Severity-Dependent Profile of the Metabolome in Hypospadias

Abstract

Background & Objective: Hypospadias, characterized by the displacement of the opening of the urethra at any point in the medial-ventral side of the penis, is classified upon severity as mild (Type I) and severe (Type II and Type III) hypospadias. Hypospadias' etiology is idiopathic in the majority of cases, and underlying causes seem of multifactorial origin. Studies regarding genetic variants support this notion. It is unknown whether downstream gene products fit this profile. This study evaluated the metabolome of hypospadias by using the emerging technology of metabolomics in the search for distinct cellular processes associated with hypospadias' etiology according to the severity of this congenital urogenital condition. Methods: Foreskin samples were collected during urethroplasty from boys with Type I, II, and III hypospadias or undergoing elective circumcision (N = 28) between 5 and 28 months of age. Samples were processed and submitted to gas chromatography-mass spectrometry (GC/MS). MetaboloAnalyst (http://www.metaboanalyst.ca/) online platform was used for bioinformatic analyses. Results: Thirty-five metabolites across experimental groups were identified by GC/MS. Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) showed that the metabolome of Type II and Type III hypospadias patients differs from the metabolome of Type I hypospadias and control patients. Of those 35, 10 amino acids were found in significantly low concentrations in severe hypospadias: aspartate, glutamate, glycine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, and tyrosine. A high concentration of the amino acid lysine was detected in mild hypospadias. Conclusions: The observed downregulation of specific amino acids in severe hypospadias provides alternative routes for future research aiming to identify disrupted networks and pathways while considering the severity of hypospadias.

A Proteomics Signature of Mild Hypospadias: A Pilot Study

“…At the mRNA and protein level, significant changes as severity increases have been observed in AR, CYR61, ATF3 , and CTGF , some of which are estrogen-responsive genes ( 30 , 41 ). In terms of downstream products, we found lower concentrations of aspartate in moderate and severe hypospadias but not in mild hypospadias ( 42 ). This finding is of interest as aspartate modulates steroidogenesis in Leydig cells ( 43 – 45 ).…”

Effects of Ferroptosis on the Metabolome in Cardiac Cells: The Role of Glutaminolysis

“…Metabolites were extracted as previously described [ 26 ] using 1 mL of cold MeOH/H 2 O (85:15). Samples were sonicated for 15 s (3x) on ice and centrifuged at 1400 rpm × 10 min at 4 °C (Rotor model: Eppendorf, FA-45-30-11).…”

“…Raw chromatography data were obtained and processed in GC-MS Labsolution Postrun Analysis software (Shimadzu Scientific Instruments, Inc., Columbia, MD) for identification of metabolites from their electron impact mass spectra by comparison to the database (NIST14/2014/EPA/NIH) [ 26 ]. Peak integration for all metabolites and multiple searches in the mass spectral library database resulted in a final database of 52 metabolic features chosen for this analysis.…”

Characterization With Gene Mutations in Han Chinese Patients With Hypospadias and Function Analysis of a Novel AR Genevariant

“…To date, over 1,000 mutations have been reported in the AR gene, and it is usually included as one of the top genes in the screening panel test for patients with hypospadias. Moreover, not only are their mutations reported in the coding region of AR, but also SNPs located in the promoter region of AR gene were involved in the development and severity of hypospadias ( Geller et al, 2014 ; Pineyro-Ruiz et al, 2020 ) however, the functional importance of these mutations remains less known. AR gene mutations also lead to several pathological situations such as androgen insensitivity syndrome (AIS), spinal and bulbar muscular atrophy (SBMA), and prostate cancer ( Shukla et al, 2016 ).…”