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