Molecular Profiling of Spermatozoa Reveals Correlations between Morphology and Gene Expression: A Novel Biomarker Panel for Male Infertility

Abstract

Choosing spermatozoa with an optimum fertilizing potential is one of the major challenges in assisted reproductive technologies (ART). This selection is mainly based on semen parameters, but the addition of molecular approaches could allow a more functional evaluation. To this aim, we used sixteen fresh sperm samples from patients undergoing ART for male infertility and classified them in the high- and poor-quality groups, on the basis of their morphology at high magnification. Then, using a DNA sequencing method, we analyzed the spermatozoa methylome to identify genes that were differentially methylated. By Gene Ontology and protein–protein interaction network analyses, we defined candidate genes mainly implicated in cell motility, calcium reabsorption, and signaling pathways as well as transmembrane transport. RT-qPCR of high- and poor-quality sperm samples allowed showing that the expression of some genes, such as AURKA, HDAC4, CFAP46, SPATA18, CACNA1C, CACNA1H, CARHSP1, CCDC60, DNAH2, and CDC88B, have different expression levels according to sperm morphology. In conclusion, the present study shows a strong correlation between morphology and gene expression in the spermatozoa and provides a biomarker panel for sperm analysis during ART and a new tool to explore male infertility.

Integration of omics studies indicates that species-dependent molecular mechanisms govern male fertility

“…In depth omics studies have recently been carried out in human and animal spermatozoa to provide a better understanding of male fertility at the genes [2][3][4][5], transcript [6][7][8], protein [9][10][11][12][13][14], and metabolite levels [6,11,[15][16][17][18]. Although these huge fertility-related omics datasets can offer prospective sources for discovering male fertility biomarkers which contribute to assess male fertility potential or sperm dysfunctions, these studies invariably contain information that is unrelated to male fertility status.…”

“…Comprehensive and comparative omics studies have recently been conducted in animals and humans to provide a better understanding of the complex multifactorial processes linked to male fertility at the gene [ 2 – 5 ], transcript [ 6 – 8 ], protein [ 9 – 14 ], and metabolite levels [ 6 , 15 – 18 ]. The development of new molecular methodologies to determine male fertility potential or sperm dysfunction has been facilitated by the accumulation of massive fertility-related omics datasets that serve as promising resources for identifying biomarkers of male fertility [ 10 , 19 ].…”

Effect of Micronutrients and L-Carnitine as Antioxidant on Sperm Parameters, Genome Integrity, and ICSI Outcomes: Randomized, Double-Blind, and Placebo-Controlled Clinical Trial

“…Abnormal sperm morphology can impact their ability to successfully fertilize an egg, influencing both natural conception and the results of procedures such as IVF and ICSI. A study conducted by Cassuto et al, 2022 involved a total of 16 couples enrolled for IVF protocol and revealed a significant correlation between morphology and gene expression in spermatozoa, a new tool to explore male infertility [51]. The antioxidant treatment can have an ameliorative effect on the sperm morphology.…”

Integrative Analysis of Small RNA and mRNA Expression Profiles Identifies Signatures Associated With Chronic Epididymitis

“…TRPM3 is involved in the regulation of inflammation through the release of neuropeptides ( 33 ). Intriguingly, a study profiled the global DNA methylation in sperm and found the aberrant modification of TRPM3 in patients with low morphology scores ( 34 ). Meanwhile, miR-22-5p, the impresser of TRPM3, was found to be upregulated in testicular tissue from patients with the non-obstructive azoospermia ( 35 ).…”