Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following Xenotransplantation
Hematopoietic reconstitution after hematopoietic stem cell transplantation (HSCT) is influenced by the number of transplanted cells. However, under certain conditions donor cell counts are limited and impair clinical outcome. Hematopoietic stem and progenitor cell (HSPC) expansion prior to HSCT is a...
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MDPI AG
2024-04-01
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| Series: | International Journal of Molecular Sciences |
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| author | Patricia M. A. Zehnle Ying Wu Naile Koleci Sheila Bohler Miriam Erlacher |
| author_facet | Patricia M. A. Zehnle Ying Wu Naile Koleci Sheila Bohler Miriam Erlacher |
| author_sort | Patricia M. A. Zehnle |
| collection | DOAJ |
| description | Hematopoietic reconstitution after hematopoietic stem cell transplantation (HSCT) is influenced by the number of transplanted cells. However, under certain conditions donor cell counts are limited and impair clinical outcome. Hematopoietic stem and progenitor cell (HSPC) expansion prior to HSCT is a widely used method to achieve higher donor cell counts and minimize transplantation-related risks such as graft failure or delayed engraftment. Still, expansion in a non-physiological environment can trigger cell death mechanisms and hence counteract the desired effect. We have shown earlier that during HSCT a relevant amount of HSPCs were lost due to apoptosis and that cell death inhibition in donor HSPCs improved engraftment in xenotransplantation experiments. Here, we assessed the effect of combined ex vivo expansion and cell death inhibition on HSPC yield and their reconstitution potential in vivo. During expansion with cytokines and the small molecule inhibitor StemRegenin 1, concomitant lentiviral overexpression of antiapoptotic BCL-X<sub>L</sub> resulted in an increased yield of transduced HSPCs. Importantly, BCL-X<sub>L</sub> overexpression enhanced the reconstitution potential of HSPCs in xenotransplantation experiments in vivo. In contrast, treatment with caspase and necroptosis inhibitors had no favorable effects on HSPC yields nor on cell viability. We postulate that overexpression of antiapoptotic BCL-X<sub>L</sub>, both during ex vivo expansion and transplantation, is a promising approach to improve the outcome of HSCT in situations with limited donor cell numbers. However, such apoptosis inhibition needs to be transient to avoid long-term sequelae like leukemia. |
| first_indexed | 2024-04-24T10:42:01Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-04-24T10:42:01Z |
| publishDate | 2024-04-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-273941eb60884fbc83c0eee8a19ceaa72024-04-12T13:20:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-04-01257410510.3390/ijms25074105Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following XenotransplantationPatricia M. A. Zehnle0Ying Wu1Naile Koleci2Sheila Bohler3Miriam Erlacher4Division of Pediatric Hematology and Oncology, University Medical Center Freiburg, 79106 Freiburg, GermanyDivision of Pediatric Hematology and Oncology, University Medical Center Freiburg, 79106 Freiburg, GermanyDivision of Pediatric Hematology and Oncology, University Medical Center Freiburg, 79106 Freiburg, GermanyDivision of Pediatric Hematology and Oncology, University Medical Center Freiburg, 79106 Freiburg, GermanyDivision of Pediatric Hematology and Oncology, University Medical Center Freiburg, 79106 Freiburg, GermanyHematopoietic reconstitution after hematopoietic stem cell transplantation (HSCT) is influenced by the number of transplanted cells. However, under certain conditions donor cell counts are limited and impair clinical outcome. Hematopoietic stem and progenitor cell (HSPC) expansion prior to HSCT is a widely used method to achieve higher donor cell counts and minimize transplantation-related risks such as graft failure or delayed engraftment. Still, expansion in a non-physiological environment can trigger cell death mechanisms and hence counteract the desired effect. We have shown earlier that during HSCT a relevant amount of HSPCs were lost due to apoptosis and that cell death inhibition in donor HSPCs improved engraftment in xenotransplantation experiments. Here, we assessed the effect of combined ex vivo expansion and cell death inhibition on HSPC yield and their reconstitution potential in vivo. During expansion with cytokines and the small molecule inhibitor StemRegenin 1, concomitant lentiviral overexpression of antiapoptotic BCL-X<sub>L</sub> resulted in an increased yield of transduced HSPCs. Importantly, BCL-X<sub>L</sub> overexpression enhanced the reconstitution potential of HSPCs in xenotransplantation experiments in vivo. In contrast, treatment with caspase and necroptosis inhibitors had no favorable effects on HSPC yields nor on cell viability. We postulate that overexpression of antiapoptotic BCL-X<sub>L</sub>, both during ex vivo expansion and transplantation, is a promising approach to improve the outcome of HSCT in situations with limited donor cell numbers. However, such apoptosis inhibition needs to be transient to avoid long-term sequelae like leukemia.https://www.mdpi.com/1422-0067/25/7/4105hematopoietic stem cellsapoptosisBCL-2BCL-X<sub>L</sub>caspases |
| spellingShingle | Patricia M. A. Zehnle Ying Wu Naile Koleci Sheila Bohler Miriam Erlacher Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following Xenotransplantation International Journal of Molecular Sciences hematopoietic stem cells apoptosis BCL-2 BCL-X<sub>L</sub> caspases |
| title | Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following Xenotransplantation |
| title_full | Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following Xenotransplantation |
| title_fullStr | Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following Xenotransplantation |
| title_full_unstemmed | Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following Xenotransplantation |
| title_short | Lentivirus-Mediated BCL-X<sub>L</sub> Overexpression Inhibits Stem Cell Apoptosis during Ex Vivo Expansion and Provides Competitive Advantage Following Xenotransplantation |
| title_sort | lentivirus mediated bcl x sub l sub overexpression inhibits stem cell apoptosis during ex vivo expansion and provides competitive advantage following xenotransplantation |
| topic | hematopoietic stem cells apoptosis BCL-2 BCL-X<sub>L</sub> caspases |
| url | https://www.mdpi.com/1422-0067/25/7/4105 |
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