Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells
Abstract Background Although oncogenic RAS mutants are thought to exert mutagenic effects upon blood cells, it remains uncertain how a single oncogenic RAS impacts non-transformed multipotent hematopoietic stem or progenitor cells (HPCs). Such potential pre-malignant status may characterize HPCs in...
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| Format: | Article |
| Language: | English |
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BMC
2024-04-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-024-03723-2 |
| _version_ | 1827277283851763712 |
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| author | Huan-Ting Lin Masatoshi Takagi Kenji Kubara Kazuto Yamazaki Fumiko Michikawa Takashi Okumura Takuya Naruto Tomohiro Morio Koji Miyazaki Hideki Taniguchi Makoto Otsu |
| author_facet | Huan-Ting Lin Masatoshi Takagi Kenji Kubara Kazuto Yamazaki Fumiko Michikawa Takashi Okumura Takuya Naruto Tomohiro Morio Koji Miyazaki Hideki Taniguchi Makoto Otsu |
| author_sort | Huan-Ting Lin |
| collection | DOAJ |
| description | Abstract Background Although oncogenic RAS mutants are thought to exert mutagenic effects upon blood cells, it remains uncertain how a single oncogenic RAS impacts non-transformed multipotent hematopoietic stem or progenitor cells (HPCs). Such potential pre-malignant status may characterize HPCs in patients with RAS-associated autoimmune lymphoproliferative syndrome-like disease (RALD). This study sought to elucidate the biological and molecular alterations in human HPCs carrying monoallelic mutant KRAS (G13C) with no other oncogene mutations. Methods We utilized induced pluripotent stem cells (iPSCs) derived from two unrelated RALD patients. Isogenic HPC pairs harboring either wild-type KRAS or monoallelic KRAS (G13C) alone obtained following differentiation enabled reliable comparative analyses. The compound screening was conducted with an established platform using KRAS (G13C) iPSCs and differentiated HPCs. Results Cell culture assays revealed that monoallelic KRAS (G13C) impacted both myeloid differentiation and expansion characteristics of iPSC-derived HPCs. Comprehensive RNA-sequencing analysis depicted close clustering of HPC samples within the isogenic group, warranting that comparative studies should be performed within the same genetic background. When compared with no stimulation, iPSC-derived KRAS (G13C)-HPCs showed marked similarity with the wild-type isogenic control in transcriptomic profiles. After stimulation with cytokines, however, KRAS (G13C)-HPCs exhibited obvious aberrant cell-cycle and apoptosis responses, compatible with "dysregulated expansion," demonstrated by molecular and biological assessment. Increased BCL-xL expression was identified amongst other molecular changes unique to mutant HPCs. With screening platforms established for therapeutic intervention, we observed selective activity against KRAS (G13C)-HPC expansion in several candidate compounds, most notably in a MEK- and a BCL-2/BCL-xL-inhibitor. These two compounds demonstrated selective inhibitory effects on KRAS (G13C)-HPCs even with primary patient samples when combined. Conclusions Our findings indicate that a monoallelic oncogenic KRAS can confer dysregulated expansion characteristics to non-transformed HPCs, which may constitute a pathological condition in RALD hematopoiesis. The use of iPSC-based screening platforms will lead to discovering treatments that enable selective inhibition of RAS-mutated HPC clones. |
| first_indexed | 2024-04-24T07:18:19Z |
| format | Article |
| id | doaj.art-41b677ec4b61423aabb1d9bf49e2ae0f |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-24T07:18:19Z |
| publishDate | 2024-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-41b677ec4b61423aabb1d9bf49e2ae0f2024-04-21T11:10:49ZengBMCStem Cell Research & Therapy1757-65122024-04-0115111810.1186/s13287-024-03723-2Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cellsHuan-Ting Lin0Masatoshi Takagi1Kenji Kubara2Kazuto Yamazaki3Fumiko Michikawa4Takashi Okumura5Takuya Naruto6Tomohiro Morio7Koji Miyazaki8Hideki Taniguchi9Makoto Otsu10Division of Regenerative Medicine, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of TokyoDepartment of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental UniversityTsukuba Research Laboratories, Eisai Co., Ltd.Tsukuba Research Laboratories, Eisai Co., Ltd.Tsukuba Research Laboratories, Eisai Co., Ltd.Division of Regenerative Medicine, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of TokyoDepartment of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental UniversityDepartment of Pediatrics and Developmental Biology, Graduate School of Medicine, Tokyo Medical and Dental UniversityDepartment of Transfusion and Cell Transplantation, Kitasato University School of MedicineDivision of Regenerative Medicine, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of TokyoDepartment of Transfusion and Cell Transplantation, Kitasato University School of MedicineAbstract Background Although oncogenic RAS mutants are thought to exert mutagenic effects upon blood cells, it remains uncertain how a single oncogenic RAS impacts non-transformed multipotent hematopoietic stem or progenitor cells (HPCs). Such potential pre-malignant status may characterize HPCs in patients with RAS-associated autoimmune lymphoproliferative syndrome-like disease (RALD). This study sought to elucidate the biological and molecular alterations in human HPCs carrying monoallelic mutant KRAS (G13C) with no other oncogene mutations. Methods We utilized induced pluripotent stem cells (iPSCs) derived from two unrelated RALD patients. Isogenic HPC pairs harboring either wild-type KRAS or monoallelic KRAS (G13C) alone obtained following differentiation enabled reliable comparative analyses. The compound screening was conducted with an established platform using KRAS (G13C) iPSCs and differentiated HPCs. Results Cell culture assays revealed that monoallelic KRAS (G13C) impacted both myeloid differentiation and expansion characteristics of iPSC-derived HPCs. Comprehensive RNA-sequencing analysis depicted close clustering of HPC samples within the isogenic group, warranting that comparative studies should be performed within the same genetic background. When compared with no stimulation, iPSC-derived KRAS (G13C)-HPCs showed marked similarity with the wild-type isogenic control in transcriptomic profiles. After stimulation with cytokines, however, KRAS (G13C)-HPCs exhibited obvious aberrant cell-cycle and apoptosis responses, compatible with "dysregulated expansion," demonstrated by molecular and biological assessment. Increased BCL-xL expression was identified amongst other molecular changes unique to mutant HPCs. With screening platforms established for therapeutic intervention, we observed selective activity against KRAS (G13C)-HPC expansion in several candidate compounds, most notably in a MEK- and a BCL-2/BCL-xL-inhibitor. These two compounds demonstrated selective inhibitory effects on KRAS (G13C)-HPCs even with primary patient samples when combined. Conclusions Our findings indicate that a monoallelic oncogenic KRAS can confer dysregulated expansion characteristics to non-transformed HPCs, which may constitute a pathological condition in RALD hematopoiesis. The use of iPSC-based screening platforms will lead to discovering treatments that enable selective inhibition of RAS-mutated HPC clones.https://doi.org/10.1186/s13287-024-03723-2Human-induced pluripotent stem cellsHematopoietic systemStem cellsLymphoproliferative disordersKRAS proteinOncogene |
| spellingShingle | Huan-Ting Lin Masatoshi Takagi Kenji Kubara Kazuto Yamazaki Fumiko Michikawa Takashi Okumura Takuya Naruto Tomohiro Morio Koji Miyazaki Hideki Taniguchi Makoto Otsu Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells Stem Cell Research & Therapy Human-induced pluripotent stem cells Hematopoietic system Stem cells Lymphoproliferative disorders KRAS protein Oncogene |
| title | Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells |
| title_full | Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells |
| title_fullStr | Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells |
| title_full_unstemmed | Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells |
| title_short | Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells |
| title_sort | monoallelic kras g13c mutation triggers dysregulated expansion in induced pluripotent stem cell derived hematopoietic progenitor cells |
| topic | Human-induced pluripotent stem cells Hematopoietic system Stem cells Lymphoproliferative disorders KRAS protein Oncogene |
| url | https://doi.org/10.1186/s13287-024-03723-2 |
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