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GEO help: Mouse over screen elements for information. |
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| Status |
Public on May 18, 2021 |
| Title |
SPAG9-JAK2 gene expression array in Ba/F3 cells |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
SPAG9-JAK2 is a novel fusion gene identified in a pediatric patient with Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL). In this study, we performed functional analysis of the SPAG9-JAK2 fusion to establish molecular targeted therapy. Ba/F3 cells expressing SPAG9-JAK2 generated by retroviral transduction (Ba/F3-SPAG9-JAK2), proliferated in the absence of IL-3, and exhibited constitutive phosphorylation of the tyrosine residues in the JAK2 kinase domain of the fusion protein and STAT3/STAT5. Mutation of tyrosine residues in the JAK2 kinase domain (SPAG9-JAK2 mut) abolished IL-3 independence, but had no influence on STAT3/STAT5 phosphorylation levels. Gene expression analysis revealed that Stat1 was significantly up-regulated in Ba/F3-SPAG9-JAK2 cells. STAT1 was also phosphorylated in Ba/F3-SPAG9-JAK2 but not SPAG9-JAK2 mut cells, suggesting that STAT1 is key for SPAG9-JAK2-mediated cell proliferation. Consistently, STAT1 induced expression of the anti-apoptotic proteins, BCL-2 and MCL-1, as did SPAG9-JAK2, but not SPAG9-JAK2 mut. Ruxolitinib abrogated Ba/F3-SPAG9-JAK2-mediated proliferation in vitro, but was insufficient in vivo. Venetoclax (a BCL-2 inhibitor) or AZD5991 (an MCL-1 inhibitor) enhanced the effects of ruxolitinib on Ba/F3-SPAG9-JAK2 in vitro. These findings suggest that activation of the JAK2-STAT1-BCL-2/MCL-1 axis contributes to SPAG9-JAK2-related aberrant growth promotion. BCL-2 or MCL-1 inhibition is a potential therapeutic option for B-ALL with SPAG9-JAK2 fusion.
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| Overall design |
Ba/F3 cells were transduced with SPAG9-JAK2 fusion protein by the Retro-X Tet-On Advanced Inducible Expression System (Takara Bio), and Ba/F3 cells expressing SPAG9-JAK2 under doxycycline (DOX) dependent manner were established. To reveal which genes or pathways were affected by SPAG9-JAK2, Ba/F3 cells expressing SPAG9-JAK2 induced by DOX (Ba/F3-SPAG9-JAK2) or uninduced Ba/F3 cells (mock Ba/F3) were cultured in triplicate, and total RNA was extracted from Ba/F3-SPAG9-JAK2 or mock Ba/F3 48 hours later.
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| Contributor(s) |
Mayumi A, Tomii T, Kanayama T, Mikami T, Tanaka K, Yoshida Y, Kato I, Kawamura M, Nakahata T, Takita J, Hosoi H, Imamura T |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
May 17, 2021 |
| Last update date |
Mar 29, 2022 |
| Contact name |
Azusa Mayumi |
| Organization name |
Kyoto Prefectural University of Medicine
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| Department |
Pediatrics
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| Street address |
465, Kajii-cho, Kawaramachi-Hirokoji, Kamigyo-ku
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| City |
Kyoto |
| ZIP/Postal code |
602-8566 |
| Country |
Japan |
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| Platforms (1) |
| GPL1261 |
[Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA730570 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE174578_RAW.tar |
22.5 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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