Environmental and Molecular Mutagenesis 00:00^00 (2014)
Research Article The Relationship Between Exposure to Air Pollution and Sperm Disomy Joanna Jurewicz,1* Michal- Radwan,2 Wojciech Sobala,1 Kinga Pola nska,1 2 3 3 Pawel- Radwan, Lucjusz Jakubowski, Anna Ula nska, and Wojciech Hanke1 1
Department of Environmental Epidemiology, Nofer Institute of Occupational Medicine, Lodz, Poland 2 Department of Gynecology and Reproduction, ”Gameta” Hospital, Rzg o w, Poland 3 Department of Medical Genetics, Polish Mother’s Memorial Hospital— Research Institute, Lodz, Poland
The causes of the chromosome abnormalities have been studied for decades. It has been suggested that exposure to various environmental agents can induce chromosomal abnormalities in germ cells. This study was designed to address the hypothesis that exposure to specific air pollutants increases sperm disomy. The study population consisted of 212 men who were attending an infertility clinic for diagnostic purposes. They represented a subset of men in a multicenter parent study conducted in Poland to evaluate environmental factors and male fertility. Sperm aneuploidy for chromosomes 13, 18, 21, X, and Y was assessed using multicolor fluorescence in situ hybridization. Air quality data were obtained from the AirBase database. After adjusting for age, smoking, alcohol consumption, temperature (90 days), season, past dis-
eases, abstinence interval, distance from the monitoring station, concentration, motility and morphology, positive associations were observed between exposure to PM2.5 and disomy Y (P 5 0.001), sex chromosome disomy (P 5 0.05) and disomy 21 (P 5 0.03). Exposure to PM10 was associated with disomy 21 (P 5 0.02). Conversely, exposure to ozone, CO, SO2, and NOx did not affect sperm aneuploidy. A separate analysis conducted among men who were nonsmokers (n 5 117) showed that the relationship between PM2.5 and disomy Y and disomy 21 remained significant (P 5 0.01, P 5 0.05, respectively). The present findings indicate that exposure to air pollution induces sperm aneuploidy. Environ. Mol. MutaC 2014 Wiley Periodigen. 00:000–000, 2014. V cals, Inc.
Key words: semen; aneuploidy; air pollution; fluorescent in situ hybridization
INTRODUCTION Aneuploidy is the most common chromosome abnormality in humans, occurring in at least 0.3% of newborns [Abruzzo and Hassold, 1995]. In comparison with other species, humans appear to have a higher incidence of chromosome anomalies and it is estimated that up to 50% of conceptions are chromosomally abnormal [Bond and Chandley, 1993; Magli et al., 2000]. Most chromosome abnormalities are lethal and are lost during embryonic development, manifesting as infertility or spontaneous abortions. Chromosomally abnormal conceptions that survive to term include trisomy 13, 18, 21, and aneuploidy for the sex chromosomes [Martin, 2008]. Human spermatozoa carry many more chromosomal abnormalities than germ cells of experimental animals [Martin, 1984]. Sperm karyotyping studies have shown C 2014 Wiley Periodicals, Inc. V
that approximately 10% of human sperm from normal men have chromosomal abnormalities [Guttenbach et al., 1997]. Based on fluorescent in situ hybridization (FISH)
Grant sponsor: National Center for Research and Development in Poland (Epidemiology of reproductive hazards—multicenter study in Poland); Grant number: PBZ-MEiN-/8/2//2006; Grant sponsor: IMP 10.19/2014; Grant number: K140/P01/2007/1.2.1.2 (Lifestyle factors and semen quality). *Correspondence to: Joanna Jurewicz; Department of Environmental Epidemiology, Nofer Institute of Occupational Medicine, 8 Teresy St, 91-362 Lodz, Poland. E-mail: [email protected] Received 6 March 2014; provisionally accepted 5 June 2014; and in final form 17 June 2014 DOI 10.1002/em.21883 Published online 00 Month 2014 in Wiley Online Library (wileyonlinelibrary.com).
Environmental and Molecular Mutagenesis. DOI 10.1002/em 2
Jurewicz et al.
methods of sperm analysis, it is estimated that healthy males have 1–4% of sperm with some type of aneuploidy, but there is high level of interindividual variation [Downie et al., 1997; Rubes et al., 2005]. It has been suggested that one reason for this interindividual variability may be male exposure to environmental agents. Exposure to potential reproductive toxicants can induce structural aberrations (breaks and translocations) or change the number of chromosomes in sperm [Robbins et al., 1993]. A few recent epidemiological studies have examined the potential relationship between environmental exposures and sperm aneuploidy. A number of synthetic chemicals have been shown to mimic endogenous hormones and affect the normal pattern of reproductive development [Tyler et al., 1998]. In humans, levels of sperm disomy can be increased by environmental and lifestyle factors, such as alcohol abuse and heavy smoking [Robbins et al., 1997b; Rubes et al., 1998; Shi and Martin, 2000; Shi et al., 2001]. Intrinsic factors, such as age and DNA polymorphisms, have also been implicated. Several studies have reported an effect of age on sperm disomy [Griffin et al., 1995; Asada et al., 2000] whereas one of the largest aging study found no evidence of a paternal age effect on aneuploidy in human sperm in a nonclinical study population [Wyrobek et al., 2006]. Also, exposure to pesticides [Padungtod et al., 1999; Xia et al., 2004; Young et al., 2013] and cancer therapeutic agents [Martin et al., 1986] may be associated with sperm aneuploidy. To our best knowledge, only two studies have examined the association between air pollution and sperm aneuploidy [Robbins et al., 1999; Rubes et al., 2005]. Both studies were performed among young men, residents of the Teplice District in Northern Bohemia, Czech Republic, exposed to high levels of air pollution generated from the combustion of high-sulfur coal used for am et al., 1996]. A local industry and home heating [Sr significant association between exposure to high levels of air pollution and YY disomy was observed among those men [Robbins et al., 1999], however, Rubes et al. [2005] found no association between exposure to high level of air pollution and sperm aneuploidy. Differences in air pollution levels between the two studies may have contributed to the contrasting results. This study was designed to address the hypothesis that exposure to specific air pollutants affects sperm disomy. Several experimental details were implemented to increase the statistical power of detecting an effect, including: increasing the participant sample size; expanding outcome measures to include chromosomes 13, 18, 21; and, localizing air pollution exposure assessment to study participants’ ZIP code of residence. Air pollutants of interests were: ozone (O3), nitrogen oxides (NOx), sulfur dioxide (SO2), carbon monoxide (CO), particular matter 0.05). We examined six types of sperm disomy in these analysis: XY sperm (sperm FISH genotype: X-Y-18), disomy X (X-X-18), disomy Y (Y-Y-18), disomy 18, 21 and 13 (X-18-18 or Y-18-18, X-21-21 or Y-21-21 and X-13-13 and Y-13-13, respectively). Additionally, total sex chromosome disomy and total disomy were recorded. The aneuploidy frequencies were expressed as percentages.
Air Pollution Data and Exposure Assessment Air quality data were obtained from the AirBase database [http:// www.eea.europa.eu/data-and-maps/data/airbase-the-european-air-qualitydatabase-6#tab-data-by-country] to assess male exposure to air pollutants. AirBase is the air quality information system maintained by the European Environmental Agency through the European topic center on air pollution and climate change mitigation. It contains air quality data delivered annually under 97/101/EC Council Decision (EoI Decision). These data include ozone, particulate matter
Research Article The Relationship Between Exposure to Air Pollution and Sperm Disomy Joanna Jurewicz,1* Michal- Radwan,2 Wojciech Sobala,1 Kinga Pola nska,1 2 3 3 Pawel- Radwan, Lucjusz Jakubowski, Anna Ula nska, and Wojciech Hanke1 1
Department of Environmental Epidemiology, Nofer Institute of Occupational Medicine, Lodz, Poland 2 Department of Gynecology and Reproduction, ”Gameta” Hospital, Rzg o w, Poland 3 Department of Medical Genetics, Polish Mother’s Memorial Hospital— Research Institute, Lodz, Poland
The causes of the chromosome abnormalities have been studied for decades. It has been suggested that exposure to various environmental agents can induce chromosomal abnormalities in germ cells. This study was designed to address the hypothesis that exposure to specific air pollutants increases sperm disomy. The study population consisted of 212 men who were attending an infertility clinic for diagnostic purposes. They represented a subset of men in a multicenter parent study conducted in Poland to evaluate environmental factors and male fertility. Sperm aneuploidy for chromosomes 13, 18, 21, X, and Y was assessed using multicolor fluorescence in situ hybridization. Air quality data were obtained from the AirBase database. After adjusting for age, smoking, alcohol consumption, temperature (90 days), season, past dis-
eases, abstinence interval, distance from the monitoring station, concentration, motility and morphology, positive associations were observed between exposure to PM2.5 and disomy Y (P 5 0.001), sex chromosome disomy (P 5 0.05) and disomy 21 (P 5 0.03). Exposure to PM10 was associated with disomy 21 (P 5 0.02). Conversely, exposure to ozone, CO, SO2, and NOx did not affect sperm aneuploidy. A separate analysis conducted among men who were nonsmokers (n 5 117) showed that the relationship between PM2.5 and disomy Y and disomy 21 remained significant (P 5 0.01, P 5 0.05, respectively). The present findings indicate that exposure to air pollution induces sperm aneuploidy. Environ. Mol. MutaC 2014 Wiley Periodigen. 00:000–000, 2014. V cals, Inc.
Key words: semen; aneuploidy; air pollution; fluorescent in situ hybridization
INTRODUCTION Aneuploidy is the most common chromosome abnormality in humans, occurring in at least 0.3% of newborns [Abruzzo and Hassold, 1995]. In comparison with other species, humans appear to have a higher incidence of chromosome anomalies and it is estimated that up to 50% of conceptions are chromosomally abnormal [Bond and Chandley, 1993; Magli et al., 2000]. Most chromosome abnormalities are lethal and are lost during embryonic development, manifesting as infertility or spontaneous abortions. Chromosomally abnormal conceptions that survive to term include trisomy 13, 18, 21, and aneuploidy for the sex chromosomes [Martin, 2008]. Human spermatozoa carry many more chromosomal abnormalities than germ cells of experimental animals [Martin, 1984]. Sperm karyotyping studies have shown C 2014 Wiley Periodicals, Inc. V
that approximately 10% of human sperm from normal men have chromosomal abnormalities [Guttenbach et al., 1997]. Based on fluorescent in situ hybridization (FISH)
Grant sponsor: National Center for Research and Development in Poland (Epidemiology of reproductive hazards—multicenter study in Poland); Grant number: PBZ-MEiN-/8/2//2006; Grant sponsor: IMP 10.19/2014; Grant number: K140/P01/2007/1.2.1.2 (Lifestyle factors and semen quality). *Correspondence to: Joanna Jurewicz; Department of Environmental Epidemiology, Nofer Institute of Occupational Medicine, 8 Teresy St, 91-362 Lodz, Poland. E-mail: [email protected] Received 6 March 2014; provisionally accepted 5 June 2014; and in final form 17 June 2014 DOI 10.1002/em.21883 Published online 00 Month 2014 in Wiley Online Library (wileyonlinelibrary.com).
Environmental and Molecular Mutagenesis. DOI 10.1002/em 2
Jurewicz et al.
methods of sperm analysis, it is estimated that healthy males have 1–4% of sperm with some type of aneuploidy, but there is high level of interindividual variation [Downie et al., 1997; Rubes et al., 2005]. It has been suggested that one reason for this interindividual variability may be male exposure to environmental agents. Exposure to potential reproductive toxicants can induce structural aberrations (breaks and translocations) or change the number of chromosomes in sperm [Robbins et al., 1993]. A few recent epidemiological studies have examined the potential relationship between environmental exposures and sperm aneuploidy. A number of synthetic chemicals have been shown to mimic endogenous hormones and affect the normal pattern of reproductive development [Tyler et al., 1998]. In humans, levels of sperm disomy can be increased by environmental and lifestyle factors, such as alcohol abuse and heavy smoking [Robbins et al., 1997b; Rubes et al., 1998; Shi and Martin, 2000; Shi et al., 2001]. Intrinsic factors, such as age and DNA polymorphisms, have also been implicated. Several studies have reported an effect of age on sperm disomy [Griffin et al., 1995; Asada et al., 2000] whereas one of the largest aging study found no evidence of a paternal age effect on aneuploidy in human sperm in a nonclinical study population [Wyrobek et al., 2006]. Also, exposure to pesticides [Padungtod et al., 1999; Xia et al., 2004; Young et al., 2013] and cancer therapeutic agents [Martin et al., 1986] may be associated with sperm aneuploidy. To our best knowledge, only two studies have examined the association between air pollution and sperm aneuploidy [Robbins et al., 1999; Rubes et al., 2005]. Both studies were performed among young men, residents of the Teplice District in Northern Bohemia, Czech Republic, exposed to high levels of air pollution generated from the combustion of high-sulfur coal used for am et al., 1996]. A local industry and home heating [Sr significant association between exposure to high levels of air pollution and YY disomy was observed among those men [Robbins et al., 1999], however, Rubes et al. [2005] found no association between exposure to high level of air pollution and sperm aneuploidy. Differences in air pollution levels between the two studies may have contributed to the contrasting results. This study was designed to address the hypothesis that exposure to specific air pollutants affects sperm disomy. Several experimental details were implemented to increase the statistical power of detecting an effect, including: increasing the participant sample size; expanding outcome measures to include chromosomes 13, 18, 21; and, localizing air pollution exposure assessment to study participants’ ZIP code of residence. Air pollutants of interests were: ozone (O3), nitrogen oxides (NOx), sulfur dioxide (SO2), carbon monoxide (CO), particular matter 0.05). We examined six types of sperm disomy in these analysis: XY sperm (sperm FISH genotype: X-Y-18), disomy X (X-X-18), disomy Y (Y-Y-18), disomy 18, 21 and 13 (X-18-18 or Y-18-18, X-21-21 or Y-21-21 and X-13-13 and Y-13-13, respectively). Additionally, total sex chromosome disomy and total disomy were recorded. The aneuploidy frequencies were expressed as percentages.
Air Pollution Data and Exposure Assessment Air quality data were obtained from the AirBase database [http:// www.eea.europa.eu/data-and-maps/data/airbase-the-european-air-qualitydatabase-6#tab-data-by-country] to assess male exposure to air pollutants. AirBase is the air quality information system maintained by the European Environmental Agency through the European topic center on air pollution and climate change mitigation. It contains air quality data delivered annually under 97/101/EC Council Decision (EoI Decision). These data include ozone, particulate matter
