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Series GSE174578 Query DataSets for GSE174578
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
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.
 
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.
 
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
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Submission date May 17, 2021
Last update date May 21, 2021
Contact name Azusa Mayumi
Organization name Kyoto Prefectural University of Medicine
Department Pediatrics
Street address 465, Kajii-cho, Kawaramachi-Hirokoji, Kamigyo-ku
City Kyoto
ZIP/Postal code 602-8566
Country Japan
 
Platforms (1)
GPL1261 [Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array
Samples (6)
GSM5320048 Ba/F3, SPAG9-JAK2(+), rep1
GSM5320049 Ba/F3, SPAG9-JAK2(+), rep2
GSM5320050 Ba/F3, SPAG9-JAK2(+), rep3
Relations
BioProject PRJNA730570

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SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE174578_RAW.tar 22.8 Mb (http)(custom) TAR (of CEL, CHP)
Raw data provided as supplementary file
Processed data included within Sample table
Processed data provided as supplementary file

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