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Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway

Intracellular reactive oxygen species (ROS) play an important role in the proliferation and differentiation of hematopoietic stem and progenitor cells (HSPCs). HSPCs are difficult to be expanded ex vivo while maintaining their stemness when they are exposed to oxidative damage after being released f...

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Autores principales: Park, Ga-Bin, Kim, Min-Jung, Vasileva, Elena A., Mishchenko, Natalia P., Fedoreyev, Sergey A., Stonik, Valentin A., Han, Jin, Lee, Ho Sup, Kim, Daejin, Jeong, Jee-Yeong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6780187/
https://www.ncbi.nlm.nih.gov/pubmed/31505769
http://dx.doi.org/10.3390/md17090526
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author Park, Ga-Bin
Kim, Min-Jung
Vasileva, Elena A.
Mishchenko, Natalia P.
Fedoreyev, Sergey A.
Stonik, Valentin A.
Han, Jin
Lee, Ho Sup
Kim, Daejin
Jeong, Jee-Yeong
author_facet Park, Ga-Bin
Kim, Min-Jung
Vasileva, Elena A.
Mishchenko, Natalia P.
Fedoreyev, Sergey A.
Stonik, Valentin A.
Han, Jin
Lee, Ho Sup
Kim, Daejin
Jeong, Jee-Yeong
author_sort Park, Ga-Bin
collection PubMed
description Intracellular reactive oxygen species (ROS) play an important role in the proliferation and differentiation of hematopoietic stem and progenitor cells (HSPCs). HSPCs are difficult to be expanded ex vivo while maintaining their stemness when they are exposed to oxidative damage after being released from the bone marrow. There have been efforts to overcome this limitation by using various cytokine cocktails and antioxidants. In this study, we investigated the effects of echinochrome A (Ech A)-a well-established and non-toxic antioxidant-on the ex vivo expansion of HSPCs by analyzing a CD34(+) cell population and their biological functions. We observed that Ech A-induced suppression of ROS generation and p38-MAPK/JNK phosphorylation causes increased expansion of CD34(+) cells. Moreover, p38-MAPK/JNK inhibitors SB203580 and SP600125 promoted ex vivo expansion of CD34(+) cells. We also demonstrated that the activation of Lyn kinase and p110δ is a novel mechanism for Ech A to enhance ex vivo expansion of CD34(+) cells. Ech A upregulated phospho-Src, phospho-Lyn, and p110δ expression. Furthermore, the Ech A-induced ex vivo expansion of CD34(+) cells was inhibited by pretreatment with the Src family inhibitor PP1 and p110δ inhibitor CAL-101; PP1 blocked p110δ upregulation and PI3K/Akt activation, whereas CAL-101 and PI3K/Akt pathway inhibitor LY294002 did not block Src/Lyn activation. These results suggest that Ech A initially induces Src/Lyn activation, upregulates p110δ expression, and finally activates the PI3K/Akt pathway. CD34(+) cells expanded in the presence of Ech A produced equal or more hematopoietic colony-forming cells than unexpanded CD34(+) cells. In conclusion, Ech A promotes the ex vivo expansion of CD34(+) cells through Src/Lyn-mediated p110δ expression, suppression of ROS generation, and p38-MAPK/JNK activation. Hence, Ech A is a potential candidate modality for the ex vivo, and possibly in vivo, expansion of CD34(+) cells.
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spelling pubmed-67801872019-10-30 Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway Park, Ga-Bin Kim, Min-Jung Vasileva, Elena A. Mishchenko, Natalia P. Fedoreyev, Sergey A. Stonik, Valentin A. Han, Jin Lee, Ho Sup Kim, Daejin Jeong, Jee-Yeong Mar Drugs Article Intracellular reactive oxygen species (ROS) play an important role in the proliferation and differentiation of hematopoietic stem and progenitor cells (HSPCs). HSPCs are difficult to be expanded ex vivo while maintaining their stemness when they are exposed to oxidative damage after being released from the bone marrow. There have been efforts to overcome this limitation by using various cytokine cocktails and antioxidants. In this study, we investigated the effects of echinochrome A (Ech A)-a well-established and non-toxic antioxidant-on the ex vivo expansion of HSPCs by analyzing a CD34(+) cell population and their biological functions. We observed that Ech A-induced suppression of ROS generation and p38-MAPK/JNK phosphorylation causes increased expansion of CD34(+) cells. Moreover, p38-MAPK/JNK inhibitors SB203580 and SP600125 promoted ex vivo expansion of CD34(+) cells. We also demonstrated that the activation of Lyn kinase and p110δ is a novel mechanism for Ech A to enhance ex vivo expansion of CD34(+) cells. Ech A upregulated phospho-Src, phospho-Lyn, and p110δ expression. Furthermore, the Ech A-induced ex vivo expansion of CD34(+) cells was inhibited by pretreatment with the Src family inhibitor PP1 and p110δ inhibitor CAL-101; PP1 blocked p110δ upregulation and PI3K/Akt activation, whereas CAL-101 and PI3K/Akt pathway inhibitor LY294002 did not block Src/Lyn activation. These results suggest that Ech A initially induces Src/Lyn activation, upregulates p110δ expression, and finally activates the PI3K/Akt pathway. CD34(+) cells expanded in the presence of Ech A produced equal or more hematopoietic colony-forming cells than unexpanded CD34(+) cells. In conclusion, Ech A promotes the ex vivo expansion of CD34(+) cells through Src/Lyn-mediated p110δ expression, suppression of ROS generation, and p38-MAPK/JNK activation. Hence, Ech A is a potential candidate modality for the ex vivo, and possibly in vivo, expansion of CD34(+) cells. MDPI 2019-09-09 /pmc/articles/PMC6780187/ /pubmed/31505769 http://dx.doi.org/10.3390/md17090526 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Park, Ga-Bin
Kim, Min-Jung
Vasileva, Elena A.
Mishchenko, Natalia P.
Fedoreyev, Sergey A.
Stonik, Valentin A.
Han, Jin
Lee, Ho Sup
Kim, Daejin
Jeong, Jee-Yeong
Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway
title Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway
title_full Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway
title_fullStr Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway
title_full_unstemmed Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway
title_short Echinochrome A Promotes Ex Vivo Expansion of Peripheral Blood-Derived CD34(+) Cells, Potentially through Downregulation of ROS Production and Activation of the Src-Lyn-p110δ Pathway
title_sort echinochrome a promotes ex vivo expansion of peripheral blood-derived cd34(+) cells, potentially through downregulation of ros production and activation of the src-lyn-p110δ pathway
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6780187/
https://www.ncbi.nlm.nih.gov/pubmed/31505769
http://dx.doi.org/10.3390/md17090526
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