e n t e r of o r R epr d uc ti v e a n e IN MEN WITH ... · Ashok Agarwal, PhD1, Ana D. Martins,...
Transcript of e n t e r of o r R epr d uc ti v e a n e IN MEN WITH ... · Ashok Agarwal, PhD1, Ana D. Martins,...
SPERM QUALITY AND PROTEOMIC PROFILE IN MEN WITH HODGKIN’S LYMPHOMA
1American Center for Reproductive Medicine, Cleveland Clinic, OH, 2Department of Microscopia, Lab of Biologia Celular/Unit for Multidisciplinary Research in Biomedicine, Inst de Ciências Biomédicas Abel Salazar-Universidade do Porto 3Center of Excellence in Genomic Medicine Research, Jeddah, Saudi Arabia, 4Cleveland Clinic, Proteomic Core Services- Lerner Research Institute, Cleveland Clinic, OH, 5Department of Urology, Cleveland Clinic, Cleveland, OH
Ashok Agarwal, PhD1, Ana D. Martins, MS1,2, Peter N. Pushparaj, PhD3, Belinda B. Willard, PhD4, Edmund Sabanegh, MD5 and Rakesh Sharma, PhD1
Introduction Results
Conclusion
• Hodgkin’s disease (HD) affects more than 45% of males of reproductive age. Sperm banking is offered for preserving fertility potential prior to cancer treatment.
• Reduced male fertility often appears even before the start of their treatment.
This study aims to analyze sperm parameters and alterations in proteomic profiles of spermatozoa from men with HD undergoing sperm banking before cancer therapy.
Experimental DesignSpermatozoa
Proven Fertile Donors(n=5)
Protein Extraction
Proteomic Study (Liquid Chromatography Tandem Mass-Spectrometry)
Bioinformatic Tools
Patients with Hodgkin’s Lymphoma
(n=6)
Identification of Differently Expressed Proteins (DEPs)
Validation of DEPs by Western Blot
Semen parameters in fertile men and patients with Hodgkin's Lymphoma The IPA analysis of DEP’s in Hodgkin’s Lymphoma
The top diseases and disorders dysregulated in spermatozoa of patient's with Hodgkin's Lymphoma
Top inhibited upstream regulators
Global proteomic analysis identified 1169 proteins
Molecular and cellular functions impaired in the spermatozoa of patients
Fertile Man
Volume ± SD (mL)
Concentration ±SD (106 /mL)
Motility ±SD (%)
Total Count ±SD (106)
Total Motile Sperm±SD (106)
3.8 ± 1.9
3.5 ± 1.8
0.50 0.01 0.44 0.02 0.02
73.8 ± 37.4
63.5 ± 64.2
58.4 ± 12.4
53.6 ± 24.5
267.8 ± 175.0
224 ± 268.7
156.7 ± 16.7
134.8 ± 30.8Patient
p-value
DEPs - 134 Proteins
35 80
16
3
Underexpressed
Overexpressed
Unique to proven fertile
Unique to HD
Categories # Molecules
Cell -To-Cell Signaling and Interaction, Reproductive System Development and Function
1.3x10-9 10
Cancer, Organismal Injury and Abnormalities 1.92x10-7 119
Cell -To-Cell Signaling and Interaction 2.26x10-6 13
Cancer, Organismal Injury and Abnormalities 3.45x10-6
p-Value
Upstream Regulator
HSF2 1.2x10-14 Inhibited
3.3x10-9 Inhibited
HSF1 1.0x10-8 Inhibited
1,2-dithiol-3-thione
p-Value of overlap
Predicted Activation
1) Semen quality was significantly decreased in men with HD relative to fertile men.
2) Proteomic data showed an altered proteome in spermatozoa of men with HD, which may explain how these alterations can potentially compromise the fertility in these men.
3) Identification of therapeutic strategies is needed to increase the quality of sperm and successful pregnancies.
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0
Cell-to-Cell Signaling and Interaction
Reproductive System Development and Function
Celluar Assembly and Organization
Protein Degradation
Protein Synthesis
Cancer
Organismal Injury and Abnormalities
Hematological System Development and Function
Immune Cell Trafficking
Inflammatory Response
Post-Translational Modification
Reproductive System Disease
Threshold
-log(B-H p-value)
Amer
ican
Center for Reproductive Medicine
Centrum Excellentiae Est.
1993
18-140835
119