# Genetic Variants in Patients with Premature Ovarian Insufficiency in a Mexican Cohort
###### Abstract  
Premature ovarian insufficiency (POI) is one of the main causes of female premature infertility. POI is a genetically heterogeneous disorder with a complex molecular etiology; as such, the genetic causes remain unknown in the majority of patients. Therefore, this study aimed to identify mutations and characterize the associated molecular contribution of gonadogenesis-determinant genes to POI. Genomic assays, including PCR-SSCP and Sanger sequencing, followed by in silico analyses were used to investigate the underpinnings of ovarian deficiency in 11 women affected by POI. Large deletions and nucleotide insertions and duplications were excluded by PCR. Thirteen genetic variants were identified in the _WT1_ (c.213G>T, c.609T>C, c.873A>G, c.1122G>A), _NR0B1_ (c.353C>T, c.425G>A), _NR5A1_ (c.437G>C, IVS4-20C>T), _LHM3_ (IVS2-12G>C, IVS3+13C>T, c.741T>C), _ZNF275_ (c.969C>T), and _NRIP1_ (c.3403C>T) genes. Seven novel genetic variants and five unpublished substitutions were identified. No genetic aberrations were detected in the _ZFP92_ and _INS13_ genes. Each variant was genotyped using PCR-SSCP in 100 POI-free subjects, and their allelic frequencies were similar to the patients. These analyses indicated that allelic variation in the _WT1_, _NR0B1_, _NR5A1_, _LHK9_, _ZFP92_, _ZNF275_, _INS13_, and _NRIP1_ genes may be a non-disease-causing change or may not contribute significantly to the genetics underlying POI disorders. Findings support the polygenic nature of this clinical disorder, with the SNVs identified representing only a probable contribution to the variability of the human genome. 0 2022 13 611 11  
these also contain multiple genes (e.g., _DIAPH2_, _XPNPEP2_, _DACH2_) associated with POI. Specifically, the region on the long arm of the X chromosome from Xq13-Xq21 to Xq23-Xq27 is associated with the POI phenotype [10; 11; 12]. However, X-structural anomalies and X-autosome translocations have not been completely useful for relating to POI; therefore, additional heretofore unknown genetic factors are likely also involved in the etiology of this disease. To date, more than 100 genes have been implicated in the loss of ovarian activity [8; 13; 14; 15], and single-gene mutations have been associated with POI [16; 17; 18]. The genes involved in the pathogenesis of POI also affect development [19; 20], DNA division and repair [21; 22], follicle development and hormonal signaling [23; 24], metabolism [25; 26], and immune regulation [27; 28]. The premutation of the fragile X mental retardation 1 (_FMR1_) gene is the most common genetic factor associated with POI. The _FMR1_ allele is located on the X chromosome at q27.3 and contains a trinucleotide repeat sequence, (CGG)n, in its 5\({}^{\prime}\) untranslated region. This gene contains 6-44 CGG repeats with AGG interspersions every 9 or 10 repeats, and having over 55 CGG repeats is associated with pathogenicity [29; 30; 31; 32; 33; 34]. Previous reports demonstrate the scarcity of knowledge concerning the molecular etiology of this ovarian disorder and highlight the importance of identifying novel genetic candidates potentially linked to the pathogenesis of POI. The genetic and molecular mechanisms underlying this disease, which could inform the development of more effective treatments to preserve fertility, have not yet been fully elucidated. To gain further insight into the contribution of gonadogenesis-determinant genes to the pathogenesis of POI, the current study examined the frequency and downstream molecular implications of _WT1_ (11p13), _NR0B1_ (Xp21.2), _NR5A1_ (11q13.1), _LHX9_ (1q31.3), _ZFP92_ (Xq28), _ZNF275_ (Xq28), _INSL3_ (19p13.11), and _NRIP1_ (21q11.2-q21.1) mutations in patients with POI.