Novel phenotype of syndromic premature ovarian insufficiency associated with <scp><i>TP63</i></scp> molecular defect

Mathorne, Stine Westergaard; Ravn, Pernille; Hansen, Dorte Gilså; Beck-Nielsen, Signe Sparre; Gjørup, Hans; Sørensen, Kristina Pilekær; Fagerberg, Christina

doi:10.1111/cge.13725

Cited by 8 publications

(7 citation statements)

References 25 publications

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“…Both the frameshift mutation p.Q568fs*3 and the nonsense mutation p.R594* fell within the SAM domain and truncated the TAp63α protein prior to the TID. Interestingly, these 2 mutations and 5 previously reported POI-related mutations were all heterozygous frameshift or nonsense mutations leading to the loss of all or part of the TID ( Figure 1A ) ( 26 – 29 ). In addition, the other 4 point mutations we identified (p.R643Q, p.L646P, p.R647C, and p.R655Q) were located in the conserved core sequence of the TID from R643 to R655 (RFTLRQTISFPPR), which is responsible for binding with the TAD ( Supplemental Figure 2 ) ( 16 ).…”

Section: Resultsmentioning

confidence: 63%

“…In previous reports, heterozygous variants in the human TP63 gene were mostly shown to impair epidermal development and to cause multiple organ malformations, including 5 syndromic and 2 nonsyndromic disorders ( 25 ). Interestingly, a relationship has been identified between C-terminal variations of the TP63 gene and POI ( 26 – 29 ). The patients in these studies with TP63 mutations presented with syndromic or isolated POI, and these variants were frameshift or nonsense mutations leading to truncated C-termini, which might disrupt the autoinhibitory state of TAp63α and induce uncontrolled oocyte death ( 30 ).…”

Section: Introductionmentioning

confidence: 99%

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TP63 gain-of-function mutations cause premature ovarian insufficiency by inducing oocyte apoptosis

Huang¹,

Zhao²,

Yang³

et al. 2023

Journal of Clinical Investigation

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The transcription factor p63 guards genome integrity in the female germline, and its mutations have been reported in patients with premature ovarian insufficiency (POI). However, the precise contribution of the TP63 gene to the pathogenesis of POI needs to be further determined. Here, in 1,030 Chinese patients with POI, we identified 6 heterozygous mutations of the TP63 gene that impaired the C-terminal transactivation-inhibitory domain (TID) of the TAp63α protein and resulted in tetramer formation and constitutive activation of the mutant proteins. The mutant proteins induced cell apoptosis by increasing the expression of apoptosis-inducing factors in vitro. We next introduced a premature stop codon and selectively deleted the TID of TAp63α in mice and observed rapid depletion of the p63 +/ Δ TID mouse oocytes through apoptosis after birth. Finally, to further verify the pathogenicity of the mutation p.R647C in the TID that was present in 3 patients, we generated p63 +/R647C mice and also found accelerated oocyte loss, but to a lesser degree than in the p63 +/ Δ TID mice. Together, these findings show that TID-related variants causing constitutive activation of TAp63α lead to POI by inducing oocyte apoptosis, which will facilitate the genetic diagnosis of POI in patients and provide a potential therapeutic target for extending female fertility.

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Section: Resultsmentioning

confidence: 63%

Section: Introductionmentioning

confidence: 99%

TP63 gain-of-function mutations cause premature ovarian insufficiency by inducing oocyte apoptosis

Huang¹,

Zhao²,

Yang³

et al. 2023

Journal of Clinical Investigation

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“…The sequencing was focused on 295 candidate genes selected from literature and our previous data. Through this approach, we could identify 9 already known variants ( 6 , 31 , 34 , 36 – 38 , 42 , 43 ) and 32 novel rare variants in genes already associated to POI etiology. Moreover, 34 novel rare variants have been found in additional genes participating in pathways important for ovarian physiology.…”

Section: Discussionmentioning

confidence: 99%

Targeted Next-Generation Sequencing Indicates a Frequent Oligogenic Involvement in Primary Ovarian Insufficiency Onset

et al. 2021

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Primary ovarian insufficiency (POI) is one of the major causes of female infertility associated with the premature loss of ovarian function in about 3.7% of women before the age of 40. This disorder is highly heterogeneous and can manifest with a wide range of clinical phenotypes, ranging from ovarian dysgenesis and primary amenorrhea to post-pubertal secondary amenorrhea, with elevated serum gonadotropins and hypoestrogenism. The ovarian defect still remains idiopathic in some cases; however, a strong genetic component has been demonstrated by the next-generation sequencing (NGS) approach of familiar and sporadic POI cases. As recent evidence suggested an oligogenic architecture for POI, we developed a target NGS panel with 295 genes including known candidates and novel genetic determinants potentially involved in POI pathogenesis. Sixty-four patients with early onset POI (range: 10–25 years) of our cohort have been screened with 90% of target coverage at 50×. Here, we report 48 analyzed patients with at least one genetic variant (75%) in the selected candidate genes. In particular, we found the following: 11/64 patients (17%) with two variants, 9/64 (14%) with three variants, 9/64 (14%) with four variants, 3/64 (5%) with five variants, and 2/64 (3%) with six variants. The most severe phenotypes were associated with either the major number of variations or a worse prediction in pathogenicity of variants. Bioinformatic gene ontology analysis identified the following major pathways likely affected by gene variants: 1) cell cycle, meiosis, and DNA repair; 2) extracellular matrix remodeling; 3) reproduction; 4) cell metabolism; 5) cell proliferation; 6) calcium homeostasis; 7) NOTCH signaling; 8) signal transduction; 9) WNT signaling; 10) cell death; and 11) ubiquitin modifications. Consistently, the identified pathways have been described in other studies dissecting the mechanisms of folliculogenesis in animal models of altered fertility. In conclusion, our results contribute to define POI as an oligogenic disease and suggest novel candidates to be investigated in patients with POI.

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“…Interestingly, one of the genetic interactors of armitage in Drosophila is the loki gene ( lok ) coding for the orthologue of the human CHEK2 , known to be an interactor of the previously identified TP63 array gene ( Bestetti et al , 2019 ) and found mutated in a new familial case of POI ( Mathorne et al , 2020 ). CHEK2 , defined as a guardian of female germline integrity ( Amelio et al , 2012 ; Xian and McKeon, 2018 ), is a multifunctional kinase involved in cell cycle checkpoint regulation, DNA repair and apoptosis ( Gebel et al , 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Targeted whole exome sequencing and Drosophila modelling to unveil the molecular basis of primary ovarian insufficiency

et al. 2021

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STUDY QUESTION Can a targeted whole exome sequencing (WES) on a cohort of women showing a primary ovarian insufficiency (POI) phenotype at a young age, combined with a study of copy number variations, identify variants in candidate genes confirming their deleterious effect on ovarian function? SUMMARY ANSWER This integrated approach has proved effective in identifying novel candidate genes unveiling mechanisms involved in POI pathogenesis. WHAT IS KNOWN ALREADY POI, a condition occurring in 1% of women under 40 years of age, affects women’s fertility leading to a premature loss of ovarian reserve. The genetic causes of POI are highly heterogeneous and several determinants contributing to its prominent oligogenic inheritance pattern still need to be elucidated. STUDY DESIGN, SIZE, DURATION WES screening for pathogenic variants of 41 Italian women with non-syndromic primary and early secondary amenorrhoea occurring before age 25 was replicated on another 60 POI patients, including 35 French and 25 American women, to reveal statistically significant shared variants. PARTICIPANTS/MATERIALS, SETTING, METHODS The Italian POI patients’ DNA were processed by targeted WES including 542 RefSeq genes expressed or functioning during distinct reproductive or ovarian processes (e.g. DNA repair, meiosis, oocyte maturation, folliculogenesis and menopause). Extremely rare variants were filtered and selected by means of a Fisher Exact test using several publicly available datasets. A case-control Burden test was applied to highlight the most significant genes using two ad-hoc control female cohorts. To support the obtained data, the identified genes were screened on a novel cohort of 60 Caucasian POI patients and the same case-control analysis was carried out. Comparative analysis of the human identified genes was performed on mouse and Drosophila melanogaster by analysing the orthologous genes in their ovarian phenotype, and two of the selected genes were fruit fly modelled to explore their role in fertility. MAIN RESULTS AND THE ROLE OF CHANCE The filtering steps applied to search for extremely rare pathogenic variants in the Italian cohort revealed 64 validated single-nucleotide variants/Indels in 59 genes in 30 out of 41 screened women. Burden test analysis highlighted 13 ovarian genes as being the most enriched and significant. To validate these findings, filtering steps and Burden analysis on the second cohort of Caucasian patients yielded 11 significantly enriched genes. Among them, AFP, DMRT3, MOV10, FYN and MYC were significant in both patient cohorts and hence were considered strong candidates for POI. Mouse and Drosophila comparative analysis evaluated a conserved role through the evolution of several candidates, and functional studies using a Drosophila model, when applicable, supported the conserved role of the MOV10 armitage and DMRT3 dmrt93B orthologues in female fertility. LARGE SCALE DATA The datasets for the Italian cohort generated during the current study are publicly available at ClinVar database (http://www.ncbi.nlm.nih.gov/clinvar/): accession numbers SCV001364312 to SCV001364375. LIMITATIONS, REASONS FOR CAUTION This is a targeted WES analysis hunting variants in candidate genes previously identified by different genomic approaches. For most of the investigated sporadic cases, we could not track the parental inheritance, due to unavailability of the parents’ DNA samples; in addition, we might have overlooked additional rare variants in novel candidate POI genes extracted from the exome data. On the contrary, we might have considered some inherited variants whose clinical significance is uncertain and might not be causative for the patients’ phenotype. Additionally, as regards the Drosophila model, it will be extremely important in the future to have more mutants or RNAi strains available for each candidate gene in order to validate their role in POI pathogenesis. WIDER IMPLICATIONS OF THE FINDINGS The genomic, statistical, comparative and functional approaches integrated in our study convincingly support the extremely heterogeneous oligogenic nature of POI, and confirm the maintenance across the evolution of some key genes safeguarding fertility and successful reproduction. Two principal classes of genes were identified: (i) genes primarily involved in meiosis, namely in synaptonemal complex formation, asymmetric division and oocyte maturation and (ii) genes safeguarding cell maintenance (piRNA and DNA repair pathways). STUDY FUNDING/COMPETING INTEREST(S) This work was supported by Italian Ministry of Health grants ‘Ricerca Corrente’ (08C621_2016 and 08C924_2019) provided to IRCCS Istituto Auxologico Italiano, and by ‘Piano Sostegno alla Ricerca’ (PSR2020_FINELLI_LINEA_B) provided by the University of Milan; M.P.B. was supported by Telethon-Italy (grant number GG14181). There are no conflicts of interest.

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Novel phenotype of syndromic premature ovarian insufficiency associated with TP63 molecular defect

Cited by 8 publications

References 25 publications

TP63 gain-of-function mutations cause premature ovarian insufficiency by inducing oocyte apoptosis

TP63 gain-of-function mutations cause premature ovarian insufficiency by inducing oocyte apoptosis

Targeted Next-Generation Sequencing Indicates a Frequent Oligogenic Involvement in Primary Ovarian Insufficiency Onset

Targeted whole exome sequencing and Drosophila modelling to unveil the molecular basis of primary ovarian insufficiency

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