The Use of Proteomics in Assisted Reproduction

Kosteria, Ioanna; Anagnostopoulos, Athanasios K.; Kanaka‐Gantenbein, Christina; Chrousos, George P.; Tsangaris, George Th.

doi:10.21873/invivo.11056

Cited by 22 publications

(24 citation statements)

References 160 publications

(117 reference statements)

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“…Although conventional semen analysis provides useful information about the sperm parameters (WHO, ), it fails to explain the differences at the molecular level. Expression levels and availability of the functional sperm proteins associated with motility, capacitation, acrosome reaction, and fertilization have an impact on the successful fertilization of spermatozoa with the oocyte (Rahman et al ., ; Kosteria et al ., ; Panner Selvam & Agarwal, ). Significant advances in proteomics‐related approaches and strategies including mass spectrometry and liquid chromatography have enabled the analysis of thousands of complex cellular proteins and peptides present in spermatozoa (Martínez‐Heredia et al ., ; Oliva et al ., ; du Plessis et al ., ; Amaral et al ., ).…”

Section: Introductionmentioning

confidence: 99%

A quantitative global proteomics approach to understanding the functional pathways dysregulated in the spermatozoa of asthenozoospermic testicular cancer patients

2019

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Background: Testicular cancer (TC) is the most common cancer diagnosed in men of reproductive age group. Sperm banking is recommended in these patients prior to cancer treatment. There is no literature describing the proteins dysregulated in the spermatozoa of TC patients with poor motility. Objective: The primary objective of this study was to compare the differences in the sperm proteome of normozoospermic (motility > 40%) and asthenozoospermic (motility < 40%) TC patients who had cryopreserved semen samples before initiating cancer therapy. Materials and methods: Pooled sperm samples from healthy fertile men (n = 8), normozoospermic (n = 20), and asthenozoospermic (n = 11) TC patients were used for quantitative global proteomic profiling by liquid chromatography-tandem mass spectrometry (LC-MS). The functional bioinformatic analysis was done by ingenuity pathway analysis software. Key differentially expressed proteins (DEPs) associated with binding of zona pellucida (CCT3), mitochondrial dysfunction (ATP5A1 and UQCRC2), sperm motility (ATP1A4), and an exosomal protein involved in the metabolic process of the spermatozoa (MMP9) were validated using Western blot analysis by comparing normozoospermic (n = 10) with asthenozoospermic (n = 10) TC patients. Statistical analysis was conducted using Mann-Whitney test. Results: A total of 813 and 957 proteins were detected in spermatozoa of normozoospermic and asthenozoospermic TC patients, respectively. On the other hand, 1139 proteins were detected in the spermatozoa of healthy fertile men. 198 proteins were identified as DEPs between TC patients with normal and abnormal semen parameters. Validation of DEPs revealed downregulation of key proteins (CCT3, ATP1A4, ATP5A1, and UQCRC2) implicated in the reproductive function in asthenozoospermic TC patients. Discussion: DEPs involved in the reproductive function pathways suggest defective spermatogenesis and maturation process in asthenozoospermic TC patients. Furthermore, dysfunctional exosomal pathway may be a possible cause of infertility in TC patients. Conclusion: The proteins associated with sperm function and fertilization process are compromised in TC patients irrespective of their semen parameters.

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Section: Introductionmentioning

confidence: 99%

A quantitative global proteomics approach to understanding the functional pathways dysregulated in the spermatozoa of asthenozoospermic testicular cancer patients

2019

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“…Qualifying and quantifying substances secreted by embryos into their surrounding media would eliminate some of the concerning aspects of embryo biopsy. Metabolomics and proteomics are promising technologies to non-invasively identify targets specific to embryo normalcy 7 ; these techniques have yet to reach the appropriate sensitivity levels due to the very small amounts of soluble analytes. The broad spectrum of proteins, both from the embryo and media, further complicate unequivocal detection of relevant molecules.…”

Section: Introductionmentioning

confidence: 99%

Human blastocysts of normal and abnormal karyotypes display distinct transcriptome profiles

Licciardi

Lhakhang²,

Kramer

et al. 2018

Sci Rep

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Unveiling the transcriptome of human blastocysts can provide a wealth of important information regarding early embryonic ontology. Comparing the mRNA production of embryos with normal and abnormal karyotypes allows for a deeper understanding of the protein pathways leading to viability and aberrant fetal development. In addition, identifying transcripts specific for normal or abnormal chromosome copy number could aid in the search for secreted substances that could be used to non-invasively identify embryos best suited for IVF embryo transfer. Using RNA-seq, we characterized the transcriptome of 71 normally developing human blastocysts that were karyotypically normal vs. trisomic or monosomic. Every monosomy and trisomy of the autosomal and sex chromosomes were evaluated, mostly in duplicate. We first mapped the transcriptome of three normal embryos and found that a common core of more than 3,000 genes is expressed in all embryos. These genes represent pathways related to actively dividing cells, such as ribosome biogenesis and function, spliceosome, oxidative phosphorylation, cell cycle and metabolic pathways. We then compared transcriptome profiles of aneuploid embryos to those of normal embryos. We observed that non-viable embryos had a large number of dysregulated genes, some showing a hundred-fold difference in expression. On the contrary, sex chromosome abnormalities, XO and XXX displayed transcriptomes more closely mimicking those embryos with 23 normal chromosome pairs. Intriguingly, we identified a set of commonly deregulated genes in the majority of both trisomies and monosomies. This is the first paper demonstrating a comprehensive transcriptome delineation of karyotypic abnormalities found in the human pre-implantation embryo. We believe that this information will contribute to the development of new pre-implantation genetic screening methods as well as a better understanding of the underlying developmental abnormalities of abnormal embryos, fetuses and children.

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“…In the last decade, several proteomic studies have been conducted using spermatozoa and seminal plasma (Amaral et al, ; Panner Selvam, & Agarwal, ; Panner Selvam, Agarwal, Sharma, & Samanta, ; Samanta et al, ) to understand the molecular pathways associated with male infertility and identification of potential protein biomarkers. High‐throughput instruments such as MALDI‐TOF and LC‐MS/MS systems are widely used to identify the proteomic profile of spermatozoa (Agarwal, Durairajanayagam, Halabi, Peng, & Vazquez‐Levin, ; Kosteria, Anagnostopoulos, Kanaka‐Gantenbein, Chrousos, & Tsangaris, ; du Plessis, Kashou, Benjamin, Yadav, & Agarwal, ). While processing the sperm proteins, it is believed that sperm proteome can be contaminated with proteins originating from round cells, such as leucocytes and epithelial somatic cells.…”

Section: Discussionmentioning

confidence: 99%

Round cells do not contaminate or mask human sperm proteome in proteomic studies using cryopreserved samples

et al. 2019

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Semen contains leucocytes and round cells, besides spermatozoa. The objective of this study was to identify whether the proteins from round cells and leucocytes affect the proteomic analysis of spermatozoa. Cryopreserved human sperm samples were divided into four groups: (1) samples with ≥1 × 106/ml leucocytes unprocessed; (2) samples with ≥1 × 106/ml leucocytes processed by 65% density centrifugation; (3) samples with round cells <1 × 106/ml unprocessed; and (4) samples with round cells <1 × 106/ml processed by 65% density centrifugation. Samples from each group (1, 2, 3 and 4) were pooled (n = 5) for quantitative proteomic analysis using liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Comparative analysis revealed nine differentially expressed proteins (DEPs) groups 1 and 2. Moreover, five DEPs were identified between groups 3 and 4. We observed that cylicin‐1, Atlastin‐1 and vesicle transport protein SFT2B are specific to spermatozoa, and none of them were associated with leucocytes. The number of DEPs in spermatozoa of processed and unprocessed cryopreserved semen samples was negligible. Our results indicate that the presence of round cells (<1 × 106/ml) in the seminal ejaculation does not interfere in the accurate detection of spermatozoa proteome by LC‐MS/MS.

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The Use of Proteomics in Assisted Reproduction

Abstract: Abstract. Despite

Cited by 22 publications

References 160 publications

A quantitative global proteomics approach to understanding the functional pathways dysregulated in the spermatozoa of asthenozoospermic testicular cancer patients

A quantitative global proteomics approach to understanding the functional pathways dysregulated in the spermatozoa of asthenozoospermic testicular cancer patients

Human blastocysts of normal and abnormal karyotypes display distinct transcriptome profiles

Round cells do not contaminate or mask human sperm proteome in proteomic studies using cryopreserved samples

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