors: a systematic review.Hum Reprod Update. 2014; 20: 279-292Crossref PubMed Scopus (72) Google Scholar However, the fertility preservation program for breast cancer is still at its initial stage in China and elsewhere.6Wan Q. Han L. Liu J. Li H. Li Y. Long Q.M. Tan L. Li J.J. Fertility preservation among young breast cancer patients: a single-center experience in China.Taiwan J Obstet Gynecol. 2021; 60: 827-830Crossref PubMed Scopus (3) Google Scholar Common adverse effects of cancer therapy include premature ovarian failure and infertility. Cryopreservation of embryos and/or oocytes is the current commonly used fertility preservation before cancer therapy. However, these protocols may increase the risk of relapse because of the hormonal therapies used.7Arecco L. Blondeaux E. Bruzzone M. Ceppi M. Latocca M.M. Marrocco C. Boutros A. Spagnolo F. Razeti M.G. Favero D. Spinaci S. Condorelli M. Massarotti C. Goldrat O. Del Mastro L. Demeestere I. Lambertini M. Safety of fertility preservation techniques before and after anticancer treatments in young women with breast cancer: a systematic review and meta-analysis.Hum Reprod. 2022; 37: 954-968Crossref PubMed Scopus (29) Google Scholar An alternative for fertility preservation for women with breast cancer is cryopreservation of ovarian tissue without the need for hyperstimulation.8Arav A. Patrizio P. Techniques of cryopreservation for ovarian tissue and whole ovary.Clin Med Insights Reprod Health. 2019; 13 (1179558119884945)Crossref PubMed Google Scholar,9Gjeterud J. Kristensen S.G. Fedder J. [Indications for cryopreservation and autotransplantation of ovarian tissue].Tidsskr Nor Laegeforen. 2021; 141 (Norwegian) (doi:10.4045/tidsskr.21.0030)PubMed Google Scholar Despite some progress, cryopreservation of ovarian tissue in humans is still being optimized, with concerns surrounding the technical aspects of the procedure.8Arav A. Patrizio P. Techniques of cryopreservation for ovarian tissue and whole ovary.Clin Med Insights Reprod Health. 2019; 13 (1179558119884945)Crossref PubMed Google Scholar,10Hossay C. Donnez J. Dolmans M.M. Whole ovary cryopreservation and transplantation: a systematic review of challenges and research developments in animal experiments and humans.J Clin Med. 2020; 9: 3196Crossref PubMed Scopus (11) Google Scholar Ovarian cryopreservation uses either ovarian cortical fragments or entire ovarian tissue,10Hossay C. Donnez J. Dolmans M.M. Whole ovary cryopreservation and transplantation: a systematic review of challenges and research developments in animal experiments and humans.J Clin Med. 2020; 9: 3196Crossref PubMed Scopus (11) Google Scholar which theoretically contains an entire follicle pool; therefore, it is an alternative to ovulation induction for preserving fertility.11Ladanyi C. Mor A. Christianson M.S. Dhillon N. Segars J.H. Recent advances in the field of ovarian tissue cryopreservation and opportunities for research.J Assist Reprod Genet. 2017; 34: 709-722Crossref PubMed Scopus (67) Google Scholar Because both the quality and the number of follicles are linked to pregnancy outcomes, it was questioned whether the grafting ovarian tissue has normal endocrine ovarian function, as well as the need for additional interventions before ovarian transplantation. It has been well established that cancer cells are under constant oxidative stress, as reflected by elevated reactive oxygen species (ROS). ROS are a group of highly reactive oxygen-containing molecules that are pathologically elevated in breast cancer.12Jelic M.D. Mandic A.D. Maricic S.M. Srdjenovic B.U. Oxidative stress and its role in cancer.J Cancer Res Ther. 2021; 17: 22-28Crossref PubMed Scopus (123) Google Scholar The tumor-derived ROS can hyperactivate phosphatidylinositol 3-kinase (PI3K)–Akt by inactivating their upstream negative regulator phosphatase regulator and tensin homolog (PTEN) in tumor cells.13Zhang T. Zhu X. Wu H. Jiang K. Zhao G. Shaukat A. Deng G. Qiu C. Targeting the ROS/PI3K/AKT/HIF-1α/HK2 axis of breast cancer cells: combined administration of polydatin and 2-deoxy-d-glucose.J Cell Mol Med. 2019; 23: 3711-3723Crossref PubMed Scopus (70) Google Scholar Furthermore, ROS can regulate the PI3K-Akt signaling pathway in metabolic disorders, such as type 2 diabetes and obesity, by follicles and M. M. The ovarian as target of and ROS in metabolic ovarian 2021; PubMed Scopus Google Scholar have oxidative in ovarian tissues of with G. M. V. S. of potential and oxidative stress in and ovarian tissue from 2014; Full Text Full Text PDF PubMed Scopus Google Scholar However, it is whether a and more whether increased ROS are the cause of ovarian in with breast The current study the need for with breast cancer to therapies to follicle health before ovarian cryopreservation. The of and of premature ovarian with breast cancer in humans and mice. The tumor-bearing follicle that an cause of these may a tumor-induced in the The study of oxidative stress in with breast cancer and in tumor-bearing mice. 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Liu L. D. K. S. Y. C. T. I. Liu K. of premature of the primordial follicle PubMed Scopus Google M. M. of follicles in and Reprod. 2019; 34: PubMed Scopus Google M. signaling in cell and their in cell and ovarian J Mol 2021; PubMed Scopus Google C. M. C. of the and follicle by ovarian cryopreservation and in Ovarian 2021; PubMed Scopus Google Scholar was in the tumor-bearing that functional from were in the tumor-bearing group compared with the to its in to regulate of of growing follicles to M.M. F. V. C.M. F. G. J. to ovarian in 2019; PubMed Scopus Google Scholar that both and were in of follicle in from both primordial follicles, was in in and follicles in the tumor-bearing was a in compared with that in the group in was in follicles in the tumor-bearing group and growth have been to in and to S. D. C. A. N. A. and and regulate via of in granulosa PubMed Scopus Google Scholar and their was increased in the tumor-bearing as by and of that were in and The of is by regulatory S. M. J. K. of the regulatory in breast cancer: in and a 2019; PubMed Scopus (11) Google Scholar and the of was by The is by of by cells and to that the disturbed regulatory network of was for abnormal in the tumor-bearing mice ROS effects on follicle and ovarian M. M. The ovarian as target of and ROS in metabolic ovarian 2021; PubMed Scopus Google Scholar of mice, that the were in the oxidative and to oxidative stress, and the were in and that functional from oxidative were in the tumor-bearing group with the of of the such as and H. T. S. M. M. S. K. M. of oxidative stress on in 2019; PubMed Scopus Google S. Y. Li J. Yu Y. M. Liu H. Y. X. L. L. P. J. Q. J. F. Liu of ovarian 2020; Full Text Full Text PDF PubMed Scopus Google Scholar and and S. Y. Li J. Yu Y. M. Liu H. Y. X. L. L. P. J. Q. J. F. Liu of ovarian 2020; Full Text Full Text PDF PubMed Scopus Google C. P. in granulosa gene with increased in PubMed Scopus Google Scholar were in the tumor-bearing group The in mitochondria of the were K. O. A. L. the impact of on fertility in women with breast cancer.J Clin Oncol. 24: PubMed Scopus Google Scholar the tumor-bearing the number of mitochondria oocytes was by compared with that in with a in and higher of abnormal mitochondria compared with which were and a mitochondria in which the of and were and were with more dysfunction is with increased ROS by the therefore, in serum ROS in mice were and were the were indicating ROS and in both serum and ovarian tissues in tumor-bearing mice whether the ROS to follicle loss and the in breast the was used to the ovarian oxidative stress and follicle in mice and The effects were by the serum and ovarian oxidative with tumor-bearing mice, was an increase in and in serum and and in ovarian tissues and was a in in serum and ovarian tissue in the indicating that the the oxidative stress of tumor-bearing mice The and of from mice were compared with primordial tumor-derived follicle loss C and of the number of the follicle in the of mice with that the number of follicles and the numbers of and follicles those in from mice of an increase in the number of primordial follicles in mice cell the of follicle increased in follicles were in the primordial follicles in the group compared with that of a in the number of not in follicles and in from the group follicles compared with mice and in was increased in the as by and Western of to the and with these Western that increased the of and compared with the group that tumor-derived follicle loss via of the pathway. cryopreservation of ovarian tissue is to young women with breast the cryopreservation for a is to normal ovarian is in of of achieve pregnancy and T. Zhu X. Wu H. Jiang K. Zhao G. Shaukat A. Deng G. Qiu C. Targeting the ROS/PI3K/AKT/HIF-1α/HK2 axis of breast cancer cells: combined administration of polydatin and 2-deoxy-d-glucose.J Cell Mol Med. 2019; 23: 3711-3723Crossref PubMed Scopus (70) Google M. J. M. the of the group for the preservation of fertility in and and therapies of the of and 2021; M. Dolmans M.M. S. D. of ovarian tissue transplantation: from 2020; Full Text Full Text PDF PubMed Scopus (72) Google Scholar ovarian follicle at the time of has the on of pregnancy and H. J. M. M. J. T. M. K. S. A. G. F. M. M. M. in women of ovarian tissue after in a fertility preservation tissue pregnancy and Reprod. PubMed Scopus Google Scholar However, the of the ovarian before cancer has not been with most on ovarian failure after a the current study for the first tumor-induced loss of primordial and growing follicles, from abnormal in breast The number of follicles were to lost to either growth or which was linked to oxidative of the pathway. with breast cancer may need to therapies to follicle health before ovarian cryopreservation. ovarian to ovarian dysfunction for women aged <40 years, with an elevated serum and of M.S. ovarian and Cell 2021; 9: PubMed Scopus (67) Google Scholar serum the of follicular and follicle K. M. Fertility in cancer survivors: preservation and the role of ovarian Reprod Update. PubMed Scopus Google Scholar the of the follicles, and has a negative on N. T. Y. T. T. T. of and of ovarian in 2014; PubMed Scopus Google Scholar young women with breast cancer experience before cancer T. Zhu X. Wu H. Jiang K. Zhao G. Shaukat A. Deng G. Qiu C. Targeting the ROS/PI3K/AKT/HIF-1α/HK2 axis of breast cancer cells: combined administration of polydatin and 2-deoxy-d-glucose.J Cell Mol Med. 2019; 23: 3711-3723Crossref PubMed Scopus (70) Google Scholar and of are diagnosed with Cancer of breast PubMed Scopus Google Scholar the current a breast cancer cell was to study the in follicular growth or loss in the grafting of with breast Yu P. J. of breast cancer cell Ther. 2020; PubMed Scopus Google Scholar the of and of premature ovarian with breast cancer in humans and mice. The tumor-bearing follicle that an cause of these may a tumor-induced in the An and increased of the primordial follicles to the of premature ovarian M.S. ovarian and Cell 2021; 9: PubMed Scopus (67) Google Scholar in tumor-bearing follicle and ovary of both primordial and growing The increase in the of follicles reflected the loss of a number in primordial follicles than growing follicles, that the primordial follicles were for follicular A. of the of ovarian increase in the of and follicles in Reprod. PubMed Scopus Google Scholar follicle is a by which primordial follicles are the growing follicle and is by the growth of as was by the number of in PI3K-Akt and signaling can in oocytes or and a has been to follicle X. Y. Y. J. Liu H. Liu J. Li J. for in of primordial follicles with and PubMed Scopus Google H. Liu K. and of the of primordial cells follicle in Reprod Update. PubMed Scopus Google Scholar signaling the of in and signaling as the the and the in of PI3K-Akt signaling H. 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I. of granulosa and cell ovarian follicle Reprod PubMed Scopus Google Scholar from follicular supports that is the cause for the increase in and the increase in of The of follicle PubMed Scopus Google Scholar PI3K-Akt and signaling may of by and growth family growth and is in the of and of in S. N. O. of that granulosa cell Cell PubMed Scopus Google Scholar the disturbed and in and can or follicle atresia. of primordial follicles is to of growing follicles in P. Liu L. D. K. S. Y. C. T. I. Liu K. of premature of the primordial follicle PubMed Scopus Google Scholar The current a in of and that the oocytes and are with in in increased follicular in both primordial and growing follicles and follicular in of tumor-bearing mice. Cancer to the network leading to ROS oxidative stress and are in serum of with breast cancer.12Jelic M.D. Mandic A.D. Maricic S.M. Srdjenovic B.U. Oxidative stress and its role in cancer.J Cancer Res Ther. 2021; 17: 22-28Crossref PubMed Scopus (123) Google S. C. S. oxidative stress, and antioxidants in breast cancer: a J Cancer PubMed Scopus Google Scholar the current were in the ovarian tissues of tumor-bearing mice. ROS can from oxidative in the to mitochondria in both the and of were to and ROS and of and pathway that oxidative was in the group in ovarian tissues of tumor-bearing mice. The of was by the of serum and ovarian oxidative stress and oxidative stress in with breast cancer and tumor-bearing mice. ROS a role in ovarian ROS can the as in the and T. Zhu X. Wu H. Jiang K. Zhao G. Shaukat A. Deng G. Qiu C. Targeting the ROS/PI3K/AKT/HIF-1α/HK2 axis of breast cancer cells: combined administration of polydatin and 2-deoxy-d-glucose.J Cell Mol Med. 2019; 23: 3711-3723Crossref PubMed Scopus (70) Google Scholar C is a that can and ROS in in as well as in a premature ovarian X. L. Li H. and in Med Cell 2019; PubMed Scopus Google S. C. H. C. Q. J. Tan J. Li H. C the potential of cells in premature ovarian Cell Res Ther. 2020; PubMed Scopus Google Scholar Fertility of vitamin C include of via regulatory signaling in in X. L. Li H. and in Med Cell 2019; PubMed Scopus Google Scholar and follicle growth in ovarian K. M.M. of on in of 20: PubMed Scopus Google Scholar breast ovarian loss of ovarian and and ROS signaling was to to and via signaling and Furthermore, current in the that supplementation with vitamin C promoted follicular quiescence, tumor-induced ovarian of the number of follicles, and normal serum and preserving ovarian and The pathway may the potential target to follicle and loss ovarian and in in C. M. C. of the and follicle by ovarian cryopreservation and in Ovarian 2021; PubMed Scopus Google Scholar Thus, inactivation of pathway by an may have additional as a for the of healthy follicles and functional ovarian tissues for with breast are to the current a the in breast cancer and ovarian has not been and are to the from the the of was the of breast loss of ovarian the health of ovarian tissue after vitamin C supplementation was not The current study demonstrates in a a breast cancer and ovarian that may by vitamin C However, a in breast cancer and ovarian in women has not been established and additional and the and the and the and serum the and the and the with of from breast cancer in mice. The of mice by The of was time with the The tumor of tumor-bearing group was of from vitamin C and mice. The tumor of mice was with in ovarian tissues of tumor-bearing mice. of gene and to the of and of for tumor group compared with fragments with of ovary of and vitamin C mice. with effects were by the serum oxidative and ovarian tissues after vitamin C are compared with the by of the and the with of phosphatase regulator and tensin homolog (PTEN) in granulosa cells in primordial and growing follicles and of in follicles in the of the the vitamin C. with serum hormonal in and vitamin C mice. are compared with the by of the and the with for and in granulosa cells in follicles at and of the of follicle of vitamin C and mice. of the are compared with the by of the the

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