Summary of Study ST003276
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002028. The data can be accessed directly via it's Project DOI: 10.21228/M8GC01 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST003276 |
| Study Title | Metabolomics analysis of human spermatozoa reveals impaired metabolic pathways in asthenozoospermia (NMR data) |
| Study Summary | Background: Infertility is a major health issue, affecting 15% of reproductive-age couples with male factors contributing to 50% of cases. Asthenozoospermia, or low sperm motility, is a common cause of male infertility with complex etiology, involving genetic and metabolic alterations, inflammation, and oxidative stress. However, the molecular mechanisms behind low motility are unclear. In this study, we used a metabolomics approach to identify metabolic biomarkers and pathways involved in sperm motility. Methods: We compared the metabolome and lipidome of spermatozoa of men with normozoospermia (n = 44) and asthenozoospermia (n = 22) using untargeted LC-MS and the metabolome of seminal fluid using 1H-NMR. Additionally, we evaluated the seminal fluid redox status to assess the oxidative stress in the ejaculate. Results: We identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of the spermatozoa’s metabolomics and lipidomics data showed a clear separation between groups. Spermatozoa of asthenozoospermic men presented lower levels of several amino acids, and increased levels of energetic substrates and lysophospholipids. However, the metabolome and redox status of the seminal fluid was not altered in asthenozoospermia. Conclusions: Our results indicate impaired metabolic pathways associated with redox homeostasis and amino acid, energy, and lipid metabolism in asthenozoospermia. Taken together, these findings suggest that the metabolome and lipidome of human spermatozoa are key factors influencing their motility and that oxidative stress exposure during spermatogenesis or sperm maturation may be in the etiology of decreased motility in asthenozoospermia. |
| Institute | University of Aveiro |
| Department | Department of Chemistry |
| Last Name | Guerra-Carvalho |
| First Name | Bárbara |
| Address | Campus Universitário de Santiago, 3810-193 Aveiro, Portugal |
| barbaraggcarvalho@gmail.com | |
| Phone | 234 370 360 |
| Submit Date | 2024-06-20 |
| Num Groups | 2 |
| Total Subjects | 57 |
| Num Males | 57 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | fid |
| Analysis Type Detail | NMR |
| Release Date | 2024-07-25 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002028 |
| Project DOI: | doi: 10.21228/M8GC01 |
| Project Title: | Metabolomics analysis of human spermatozoa reveals impaired metabolic pathways in asthenozoospermia |
| Project Summary: | Background: Infertility is a major health issue, affecting 15% of reproductive-age couples with male factors contributing to 50% of cases. Asthenozoospermia, or low sperm motility, is a common cause of male infertility with complex etiology, involving genetic and metabolic alterations, inflammation, and oxidative stress. However, the molecular mechanisms behind low motility are unclear. In this study, we used a metabolomics approach to identify metabolic biomarkers and pathways involved in sperm motility. Methods: We compared the metabolome and lipidome of spermatozoa of men with normozoospermia (n = 44) and asthenozoospermia (n = 22) using untargeted LC-MS and the metabolome of seminal fluid using 1H-NMR. Additionally, we evaluated the seminal fluid redox status to assess the oxidative stress in the ejaculate. Results: We identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of the spermatozoa’s metabolomics and lipidomics data showed a clear separation between groups. Spermatozoa of asthenozoospermic men presented lower levels of several amino acids, and increased levels of energetic substrates and lysophospholipids. However, the metabolome and redox status of the seminal fluid was not altered in asthenozoospermia. Conclusions: Our results indicate impaired metabolic pathways associated with redox homeostasis and amino acid, energy, and lipid metabolism in asthenozoospermia. Taken together, these findings suggest that the metabolome and lipidome of human spermatozoa are key factors influencing their motility and that oxidative stress exposure during spermatogenesis or sperm maturation may be in the etiology of decreased motility in asthenozoospermia. |
| Institute: | University of Aveiro |
| Department: | Department of Chemistry |
| Last Name: | Guerra-Carvalho |
| First Name: | Bárbara |
| Address: | Campus Universitário de Santiago, 3810-193 Aveiro, Portugal |
| Email: | barbaraggcarvalho@gmail.com |
| Phone: | 234 370 360 |
| Project Comments: | Study part 1 of 2 |
Subject:
| Subject ID: | SU003396 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 20 to 49 years old |
| Gender: | Male |
| Human Inclusion Criteria: | Participants enrolled in this study were randomly recruited among men who attended fertility consultations at the Centre for Reproductive Genetics Professor Alberto Barros (Porto, Portugal) and the Centro Hospitalar Universitário de Santo António (Porto, Portugal) from March to June 2021. Adult men (age ≥ 18 years) diagnosed with asthenozoospermia (AS, sperm total motility < 42%) were recruited for AS group and men with normal sperm parameters (sperm concentration ≥ 15 million cells/mL, sperm count ≥ 39 million cells and sperm total motility ≥ 42%) were recruited for the control group (normozoospermia (NZ) group). |
| Human Exclusion Criteria: | Men with a history of abusing drugs or alcoholism, under treatment or exposed to drugs known to interfere with fertility, diagnosed with acute genital inflammatory disease or with systemic disease known to affect fertility before sperm collection, survivors of cancer treatment, or men diagnosed with leukocytospermia were excluded from this study. Smoking was not included as an exclusion criterion for this study. |
Factors:
Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Group | Sample source |
|---|---|---|---|
| SA354646 | N39 | Asthenozoospermia | Human seminal fluid |
| SA354647 | N4 | Asthenozoospermia | Human seminal fluid |
| SA354648 | N64 | Asthenozoospermia | Human seminal fluid |
| SA354649 | N58 | Asthenozoospermia | Human seminal fluid |
| SA354650 | N56 | Asthenozoospermia | Human seminal fluid |
| SA354651 | N53 | Asthenozoospermia | Human seminal fluid |
| SA354652 | N49 | Asthenozoospermia | Human seminal fluid |
| SA354653 | N44 | Asthenozoospermia | Human seminal fluid |
| SA354654 | N43 | Asthenozoospermia | Human seminal fluid |
| SA354655 | N3 | Asthenozoospermia | Human seminal fluid |
| SA354656 | N34 | Asthenozoospermia | Human seminal fluid |
| SA354657 | N14 | Asthenozoospermia | Human seminal fluid |
| SA354658 | N29 | Asthenozoospermia | Human seminal fluid |
| SA354659 | N28 | Asthenozoospermia | Human seminal fluid |
| SA354660 | N27 | Asthenozoospermia | Human seminal fluid |
| SA354661 | N26 | Asthenozoospermia | Human seminal fluid |
| SA354662 | N24 | Asthenozoospermia | Human seminal fluid |
| SA354663 | N22 | Asthenozoospermia | Human seminal fluid |
| SA354664 | N15 | Asthenozoospermia | Human seminal fluid |
| SA354665 | N33 | Asthenozoospermia | Human seminal fluid |
| SA354666 | N42 | Normozoospermia | Human seminal fluid |
| SA354667 | N45 | Normozoospermia | Human seminal fluid |
| SA354668 | N46 | Normozoospermia | Human seminal fluid |
| SA354669 | N47 | Normozoospermia | Human seminal fluid |
| SA354670 | N48 | Normozoospermia | Human seminal fluid |
| SA354671 | N50 | Normozoospermia | Human seminal fluid |
| SA354672 | N59 | Normozoospermia | Human seminal fluid |
| SA354673 | N51 | Normozoospermia | Human seminal fluid |
| SA354674 | N52 | Normozoospermia | Human seminal fluid |
| SA354675 | N55 | Normozoospermia | Human seminal fluid |
| SA354676 | N57 | Normozoospermia | Human seminal fluid |
| SA354677 | N38 | Normozoospermia | Human seminal fluid |
| SA354678 | N61 | Normozoospermia | Human seminal fluid |
| SA354679 | N63 | Normozoospermia | Human seminal fluid |
| SA354680 | N68 | Normozoospermia | Human seminal fluid |
| SA354681 | N70 | Normozoospermia | Human seminal fluid |
| SA354682 | N40 | Normozoospermia | Human seminal fluid |
| SA354683 | N23 | Normozoospermia | Human seminal fluid |
| SA354684 | N36 | Normozoospermia | Human seminal fluid |
| SA354685 | N12 | Normozoospermia | Human seminal fluid |
| SA354686 | N2 | Normozoospermia | Human seminal fluid |
| SA354687 | N5 | Normozoospermia | Human seminal fluid |
| SA354688 | N6 | Normozoospermia | Human seminal fluid |
| SA354689 | N7 | Normozoospermia | Human seminal fluid |
| SA354690 | N8 | Normozoospermia | Human seminal fluid |
| SA354691 | N10 | Normozoospermia | Human seminal fluid |
| SA354692 | N11 | Normozoospermia | Human seminal fluid |
| SA354693 | N13 | Normozoospermia | Human seminal fluid |
| SA354694 | N35 | Normozoospermia | Human seminal fluid |
| SA354695 | N18 | Normozoospermia | Human seminal fluid |
| SA354696 | N19 | Normozoospermia | Human seminal fluid |
| SA354697 | N20 | Normozoospermia | Human seminal fluid |
| SA354698 | N21 | Normozoospermia | Human seminal fluid |
| SA354699 | N25 | Normozoospermia | Human seminal fluid |
| SA354700 | N30 | Normozoospermia | Human seminal fluid |
| SA354701 | N31 | Normozoospermia | Human seminal fluid |
| SA354702 | N16 | Normozoospermia | Human seminal fluid |
| Showing results 1 to 57 of 57 |
Collection:
| Collection ID: | CO003389 |
| Collection Summary: | Human seminal samples were obtained through masturbation into a specific sterile container after 2 to 4 days of sexual abstinence. After collection, samples were left at 37°C until complete liquefaction and sperm parameters (semen pH, ejaculate volume, sperm concentration, sperm count and sperm motility) were analyzed according to the WHO guidelines. After routine semen analysis, spermatozoa were separated from the seminal fluid through centrifugation at 500.g for 5 min. Seminal fluid was collected into new centrifuge tubes and stored at -80°C until further use. |
| Sample Type: | Seminal plasma |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003405 |
| Treatment Summary: | Participants enrolled in this study were randomly recruited among men who attended fertility consultations at the Centre for Reproductive Genetics Professor Alberto Barros (Porto, Portugal) and the Centro Hospitalar Universitário de Santo António (Porto, Portugal) from March to June 2021. Adult men (age ≥ 18 years) diagnosed with asthenozoospermia (AS, sperm total motility < 42%) were recruited for AS group and men with normal sperm parameters (sperm concentration ≥ 15 million cells/mL, sperm count ≥ 39 million cells and sperm total motility ≥ 42%) were recruited for the control group (normozoospermia (NZ) group). |
Sample Preparation:
| Sampleprep ID: | SP003403 |
| Sampleprep Summary: | Seminal fluid samples were allowed to thaw at room temperature and homogenized using a vortex mixer. Then, samples were centrifuged at 20000.g for 20 min at 4°C to remove any insoluble particles. After centrifugation, 45 µL of seminal fluid was diluted in 135 µL of deuterated water and 45 µL of a 10 mM sodium fumarate in 0.2 M phosphate buffer (pH 7.0) deuterated water (99.9%) solution. |
Analysis:
| Analysis ID: | AN005364 |
| Analysis Type: | NMR |
| Num Factors: | 2 |
| Num Metabolites: | 23 |
| Units: | normalized areas |
NMR:
| NMR ID: | NM000284 |
| Analysis ID: | AN005364 |
| Instrument Name: | 500 MHz Bruker Avance III HD |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D-1H |
| NMR Comments: | The samples were transferred to 3-mm NMR tubes (Norell, Landisville, NJ, USA) and analyzed using the NOR5X3INSB optimizer inserts (Norell, Landisville, NJ, USA) on a 500 MHz Bruker Avance III HD spectrometer equipped with a 5 mm TXI probe (Bruker Corporation, Billerica, MA, USA), working at 298K. Solvent-suppressed 1D-1H-NMR spectra were acquired using a zgpr pulse sequence, with a sweep width of 7 kHz, relaxation delay of 7 s, pulse angle of 30°, acquisition time of 2.3 s and 64 scans. Spectra were processed by applying a line broadening of 0.2 Hz before Fourier transformation, and manually phased and baseline corrected. Sodium fumarate (6.50 ppm) was used as an internal reference for chemical shifts and metabolite quantification. The NUTS-Pro NMR software (Acorn NMR, Inc, Fremont, CA, USA) was used for the processing of the spectra and the quantification of the metabolites. Peaks were assigned by comparing the acquired spectra with reference spectra in Chenomx (Chenomx Inc. Edmonton, Canada) and the Human Metabolome Database (HMDB) V 5.0. 1H-NMR metabolomics data was analyzed using Metaboanalyst 5.0 online (https://www.metaboanalyst.ca/). Missing data was replaced by feature mean and data was log-transformed. |
| Spectrometer Frequency: | 500 MHz |
| NMR Probe: | 5 mm TXI probe |
| NMR Solvent: | D2O |
| NMR Tube Size: | 3mm |
| Pulse Sequence: | zgpr |
| Water Suppression: | Yes |
| Receiver Gain: | 203 |
| Offset Frequency: | 4.696 ppm |
| Presaturation Power Level: | 50 dB |
| Temperature: | 25 |
| Number Of Scans: | 64 |
| Dummy Scans: | 2 |
| Acquisition Time: | 2.3 sec |
| Relaxation Delay: | 7 sec |
| Spectral Width: | 14 ppm |
| Line Broadening: | 0.2 Hz |
| Chemical Shift Ref Std: | Fumarate (doublet, 6.50 ppm) |
