Document text
Principal Investigator: Ketan Shrestha
Organization: UNIVERSITY OF KENTUCKY
Fiscal Year: 2024
Award: $154,000
Funding agency: Eunice Kennedy Shriver National Institute of Child Health and Human Development
TITLE: Role of Long non-coding RNAs in the Ovulatory Process.
ABSTRACT
Women’s reproductive health plays a pivotal role in regulating overall well-being. Unfortunately, about 12%
of reproductive-age women experience infertility. One of the most common causes of infertility is anovulation or
oligo-ovulation, supporting the paramount role of the ovulatory process in female reproductive success. Although
a few key ovulatory mediators induced by LH have been established, new and innovative research techniques
continue to discover new players in the ovulatory process. Using RNA sequencing, we have identified several
long non-coding RNAs (lncRNAs, transcripts >200 nucleotides) that are regulated during the periovulatory period
in mouse granulosa cells. In past, these lncRNAs were considered as junk DNA and believed to be transcriptional
noise and functionless. But, in recent years, evidence has accumulated showing that lncRNAs are widely
expressed, are tissue-specific, and have key roles in gene regulation by interacting with DNA, RNA, and proteins.
However, the role of lncRNAs in the ovary and specifically the ovulatory process has not been well explored.
From our novel RNA sequencing data, we have found 10 lncRNAs transcripts that are increased in granulosa
cells collected at 4h and 12h after human chorionic gonadotropin (hCG: LH analog) administration. Amongst
these 10 lncRNAs, Gm12648, a 656-nucleotide lncRNA was the most highly induced by hCG (160-fold).
Although very little is known about Gm12648, it is located directly upstream of cyclin dependent kinase inhibitor
2A which encodes the cell-cycle inhibitor p16 in the mouse. Moreover, the RNA-RNA interactome shows that
Gm12648 can interact with transcription factors (TFs) that are essential during the ovulatory process. Thus, it is
possible that Gm12648 may exert some regulatory control of genes and TFs and thereby possibly contribute to
ovarian cell fate during the periovulatory period. Similarly, other hCG upregulated lncRNAs could also exert their
regulatory role on adjacent protein-coding genes. Thus, our goal is to identify the top 5 most highly LH/hCG-
regulated candidate lncRNAs and investigate the function of these lncRNAs in the ovary, specifically their role
in the ovulatory process. To achieve our goal, mouse ovaries, granulosa cells, and COCs collected across the
periovulatory period will be utilized to characterize the expression of lncRNAs that were discovered in our
preliminary study (Aim 1), determine the target genes of Gm12648 in the granulosa cells using a RNA-seq and
RNA-RNA interaction assay approach after knocking down the expression of Gm12648 (Aim 2), and begin to
elucidate the function of Gm12648 by characterizing the proteins coded by the target genes, and identify post-
translational modifications such as phosphorylation, acetylation, methylation, and ubiquitination (Aim 3) in the
ovulation process. We will use a high-impact, translational mouse model in this study. The proposed project will
lay a foundation for future studies examining lncRNA function in the human ovary. These discoveries will further
improve our understanding of the ovulatory process which provides a fundamental platform to improve infertility
treatments and/or to develop the contraceptive measures that specifically target ovulation inhibition.
Terms: <Acetylation><Affect><Age><Anovulation><Apoptosis><Apoptosis Pathway><Assay><Basal Transcription Factor><Basal transcription factor genes><Bioassay><Biological Assay><Body Tissues><CDK4 Inhibitor><CDKN2A Protein><Candidate Disease Gene><Candidate Gene><Cannot achieve a pregnancy><Cdk4-Associated Protein p16><Cell Body><Cell Cycle><Cell Cycle Negative Regulator Beta><Cell Division Cycle><Cell Function><Cell Physiology><Cell Process><Cells><Cellular Function><Cellular Physiology><Cellular Process><Code><Coding System><Collecting Cell><Collection><Complex><Contraception><Contraceptive Agents><Contraceptive methods><Contraceptives><Cyclin-Dependent Kinase Inhibitor 2A><Cyclin-Dependent Kinase Inhibitor p12><Cyclin-Dependent Kinase Inhibitor p16><DNA><Data><Defect><Deoxyribonucleic Acid><Difficulty conceiving><Fecundability><Fecundity><Female><Female infertility><Fertility><Fertility Control><Foundations><Functional RNA><Future><Gene Action Regulation><Gene Expression><Gene Expression Regulation><Gene Regulation><Gene Regulation Process><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genes><Genetic Transcription><Goals><Human><Human Chorionic Gonadotropin><INK4A Gene Product><INK4A Protein><Infertility><Inhibition of Fertilization><Interstitial Cell Stimulating Hormone><Interstitial Cell-Stimulating Hormone><Junk DNA><Knock-out><Knockout><Leuteinizing Hormone><Liver><Luteinizing Hormone><Lutropin><Measures><Mediator><Methylation><Mice><Mice Mammals><Modern Man><Multiple Tumor Suppressor-1><Murine><Mus><Noise><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Nucleotides><Oocytes><Ovarian><Ovary><Ovocytes><Ovulation><Ovulation Inhibition><Ovulation Suppression><Pathway interactions><Personal Satisfaction><Phosphorylation><Pituitary Lutenizing Hormone><Play><Polycystic Ovarian Disease><Polycystic Ovarian Syndrome><Polycystic Ovary Syndrome><Post-Translational Modification Protein/Amino Acid Biochemistry><Post-Translational Modifications><Post-Translational Protein Modification><Post-Translational Protein Processing><Posttranslational Modifications><Posttranslational Protein Processing><Premature Ovarian Failure><Process><Programmed Cell Death><Protein CDKN2><Protein MTS1><Protein Modification><Protein Phosphorylation><Protein p16><Proteins><RNA><RNA Expression><RNA Gene Products><RNA Seq><RNA Sequences><RNA sequencing><RNAseq><Recombinant Luteinizing Hormone><Regulation><Reporting><Research Technics><Research Techniques><Residual><Residual state><Ribonucleic Acid><Role><Rupture><Sclerocystic Ovarian Degeneration><Sclerocystic Ovary Syndrome><Short interfering RNA><Small Interfering RNA><Sterility><Structure><Subcellular Process><Techniques><Testing><Therapeutic LH><Time><Tissues><Transcript><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Ubiquitilation><Ubiquitination><Ubiquitinoylation><Untranslated RNA><Woman><ages><diagnosis among females><diagnosis among women><diagnosis in females><diagnosis in women><diagnosis within females><diagnosis within women><differential expression><differentially expressed><experience><female diagnosis><female fertility><female reproductive health><fertility assistance><fertility cessation><fertility interventions><fertility loss><fertility treatment><gene desert><granulosa cell><hCG><hepatic body system><hepatic organ system><hormone analog><improved><improved outcome><infertile><infertility in women><infertility treatment><inhibitor><innovate><innovation><innovative><knock-down><knockdown><mouse model><murine model><noncoding><novel><oligo ovulation><oligoovulation><p16(INK4A)><p16-INK4><p16INK4 Protein><p16INK4A Protein><pathway><polycystic ovary><polycystic ovary disease><polycystic ovary disorder><premature ovarian aging><reproductive><reproductive health among females><reproductive health among women><reproductive health in females><reproductive health in women><reproductive success><siRNA><social role><sterile><transcription factor><transcriptional differences><transcriptome sequencing><transcriptomic sequencing><ubiquination><ubiquitin conjugation><unable to bear children><well-being><wellbeing><women's diagnosis><women's reproductive health>