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An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq

Alexander Kolliari-Turner Orcid Logo, Giscard Lima Orcid Logo, Guan Wang Orcid Logo, Fernanda Rossell Malinsky, Antonia Karanikolou, Gregor Eichhorn, Kumpei Tanisawa Orcid Logo, Jonathan Ospina-Betancurt Orcid Logo, Blair Hamilton Orcid Logo, Paulette Y.O. Kumi Orcid Logo, Jonathan Shurlock Orcid Logo, Vasileios Skiadas, Richard Twycross-Lewis Orcid Logo, Liam Kilduff Orcid Logo, Renan Paulo Martin Orcid Logo, Garrett I. Ash Orcid Logo, Cynthia Potter, Fergus M. Guppy Orcid Logo, Jane T. Seto Orcid Logo, Chiara Fossati Orcid Logo, Fabio Pigozzi Orcid Logo, Paolo Borrione Orcid Logo, Yannis Pitsiladis Orcid Logo

BMC Medical Genomics, Volume: 16, Issue: 1

Swansea University Author: Liam Kilduff Orcid Logo

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DOI (Published version): 10.1186/s12920-023-01512-z

Abstract

BackgroundThe effects of Anabolic Androgenic Steroids (AAS) are largely illustrated through Androgen Receptor induced gene transcription, yet RNA-Seq has yet to be conducted on human whole blood and skeletal muscle. Investigating the transcriptional signature of AAS in blood may aid AAS detection an...

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ISSN: 1755-8794
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fullrecord <?xml version="1.0" encoding="utf-8"?><rfc1807 xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:xsd="http://www.w3.org/2001/XMLSchema"><bib-version>v2</bib-version><id>63140</id><entry>2023-04-13</entry><title>An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq</title><swanseaauthors><author><sid>972ed9a1dda7a0de20581a0f8350be98</sid><ORCID>0000-0001-9449-2293</ORCID><firstname>Liam</firstname><surname>Kilduff</surname><name>Liam Kilduff</name><active>true</active><ethesisStudent>false</ethesisStudent></author></swanseaauthors><date>2023-04-13</date><deptcode>STSC</deptcode><abstract>BackgroundThe effects of Anabolic Androgenic Steroids (AAS) are largely illustrated through Androgen Receptor induced gene transcription, yet RNA-Seq has yet to be conducted on human whole blood and skeletal muscle. Investigating the transcriptional signature of AAS in blood may aid AAS detection and in muscle further understanding of AAS induced hypertrophy.MethodsMales aged 20–42 were recruited and sampled once: sedentary controls (C), resistance trained lifters (RT) and resistance trained current AAS users (RT-AS) who ceased exposure ≤ 2 or ≥ 10 weeks prior to sampling. RT-AS were sampled twice as Returning Participants (RP) if AAS usage ceased for ≥ 18 weeks. RNA was extracted from whole blood and trapezius muscle samples. RNA libraries were sequenced twice, for validation purposes, on the DNBSEQ-G400RS with either standard or CoolMPS PE100 reagents following MGI protocols. Genes were considered differentially expressed with FDR &lt; 0.05 and a 1.2- fold change.ResultsCross-comparison of both standard reagent whole blood (N = 55: C = 7, RT = 20, RT-AS ≤ 2 = 14, RT-AS ≥ 10 = 10, RP = 4; N = 46: C = 6, RT = 17, RT-AS ≤ 2 = 12, RT-AS ≥ 10 = 8, RP = 3) sequencing datasets, showed that no genes or gene sets/pathways were differentially expressed between time points for RP or between group comparisons of RT-AS ≤ 2 vs. C, RT, or RT-AS ≥ 10. Cross-comparison of both muscle (N = 51, C = 5, RT = 17, RT-AS ≤ 2 = 15, RT-AS ≥ 10 = 11, RP = 3) sequencing (one standard &amp; one CoolMPS reagent) datasets, showed one gene, CHRDL1, which has atrophying potential, was upregulated in RP visit two. In both muscle sequencing datasets, nine differentially expressed genes, overlapped with RT-AS ≤ 2 vs. RT and RT-AS ≤ 2 vs. C, but were not differentially expressed with RT vs. C, possibly suggesting they are from acute doping alone. No genes seemed to be differentially expressed in muscle after the long-term cessation of AAS, whereas a previous study found long term proteomic changes.ConclusionA whole blood transcriptional signature of AAS doping was not identified. However, RNA-Seq of muscle has identified numerous differentially expressed genes with known impacts on hypertrophic processes that may further our understanding on AAS induced hypertrophy. Differences in training regimens in participant groupings may have influenced results. Future studies should focus on longitudinal sampling pre, during and post-AAS exposure to better control for confounding variables.</abstract><type>Journal Article</type><journal>BMC Medical Genomics</journal><volume>16</volume><journalNumber>1</journalNumber><paginationStart/><paginationEnd/><publisher>Springer Science and Business Media LLC</publisher><placeOfPublication/><isbnPrint/><isbnElectronic/><issnPrint/><issnElectronic>1755-8794</issnElectronic><keywords>Anabolic androgenic steroids, Doping, Skeletal muscle, Whole blood, RNA-Seq, Gene expression, Hypertrophy.</keywords><publishedDay>0</publishedDay><publishedMonth>0</publishedMonth><publishedYear>0</publishedYear><publishedDate>0001-01-01</publishedDate><doi>10.1186/s12920-023-01512-z</doi><url>http://dx.doi.org/10.1186/s12920-023-01512-z</url><notes/><college>COLLEGE NANME</college><department>Sport and Exercise Sciences</department><CollegeCode>COLLEGE CODE</CollegeCode><DepartmentCode>STSC</DepartmentCode><institution>Swansea University</institution><apcterm>External research funder(s) paid the OA fee (includes OA grants disbursed by the Library)</apcterm><funders>The research and publication costs were funded by a research grant (16E11FP) from the World Anti-Doping Agency. This sponsor did not have a role in the design of the study and in collection, analysis, and interpretation of data, or in writing the manuscript.</funders><projectreference/><lastEdited>2023-05-18T14:32:35.5260908</lastEdited><Created>2023-04-13T11:13:50.7519477</Created><path><level id="1">Faculty of Science and Engineering</level><level id="2">School of Engineering and Applied Sciences - Sport and Exercise Sciences</level></path><authors><author><firstname>Alexander</firstname><surname>Kolliari-Turner</surname><orcid>0000-0002-2469-7645</orcid><order>1</order></author><author><firstname>Giscard</firstname><surname>Lima</surname><orcid>0000-0003-3781-9522</orcid><order>2</order></author><author><firstname>Guan</firstname><surname>Wang</surname><orcid>0000-0002-7432-7933</orcid><order>3</order></author><author><firstname>Fernanda Rossell</firstname><surname>Malinsky</surname><order>4</order></author><author><firstname>Antonia</firstname><surname>Karanikolou</surname><order>5</order></author><author><firstname>Gregor</firstname><surname>Eichhorn</surname><order>6</order></author><author><firstname>Kumpei</firstname><surname>Tanisawa</surname><orcid>0000-0002-9334-7104</orcid><order>7</order></author><author><firstname>Jonathan</firstname><surname>Ospina-Betancurt</surname><orcid>0000-0001-7999-9221</orcid><order>8</order></author><author><firstname>Blair</firstname><surname>Hamilton</surname><orcid>0000-0001-7412-1188</orcid><order>9</order></author><author><firstname>Paulette Y.O.</firstname><surname>Kumi</surname><orcid>0000-0002-5057-995x</orcid><order>10</order></author><author><firstname>Jonathan</firstname><surname>Shurlock</surname><orcid>0000-0001-8681-6534</orcid><order>11</order></author><author><firstname>Vasileios</firstname><surname>Skiadas</surname><order>12</order></author><author><firstname>Richard</firstname><surname>Twycross-Lewis</surname><orcid>0000-0001-9564-333x</orcid><order>13</order></author><author><firstname>Liam</firstname><surname>Kilduff</surname><orcid>0000-0001-9449-2293</orcid><order>14</order></author><author><firstname>Renan Paulo</firstname><surname>Martin</surname><orcid>0000-0002-6747-8784</orcid><order>15</order></author><author><firstname>Garrett I.</firstname><surname>Ash</surname><orcid>0000-0002-8655-7525</orcid><order>16</order></author><author><firstname>Cynthia</firstname><surname>Potter</surname><order>17</order></author><author><firstname>Fergus M.</firstname><surname>Guppy</surname><orcid>0000-0002-8526-9169</orcid><order>18</order></author><author><firstname>Jane T.</firstname><surname>Seto</surname><orcid>0000-0003-2864-6199</orcid><order>19</order></author><author><firstname>Chiara</firstname><surname>Fossati</surname><orcid>0000-0002-2870-7185</orcid><order>20</order></author><author><firstname>Fabio</firstname><surname>Pigozzi</surname><orcid>0000-0001-5808-9405</orcid><order>21</order></author><author><firstname>Paolo</firstname><surname>Borrione</surname><orcid>0000-0003-0506-169x</orcid><order>22</order></author><author><firstname>Yannis</firstname><surname>Pitsiladis</surname><orcid>0000-0001-6210-2449</orcid><order>23</order></author></authors><documents><document><filename>63140__27429__9a03bde21c154ff685f630cd88d40a7e.pdf</filename><originalFilename>63140.pdf</originalFilename><uploaded>2023-05-11T11:39:55.9767576</uploaded><type>Output</type><contentLength>2422648</contentLength><contentType>application/pdf</contentType><version>Version of Record</version><cronfaStatus>true</cronfaStatus><documentNotes>Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material.</documentNotes><copyrightCorrect>true</copyrightCorrect><language>eng</language><licence>http://creativecommons.org/licenses/by/4.0/</licence></document></documents><OutputDurs/></rfc1807>
spelling v2 63140 2023-04-13 An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq 972ed9a1dda7a0de20581a0f8350be98 0000-0001-9449-2293 Liam Kilduff Liam Kilduff true false 2023-04-13 STSC BackgroundThe effects of Anabolic Androgenic Steroids (AAS) are largely illustrated through Androgen Receptor induced gene transcription, yet RNA-Seq has yet to be conducted on human whole blood and skeletal muscle. Investigating the transcriptional signature of AAS in blood may aid AAS detection and in muscle further understanding of AAS induced hypertrophy.MethodsMales aged 20–42 were recruited and sampled once: sedentary controls (C), resistance trained lifters (RT) and resistance trained current AAS users (RT-AS) who ceased exposure ≤ 2 or ≥ 10 weeks prior to sampling. RT-AS were sampled twice as Returning Participants (RP) if AAS usage ceased for ≥ 18 weeks. RNA was extracted from whole blood and trapezius muscle samples. RNA libraries were sequenced twice, for validation purposes, on the DNBSEQ-G400RS with either standard or CoolMPS PE100 reagents following MGI protocols. Genes were considered differentially expressed with FDR < 0.05 and a 1.2- fold change.ResultsCross-comparison of both standard reagent whole blood (N = 55: C = 7, RT = 20, RT-AS ≤ 2 = 14, RT-AS ≥ 10 = 10, RP = 4; N = 46: C = 6, RT = 17, RT-AS ≤ 2 = 12, RT-AS ≥ 10 = 8, RP = 3) sequencing datasets, showed that no genes or gene sets/pathways were differentially expressed between time points for RP or between group comparisons of RT-AS ≤ 2 vs. C, RT, or RT-AS ≥ 10. Cross-comparison of both muscle (N = 51, C = 5, RT = 17, RT-AS ≤ 2 = 15, RT-AS ≥ 10 = 11, RP = 3) sequencing (one standard & one CoolMPS reagent) datasets, showed one gene, CHRDL1, which has atrophying potential, was upregulated in RP visit two. In both muscle sequencing datasets, nine differentially expressed genes, overlapped with RT-AS ≤ 2 vs. RT and RT-AS ≤ 2 vs. C, but were not differentially expressed with RT vs. C, possibly suggesting they are from acute doping alone. No genes seemed to be differentially expressed in muscle after the long-term cessation of AAS, whereas a previous study found long term proteomic changes.ConclusionA whole blood transcriptional signature of AAS doping was not identified. However, RNA-Seq of muscle has identified numerous differentially expressed genes with known impacts on hypertrophic processes that may further our understanding on AAS induced hypertrophy. Differences in training regimens in participant groupings may have influenced results. Future studies should focus on longitudinal sampling pre, during and post-AAS exposure to better control for confounding variables. Journal Article BMC Medical Genomics 16 1 Springer Science and Business Media LLC 1755-8794 Anabolic androgenic steroids, Doping, Skeletal muscle, Whole blood, RNA-Seq, Gene expression, Hypertrophy. 0 0 0 0001-01-01 10.1186/s12920-023-01512-z http://dx.doi.org/10.1186/s12920-023-01512-z COLLEGE NANME Sport and Exercise Sciences COLLEGE CODE STSC Swansea University External research funder(s) paid the OA fee (includes OA grants disbursed by the Library) The research and publication costs were funded by a research grant (16E11FP) from the World Anti-Doping Agency. This sponsor did not have a role in the design of the study and in collection, analysis, and interpretation of data, or in writing the manuscript. 2023-05-18T14:32:35.5260908 2023-04-13T11:13:50.7519477 Faculty of Science and Engineering School of Engineering and Applied Sciences - Sport and Exercise Sciences Alexander Kolliari-Turner 0000-0002-2469-7645 1 Giscard Lima 0000-0003-3781-9522 2 Guan Wang 0000-0002-7432-7933 3 Fernanda Rossell Malinsky 4 Antonia Karanikolou 5 Gregor Eichhorn 6 Kumpei Tanisawa 0000-0002-9334-7104 7 Jonathan Ospina-Betancurt 0000-0001-7999-9221 8 Blair Hamilton 0000-0001-7412-1188 9 Paulette Y.O. Kumi 0000-0002-5057-995x 10 Jonathan Shurlock 0000-0001-8681-6534 11 Vasileios Skiadas 12 Richard Twycross-Lewis 0000-0001-9564-333x 13 Liam Kilduff 0000-0001-9449-2293 14 Renan Paulo Martin 0000-0002-6747-8784 15 Garrett I. Ash 0000-0002-8655-7525 16 Cynthia Potter 17 Fergus M. Guppy 0000-0002-8526-9169 18 Jane T. Seto 0000-0003-2864-6199 19 Chiara Fossati 0000-0002-2870-7185 20 Fabio Pigozzi 0000-0001-5808-9405 21 Paolo Borrione 0000-0003-0506-169x 22 Yannis Pitsiladis 0000-0001-6210-2449 23 63140__27429__9a03bde21c154ff685f630cd88d40a7e.pdf 63140.pdf 2023-05-11T11:39:55.9767576 Output 2422648 application/pdf Version of Record true Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. true eng http://creativecommons.org/licenses/by/4.0/
title An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq
spellingShingle An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq
Liam Kilduff
title_short An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq
title_full An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq
title_fullStr An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq
title_full_unstemmed An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq
title_sort An observational human study investigating the effect of anabolic androgenic steroid use on the transcriptome of skeletal muscle and whole blood using RNA-Seq
author_id_str_mv 972ed9a1dda7a0de20581a0f8350be98
author_id_fullname_str_mv 972ed9a1dda7a0de20581a0f8350be98_***_Liam Kilduff
author Liam Kilduff
author2 Alexander Kolliari-Turner
Giscard Lima
Guan Wang
Fernanda Rossell Malinsky
Antonia Karanikolou
Gregor Eichhorn
Kumpei Tanisawa
Jonathan Ospina-Betancurt
Blair Hamilton
Paulette Y.O. Kumi
Jonathan Shurlock
Vasileios Skiadas
Richard Twycross-Lewis
Liam Kilduff
Renan Paulo Martin
Garrett I. Ash
Cynthia Potter
Fergus M. Guppy
Jane T. Seto
Chiara Fossati
Fabio Pigozzi
Paolo Borrione
Yannis Pitsiladis
format Journal article
container_title BMC Medical Genomics
container_volume 16
container_issue 1
institution Swansea University
issn 1755-8794
doi_str_mv 10.1186/s12920-023-01512-z
publisher Springer Science and Business Media LLC
college_str Faculty of Science and Engineering
hierarchytype
hierarchy_top_id facultyofscienceandengineering
hierarchy_top_title Faculty of Science and Engineering
hierarchy_parent_id facultyofscienceandengineering
hierarchy_parent_title Faculty of Science and Engineering
department_str School of Engineering and Applied Sciences - Sport and Exercise Sciences{{{_:::_}}}Faculty of Science and Engineering{{{_:::_}}}School of Engineering and Applied Sciences - Sport and Exercise Sciences
url http://dx.doi.org/10.1186/s12920-023-01512-z
document_store_str 1
active_str 0
description BackgroundThe effects of Anabolic Androgenic Steroids (AAS) are largely illustrated through Androgen Receptor induced gene transcription, yet RNA-Seq has yet to be conducted on human whole blood and skeletal muscle. Investigating the transcriptional signature of AAS in blood may aid AAS detection and in muscle further understanding of AAS induced hypertrophy.MethodsMales aged 20–42 were recruited and sampled once: sedentary controls (C), resistance trained lifters (RT) and resistance trained current AAS users (RT-AS) who ceased exposure ≤ 2 or ≥ 10 weeks prior to sampling. RT-AS were sampled twice as Returning Participants (RP) if AAS usage ceased for ≥ 18 weeks. RNA was extracted from whole blood and trapezius muscle samples. RNA libraries were sequenced twice, for validation purposes, on the DNBSEQ-G400RS with either standard or CoolMPS PE100 reagents following MGI protocols. Genes were considered differentially expressed with FDR < 0.05 and a 1.2- fold change.ResultsCross-comparison of both standard reagent whole blood (N = 55: C = 7, RT = 20, RT-AS ≤ 2 = 14, RT-AS ≥ 10 = 10, RP = 4; N = 46: C = 6, RT = 17, RT-AS ≤ 2 = 12, RT-AS ≥ 10 = 8, RP = 3) sequencing datasets, showed that no genes or gene sets/pathways were differentially expressed between time points for RP or between group comparisons of RT-AS ≤ 2 vs. C, RT, or RT-AS ≥ 10. Cross-comparison of both muscle (N = 51, C = 5, RT = 17, RT-AS ≤ 2 = 15, RT-AS ≥ 10 = 11, RP = 3) sequencing (one standard & one CoolMPS reagent) datasets, showed one gene, CHRDL1, which has atrophying potential, was upregulated in RP visit two. In both muscle sequencing datasets, nine differentially expressed genes, overlapped with RT-AS ≤ 2 vs. RT and RT-AS ≤ 2 vs. C, but were not differentially expressed with RT vs. C, possibly suggesting they are from acute doping alone. No genes seemed to be differentially expressed in muscle after the long-term cessation of AAS, whereas a previous study found long term proteomic changes.ConclusionA whole blood transcriptional signature of AAS doping was not identified. However, RNA-Seq of muscle has identified numerous differentially expressed genes with known impacts on hypertrophic processes that may further our understanding on AAS induced hypertrophy. Differences in training regimens in participant groupings may have influenced results. Future studies should focus on longitudinal sampling pre, during and post-AAS exposure to better control for confounding variables.
published_date 0001-01-01T14:32:34Z
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