Molecular networks underlying ovarian aging for clinical applications

YoungHo Roh,SookRyung Kim,EunJung Choi,YoungJoo Kim,HyangGi Park,PureunNarae Kang,DaJung Chung,NaYoung Kim,MinJae Kim,AeRa Han,KwangRae Kim,Chan Park,YongPil Cheon,YoungJin Lee 한국발생생물학회 2017 한국발생생물학회 학술발표대회 Vol.2017 No.8

Due to modern trends with postponing child-bearing and getting worse living environment in women, an ovarian aging increased pregnancy failure and other complications with menopause or premature ovarian failure. Although several theories have been suggested such as mitochondrial malfunction, DNA damage/repair/methylation, caloric restriction, studies regarding ovarian aging-related molecular mechanisms for development of therapeutic methods are insufficient so far. Our objective is to determine molecular pathways of ovarian aging that result in pregnancy failure and other complications in women health to develop treatment strategies. This study is consisted of two parts: in Phase I stage, we analyzed distinct gene expression profile between young and aged mouse ovaries, and in Phase II stage several preferentially expressed genes in both ovaries were selected and analyzed their physiological functions and involved molecular networks related to ovarian aging for development of diagnostic markers and therapeutic methods. Ovaries from 10 week and 11 month-old FVB/NJ female mice with synchronized estrus cycle were collected for this study. A half of each ovary was used for RNA preparation and the other half for histological analysis. Using the Illumina HiSeq 2000 System, preferentially expressed genes were identified. Functional annotation database-based gene-set enrichment analyses and Pathway Studio® were employed to evaluate aging-related molecular networks. These findings were confirmed through qRT-PCR and immunohistochemistry. To validate RNA-Seq data, we examined expression patterns of marker genes (Amh, Bmp15 and Nobox) that were wellknown to be decreased in ovarian aging process. In young or aged ovary, preferentially expressed 876 genes were identified and extracellular matrix (ECM; p<0.001) and chromatin/nucleosome-related (p<0.001) protein-coded genes have the majority in these genes by GOTERM analysis. Amongthem, we selected several candidate genes and confirmed their expression profiles by qRT-PCR and immunohistochemistry followed by molecular network analysis. Regarding molecular interactions in these genes, PathwayStudio® was employed to predict aging-involved molecular networks in mouse ovary. Here we report a couple of candidate molecular networks and medicines (chemicals) for targeting these preferentially expressed genes/proteins. Further analyses are scheduled to produce transgenic animal models and with human ovarian tissues/cell lines.

Combined panel of serum human tissue kallikreins and CA-125 for the detection of epithelial ovarian cancer

Stephen Chee Liang Koh,Chan Yiong Huak,Delfi Lutan,Johny Marpuang,Suwiyoga Ketut,Nyoma Gede Budiana,Agustria Zainu Saleh,Mohamad Farid Aziz,Hariyono Winarto,Heru Pradjatmo,Nguyen Khac Han Hoan,Pham Vi 대한부인종양학회 2012 Journal of Gynecologic Oncology Vol.23 No.3

Objective: To determine the predictive accuracy of the combined panels of serum human tissue kallikreins (hKs) and CA-125 for the detection of epithelial ovarian cancer. Methods: Serum specimens collected from 5 Indonesian centers and 1 Vietnamese center were analyzed for CA-125, hK6, and hK10 levels. A total of 375 specimens from patients presenting with ovarian tumors, which include 156 benign cysts, 172 epithelial ovarian cancers (stage I/II, n=72; stage III/IV, n=100), 36 germ cell tumors and 11 borderline tumors, were included in the study analysis. Receiver operating characteristic analysis were performed to determine the cutoffs for age, CA-125, hK6, and hK10. Sensitivity, specificity, negative, and positive predictive values were determined for various combinations of the biomarkers. Results: The levels of hK6 and hK10 were significantly elevated in ovarian cancer cases compared to benign cysts. Combination of 3 markers, age/CA-125/hk6 or CA-125/hk6/hk10, showed improved specificity (100%) and positive predictive value (100%) for prediction of ovarian cancer, when compared to the performance of single markers having 80-92% specificity and 74-87% positive predictive value. Four-marker combination, age/CA-125/hK6/hK10 also showed 100% specificity and 100% positive predictive value, although it demonstrated low sensitivity (11.9%) and negative predictive value (52.8%). Conclusion: The combination of human tissue kallikreins and CA-125 showed potential for improving prediction of epithelial ovarian cancer in patients presenting with ovarian tumors. Objective: To determine the predictive accuracy of the combined panels of serum human tissue kallikreins (hKs) and CA-125 for the detection of epithelial ovarian cancer. Methods: Serum specimens collected from 5 Indonesian centers and 1 Vietnamese center were analyzed for CA-125, hK6, and hK10 levels. A total of 375 specimens from patients presenting with ovarian tumors, which include 156 benign cysts, 172 epithelial ovarian cancers (stage I/II, n=72; stage III/IV, n=100), 36 germ cell tumors and 11 borderline tumors, were included in the study analysis. Receiver operating characteristic analysis were performed to determine the cutoffs for age, CA-125, hK6, and hK10. Sensitivity, specificity, negative, and positive predictive values were determined for various combinations of the biomarkers. Results: The levels of hK6 and hK10 were significantly elevated in ovarian cancer cases compared to benign cysts. Combination of 3 markers, age/CA-125/hk6 or CA-125/hk6/hk10, showed improved specificity (100%) and positive predictive value (100%) for prediction of ovarian cancer, when compared to the performance of single markers having 80-92% specificity and 74-87% positive predictive value. Four-marker combination, age/CA-125/hK6/hK10 also showed 100% specificity and 100% positive predictive value, although it demonstrated low sensitivity (11.9%) and negative predictive value (52.8%). Conclusion: The combination of human tissue kallikreins and CA-125 showed potential for improving prediction of epithelial ovarian cancer in patients presenting with ovarian tumors.

Decreased Expression of Inhibitor of DNA-binding (Id) Proteins and Vascular Endothelial Growth Factor and Increased Apoptosis in Ovarian Aging

Min Jung Park,Sea Hee Park,문성은,Ja Seong Koo,Hwa Sook Moon,주보선 한국발생생물학회 2013 발생과 생식 Vol.17 No.1

This study examined the expression of inhibitor of DNA-binding (Id) proteins and vascular endothelial growth factor (VEGF) in the ovary according to female age using a mice model as the first step in investigating the potential role of Ids and VEGF in ovarian aging. C57BL inbred female mice of three age groups (6-9, 14-16, and 23-26weeks) were injected with 5 IU pregnant mare’s serum gonadotropin (PMSG) in order to synchronize the estrus cycle. After 48 h, ovarian expression of Ids and VEGF was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR),western blot and immunohistochemistry. Ovarian apoptosis was examined by ovarian expression of Bcl-2 and Bcl-xL. Expression of Id-1 and VEGF was decreased with advancing female age, but not Id-2, Id-3, and Id-4. In particular, their expressions were significantly decreased in aged mice of 23-26 weeks compared with the young mice of 6-9 weeks (p < 0.05). In contrast, ovarian apoptosis was greatly increased in the aged mice compared to the young mice. This result suggests that Id-1 may have an implicated role in ovarian aging by associating with VEGF.

Decreased Expression of Inhibitor of DNA-binding (Id) Proteins and Vascular Endothelial Growth Factor and Increased Apoptosis in Ovarian Aging

Park, Min Jung,Park, Sea Hee,Moon, Sung Eun,Koo, Ja Seong,Moon, Hwa Sook,Joo, Bo Sun The Korean Society of Developmental Biology 2013 발생과 생식 Vol.17 No.1

This study examined the expression of inhibitor of DNA-binding (Id) proteins and vascular endothelial growth factor (VEGF) in the ovary according to female age using a mice model as the first step in investigating the potential role of Ids and VEGF in ovarian aging. C57BL inbred female mice of three age groups (6-9, 14-16, and 23-26 weeks) were injected with 5 IU pregnant mare's serum gonadotropin (PMSG) in order to synchronize the estrus cycle. After 48 h, ovarian expression of Ids and VEGF was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR), western blot and immunohistochemistry. Ovarian apoptosis was examined by ovarian expression of Bcl-2 and Bcl-xL. Expression of Id-1 and VEGF was decreased with advancing female age, but not Id-2, Id-3, and Id-4. In particular, their expressions were significantly decreased in aged mice of 23-26 weeks compared with the young mice of 6-9 weeks (p < 0.05). In contrast, ovarian apoptosis was greatly increased in the aged mice compared to the young mice. This result suggests that Id-1 may have an implicated role in ovarian aging by associating with VEGF.

Reproductive Aging in Female Rodents

이성호 한국발생생물학회 2007 발생과 생식 Vol.11 No.1

암컷 포유동물의 노화 과정에서 생식계는 체내 여러 시스템 가운데 가장 먼저 기능 저하가 나타난다. 생식 노화(reproductive aging) 과정 중인 암컷 포유동물은 비록 종 특이성이 있지만 주기성(cyclicity)의 소실과 같은 생식 능력의 감소와 함께 여러 기능들의 변화가 동반되면서 궁극적으로 인간의 폐경 현상과 같은 생식 능력 상실이 나타난다. 본 증설은 암컷 포유동물의 생식호르몬 축, 특히 신경내분비 회로의 노화에 대한 정보들과 노화 연구 In all female mammals, reproductive system is one of the first biological systems to show age-related decline. Female mammals in reproductive aging, though the phenomena is somewhat species-specific, start to show declining fertility and changes of numerous physiological functions. This review will present a current information on the aging of the female reproductive hormonal axis and introduce three useful rodent models for studying this field. Middle age( months old) in female rats and mice is comparable to the stage prior to the entry of menopause in human. In this period pulsatile and surge GnRH secretion from hypothalamus gradually attenuated, then reduced pulsatile and surge LH secretion is followed consequently. This age-related defects in GnRH-LH neuroendocrine axis seem to be highly correlated with the defects in brain signals which modulate the activities of GnRH neuron. Many researchers support the idea which the age-related hypothalamic defects are the main cause of reproductive aging, but some ovarian factors such as inhibin response also could contribute to the induction of reproductive senescence. Some rodent models are quite valuable in studying the reproductive aging. The follitropin receptor knockout(FORKO) mice, both of null and haploinsufficient state, could produce depletion of oocyte/follicle with age. Dioxin/aryl hydrocarbon receptor(AhR) knockout mice also show severe ovarian defects and poor reproductive success early in their life compared to the age-matched normal mice. Further studies on the reproductive aging will be a great help to evaluate the benefits and risks of hormone replacement therapy(HRT) and to improve the safety of HRT.

Gonadotropins Regulate the mRNA Expression of Gonadotropin-Releasing Hormone and Its Receptors in the Mouse Ovary and Uterus

문소은,석은지,하진아,양현원,윤보경,이민주 한국발생생물학회 2024 발생과 생식 Vol.28 No.1

Gonadotropin-releasing hormone (GnRH), a critical hormone produced in the hypothalamus, is essential for regulating reproductive processes. It has also been demonstrated the presence of GnRH and its receptors (GnRHR) in ovarian and uterine tissues, but little was known about the regulation mechanism of their expression in these organs and ovarian aging. Therefore, the aim of this study was to investigate the expression of GnRHR in the ovary and uterus of mice, particularly after high-dose gonadotropin treatments and in relation to aging. Quantitative realtime-PCR (qRT-PCR) revealed that pituitary gland had the highest GnRHR expression in both young and aged mice. In addition, liver expression was higher in young mice, whereas thymus expression was higher in aged mice. GnRHR mRNA was present in the ovaries of both young and aged mice but nearly undetectable in the uterus of aged mice. We next examined the expression of GnRHR in the ovary and uterus in response to high-dose administration of pregnant mare serum gonadotropin (PMSG). After PMSG administration, GnRH mRNA levels were significantly decreased in the ovary but increased in the uterus. The expression of GnRH mRNA in these organs showed opposite trends to that of GnRHR expression. These results suggest the involvement of GnRH in age-related reproductive decline and the potential effects of high-dose gonadotropin treatments on reproductive organ function.

Intravenous human endothelial progenitor cell administration into aged mice enhances embryo development and oocyte quality by reducing inflammation, endoplasmic reticulum stress and apoptosis

KIM, Geon A,LEE, Yeonjae,KIM, Hyun Jin,OH, Hyun Ju,KANG, Sung Keun,RA, Jeong Chan,LEE, Byeong Chun The Japanese Society of Veterinary Science 2018 The Journal of veterinary medical science Vol.80 No.12

<P>Stem cell therapy has been proposed to restore the function and structure of injured tissues. In the present study, we investigated the ability of human endothelial progenitor cells (hEPCs) to attenuate ovarian aging and dysfunction. Female ICR mice aged 4 and 6 months were injected with cultured hEPCs. Cultured hEPCs were injected intravenously twice with 5 × 10<SUP>4</SUP> cells with a 4 day interval. After pregnant mare serum gonadotropin and human chorionic gonadotropin stimulation, oocytes and ovaries of aged mice were collected, cumulus-free oocytes were activated by SrCl<SUB>2</SUB> and gene expression levels related to inflammation, apoptosis, follicle development and endoplasmic reticulum (ER) stress in ovaries were compared. Administration of hEPCs attenuated the level of inflammatory cytokines and adverse apoptotic factor, as well as reducing ER stress in the ovaries. Increased cleavage and blastocyst formation rates and cell numbers in blastocysts from hEPCs-treated aged mice vs. same aged control mice demonstrated a protective function of hEPCs against reproductive aging. Based on these data, we suggest that treatment with hEPCs attenuates reproductive aging and dysfunction potentially via regulation of inflammation, apoptosis and ER stress.</P>

Age at Menopause and Suicidal Ideation in Menopausal Women: A Study of Korea National Health and Nutrition Examination Survey Data

Ryu Ki-Jin,Park Hyuntae,Jeong Yujin,Nam Seunghyun,Jeong Hye Gyeong,Kim Tak 대한의학회 2022 Journal of Korean medical science Vol.37 No.45

Background: Although menopause is considered a risk factor for depression, no association has been established between the risk of suicidal ideation and age at menopause. This study aimed to evaluate the association between age at menopause and suicidal ideation in middleaged menopausal Korean women. Methods: This cross-sectional study used data from the Korea National Health and Nutrition Examination Survey (2013–2018). Women aged 40–65 years were divided into the following three categories: primary ovarian insufficiency (POI), early menopause, and menopause, according to age at natural menopause (< 40, 40–45, and > 45 years, respectively). Depressive symptoms were assessed using the Patient Health Questionnaire-9 (PHQ-9). Results: Among 2,232 menopausal women, 25 (1.1%) experienced POI and 114 (5.1%) experienced early menopause. The PHQ-9 items that pertained to low self-esteem and suicidal ideation scored higher in women with POI than in those who experienced menopause after 45 years of age. The prevalence of suicidal ideation differed significantly according to age at menopause (POI, 30.0%; early menopause, 12.7%; menopause, 8.0%; P = 0.016). Logistic regression analysis revealed that POI was significantly associated with suicidal ideation after the adjustment for age, body mass index, and education, household income, and walking levels (odds ratio, 4.2; 95% confidence interval, 1.0–17.7). Conclusion: Korean middle-aged women with POI were more likely to have suicidal ideation than those who experienced menopause at 45 years or above, despite not being diagnosed with major depressive disorder.

Time Trends of Ovarian Cancer Incidence in China

Wang, Bing,Liu, Shu-Zheng,Zheng, Rong-Shou,Zhang, Fang,Chen, Wan-Qing,Sun, Xi-Bin Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.1

The aim of this study was to examine the trend of ovary cancer incidence from 1999 to 2010 in China and predict the burden up to 2020. Crude incidence, age specific incidence and age-adjusted incidence rates were calculated. Joinpoint regression was performed to obtain estimated annual percentages and Bayesian age-period-cohort modeling was used to predict the incidence rate until the year 2020. In China, the crude rate of ovary cancer was 7.91/100,000 and the age-adjusted rate was 5.35/100,000 overall during period 1999-2010. The rates in urban regions were higher than in rural regions. A significant rising trend during 1999-2006 was followed by a drop during 2006-2010 in age-adjusted rates for urban females. In contrast, constant rise was observed in rural women. The decrease in ovary cancer of urban areas tended to be restricted to women aged 50 years and younger. In contrast, increases of ovary cancer in rural areas appeared in virtually all age groups. Although the age-adjusted incidence rate for ovary cancer was predicted to be reduced after year 2011, the crude rate was likely to be relative stable up to 2020. The burden of ovary cancer in China will continue to be relative stable due to the aging population.

Age-Adjusted Prevalence and Characteristics of Women with Polycystic Ovarian Syndrome in Korea: A Nationwide Population-Based Study (2010–2019)

김주희,정민형,홍세화,문나래,강대룡 연세대학교의과대학 2022 Yonsei medical journal Vol.63 No.8

Polycystic ovarian syndrome (PCOS) is a common endocrine disorder in women of reproductive age and is associated with anincreased risk of obesity, compensatory hyperinsulinemia, dyslipidemia, metabolic syndrome, and endometrial cancer. Thisstudy analyzed 544619 women using the Korean Informative Classification of Disease, version 10, codes E28.0–E28.9 in the popu lation-based National Health Information Databases from 2010 to 2019. The age-adjusted incidence and prevalence rates of PCOSover 10 years among Korean women were 2.8% and 4.3%, respectively; and they increased in the late teens, peaked in the 20s, andbegan to decrease at the age of 30. We also found that the body mass index, levels of fasting blood glucose, and high-density lipo protein values in the recent two years (2018–2019) were higher in women with PCOS compared to the general population. This is thefirst study to investigate the prevalence of PCOS in a nationwide population of reproductive-aged Korean women. Further researchis needed to examine the short- and long-term health risks and psychological problems associated with PCOS.