Search bioRxivSearch

bioRxiv · 10.1101/748277

Low frequency of pathogenic allelic variants in the 46,XY differences of sex development (DSD)-related genes in small for gestational age children with hypospadias

Abstract

BackgroundHypospadias is a congenital disorder of male genital formation where the urinary opening is not on the head of the penis and genetic factors play an important role in the incidence of this early developmental defect in 46,XY individuals, in both isolated and syndromic forms. Children born small for gestational age (SGA) present a high frequency of hypospadias of undetermined etiology, ranging from 15 to 30%, but the detection of hypospadias etiology remains low.\n\nPatients and methodsfrom a cohort of 46,XY DSD patients, we identified 25 SGA children with medium or proximal hypospadias; four of them with associated syndromic characteristics. DNA samples from subjects were studied by massively parallel targeted sequencing (MPTS) using a targeted panel. MLPA was used for molecular diagnosis in two children with clinical phenotype of Silver Russel syndrome.\n\nResultsLoss of DNA methylation (11p15 LOM) at ICR1 was identified in two out of four syndromic children. The other syndromic patient had 3M syndrome phenotype and two novel likely-pathogenic variants in compound heterozygous state were found in CUL7 gene. The last syndromic subject had Mulibrey nanism and, a homozygous variant in TRIM37 was identified in the patient and confirmed in heterozygous state in the mother. Among non-syndromic children seven rare heterozygous variants with uncertain significance in six DSD-related genes were identified: two children had DHX37 variants associated with GATA4 and WWOX variants, respectively; three children had heterozygous variants, in WT1, IGF1R, and BMP8B genes.\n\nConclusionPathogenic or likely-pathogenic variants in DSD-related genes were not identified in non-syndromic SGA children with hypospadias, suggesting that multi-factorial causes, unknown genes or unidentified environmental factors (epigenetic defects), may be involved in the etiology of this condition.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Leitao Braga, B., Lisboa Gomes, N., YUMIE NISHI, M., LUCHEZE FREIRE, B., LOCH BATISTA, R., FERREIRA DE ASSIS FUNARI, M., FRADE COSTA, E. M., MARCONDES LERARIO, A., D. FARIA JUNIOR, J. A., DOMENICE, S., LIMA JORGE, L. A., BILHARINHO MENDONCA, B.. 2019-08-28. Low frequency of pathogenic allelic variants in the 46,XY differences of sex development (DSD)-related genes in small for gestational age children with hypospadias. https://doi.org/10.1101/748277

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

The histone demethylase Kdm5 and the ARGONAUTE proteins Piwi and Aubergine regulate female abdominal pigmentation in Drosophila melanogaster

Insect pigmentation is an ecologically critical trait influencing many physiological processes. In Drosophila melanogaster, abdominal pigmentation is sexually dimorphic: males have fully pigmented posterior segments, while females exhibit a posterior melanin stripe. Pigmentation relies on the expression of pigmentation genes that encode enzymes involved in pigment synthesis. These genes are tightly regulated during pupal and young adult stages. To expand the gene regulatory network of pigmentation genes, we conducted an RNAi screen using the yellow-Gal4 driver, expressed during the pupal stage in abdominal epidermis. One of the candidates from this screen, Kdm5, encodes a histone demethylase erasing the H3K4me3 histone mark catalyzed by the histone methyl-transferase Trithorax (Trx). We show that Kdm5 down-regulation reduces abdominal pigmentation, mimicking trx down-regulation. Kdm5 activates melanin production through regulation of the pigmentation gene tan. Transcriptomic analyses reveal that Kdm5 and Trx share many targets in pupal abdominal epidermis, including piRNA pathway components such as piwi and aubergine. These piRNA components, originally associated with transposon silencing in the germline, also function in some somatic tissues such as the nervous system, the fat body or the gut. We demonstrate that Piwi and Aubergine participate in female abdominal pigmentation establishment, without evident piRNA production. We also show that Kdm5 and Piwi act not only in pupal abdominal epidermis but also in pupal fat body. This study therefore expands the regulatory network of pigmentation genes. It identifies a new somatic function for Kdm5 and Piwi and reveals a role for pupal fat body in female abdominal pigmentation regulation.

genetics

Genetic diversity within and between polyploid sugarcane (Saccharum spp.) families obtained via caryopsis using microsatellite markers and multicategory model

Genetic diversity analyses are essential for sugarcane (Saccharum spp.) breeding programs. Crossbreeding, based on genetic distances between parental plants, is a tool used to increase genetic variability and enhance plant selection; however, quantifying variation in highly polyploid species remains a challenge. The present study aimed to evaluate the diversity within and between 12 families of sugarcane derived from caryopses, analyzing 120 individual seedlings arranged in an augmented block design. Genotyping was performed using primers for 16 microsatellite loci, five simple sequence repeat (SSR) loci, and 11 expressed sequence tag-SSR (EST-SSR) loci. To accurately account for polyploidy, similarity calculations were performed using Bruvos distances among individuals and RST distances among the families. Analysis of molecular variance (AMOVA) indicated that most of the genetic variability was within families (72%), with only 28% found between them. This high level of intra-family variation demonstrates that a significant reservoir of genetic diversity remains available within the crosses. The highest genetic similarity was observed between the families RB986952 x RB986960 and RB036122 x RB03611, whereas the lowest genetic similarity was observed between the families RB97319 x RB966928 and RB106802 x RB855036. Although the evaluated families shared high genetic similarity, the pronounced genetic variation within them demonstrates a robust recombination potential, indicating that the genetic basis of sugarcane can be better explored using the high variability that already exists in the selection of desirable morpho-agronomic characteristics within the families. Furthermore, this study highlights the importance of using appropriate distances for diversity studies with codominant markers, such as microsatellites, in polyploid species.

genetics

Optimizing DNA extraction from environmentally degraded bone samples for molecular identification of cetacean species

Molecular identification of cetacean bone remains can be limited by DNA degradation and the presence of PCR inhibitors. Here, we present an optimized DNA extraction protocol based on a total demineralization method for environmentally exposed cetacean bones. The protocol uses 100 mg of bone powder, 24 h digestion with EDTA, N-lauroylsarcosine, and proteinase K, followed by a modified silica-column purification. Nine environmentally degraded bone samples representing eight individuals were processed. DNA concentrations ranged from 7.3 to 57.1 ng/uL (mean SD = 25.91- 13.91 ng/uL). The mitochondrial cytochrome b gene was successfully amplified from all samples using conventional PCR, and five samples (55.6%) yielded sequences suitable for downstream analysis. BLASTn identified Balaenoptera physalus as the closest database match for all recovered sequences, and phylogenetic analysis further supported their association with B. physalus reference sequences. These results demonstrate that the proposed protocol provides a practical approach for recovering amplifiable and molecularly informative mitochondrial DNA from environmentally degraded cetacean bone material, facilitating molecular identification from challenging skeletal remains.

genetics