Novel and recurrent genetic variants associated with male and female infertility

doi:10.21203/rs.3.rs-4711926/v1

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Novel and recurrent genetic variants associated with male and female infertility

https://doi.org/10.21203/rs.3.rs-4711926/v1

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Recently, the knowledge of the genetic basis of fertility disorders has expanded enormously, mainly thanks to the use of next generation sequencing (NGS). However, the genetic cause of the infertility in majority patients is still undefined.

Aim: The aim was to identify genetic variants associated with infertility disorders using targeting NGS technique and to describe phenotype - genotype correlation.

Methods: We have enrolled 41 patients (36 males and 5 females) with infertility problems or delayed puberty including the patients with hypogonadism hypogonadotropic (HH, n = 12), primary hypogonadism (n = 15), abnormal semen parameters or gonadal dysgenesis (n = 11), androgen insensitivity (n = 3). Genetic tests were performed using NGS panel of 35 genes.

Results: Overall, 14 pathogenic (P) or likely pathogenic (LP) variants including 3 novel and 11 recurrent variants were identified. Novel variants were found in genes associated with HH (FGF8, FGFR1, SEMA3). The genetic cause of the HH was determined in 58% (7/12) of the cases. Overall, The genetic testing enabled identification of the cause of the clinical phenotype in 26% (11/41) of the patients.

Conclusions: Our study expands the knowledge of the genetic basis of the infertility disorders and highlights the importance of genetic testing for proper diagnosis making and genetic counselling.

human infertility

genetic variants

genetic diagnostics

Infertility, defined (according to WHO) as the failure of reproduction after one year of regular intercourse without the use of contraception, is one of the major global health and social problems that affects 10–15% of couples of reproductive age (in Poland: 20% of couples, 1.5 million) [1, 2].

It is estimated that in 30–50% of cases, the causes of infertility are identified in women, 20–30% in men, while the problem affect both partners in approximately 20–30% of the couples [3]. Infertility is a multifactorial disease that can be caused by a number of reasons, including: abnormal gametogenesis, disorders of the structure and function of the reproductive system systemic diseases, including immunological disorders, infections, environmental factors, mental and genetic factors. Idiopathic infertility affects approximately 20–40% of couples. It is estimated that genetic disorders are the cause of approximately 50% of them, although the exact role of genetic factors in infertility is still unknown [3].

In both men and women, standard genetic tests towards infertility include karyotype assessment and, in the case of men with azoospermia/oligozoospermia, also analysis of the deletion of the AZF region of the Y chromosome and examination of pathogenic variants in CFTR gene [3]. Deletions of the AZF region of the Y chromosome are identified in 5–10% of men with azoospermia/oligozoospermia, while pathogenic variants of the CFTR gene in 6% of all infertile men and 60–70% of men with CBAVD [4]. However, according to literature data, about 30 genes are known to be associated with abnormal semen parameters in men (i.e. azoospermia, oligozoospermia, asthenozoospermia, or teratozoospermia) [5]. This number will certainly increase in the future, taking into account that approximately 2,000 genes are involved in the process of spermatogenesis, 600–900 of which are expressed in male sex cells exclusively [6].

Abnormal human spermatogenesis can be also caused by defects in androgen action and androgen insensitivity. Androgen Insensitivity Syndrome (AIS) is a disorder of male sexual development resulting in a wide range of clinical phenotypes and typically classified into three main types based on the degree of androgen receptor dysfunction and the resulting phenotypic characteristics. These types include complete androgen insensitivity syndrome (CAIS), partial androgen insensitivity syndrome (PAIS), and mild or minimal androgen insensitivity syndrome (MAIS). A variety of pathogenic variants have been described in the human androgen receptor gene (AR) associated with male infertility. Due to these molecular defects the androgen receptor becomes less responsive or completely unresponsive to androgens affecting sexual development and spermatogenesis [7].

The infertility can be also associated with endocrine disorders such as hypogonadism, in which the gonads do not function properly and produce inadequate amount of sex hormones There are two main categories of hypogonadism: hypogonadotropic and hypergonadotropic. Hypogonadotropic hypogonadism is characterized by low levels of sex hormones in males or females due to insufficient stimulation of the gonads by the pituitary gland and hypothalamus. Genetic causes of hypogonadotropic hypogonadism can result from mutations in genes involved in the development and functioning of these regulatory systems. Congenital hypogonadotropic hypogonadism (CHH) is a rare disorder (occurs in 1–10/100,000 infertile men) caused by the failure of the normal episodic gonadotropin-releasing hormone (GnRH) secretion, leading to delayed puberty and infertility [10]. Most of the genes mutated in CHH encode receptor-ligand pairs including GNRHR (gonadotropin-releasing hormone receptor) and GNRH1 (gonadotropin-releasing hormone 1), PROKR2 (Prokineticin receptor2) and PROK2 (Prokineticin 2), KISS1R (KiSS1 receptor) and KISS1, TACR3 (Tachykinin Receptor 3) and TAC3 (Tachykinin 3), FGFR1 (Fibroblast Growth Factor Receptor 1) and FGF8 (Fibroblast Growth Factor 8) suggesting that multiple receptor-ligand-encoding gene networks are involved in the molecular pathology of the disease. Part of the cases with CHH demonstrate an impairment sense of smell, called also Kallmann syndrome, in which pathogenic variants in genes such as ANOS1 (KAL1) (Anosmin 1), HS6ST1 (Heparan Sulfate 6-O-Sulfotransferase 1), SPRY4 (Sprouty RTK Signaling Antagonist 4), DUSP6 (Dual Specificity Phosphatase 6) are also identified [8]. The CHH is one of the few treatable causes of male infertility. Diagnosis of CHH is, however, challenging, especially in early adolescence where the clinical features mimics that of constitutional delay of growth and puberty. Timely diagnosis is essential for a proper treatment. Pathogenic variants, in more than 30 genes, has been associated with CHH, acting either alone or in combination [5].

Hypergonadotropic hypogonadism, also known as primary hypogonadism, is characterized by low levels of sex hormones accompanied by elevated levels of gonadotropins (FSH and LH). This condition can result from various genetic defects that directly affect the gonads. Some of the main genetic causes of hypergonadotropic hypogonadism involve mutations in genes such as FSHR (Follicle-Stimulating Hormone Receptor), LHR (Luteinizing Hormone Receptor), INSL3 (Insulin-Like 3), NR5A1 (Nuclear Receptor Subfamily 5 Group A Member 1) [5].

Identification of the causes of infertility is of paramount importance for each couple, due to several of the following reasons: the choice of therapy, genetic counselling, including determination of the risk of having offspring with cystic fibrosis (patients with CBAVD) or with deletion of the AZF region of the chromosome Y, improving mental comfort related to regaining hope for further effective treatment. Sometimes it may help to make decisions such as discontinue therapy and choosing alternative options (use of donor sperm, adoption, etc.).

In recent years, thanks to the use of next generation sequencing (NGS), more and more genes have been described in the context of fertility disorders. However, still in only 4% of all infertile men the genetic cause is established, and majority of the cases are classified as unexplained. This is partly due to a delay to adopt NGS techniques in the field of diagnostics and the lack of genotype–phenotype correlations data [5]. Therefore, the objective of the study was to identify genetic variants associated with isolated infertility using NGS technique. Secondary aim was to describe genotype-phenotype correlations.

Patients

The patients with fertility problems or delayed puberty were referred for genetic testing from Department of Endocrinology, Bielanski Hospital, Genetic counselling unit of the Department of Medical Genetics, Institute of Mother and Child, Oviclinic Infertility Center and nOvum Fertility Clinic in Warsaw. Women with endometriosis and PCOS were excluded. Phenotypical information was based on clinical interview, physical and laboratory examinations. All patients had karyotype analysis preformed before they were subjected to molecular analysis. All patients gave their informed consent for the genetic testing. The study was also approved by bioethical committee of the Institute of Mother and Child, Nr 22/2019.

Genetic analysis

AZF microdeletions analyses

To exclude AZF microdeletions on the chromosome Y, all males were subjected to analysis of 6 loci in the AZF by PCR. The panel of analyzed loci, including AZF c (sY254, sY255), AZF b(sY134, sY127), AZF a(sY86,S84), is consistent with the recommendations of the European Molecular Quality Network: "EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions: state-of-the-art 2013”.

NGS analysis

Genetic tests were performed by targeted next generation sequencing. The NGS panel included 35 genes with well-established association with hypogonadotropic hypogonadism (ANOS1, CHD7, DUSP6, FEZF1, FGF17, FGF8, FGFR1, FLRT3,FSHB, GNRH1, GNRHR, IL17RD, KISS1, KISS1R, LHB, NSMF, PROK2, PROKR2, SEMA3A, SOX10,SPRY4, WDR11, HS6ST1, TAC3) hypergonadotropic hypogonadism (NR5A1, INSL3,, LHCGR, LEP, GATA4, FSHR), spermatogenesis disorders (SPATA16, CFTR,), androgen insensitivity (AR), ovarian dysgenesis (BMP15) and sex reversal (SRY). Genes relevant to these phenotypes where included bases on OMIM and Human Phenotype Ontology (HPO) databases.

The DNA library was prepared using the SureSelect kit from Agilent. Sequencing was preformed using The NextSeq (Illumina). Only the coding fragments of genes (exons) and exon-intron junctions +/- 15 nucleotides were analyzed. The sequences were mapped to the human reference genome version GRCh38/hg38. The results were interpreted using internal bioinformatics tools based on the VEP program (Ensembl), and in relation to the following databases: GnomAD v3, HGMD Professional, NCBI, Ensembl, OMIM, internal database of IMiD variants. The variants classification was preformed according to ACMG (American College of Medical Genetics) recommendations using Varsome or Franklin platform. The presence of the detected genetic variants was confirmed by Sanger sequencing (the primers available on the request). The detected variants were named according to HGVS nomenclature and canonical transcript (NCBI RefSeq).

Patients

The study group consisted of 41 patients with following clinical diagnosis: hypogonadism hypogonadotropic (n = 12), hypogonadism hypergonadotropic (n = 15), abnormal semen parameters (oligozoospermia, astenozoospermia, teratozoospermia or azoospermia) (n = 10), androgen insensitivity (n = 3), gonadal dysgenesis (n = 1).

Genetic results

No deletion in the AZF a,b,c regions of the Y chromosome was detected in the examined males. One of 41 patients showed karyotype abnormality. This was a patient with typical female external genitalia, however, the structures resembling testes were found during inguinal hernias surgery. The karyotype analysis revealed presence of XY indicating sex reversal in this patient. The rest of patient had normal karyotype results.

The NGS gene panel analysis revealed 18 unique, missense or nonsense variants in 16/42 cases, including 11 known pathogenic (P) or likely pathogenic (LP) variants described in clinical databases (HGMD/ClinVar) and 4 novel or very rare variants (classified as LP or VUS according to ACMG recommendation), not reported in clinical databases and literature (Table 1).

Table 1

Genotype- phenotype correlation in patients with infertility problems or delayed puberty.
Patient Nr		Gene	Inheritance OMIM		Variant protein			HGMD /ClinVar		ACMG &
1	M	SPATA16	AR	c.848G > A	p.Arg283Gln	het	0.0004	DM/P (Globozoospermia)		LP	nHH	OAT, bilateral cryptorchidism, 6 toes on one foot
1	M	FGF8	AD, Oligo	c.77C > T	p.Pro26Leu	het	0.00115	DM/P (idiopathic HH)	LP		nHH	OAT, bilateral cryptorchidism, 6 toes on one foot
2	M	DUSP6	AD	c.566A > G	p.Asn189Ser	het	0.00104	DM/P (Kallmann syndrome)	LP		nHH	cryptozoospermiahypopituitarism (empty sella syndrome)
3	M	SPRY4	AD, Oligo	c.530A > G	p.Lys177Arg	het	0.00227	DM / Conflicting (Kallmann syndrome )	VUS/LP		nHH	OAT
4	M	AR	XLR	c.2395C > G	p.Gln799Glu	hemi	0.0015	DM/Confliciting (AIS)	P		PAIS	testicular hypoplasia, azooseprmia, decreased testosterone
5	F (k.XY)	AR	XLR	c.2287C > G	p.Leu763Val	hemi	0	DM (AIS)	P		CAIS
6	M	GNRHR	AR, Oligo	c.416G > A	p.Arg139His	het	0.00015	DM/P (HH)	P		nHH
7	M	NR5A1	AD	c.776C > T	p.Pro259Leu	het	0.00005	DM?( XY gonadal dysgenesis)/nd	P		Gonadal dysgenesis	epididymal cysts, microcalcifications, varicocele
8	M	DUSP6	AD	c.215C > A	p.Pro72Gln	het	0.00013	nd/VUS	VUS		nHH	tetratozoospermia (0% of normal sperm)
9	M	HS6ST1	AD	c.1132C > T	p.Arg378Cys	het	0.00003	nd/VUS	VUS		primary hypogonadism	severe oligozoospermia, very high level of FSH
10	F	FGFR1	AD	c.1270C > T	p.Arg424Cys	het	0.00001	nd/VUS	VUS		nHH
11	M	FGFR1	AD, Oligo	c.246_247del	p.Glu84GlyfsTer26	het	0	DM/ P (GnRH deficiency)	P		nHH
12	M	FGFR1	AD, Oligo	c.863A > T	p.Gln288Leu	het	0	nd/VUS	LP		Kallman syndrome	azoospermia
12	M	FGF8	AD, Oligo	c.439G > A	p.Ala147Thr	het	0.00001	nd/nd	LP		Kallman syndrome	azoospermia
13	M	SEMA3	AD	c.1630C > T	p.Arg544Cys	het	0.000001	nd/nd	LP		nHH	tetrazoospermia
14	F	FGFR1	AD	c.420_423del	p.Thr141IlefsTer10	het	0	nd/nd	LP		nHH	delayed puberty, primary amenorrhea
15	M	NSMF	AD	c.725A > G	p.Tyr242Cys	het	0.000001	nd/nd	VUS (PM2)		azoospermia	high level of FSH but normal level of testosterone
M - male; F- female; AD - autosomal dominant inheritance; AR - autosomal recessive inheritance; Oligo - oligogenic inheritance; nd - not described; Zyg -zygosity, MAF- allele minor frequency, DM- damaging, P-pathogenic, LP- likely pathogenic, VUS- variant of uncertain significance, PAIS - partial androgen insensitivity syndrome; AIS- androgen insensitivity, CAIS - complete androgen insensitivity syndrome; HH hypogonadotropic hypogonadism, nHH - normosomic hypogonadotropic hypogonadism; OAT- oligoasthenoteratozoospermia. k.-karyotype. & - ACMG classification according to .Brandt et al.2020, Genet Med. Bolded are novel variants (not found in clinical databases nor literature and absent/ very rare in gnomAD)

The genotype-phenotype correlation is described in Table 1. In patients with hypogonadotropic hypogonadism (HH), we have identified P/LP and VUS variants in 58% (7/12) and 25% (3/12) of the cases, respectively. One of these cases, diagnosed with Kallmann syndrome, harboured a very rare, heterozygous, LP variants in two different genes: p.Gln288Leu in FGFR1 and p.Ala147Thr in FGF8. In two of the CHH cases without anosmia, we found novel p.Thr141IlefsTer10 (LP) variant and a very rare, heterozygous VUS, p.Arg424Cys in FGFR1 gene.

In 1/15 cases with hypergonadotropic hypogonadism, we found one very rare VUS variant in HS6ST1 gene. In 2/3 of the cases with androgen insensitivity, we have detected a very rare, hemizygous, pathogenic variants in AR gene including p.Leu763Val identified in a patient with sex reversal (female phenotype but XY karyotype) and p.Leu763Val variant found in a male patient with partial androgen insensitivity (PAIS). One patient with gonadal dysgenesis harbored p.Arg139His, known pathogenic variant in one allele of the GNRHR gene. In 1/10 cases with abnormal semen parameters, we detected a very rare p.Tyr242Cys variant in NSMF gene classified as LP.

In this study, we have found 11 known, pathogenic/likely pathogenic variants (P/PL), 3 novel, likely pathogenic variants (LP) and 4 variants of uncertain significance (including one novel) in the group of patients with infertility or delayed puberty. The genetic testing enabled identification of the cause of the infertility or delayed puberty in 26% of the patients. We have examined patients with different clinical features such as hypogonadotropic and hypergonadotropic hypogonadism, abnormal semen parameters, androgen insensitivity and gonadal dysgenesis. We have described observed genotype-phenotype correlations.

Hypogonadism

Genetic studies on congenital hypogonadotropic hypogonadism (CHH) have led to discovery of pathogenic variants in various genes implicated in specification and proliferation of GnRH neurons, their migration to the hypothalamus during embryonic development and the response of pituitary gonadotropes to GnRH stimulation [9]. The genetic complexity of the disease is reflected by different models of inheritance: X-linked, autosomal dominant and autosomal recessive as well as incomplete penetrance and variable phenotypic expression as well as oligogenic inheritance pattern in the certain cases [9, 10]. Using our NGS panel, we identified P/LP in 58% (7/12) of the CHH (including one case of Kallmann syndrome) patients which is higher than in previous reports showing around 31–35% of the pathogenic variants detection rate [8]. Our results are consistent with the study by Cassatella D et al., which detected 51% of pathogenic variants in CHH patients using similar gene panel [11].

Multiple genes encoding components of the FGF pathway are known to be mutated in CHH and IHH eg. FGFR1, FGF8,, HS6ST1 DUSP6 and SPRY4 [8]. In our CHH patients, FGFR1 and FGF8 were the most frequently mutated genes with the same frequency of 33% (4/12), each. These results are consistent with previous studies showing very important role of the two genes in the development of CHH [8, 12, 13].

In FGFR1, we have detected one novel p.Thr141IlefsTer10 (LP) variant, one known pathogenic variant p.Glu84GlyfsTer26 and two very rare variants including p.Gln288Leu (LP) and p.Arg424Cys (VUS). The novel p.Thr141IlefsTer10 was found in a girl with hypogonadotropic hypogonadism, delayed puberty and primary amenorrhea. The p.Arg424Cys variant (VUS) was harboured by a girl with HH.Importantly, different amino acid substitution, Arg to His (p.Arg424His, c.1271G > A), was described in the same codon in a patient with HH (HGMD no.: CM2115069) suggesting that substations in this codon may be likely disruptive.

In FGF8, we found 2 variants including 1 know pathogenic p.Pro26Leu and 1 novel, LP p.Ala147Thr variant. Previously, Falardeau et al. described pathogenic variants in FGF8, including p.Pro26Leu, in IHH probands with variable olfactory phenotypes. These patients exhibited varied degrees of GnRH deficiency, including the rare adult-onset form of hypogonadotropic hypogonadism [13].

Pathogenic variants in DUSP6 (OMIM #615269) and SPRY4 (OMIM#615266) gene cause CHH with normal smell or anosmia and both are inherited in autosomal dominant manner. In DUSP6 gene, we detected variants in 2 of the 12 cases with HH including a known pathogenic variant p.Asn189Ser and p.Pro72Gln classified as VUS. The p.Asn189Ser variant was previously reported in case with Kallman syndrome [8]. Our patient showed a normal sense of smell but had hypopituitarism (empty sella syndrome) and cryptozoospermia. The other case with p.Pro72Gln (VUS) had teterathozoospermia and HH. Pheotypic variability is described both in a group of unrelated patients and within members of one family with the same pathogenic variant in DUSP6 gene. In some patients, variable phenotypic expression, maybe related to the simultaneous occurrence of pathogenic variants in other genes associated with hypogonadotropic hypogonadism [14]. We also have identified known pathogenic variant p.Lys177Arg in SPRY gene in one patient with hypogonadism and oligoasthenospermia. Previously, Miraoui et al. identified heterozygous missense variants in the SPRY4 gene in 14 unrelated individuals with congenital CHH), including p.Lys177Arg variant which was detected in patients with Kallmann syndrome [8].

The genetic complexity and phenotypic variability of CHH is exemplified by oligogenesity and involvement of pleotropic genes implicated in the disease. In our study, oligogenic interactions defined as ≥ 2 genes mutated were found in 2 out of 12 CHH cases. In one patient with Kallman syndrome, we detected simultaneous occurrence of p.Gln288Leu in FGFR1 and a novel Ala147Thr variant in FGF8. The pathogenic variants in FGFR1 and FGF8 genes are inherited in autosomal dominant manner, however, oligogenic interactions has been also reported [9, 15]. For instance, study by Sykiotis GP et al, investigating a large cohort of 397 patients with idiopathic HH, found that 2.5% of the patients show oligogenic interactions including genes such as FGFR1 and FGF8 [9]. The other digenic interaction was observed in a male patients with hypogonadotropic hypogonadism, oligoasthenoteratozoospermia (OAT) and bilateral cryptorchidism. This patient harbored known pathogenic variants such as p.Pro26Leu and p.Arg283Gln in FGF8 and SPATA16 gene, respectively. Pathogenic variants in FGF8 gene are associated with hypogonadotropic hypogonadism with or without ansomia. The literature describes patients with the p.Pro26Leu variant in FGF8 who were diagnosed with Kalman syndrome with partial loss of smell (hyposomia) [13, 16]. The phenotype is, however, variable even within members of one family with the same variant [13]. In our patient, no anosmia or hyposomia was present. Interestingly, this patient showed foot malformation (six toes) confirming a pleotropic role of the FGF8 gene [17]. Interestingly, FGFR1 pathogenic variants occur in 87% of patients with both CHH and split hand/food malformation [17]. Our patient also harbored heterozygous p.Arg283Gln variant in one allele of SPATA16, known to disturb splicing [18]. The pathogenic variants SPATA16 are the cause of defects in spermatogenesis (including abnormal sperm morphology) inherited in an autosomal recessive manner. In this case, however, no variants in the second allele of SPATA16 gene was identified.

NSMF encodes protein involved in guidance of olfactory axon projections and migration of luteinizing hormone-releasing hormone neurons. Defects in this gene are a cause of idiopathic hypogonadotropic hypogonadism (IHH) inherited in autosomal dominant manner (OMIM#614838). We have identified two variants in NSMF gene including a rare, LP, p.Pro213Leu and a novel p.Tyr242Cys (VUS) variant. The p.Pro213Leu variant, was present in one patient with hypogonadotropic hypogonadism and p.Tyr242Cys was found in a patient with azoospermia and high level of FSH but normal level of testosterone (not fulfilling criteria of hypogonadism).

In our study, we studied 15 cases with hypergonadotropic hypogonadism (primary hypogonadyzm) but no P/LP was detected in this group using our NGS panel. We have detected one VUS - p.Arg378Cys variant - in HS6ST1 gene in a patient with hypergonadotropic hypogonadism and oligozoospermia. The low frequency of detected variants in this group of patients could be due to limited number genes associated with primary hypogonadism included in our panel.

Androgen Insensitivity

The androgen insensitivity syndrome (AIS) is a disorder of male sexual development resulting in a wide range of clinical phenotypes. It affects-XY males but phenotypically is characterized by a female phenotype. AIS is classified into two phenotypic forms: complete (CAIS) and partial (PAIS). We have detected a very rare hemizygous variant p.Leu763Val in AR gene in patient with XY karyotype, female phenotype and complete androgen insensitivity syndrome (CAIS). This patient with typical female external genitalia was referred to the genetic counselling unit because during inguinal hernias surgery, structures resembling testes were found. Ultrasound examination demonstrated presence of two structures corresponded to testes localized along iliac blood vessels in absence of müllerian structures as fallopian tubes and uterus and absence of extragenital abnormalities apart from 3 cafe-au-lait spots. Very recently the p.Leu763Val variant has been described in an Iranian family with two female patients with normal external genitalia and 46, XY karyotype and CAIS phenotype [19]. Thus, we have described the second case in the world with the p.Leu763Val variant in AR gene. A pathogenic variant in the same 763 codon leading to Phe amino acid substitution (p.Leu763Phe and causing the loss of androgen binding receptor function was described as well [20]. We also identified a known pathogenic p.Gln799Glu variant in AR gene in one patient with PAIS and clinical features such as hypogonadotropic hypogonadism, testicular hypoplasia and azoospermia. In the literature, the p.Gln799Glu variant was described in patients partial androgen insensitivity, hypospadia and oligoasthenoteratozoospermia [21–23].

In summary, we have identified several new and recurrent pathogenic/likely pathogenic genetic variants associated with hypogonadism, androgen insensitivity and described observed genotype-phenotype correlations what expands the knowledge of the genetic basis of these disorders. Importantly, our results are in line with other studies. They provide further evidence for the heterogeneity of infertility disorders, especially in CHH, indicating that multigene panels are the best method of choice for genetic analysis in this group of patients. Further, this study highlights the importance of genetic testing for proper diagnosis making. Genetic diagnostics may also be very helpful in treating causes of infertility such as hypogonadism but requires the assistance of a clinician endocrinologist or andrologist, as well as a geneticist [24]. Moreover, the possibility of getting genetic counselling is also of high importance for the couple with infertility problems.

Acknowledgements: We are grateful to all patients who participated in this study.

Author’s Contributions:

Conceptualization, AMR and KKJ; Methodology (Molecular Analysis): KŚ, AD, KD, JS; Formal analysis: AMR, KŚ, AD, KD, JS, KWT(molecular analysis), KKJ., AKK, JKW, JK, PL (clinical analysis); Writing – Original Draft Preparation: AMR.; Writing - Review & Editing KKJ, AKK, JKW, PL, KWT. Visualization, KŚ.; Supervision: AMR.; Funding Acquisition, AMR.”, All authors have read and agreed to the published version of the manuscript.

Ethics approval

This study complies with the latest Declaration of Helsinki and was approved by bioethical committee of the Institute of Mother and Child, Warsaw, Nr 22/2019.

Consent to participate:

Informed consent to participate was obtained from all subjects involved in the study.

Consent for publication:

Consent for publication was received from all the patients enrolled in the study.

Competing interest:

The authors declare no conflicts of interest.

Funding:

This research was funded by Ministry of Science and Higher Education, Poland , project nr 510-18-61 granted to Institute of Mother and Child ( A.M. Rygiel)

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Editorial decision: Major Revisions Needed
08 Sep, 2024
Reviewers agreed at journal
20 Jul, 2024
Reviewers invited by journal
13 Jul, 2024
Editor assigned by journal
11 Jul, 2024
First submitted to journal
09 Jul, 2024

You are reading this latest preprint version

Novel and recurrent genetic variants associated with male and female infertility

Status:

Version 1

Abstract

Introduction

Materials and Methods

Patients

Genetic analysis

AZF microdeletions analyses

NGS analysis

Results

Patients

Genetic results

Discussion

Hypogonadism

Androgen Insensitivity

Conclusions

Declarations

References

Status:

Version 1