Targeting TRAF2 and NCK-interacting protein kinase restores sensitivity to chemotherapy in mesenchymal small cell lung cancer
Azusa Tanimoto 1 , Benjamin B Morris 1 , Kavya Ramkumar 1 , Robert J. Cardnell 1 , ShenLi 2 , QiWang 2 , C. Allison Stewart 1 , Carl M. Gay 1 , JingWang 2 , Lauren Averett Byers 1
Title of the Poster Presentation Goes Here Authors of the Poster Presentation Goes Here Institutional and/or Graduate School of Biomedical Sciences Affiliation Goes Here 1 The University of Texas MD Anderson Cancer Center Department of Thoracic/Head & Neck Medical Oncology; 2 The University of Texas MD Anderson Cancer Center of Bioinformatics & Computational Biology;
Background
Finding 1 : TNIK expression is upregulated in SCLC cell lines.
Small cell lung cancer (SCLC) is a highly lethal malignancy, with rapidly acquired chemotherapy resistance. Some studies have reported that Wnt signaling pathway activation promoted cell proliferation and was correlated with chemo-resistance in SCLC (1-3). Additionally, our group has demonstrated that cisplatin relapsed model generated high mesenchymal subgroup using single-cell analysis (4). None of the therapies targeting Wnt pathway components of the transmembrane and the cytoplasm have been successful in a clinical application due to toxicity and insufficient efficacy (5). However, targeting Wnt signaling inside the nucleus has been drawing increasing attention as cancer therapeutics. TRAF2 and NCK-interacting protein kinase (TNIK), which interacts with downstream effectors, TCF4/β -catenin transcriptional complex, is an essential activator of Wnt target genes (6). TINIK is highly expressed in several cancers for cell proliferation, thus TNIK is expected as a novel druggable target (7-11). On the other hand, the question remains whether TNIK is a critical target in SCLC, meanwhile there is a lack of research concerning how TNIK contributes to chemo-resistance in SCLC.
Fig 3. TNIK inhibition overcame resistance to cisplatin (CDDP) in SCLC cells. (A) Cell viability in CDDP-resistant SCLC cells treated with either scrambled siRNA or siRNA targeting TNIK. The cells were treated with DMSO, CDDP (1 µmol/L) and CDDP (3 µmol/L) for 96 h. *, P<0.05, **, P<0.01 (B) Western blotting of the SCLC cells with either β -catenin or TNIK which were treated with CDDP ( 1 µmol/L ) for 48h for 48 h. (C) Western blotting of the SCLC cells expressing mesenchymal markers treated with TNIK knockdown. (D) Western blotting of the cells treated with DMSO, CDDP ( 1 µmol/L ) and/or NCB- 0846 (500 nmol/L) for 48 h. (E) Western blotting of the SCLC cells expressing mesenchymal markers treated with escalating doses of NCB-0846 for 48 h.
Fig 1. Enriched expression of genes associated with Wnt and EMT in SCLC. Violin plots of Wnt ( A ) and EMT ( B )-related genes between NSCLC and SCLC. ( C ) Spearman correlation between TNIK and mesenchymal markers. *, P<0.05, ***, P<0.001, ****, P<0.0001
Summary and Future Directions
Outline of Wnt/ β -catenin pathway Wnt ligand binds to its receptors (Frizzled and LRP5/6), followed by dishevelled (Dsh) protein becomes
• Our preclinical results indicate that NCB-0846 enhanced sensitivity to cisplatin and decreased mesenchymal markers (ZEB1 and N-Cadherin) in SCLC cells resistant to cisplatin. • Findings support a promising rational combination therapy of NCB-0846 plus cisplatin for mesenchymal phenotype SCLC resistant to cisplatin. • Further studies should address the best schedule of the combination to maximize these effects.
activated. Dsh inhibits GSK- 3β/ Axin/APC complex and
Finding 2 : A TNIK inhibitor significantly reduces cell viability of SCLC-N and SCLC-P subtype.
subsequently dephosphorylates β - catenin. Accumulated β -catenin in cytoplasm moves to nucleus and binds to TCF4. TNIK is required for activation of TCF4/β -catenin complex and initiates transcription of Wnt target genes.
Fig 5. Schematic diagram of overcoming cisplatin resistance by TNIK inhibition in SCLC
Hypothesis
We hypothesize that TNIK activation confers resistance to chemotherapy in SCLC and the novel TNIK inhibitor may overcome the resistance via change of EMT status.
References
Fig 2. A novel TNIK inhibitor, NCB-0846 showed potent activity in non- neuroendocrine SCLC cell lines. (A) IC50 values of NCB-0846 after 96h treatment in all subtypes of SCLC cell lines. (B) Boxplots for IC50 values of NCB-0846 by subtype. *, P<0.05, **, P<0.01
Experimental Design
1. Tenjin Y, et al. Lab Invest. 99(11):1622-35. (2019) 2. Kim KB, et al. Cancer research. (2022). 3. Wagner AH, et al. Nature communications. 9(1):3787. (2018) 4. Gay CM, et al. Cancer cell. 39(3):346-60.e7. (2021) 5. Yang P, et al. European journal of medicinal chemistry. 243:114789. (2022) 6. MahmoudiT, et al. Embo j. 28(21):3329-40. (2009) 7. Yamada T, et al. Cancer science. 108(5):818-23. (2017) 8. Masuda M, et al. Nature communications. 7:12586. (2016) 9. Torres-Ayuso P, et al. Cancer discovery. 11(6):1411-23. (2021) 10. HirozaneT, et al. JCI Insight. 6(3). (2021) 11. Shitashige M, et al. Cancer research. 2010;70(12):5024-33. (2010)
• We compared RNA expression of Wnt and EMT-related genes between NSCLC and SCLC cell lines using the cancer cell line encyclopedia (CCLE) dataset. • We determined the correlation of RNA expression between TNIK and mesenchymal phenotype genes using CCLE dataset. • We evaluated susceptibility to a TNIK inhibitor, NCB-0846 in 29 human-derived SCLC cell lines using 96-hour proliferation assays. • We reduced TNIK and β -catenin expression using siRNA against either TNIK or β -catenin in human SCLC cell lines. • We assayed DNA damage protein and EMT-related proteins changed by NCB-0846 using western blot in human SCLC cell lines.
Finding 3 : TNIK inhibition reinforces sensitivity to cisplatin and suppresses mesenchymal markers in cisplatin-resistant SCLC cells.
Acknowledgements
This work was supported by: Lung SPORE P50-CA070907, NCI/NIH U01- CA213273, NCI/NIH R01-CA207295,
COI : I have no financial relationships to disclose.
Page 1Made with FlippingBook - Online magazine maker