Abstract 5489: Characterization of mitochondrial STAT3 (mitostat3) function and the mitoStat3 interactome as a therapeutic strategy for multiple myeloma
Bibliographic record
Abstract
Abstract The oncogenic transcription factor STAT3 is an appealing therapeutic target in cancer including multiple myeloma (MM) where its inhibition has the potential to downregulate aberrant signaling from various upstream molecules. STAT3 has been studied extensively as a transcription factor however, much less is known about its non-classical functions in the mitochondria and in turn, the role of mitochondrial STAT3 (mitoStat3) in cancer biology. Data suggests that mitoStat3 is critical for mediating Ras-induced oncogenic transformation. As activating mutations of the MAPK pathway are reported in over 50% of myeloma patients, we sought to characterize the function of mitoStat3 in MM. We first confirmed the presence of STAT3 in the mitochondria of myeloma cell lines, AMO1 and XG6 by Western blot analysis. We next used CRISPR/CAS9 to knockdown STAT3 in XG6 and AMO1 cells, which resulted in inhibition of cell proliferation (XG6 and AMO1), apoptosis (XG6) and reduced oxygen consumption rate as measured by extracellular flux analysis (Seahorse assay). To better define the role of mitoStat3 and to identify indirect strategies to interfere with STAT3 activity, we employed a proximity-dependent Biotin Identification (BioID) method to discover mitoStat3 interacting proteins. BioID is a unique method to screen for physiologically relevant protein interactions that occur in living cells. First, wild-type STAT3 was fused to a mitochondrial localization Signal (MLS-Stat3), cloned into pcDNA5 FRT/TO [MCS]-BirAR118G-FLAG plasmid and transfected into HEK293 TRE-x Flp-In cells. BioID identified 225 high confidence mitoStat3-interacting partners in MLS-Stat3 transfected cells. The most abundant proteins in the mitoStat3 interactome included those with roles in mitochondrial translation and mitochondrial electron transport. Futhermore, mitoStat3 was found to interact with the translocase of inner membrane protein Tim44, which together with Tim23 complex facilitates translocation of proteins into the mitochondrial matrix. Genetic knockdown of Tim23 in XG6 cells resulted in decreased mitoStat3 suggesting that this complex may be involved in the import of STAT3 into the mitochondria. Additional mitoStat3 interacting proteins identified by BioID are currently being validated and will be presented. Taken together, results support the further exploration of the role of mitoStat3 in myeloma cells. Further, identification of mitoStat3 interacting proteins including Tim44 and others may inform alternative strategies to therapeutically target oncogenic STAT3. Citation Format: Serges P. Tsofack, Aaron Botham, Zhihua Li, Ellen Nong Wei, Danielle Croucher, Ioulia Jitkova, Neil MacLean, Aaron D. Schimmer, Brian Raught, Suzanne Trudel. Characterization of mitochondrial STAT3 (mitostat3) function and the mitoStat3 interactome as a therapeutic strategy for multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5489.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.002 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".