Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector

Version 1 : Received: 1 July 2021 / Approved: 1 July 2021 / Online: 1 July 2021 (14:30:04 CEST)

How to cite: Sugasawa, T.; Nakano, T.; Fujita, S.; Matsumoto, Y.; Ishihara, G.; Aoki, K.; Yanazawa, K.; Ono, S.; Tamai, S.; Manevich, L.; Ueda, H.; Ishibashi, N.; Tamai, K.; Kanki, Y.; Yoshida, Y.; Watanabe, K.; Takemasa, T.; Kawakami, Y.; Takekoshi, K. Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector. Preprints 2021, 2021070034 (doi: 10.20944/preprints202107.0034.v1). Sugasawa, T.; Nakano, T.; Fujita, S.; Matsumoto, Y.; Ishihara, G.; Aoki, K.; Yanazawa, K.; Ono, S.; Tamai, S.; Manevich, L.; Ueda, H.; Ishibashi, N.; Tamai, K.; Kanki, Y.; Yoshida, Y.; Watanabe, K.; Takemasa, T.; Kawakami, Y.; Takekoshi, K. Proof of Gene Doping in a Mouse Model with a Human Erythropoietin Gene Transferred Using an Adenoviral Vector. Preprints 2021, 2021070034 (doi: 10.20944/preprints202107.0034.v1).

Abstract

The World Anti-Doping Agency (WADA) has prohibited gene doping in the context of progress in gene therapy. In addition, there is a risk of the EPO gene being applied in gene doping among athletes. Along with this, development of a gene-doping test has been underway in worldwide. Here, we had two purposes: to develop a robust gene doping mouse model using the human EPO gene (hEPO) transferred using recombinant adenovirus (rAdV) as a vector and to develop a detection method to prove gene doping using this model. The rAdV including the hEPO gene were injected intravenously to transfer the gene to the liver. After injection, the mice developed significantly increased red blood cell counts in whole blood and increased gene expressions of hematopoietic markers in the spleen, indicating successful development of the gene doping model. Next, we detected direct and indirect proof of gene doping in whole blood DNA and RNA using qPCR assay and RNA sequencing. Proof was detected in one drop of whole blood DNA and RNA over a long period; furthermore, the overall RNA expression profiles significantly changed. Therefore, we have advanced detection of hEPO gene doping in humans.

Subject Areas

Gene doping; Gene therapy; Erythropoietin; Adenoviral vector; Sports; Athlete; RNA sequencing

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