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In vitro effect of granulocyte–macrophage colony-stimulating factor (GM-CSF) on the expression of genes related to sperm motility and energy metabolism and intracytoplasmic sperm injection outcomes in obstructive azoospermic patients

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Abstract

Background

The presence of granulocyte–macrophage colony-stimulating factor (GM-CSF) and its receptor in various testicular cells and spermatozoa suggests a potential role in enhancing spermatogonial and postmeiotic cell development. Moreover, GM-CSF activates the pivotal pathways implicated in sperm motility regulation and glucose metabolism. However, the impact of GM-CSF on testicular biopsies from patients with obstructive azoospermia (OA) remains unexplored. Therefore, this study aimed to investigate the in vitro effects of GM-CSF on the expression of genes related to glucose transporters and signaling pathways, sperm motility, and viability in testicular biopsies.

Methods and results

Following testicular sperm extraction from 20 patients diagnosed with OA, each sample was divided into two parts: the experimental samples were incubated with medium containing 2 ng/ml GM-CSF at 37 °C for 60 min, and the control samples were incubated with medium without GM-CSF. Subsequently, the oocytes retrieved from the partner were injected with sperm from the treatment and control groups. The sperm parameters (motility and viability), the expression levels of sperm motility-related genes (PIK3R1, PIK3CA, and AKT1), and the expression levels of sperm energy metabolism-related genes (GLUT1, GLUT3, and GLUT14) were assessed. Furthermore, the fertilization and day 3 embryo development rate and embryo quality were evaluated. Compared with those in the nontreated group, the motility parameters and the mRNA expression levels of PIK3R1, AKT1, and GLUT3 in testicular sperm supplemented with GM-CSF were significantly greater (p < 0.05). However, no significant differences in the mRNA expression of PIK3CA, GLUT1, or GLUT14 were detected. According to the ICSI results, compared with the control group, the GM-CSF treatment group exhibited significantly greater fertilization rates (p = 0.027), Day 3 embryo development rate (p = 0.001), and proportions of good-quality embryos (p = 0.002).

Conclusions

GM-CSF increased the expression of genes related to motility and the energy metabolism pathway and effectively promoted the motility of testis-extracted spermatozoa, consequently yielding positive clinical outcomes.

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Availability of data and materials

All data generated or analyzed during this study are included in this published article.

Abbreviations

GM-CSF:

Granulocyte–macrophage colony-stimulating factor

ICSI:

Intracytoplasmic sperm injection

ART:

Assisted reproductive technology

TESE:

Testicular sperm extraction

PTF:

Pentoxifylline

GLUTs:

Glucose transporters

GLUT1:

Glucose transporter 1

PI3K/AKT:

Phosphoinositide-3-kinase/protein kinase B

OAT:

Oligoasthenoteratospermia

GnRH:

Gonadotropin-releasing hormone

hCG:

Human chorionic gonadotrophin

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Acknowledgements

 The authors would like to thank the Clinical Research Development Unit of Ayatollah Mousavi Hospital, Zanjan University of Medical Sciences, Zanjan, Iran for their cooperation and assistance throughout the period of study.

Funding

This study was financially supported by a grant from the Zanjan University of Medical Sciences (Grant number: A-11-1314-1).

Author information

Authors and Affiliations

  1. Department of Anatomical Sciences and Pathology, School of Medicine, North Khorasan University of Medical Sciences, Bojnurd, Iran

    Fatemeh Tanhaye Kalate Sabz, Fatemeh Sadat Amjadi & Zahra Zandieh

  2. Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Fatemeh Tanhaye Kalate Sabz

  3. Department of Obstetrics and Gynecology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    Elham Hosseini, Masoud Mohammadian & Farnaz Mohammadian

  4. Zanjan Metabolic Diseases Research Center, Zanjan University of Medical Sciences, Zanjan, Iran

    Elham Hosseini

  5. Shahid Akbarabadi Clinical Research Development Unit (ShACRDU), Iran University of Medical Sciences, Tehran, Iran

    Fatemeh Sadat Amjadi, Zahra Zandieh & Mahnaz Ashrafi

  6. Department of Microbiology, Biology Research Center, Islamic Azad University, Zanjan Branch, Zanjan, Iran

    Masoud Mohammadian

  7. Department of Biology, Faculty of Basic Sciences, University of Maragheh, Maragheh, Iran

    Raheleh Kafaeinezhad

  8. Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    Mahnaz Ashrafi

  9. Mousavi Hospital, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    Masoud Mohammadian

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  1. Fatemeh Tanhaye Kalate Sabz
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Contributions

FTKS: Original draft preparation. EH., FSA., ZZ., and MM: Conceptualization and Methodology. EH., and FSA: Supervision. MA., and FM: patient recruitment. FTKS., and RK: Performing laboratory work, collecting the data and analysis. All the authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Elham Hosseini or Fatemeh Sadat Amjadi.

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The authors declare that they have no conflict of interest.

Ethics approval and consent to participate

The experimental protocol was approved by the ethical committee of Zanjan University of Medical Sciences, Zanjan, Iran, (IR.ZUMS.REC.1398.357). All participants signed informed consent to participate.

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Tanhaye Kalate Sabz, F., Hosseini, E., Amjadi, F.S. et al. In vitro effect of granulocyte–macrophage colony-stimulating factor (GM-CSF) on the expression of genes related to sperm motility and energy metabolism and intracytoplasmic sperm injection outcomes in obstructive azoospermic patients. Mol Biol Rep 51, 727 (2024). https://doi.org/10.1007/s11033-024-09676-2

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