Primary Ovarian Insufficiency Induced by Fanconi Anemia E Mutation in a Mouse Model

Abstract More than 50% of cases of primary ovarian insufficiency (POI) and nonobstructive azoospermia in humans are classified as idiopathic infertility. Meiotic defects may relate to at least some of these cases. Mutations in genes coding for synaptonemal complex (SC) components have been identified in humans, and hypothesized to be causative for the observed infertile phenotype. Mutation SYCE1 c.721C>T (former c.613C>T)—a familial mutation reported in two sisters with primary amenorrhea—was the first such mutation found in an SC central element component-coding gene. Most fundamental mammalian oogenesis events occur during the embryonic phase, and eventual defects are identified many years later, thus leaving few possibilities to study the condition’s etiology and pathogenesis. Aiming to validate an approach to circumvent this difficulty, we have used the CRISPR/Cas9 technology to generate a mouse model with an SYCE1 c.721C>T equivalent genome alteration. We hereby present the characterization of the homozygous mutant mice phenotype, compared to their wild type and heterozygous littermates. Our results strongly support a causative role of this mutation for the POI phenotype in human patients, and the mechanisms involved would relate to defects in homologous chromosome synapsis. No SYCE1 protein was detected in homozygous mutants and Syce1 transcript level was highly diminished, suggesting transcript degradation as the basis of the infertility mechanism. This is the first report on the generation of a humanized mouse model line for the study of an infertility-related human mutation in an SC component-coding gene, thus representing a proof of principle.

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Heterozygous Eif4nif1 Stop Gain Mice Replicate the Primary Ovarian Insufficiency Phenotype

Journal of the Endocrine Society ◽

10.1210/jendso/bvab048.1576 ◽

2021 ◽

Vol 5 (Supplement_1) ◽

pp. A774-A775

Author(s):

Mika Moriwaki ◽

Corrine Kolka Welt

Keyword(s):

Mouse Model ◽

Litter Size ◽

Average Length ◽

Reproductive Function ◽

Protein Translation ◽

Antral Follicle ◽

Primary Ovarian Insufficiency ◽

Primary Follicle ◽

Ovarian Insufficiency ◽

Exon 10

Abstract We identified a stop-gain mutation in eIF4ENIF1 in a family in which multiple women developed primary ovarian insufficiency (POI) at approximately age 30 years. We hypothesized that the same mutation in a mouse model would replicate POI. Methods: The Eif4enif1 C57/Bl6 transgenic mouse model contains a floxed exon 10-19 cassette and a conditional knock-in cassette containing exon 10 with the c.1286C>G stop-gain mutation causing familial POI and WT exons 11-19 (Eif4enif1WT/flx). The hybrid offspring of CMV-Cre mice with Eif4enif1WT/flx mice were designated Eif4enif1WT/Δ for simplicity. Follicles were counted in fixed H&E stained ovaries from mice age days 1-5 (primordial and primary follicles), day 10, day 22 (first wave of growing follicles from small preantral to small antral follicles), week 20 (peak fertility), then every 2 months from 10 months to 26 months (follicle exhaustion). Litter frequency, pup number and genotype were recorded. Serum FSH levels were measured by the University of Virginia Ligand Assay and Analysis Core. Results: The heterozygotes have no outward or internal phenotypic differences compared to WT (Eif4enif1WT/flx), with the exception of reproductive organs in females and males. A subset of female heterozygotes (Eif4enif1WT/Δ) had no litters for 20 weeks (2 of 18; 11%). In those with litters, the average length of time between litters was not different but the final litter was earlier (5.6±2.7 vs. 10.5±0.7 months; p=0.02). Heterozygous breeding pair (Eif4enif1WT/Δx Eif4enif1WT/Δ) litter size was 60% of WT litter size (3.9±2.3 vs. 7.2±2.1 pups/litter; 0<0.001). The genotypes were 35% Eif4enif1WT/flx and 65% Eif4enif1WT/Δ, with no homozygotes. The number of follicles in ovaries from Eif4enif1WT/Δ mice was lower starting at the primordial (499±290 vs. 1445±381) and primary follicle stage (1069±346 vs. 1450±193) on day 10 (p<0.05). The preantral follicle number was lower starting on day 21 (213±86 vs. 522±227; p<0.01) and the antral follicle count was lower starting on week 20 (78±38 vs. 119±18; p<0.01). The FSH level in 12-month old mice during estrus was higher in a heterozygote compared to WT (25.0 vs. 12.1 ng/mL). Conclusions: Heterozygous Eif4enif1 stop-gain mutants have follicle loss documented by day 10, decreased pup number with no homozygotes, earlier end of reproductive function and elevated FSH levels. These mice replicate the POI phenotype in women. eIF4ENIF1 regulates protein translation by binding and storing eIF4E bound mRNA. Therefore, the unique mouse model provides a platform to study temporal and spatial regulation of protein translation across oocyte and embryo development in mammals. Further studies will determine whether follicle loss results from premature protein translation in oocytes.

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A homozygous hypomorphic BRCA2 variant causes primary ovarian insufficiency without cancer or Fanconi anemia traits

10.1101/751644 ◽

2019 ◽

Cited By ~ 2

Author(s):

Sandrine Caburet ◽

Abdelkader Heddar ◽

Elodie Dardillac ◽

Helene Creux ◽

Marie Lambert ◽

...

Keyword(s):

Fanconi Anemia ◽

Missense Variant ◽

Strand Break ◽

Primary Ovarian Insufficiency ◽

Ovarian Insufficiency ◽

Her Family ◽

Major Player ◽

Chromosomal Breaks ◽

Turkish Family ◽

Radiation Induced

ABSTRACTPrimary Ovarian insufficiency (POI) affects 1% of women under forty. We studied a patient with a non-syndromic POI, from a consanguineous Turkish family. Exome sequencing identified a homozygous missense variant c.8524C>T/p.R2842C in BRCA2. BRCA2 is a major player in homologous recombination (HR). BRCA2 deficiency induces cancer predisposition and Fanconi Anemia (FA). Remarkably, neither the patient nor her family exhibit somatic pathologies. The patient’s somatic cells presented intermediate levels of chromosomal breaks, cell proliferation and radiation-induced RAD51 foci formation when compared to controls, the heterozygous mother’s and FA cells. R2842C-BRCA2 partially complemented BRCA2 depletion for double-strand break-induced HR. The residual HR function in patient’s cells could explain the absence of somatic pathology. BRCA2 is expressed in human fetal ovaries in pachytene stage oocytes, when meiotic HR occurs. This study has a major impact on the understanding of genome maintenance in somatic and meiotic cells and on the management of POI patients.

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EFFECTS OF INTRAOVARIAN INJECTION OF PLATELET RICH PLASMA (PRP) ON A PRIMARY OVARIAN INSUFFICIENCY (POI) MOUSE MODEL

Fertility and Sterility ◽

10.1016/j.fertnstert.2021.07.124 ◽

2021 ◽

Vol 116 (3) ◽

pp. e43

Author(s):

Mauro Cozzolino ◽

Sonia Herraiz ◽

Emre Seli

Keyword(s):

Mouse Model ◽

Platelet Rich Plasma ◽

Primary Ovarian Insufficiency ◽

Ovarian Insufficiency

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Allelic reduction of Dlx5 and Dlx6 results in early follicular depletion: a new mouse model of primary ovarian insufficiency

Human Molecular Genetics ◽

10.1093/hmg/ddr166 ◽

2011 ◽

Vol 20 (13) ◽

pp. 2642-2650 ◽

Cited By ~ 12

Author(s):

K. Bouhali ◽

A. Dipietromaria ◽

A. Fontaine ◽

S. Caburet ◽

O. Barbieri ◽

...

Keyword(s):

Mouse Model ◽

Primary Ovarian Insufficiency ◽

Ovarian Insufficiency

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Granulosa cell and oocyte mitochondrial abnormalities in a mouse model of fragile X primary ovarian insufficiency

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gaw023 ◽

2016 ◽

Vol 22 (6) ◽

pp. 384-396 ◽

Cited By ~ 29

Author(s):

Carola Conca Dioguardi ◽

Bahar Uslu ◽

Monique Haynes ◽

Meltem Kurus ◽

Mehmet Gul ◽

...

Keyword(s):

Mouse Model ◽

Granulosa Cell ◽

Fragile X ◽

Primary Ovarian Insufficiency ◽

Ovarian Insufficiency ◽

Mitochondrial Abnormalities

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Human BM-MSC secretome enhances human granulosa cell proliferation and steroidogenesis and restores ovarian function in primary ovarian insufficiency mouse model

Scientific Reports ◽

10.1038/s41598-021-84216-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Hang-soo Park ◽

Rishi Man Chugh ◽

Abdeljabar El Andaloussi ◽

Elie Hobeika ◽

Sahar Esfandyari ◽

...

Keyword(s):

Cell Proliferation ◽

Mouse Model ◽

Granulosa Cell ◽

Granulosa Cells ◽

Cellular Proliferation ◽

Ovarian Function ◽

Human Mesenchymal Stem Cells ◽

Primary Ovarian Insufficiency ◽

Ovarian Insufficiency ◽

And Function

AbstractPrimary ovarian insufficiency (POI) is defined as the loss of ovarian function before 40 years of age. It clinically manifests as amenorrhea, infertility, and signs of estrogen insufficiency. POI is frequently induced by chemotherapy. Gonadotoxic chemotherapy reagents damage granulosa cells, which are essential for follicular function and development. Our recently published studies demonstrated that intraovarian transplantation of human mesenchymal stem cells (hMSCs) can restore fertility in a chemotherapy-induced POI mouse model. However, the regenerative mechanism underlying the hMSC effect in POI mice is not fully understood. Here, we report that the hMSC secretome increased the proliferation of human granulosa cells (HGrC1). We showed by FACS analysis that treatment of HGrC1 cells with hMSC-conditioned media (hMSC CM) stimulates cellular proliferation. We also demonstrated that the expression of steroidogenic enzymes involved in the production of estrogen, CYP19A1 and StAR, are significantly elevated in hMSC CM-treated HGrC1 cells. Our data suggest that hMSC CM stimulates granulosa cell proliferation and function, which may explain the therapeutic effect of hMSCs in our chemotherapy-induced POI animal model. Our findings indicate that the hMSC secretome may be a novel treatment approach for restoring granulosa cell and ovarian function in patients with POI.

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Establishment and Mechanism Study of a Primary Ovarian Insufficiency Mouse Model using Lipopolysaccharide

10.21203/rs.3.rs-545549/v1 ◽

2021 ◽

Author(s):

Si-Ji Lv ◽

Shu-Hui Hou ◽

Lei Gan ◽

Jing Sun

Keyword(s):

Mouse Model ◽

Granulosa Cell ◽

Cell Apoptosis ◽

Low Dose ◽

Ovarian Function ◽

Histopathological Examination ◽

Primary Ovarian Insufficiency ◽

High Dose ◽

Ovarian Insufficiency ◽

Primordial Follicles

Abstract Background: This study aimed to establish a lipopolysaccharide (LPS)-induced primary ovarian insufficiency (POI) mouse model and to investigate the underlying mechanism.Methods: C57BL/6N female mice were intraperitoneally injected with low-dose LPS (0.5 mg/kg) once daily for 14 days, high-dose LPS (2.5 mg/kg) twice weekly for 2 weeks, and cyclophosphamide (CTX; 150 mg/kg) once weekly for 2 weeks. Ovarian function was assessed by measuring the length of the estrous cycle, the number of primordial follicles, and the levels of serum pituitary/ovarian hormones. Expression and production of interleukin 1β (IL-1β) were determined to evaluate ovarian inflammation. Histopathological examination was performed to examine ovarian fibrosis. TUNEL assay was carried out to evaluate granulosa cell apoptosis. Western blotting was performed to measure the levels of inflammation-, fibrosis-, and apoptosis-related proteins in mouse ovaries.Results: Like CTX, both low- and high-dose LPS administration significantly impaired ovarian functions in mice, as evidenced by extended lengths of estrous cycles, reduced counts of primordial follicles, and alterations in the levels of serum hormones. Also, LPS administration promoted granulosa cell apoptosis and ovarian fibrosis in mice. However, LPS but not CTX significantly promoted IL-1β expression and production in mice. Moreover, LPS treatment but not CTX significantly enhanced TLR, p-p65, p65, and MyD88 protein expression in mouse ovaries, suggesting that LPS differs from CTX in triggering ovarian inflammation. In general, continuous low-dose LPS stimulation was less potent than high-dose LPS stimulation in the above-mentioned effects.Conclusions: LPS induces ovarian inflammation, fibrosis, and granulosa cell apoptosis and can be used to establish a POI model in mice.

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