L Simon1, K Murphy1, M B Shamsi1, L Liu1, B Emery1, K I Aston1, J Hotaling1, D T Carrell2. 1. Andrology and IVF Laboratory, Department of Surgery (Urology), University of Utah, Salt Lake City, UT 84108, USA. 2. Andrology and IVF Laboratory, Department of Surgery (Urology), University of Utah, Salt Lake City, UT 84108, USA Department of Obstetrics and Gynecology, University of Utah, Salt Lake City, UT 84108, USA Department of Human Genetics, University of Utah, Salt Lake City, UT 84108, USA douglas.carrell@hsc.utah.edu.
Abstract
STUDY QUESTION: Does sperm DNA damage affect early embryonic development? SUMMARY ANSWER: Increased sperm DNA damage adversely affects embryo quality starting at Day 2 of early embryonic development and continuing after embryo transfer, resulting in reduced implantation rates and pregnancy outcomes. WHAT IS KNOWN ALREADY: Abnormalities in the sperm DNA in the form of single and double strand breaks can be assessed by an alkaline Comet assay. Some prior studies have shown a strong paternal effect of sperm DNA damage on IVF outcome, including reduced fertilization, reduced embryo quality and cleavage rates, reduced numbers of embryos developing into blastocysts, increased percentage of embryos undergoing developmental arrest, and reduced implantation and pregnancy rates. STUDY DESIGN, SIZE, DURATION: A cross-sectional study of 215 men from infertile couples undergoing assisted reproduction techniques at the University of Utah Center for Reproductive Medicine. PARTICIPANTS/MATERIALS, SETTING, METHODS: Sperm from men undergoing ART were analyzed for DNA damage using an alkaline Comet assay and classified into three groups: 'low damage' (0-30%), 'intermediate damage' (31-70%) and 'high damage' (71-100%). The cause of couples' infertility was categorized into one of the three types (male, female or unexplained). Each embryo was categorized as 'good', 'fair' or 'poor' quality, based on the number and grade of blastomeres. The influence of sperm DNA damage on early embryonic development was observed and classified into four stages: peri-fertilization effect (fertilization rate), early paternal effect (embryonic days 1-2), late paternal effect (embryonic days 3-5) and implantation stage effect. MAIN RESULTS AND THE ROLE OF CHANCE: The paternal effect of sperm DNA damage was observed at each stage of early embryonic development. The peri-fertilization effect was higher in oocytes from patients with female infertility (20.85%) compared with male (8.22%; P < 0.001) and unexplained (7.30%; P < 0.001) infertility factors. In both the early and late paternal effect stages, the low DNA damage group had a higher percentage of good quality embryos (P < 0.05) and lower percentage of poor quality embryos (P < 0.05) compared with the high DNA damage group. Implantation was lower in the high DNA damage (33.33%) compared with intermediate DNA damage (55.26%; P < 0.001) and low DNA damage (65.00%; P < 0.001) groups. The implantation rate was higher following blastocyst transfer (58.33%), when compared with early stage blastocyst (53.85%; P = 0.554) and cavitating morula transfers (34.40%; P < 0.001). Implantation was higher when the female partner age was ≤35 years when compared with >35 year age group (52.75 versus 35.44%; P = 0.008). LIMITATIONS, REASONS FOR CAUTION: A potential limitation of this study is that it is cross-sectional. Generally in such studies more than one variable could affect the outcome. Analyzing sperm is one part of the equation but a number of environmental and female factors also have the potential to influence embryo development and implantation. Furthermore, the selection of morphologically normal and physiologically motile sperm may result in isolation of sperm with reduced DNA damage. Therefore, selecting the best available sperm for ICSI may lead to experimental bias, as the selected sperm do not represent the overall sperm population in which the DNA damage is measured. Similar studies on selected sperm and with a larger sample size are now required. WIDER IMPLICATIONS OF THE FINDINGS: The paternal influence of damaged chromatin is more prominent after zygotic transcriptional activation. A prolonged paternal effect on the developing embryo may be due to the active repair mechanism present in oocytes that tends to overcome the damaged paternal chromatin. The probability of eliminating an embryo fertilized by a sperm with damaged DNA is higher at the blastocyst stage than the cleavage stage; therefore blastocyst transfer could be recommended for better implantation success. Finally, we recommend ICSI treatment for patients with a higher percentage of sperm with DNA damage as well as additional studies with a larger sample size aimed at assessing DNA damage analysis as a diagnostic tool for IVF. STUDY FUNDING/COMPETING INTERESTS: This work was supported by the University of Utah internal funds. The authors declare no competing interests. TRIAL REGISTRATION NUMBER: N/A.
STUDY QUESTION: Does sperm DNA damage affect early embryonic development? SUMMARY ANSWER: Increased sperm DNA damage adversely affects embryo quality starting at Day 2 of early embryonic development and continuing after embryo transfer, resulting in reduced implantation rates and pregnancy outcomes. WHAT IS KNOWN ALREADY: Abnormalities in the sperm DNA in the form of single and double strand breaks can be assessed by an alkaline Comet assay. Some prior studies have shown a strong paternal effect of sperm DNA damage on IVF outcome, including reduced fertilization, reduced embryo quality and cleavage rates, reduced numbers of embryos developing into blastocysts, increased percentage of embryos undergoing developmental arrest, and reduced implantation and pregnancy rates. STUDY DESIGN, SIZE, DURATION: A cross-sectional study of 215 men from infertile couples undergoing assisted reproduction techniques at the University of Utah Center for Reproductive Medicine. PARTICIPANTS/MATERIALS, SETTING, METHODS: Sperm from men undergoing ART were analyzed for DNA damage using an alkaline Comet assay and classified into three groups: 'low damage' (0-30%), 'intermediate damage' (31-70%) and 'high damage' (71-100%). The cause of couples' infertility was categorized into one of the three types (male, female or unexplained). Each embryo was categorized as 'good', 'fair' or 'poor' quality, based on the number and grade of blastomeres. The influence of sperm DNA damage on early embryonic development was observed and classified into four stages: peri-fertilization effect (fertilization rate), early paternal effect (embryonic days 1-2), late paternal effect (embryonic days 3-5) and implantation stage effect. MAIN RESULTS AND THE ROLE OF CHANCE: The paternal effect of sperm DNA damage was observed at each stage of early embryonic development. The peri-fertilization effect was higher in oocytes from patients with female infertility (20.85%) compared with male (8.22%; P < 0.001) and unexplained (7.30%; P < 0.001) infertility factors. In both the early and late paternal effect stages, the low DNA damage group had a higher percentage of good quality embryos (P < 0.05) and lower percentage of poor quality embryos (P < 0.05) compared with the high DNA damage group. Implantation was lower in the high DNA damage (33.33%) compared with intermediate DNA damage (55.26%; P < 0.001) and low DNA damage (65.00%; P < 0.001) groups. The implantation rate was higher following blastocyst transfer (58.33%), when compared with early stage blastocyst (53.85%; P = 0.554) and cavitating morula transfers (34.40%; P < 0.001). Implantation was higher when the female partner age was ≤35 years when compared with >35 year age group (52.75 versus 35.44%; P = 0.008). LIMITATIONS, REASONS FOR CAUTION: A potential limitation of this study is that it is cross-sectional. Generally in such studies more than one variable could affect the outcome. Analyzing sperm is one part of the equation but a number of environmental and female factors also have the potential to influence embryo development and implantation. Furthermore, the selection of morphologically normal and physiologically motile sperm may result in isolation of sperm with reduced DNA damage. Therefore, selecting the best available sperm for ICSI may lead to experimental bias, as the selected sperm do not represent the overall sperm population in which the DNA damage is measured. Similar studies on selected sperm and with a larger sample size are now required. WIDER IMPLICATIONS OF THE FINDINGS: The paternal influence of damaged chromatin is more prominent after zygotic transcriptional activation. A prolonged paternal effect on the developing embryo may be due to the active repair mechanism present in oocytes that tends to overcome the damaged paternal chromatin. The probability of eliminating an embryo fertilized by a sperm with damaged DNA is higher at the blastocyst stage than the cleavage stage; therefore blastocyst transfer could be recommended for better implantation success. Finally, we recommend ICSI treatment for patients with a higher percentage of sperm with DNA damage as well as additional studies with a larger sample size aimed at assessing DNA damage analysis as a diagnostic tool for IVF. STUDY FUNDING/COMPETING INTERESTS: This work was supported by the University of Utah internal funds. The authors declare no competing interests. TRIAL REGISTRATION NUMBER: N/A.
Authors: Anton Neyer; Martin Zintz; Astrid Stecher; Magnus Bach; Barbara Wirleitner; Nicolas H Zech; Pierre Vanderzwalmen Journal: J Assist Reprod Genet Date: 2015-08-19 Impact factor: 3.412
Authors: Sandro C Esteves; Armand Zini; Robert Matthew Coward; Donald P Evenson; Jaime Gosálvez; Sheena E M Lewis; Rakesh Sharma; Peter Humaidan Journal: Andrologia Date: 2020-10-27 Impact factor: 2.775