RESUMEN
Premature Ovarian Insufficiency (POI) is a highly heterogeneous condition characterized by ovarian dysfunction in women occurring before the age of 40, representing a significant cause of female infertility. It manifests through primary or secondary amenorrhea. While more than half of POI cases are idiopathic, genetic factors play a pivotal role in all instances with known causes, contributing to approximately 20-25% of cases. This article comprehensively reviews the genetic factors associated with POI, delineating the primary candidate genes. The discussion delves into the intricate relationship between these genes and ovarian development, elucidating the functional consequences of diverse mutations to underscore the fundamental impact of genetic effects on POI. The identified genetic factors, encompassing gene mutations and chromosomal abnormalities, are systematically classified based on whether the resulting POI is syndromic or non-syndromic. Furthermore, this paper explores the genetic interplay between mitochondrial genes, such as Required for Meiotic Nuclear Division 1 homolog Gene (RMND1), Mitochondrial Ribosomal Protein S22 Gene (MRPS22), Leucine-rich Pentapeptide Repeat Gene (LRPPRC), and non-coding RNAs, including both microRNAs and Long non-coding RNAs, with POI. The insights provided serve to consolidate and enhance our understanding of the etiology of POI, contributing to establishing a theoretical foundation for diagnosing and treating POI patients, as well as for exploring the mechanisms underlying the disease.
Asunto(s)
Insuficiencia Ovárica Primaria , Insuficiencia Ovárica Primaria/genética , Humanos , Femenino , Mutación , Predisposición Genética a la EnfermedadRESUMEN
PURPOSE: To evaluate the effects of atosiban on clinical outcomes in patients undergoing frozen-thawed embryo transfer. METHODS: The clinical data of 1093 infertile patients who underwent frozen-thawed embryo transfer in our center from January 2019 to December 2020 were retrospectively analyzed (control, 418; atosiban, 675). Propensity score matching (PSM) analysis identified 400 matched pairs of patients. The implantation rate, clinical pregnancy rate, live birth rate, biochemical pregnancy rate, abortion rate, multiple pregnancy rate, and ectopic pregnancy rate between the two groups were compared. RESULTS: Before PSM, patients differed by infertility factors, number of transferred embryos, and endometrial preparation protocol (P < 0.05). After PSM, characteristics were similar in corresponding patients of the atosiban and control groups. After propensity score matching, we found that there was no significant difference in the implantation rate, clinical pregnancy rate, live birth rate, biochemical pregnancy rate, abortion rate, multiple pregnancy rate, and ectopic pregnancy rate in atosiban and control group (P > 0.05). CONCLUSION: Atosiban did not improve the clinical outcomes of infertile patients with frozen-thawed embryo transfer.
Asunto(s)
Infertilidad , Embarazo Ectópico , Vasotocina/análogos & derivados , Embarazo , Femenino , Humanos , Estudios Retrospectivos , Puntaje de Propensión , Criopreservación , Transferencia de Embrión/métodos , Implantación del Embrión , Índice de EmbarazoRESUMEN
PURPOSE: Can small RNA derived from embryos in conditioned embryo culture medium (ECM) influence embryo implantation? METHODS: We employed small RNA sequencing to investigate the expression profiles of transfer RNA-derived small RNA (tsRNA) and microRNA (miRNA) in ECM from high-quality and low-quality embryos. Quantitative real-time PCR was employed to validate the findings of small RNA sequencing. Additionally, we conducted bioinformatics analysis to predict the potential functions of these small RNAs in embryo implantation. To establish the role of tiRNA-1:35-Leu-TAG-2 in embryonic trophoblast cell adhesion, we utilized co-culture systems involving JAR and Ishikawa cells. RESULTS: Our analysis revealed upregulation of nine tsRNAs and four miRNAs in ECM derived from high-quality embryos, whereas 37 tsRNAs and 12 miRNAs exhibited upregulation in ECM from low-quality embryos. The bioinformatics analysis of tsRNA, miRNA, and mRNA pathways indicated that their respective target genes may play pivotal roles in both embryo development and endometrial receptivity. Utilizing tiRNA mimics, we demonstrated that the prominently expressed tiRNA-1:35-Leu-TAG-2 in the low-quality ECM group can be internalized by Ishikawa cells. Notably, transfection of tiRNA-1:35-Leu-TAG-2 into Ishikawa cells reduced the attachment rate of JAR spheroids. CONCLUSION: Our investigation uncovers significant variation in the expression profiles of tsRNAs and miRNAs between ECM derived from high- and low-quality embryos. Intriguingly, the release of tiRNA-1:35-Leu-TAG-2 by low-quality embryos detrimentally affects embryo implantation and endometrial receptivity. These findings provide fresh insights into understanding the molecular foundations of embryo-endometrial communication.
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MicroARNs , Humanos , Femenino , MicroARNs/genética , MicroARNs/metabolismo , Implantación del Embrión/genética , Embrión de Mamíferos/metabolismo , Técnicas de Cocultivo , Desarrollo Embrionario/genética , Endometrio/metabolismoRESUMEN
Craniosynostosis is one of the most common congenital craniofacial birth defects. The genetic etiology is complex, involving syndromic developmental diseases, chromosomal abnormalities, and monogenic non-syndromic diseases. Herein, we presented a proband of craniosynostosis, who firstly displayed structural abnormalities. This research conducted dynamic ultrasound monitoring a fetus with gradually developing intrauterine growth retardation (IUGR). A novel de novo variant c.41G > A: p.W14* in SMAD6 was identified by pedigree analysis and genetic examination approaches. Recombinant plasmid carrying wild-type sequence and mutant that carries c.41G > A in SMAD6 were constructed and transfected into HEK293T cells. mRNA and protein expression of SMAD6 were reduced in SMAD6 mutants compared to the wild type. Cycloheximide (CHX) treatment and si-UPF1 transfection rescued the SMAD6 mRNA expression in the mutant construct, indicating that c.41G > A: p.W14* in SMAD6 triggered nonsense-mediated mRNA degradation (NMD) process and thus led to haploinsufficiency of the protein product. Our study demonstrated that whole-exome sequencing (WES) was a powerful tool for further diagnosis and etiological identification once fetal malformation was detected by ultrasound. Novel de novo c.41G > A: p.W14* in SMAD6 is pathogenic and potentially leads to craniosynostosis via NMD process.
Asunto(s)
Craneosinostosis , Embarazo , Femenino , Humanos , Células HEK293 , Craneosinostosis/diagnóstico por imagen , Craneosinostosis/genética , Feto , ARN Mensajero/genética , China , Proteína smad6/genética , Transactivadores , ARN HelicasasRESUMEN
Genomic abnormalities are strongly associated with cancer and infertility. In this study, we develop a simple and efficient method - multiple genetic abnormality sequencing (MGA-Seq) - to simultaneously detect structural variation, copy number variation, single-nucleotide polymorphism, homogeneously staining regions, and extrachromosomal DNA (ecDNA) from a single tube. MGA-Seq directly sequences proximity-ligated genomic fragments, yielding a dataset with concurrent genome three-dimensional and whole-genome sequencing information, enabling approximate localization of genomic structural variations and facilitating breakpoint identification. Additionally, by utilizing MGA-Seq, we map focal amplification and oncogene coamplification, thus facilitating the exploration of ecDNA's transcriptional regulatory function.
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Variaciones en el Número de Copia de ADN , Oncogenes , Genómica/métodos , Regulación de la Expresión Génica , ADNRESUMEN
BACKGROUND: Diminished ovarian reserve (DOR) is a common cause of female infertility, with genetic factors being a significant contributor. However, due to high genetic heterogeneity, the etiology of DOR in many cases remains unknown. In this study, we analyzed the phenotype of a young woman with primary infertility and performed molecular genetic analysis to identify the genetic cause of her condition, thus providing important insights for genetic counseling and reproductive guidance. METHODS: We collected the patient's basic information, clinical data, as well as diagnostic and therapeutic history and performed whole-exome sequencing on her peripheral blood. Candidate pathogenic variants were validated by Sanger sequencing in family members, and the pathogenicity of variants was analyzed using ACMG guidelines. We used bioinformatics tools to predict variant effects on splicing and protein function, and performed in vitro experiments including minigene assay and expression analysis to evaluate their functional effects on HEK293T. RESULTS: We identified biallelic MSH4 variants, c.2374 A > G (p.Thr792Ala) and c.2222_2225delAAGA (p.Lys741Argfs*2) in the DOR patient. According to ACMG guidelines, the former was classified as likely pathogenic, while the latter was classified as pathogenic. The patient presented with poor oocyte quantity and quality, resulting in unsuccessful in vitro fertilization cycles. Bioinformatics and in vitro functional analysis showed that the c.2374 A > G variant altered the local conformation of the MutS_V domain without decreasing MSH4 protein expression, while the c.2222_2225delAAGA variant led to a reduction in MSH4 protein expression without impacting splicing. CONCLUSIONS: In this study, we present evidence of biallelic variants in MSH4 as a potential cause of DOR. Our findings indicate a correlation between MSH4 variants and reduced oocyte quality, as well as abnormal morphology of the first polar body, thereby expanding the phenotypic spectrum associated with MSH4 variants. Furthermore, Our study emphasizes the importance of utilizing whole-exome sequencing and functional analysis in diagnosing genetic causes, as well as providing effective genetic counseling and reproductive guidance for DOR patients.
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Enfermedades del Ovario , Reserva Ovárica , Femenino , Humanos , Células HEK293 , Reserva Ovárica/genética , Bioensayo , Biología Computacional , Familia , Proteínas de Ciclo CelularRESUMEN
An association between endometriosis and luteinized unruptured follicle syndrome (LUFs) has long been identified. Although inactivating mutation of luteinizing hormone/choriogonadotropin receptor (LHGCR) results in LUFs, whether LHCGR contributes to promoting LUFs in endometriosis remains elusive. To investigate the effect of LHCGR signaling in the development of endometriosis-associated LUFs and dissect the underlying mechanism in vivo mouse endometriosis model was established to measure the effect on ovarian folliculogenesis. In vitro cultures of primary human GCs collected from patients undergoing in vitro fertilization were performed and treated with human chorionic gonadotropin (hCG), dibutyryl cyclic-AMP (db-cAMP), LHCGR or CCAAT/enhancer binding protein-α (C/EBPα) small interfering RNA to identify the potential mechanisms. KGN cell line was used to investigate the mechanistic features of transcriptional regulation. Results showed an increased incidence of LUFs was observed in mice with endometriosis. The expression of LHCGR was decreased in the GCs of endometriosis mice. In in vitro cell models, LHCGR signaling increased the expression of C/EBPα and cyclooxygenase-2(COX-2), while inhibiting C/EBPα mitigated the induced COX-2 expression. Mechanically, C/EBPα bounded to the promoter region of COX-2 and increased the transcriptional activity under the stimulation of hCG or db-cAMP. Taken together, this study demonstrated that the LHCGR signaling was reduced in GCs of endometriosis and resulted in a decrease in gonadotropin-induced COX-2 expression. Our study might provide new insights into the dysfunction of GCs in endometriosis.
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Ciclooxigenasa 2 , Endometriosis , Receptores de HL , Animales , Gonadotropina Coriónica/uso terapéutico , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Regulación hacia Abajo , Endometriosis/genética , Femenino , Regulación de la Expresión Génica , Humanos , Ratones , Receptores de HL/genéticaRESUMEN
Meckel-Gruber syndrome (MKS) is a rare lethal autosomal recessive inherited disorder. Missed diagnosis might happen in clinical works due to an unclear genotype-phenotype correlation. We analyzed two families visiting our center; the parents are normal; each of the family aborted a fetus at 12WG. Following ultrasonography and pathological examination, both were diagnosed as MKS. Whole exome sequencing identified a compound heterozygous of two novel variants of CEP290 and a heterozygous of a novel variant of CC2D2A. Frameshift mutations in ZNF77 were also detected. Western blot analyzing whole-brain tissue showed that the expression of ZNF77, CC2D2A, and CEP290 was enhanced. HEK293T transfected with over-expression wildtype/mutated ZNF77 plasmid showed that SHH was increased in wildtype ZNF77 cells, while SHH and CC2D2A were increased in mutated ZNF77 cells. Our research provided two novel pathogenic variants of CEP290 and CC2D2A and suggested that ZNF77 might promote the expression of CC2D2A and regulate the amount of SHH.
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Trastornos de la Motilidad Ciliar , Proteínas del Citoesqueleto , Encefalocele , Enfermedades Renales Poliquísticas , Retinitis Pigmentosa , Antígenos de Neoplasias/genética , Proteínas de Ciclo Celular/genética , Trastornos de la Motilidad Ciliar/genética , Proteínas del Citoesqueleto/genética , Encefalocele/genética , Feto , Células HEK293 , Proteínas Hedgehog/genética , Humanos , Mutación , Enfermedades Renales Poliquísticas/genética , Retinitis Pigmentosa/genética , Transducción de SeñalRESUMEN
Lead (Pb) is a common environmental pollutant. It has been demonstrated that long-term exposure to Pb at environmental levels may cause severe and irreversible damage to the male reproductive system. Of note, the impairments may originate from environmental Pb exposure at puberty. However, the underlying mechanisms remain unclear. In this study, we administrated male ICR mice with 200 mg/L Pb through the drinking water for 30-, 60-, 90-day from postnatal day 28. RNA sequencing was performed in the control group and the 90-day Pb exposure group. It was found that Pb exposure induced testicular damage, increased oxidative stress levels and poor sperm quality. Bioinformatic analysis displayed 199 genes up-regulated (such as GLUT1 and MCT4 genes) and 156 genes down-regulated (such as GLUT3, PFK1, LDH, CD147 and AMPK genes) in the Pb exposure group compared to the control group. Gene ontology (GO) terms enrichment analysis showed differentially expressed genes (DEGs) are involved in the protein catabolic, cellular catabolic and triglyceride catabolic processes. KEGG pathways enrichment analysis indicated glycerolipid metabolism and AMPK signaling were significantly enriched. Furthermore, experimental verification showed that Pb exposure induces energy dysmetabolism and decreases glycolysis products in mice testicular tissue. The AMPK signaling pathway was found to be deactivated after Pb exposure. The GLUT1, GLUT3, PFK1 and LDH proteins, which play a critical role in the cell glycolysis process, also were decreased. Besides, the expression of CD147 was decreased and the location of CD147 was altered upon Pb exposure. Together, these findings indicated the implication of the AMPK signaling pathway in Pb exposure induced pubertal testicular damage and poor sperm quality by inhibiting cell glycolysis and disordering lactate transportation in testicular cells.
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Proteínas Quinasas Activadas por AMP , Plomo , Proteínas Quinasas Activadas por AMP/genética , Animales , Glucólisis , Plomo/toxicidad , Masculino , Ratones , Ratones Endogámicos ICR , Maduración SexualRESUMEN
Lead (Pb) is a toxic metal that affects the male reproductive system. This study aimed to investigate the effects of zinc (Zn) intake between recommended dietary allowances (RDAs) and tolerable upper intake levels (ULs) in preventing male testis damage induced by low-dose Pb. Forty-five mice were randomly divided into control, Pb, and Pb + Zn groups. They were given distilled water ad libitum with 0, 200 mg/L Pb2+, or 15 mg/L Zn2+ mixed with 200 mg/L Pb2+ for 90 consecutive days. The Zn levels in the blood and testis of the Pb group were significantly lower than those of the control group. The Pb levels in the blood and testis of the Pb + Zn group were significantly lower than those of the Pb group. Additionally, a significant decrease in sperm density and viability, with a significant increase in sperm abnormality rate and DNA fragmentation index, was observed in the Pb group. Zn supplementation significantly improved the above sperm parameters. Moreover, Zn supplementation decreased low-dose Pb-induced lipid peroxidation and increased glutathione, total superoxide dismutase (SOD), and copper/Zn-SOD levels. Furthermore, Zn treatment improved glycolysis products and lactate transporters in Pb-treated mouse testes. Our findings suggest that Zn intake between RDAs and UL can act as a therapeutic agent in protecting against the reproductive impairments associated with Pb exposure.
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Glucólisis/efectos de los fármacos , Plomo/toxicidad , Testículo/efectos de los fármacos , Zinc/farmacología , Animales , Suplementos Dietéticos , Glutatión/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos ICR , Espermatozoides/efectos de los fármacos , Superóxido Dismutasa/metabolismo , Zinc/administración & dosificaciónRESUMEN
OBJECTIVE: To explore the genetic basis for a fetus with hydrocephalus. METHODS: The fetus was found to have hydrocephalus upon ultrasonography duringthe second trimester. Following induced abortion, fetal tissue was collected for the extraction of DNA and whole exome sequencing.Sanger sequencing was used to verify the suspected variants in the family. RESULTS: The fetus was found to harbor a hemizygous c.620A>G (p.Tyr207Cys) variant of the L1CAM gene (OMIM 308840),for which his mother and sister were heterozygous carriers. The same variant was not found in his father, uncle and grandparents.Based on the standards and guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be likely pathogenic (PM1+PM2+PP3+PP4). CONCLUSION: The hemizygous c.620A>G (p.Tyr207Cys) variant of the L1CAM gene probably underlay the hydrocephalus in this fetus.
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Hidrocefalia , Mutación , Molécula L1 de Adhesión de Célula Nerviosa , Adulto , Femenino , Heterocigoto , Humanos , Hidrocefalia/genética , Masculino , Molécula L1 de Adhesión de Célula Nerviosa/genética , Linaje , Embarazo , Secuenciación del ExomaRESUMEN
This study aimed to investigate the mechanism of the lead exposure-induced oxidative stress and apoptosis of renal tubular epithelial cells. We explored the effects of lead acetate (PbAc) on the oxidation and apoptosis of renal proximal tubular cells (NRK-52E) through in vitro experiments. Results showed that PbAc induced dose-dependent reactive oxygen species (ROS) accumulation in NRK-52E cells, and the activities of superoxide dismutase (SOD) and glutathione (GSH) decreased, whereas the malondialdehyde (MDA) content increased. Under the exposure of 40 and 80 µM PbAc, the mRNA level of B cell lymphoma-2 (Bcl-2) in the cells decreased, the mRNA levels of Bcl-2-associated X protein (Bax) and caspase-3 increased, and apoptosis was obvious. Furthermore, the nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4) activity was enhanced by PbAc in a dose-dependent manner. The mRNA levels of protein kinase A (PKA) were upregulated by PbAc. H-89, a PKA inhibitor, suppressed PKA activation, ROS accumulation, and Nox4 activity in NRK-52E cells. Our results indicated that PbAc potentially stimulated oxidative stress and apoptosis in NRK-52E cells by increasing Nox4-dependent ROS production via the PKA signaling pathway.