TY - JOUR
T1 - Therapeutic targeting of nemo-like kinase in primary and acquired endocrine-resistant breast cancer
AU - Wang, Xian
AU - Veeraraghavan, Jamunarani
AU - Liu, Chia Chia
AU - Cao, Xixi
AU - Qin, Lanfang
AU - Kim, Jin Ah
AU - Tan, Ying
AU - Loo, Suet Kee
AU - Hu, Yiheng
AU - Lin, Ling
AU - Lee, Sanghoon
AU - Shea, Martin J.
AU - Mitchell, Tamika
AU - Li, Shunqiang
AU - Ellis, Matthew J.
AU - Hilsenbeck, Susan G.
AU - Schiff, Rachel
AU - Wang, Xiao Song
N1 - Funding Information:
This study was supported by NIH grant R01CA181368 (to X.-S. Wang), R01CA183976 (to X.-S. Wang), Susan G. Komen Foundation PDF15333523 (to X. Wang), Congressionally Directed Medical Research Program W81XWH- 12-1-0166 (to X.-S. Wang), W81XWH-12-1-0167 (to R. Schiff), W81XWH-13-1- 0201 (to X.-X. Cao), and Nancy Owens Foundation. This study was also supported, in part, by NIH grant R21CA237964 (to X.-S. Wang); CDMRP W81XWH-13-1-0431 (to J. Veeraraghavan); Susan G. Komen Foundation PDF12231561 (to J.-A. Kim); the Breast Cancer Research Foundation grants BCRF-16-142, 17-143, 18-145 (to R. Schiff), Stand Up To Cancer-American Association for Cancer Research Dream Team Translational Research Grant, grant number SU2C-AACR-DT0409 (to R. Schiff); Commonwealth of PA Tobacco Phase 15 Formula Fund (to X.-S. Wang), the Shear Family Foundation, and the Hillman Foundation (to X.-S. Wang); and U24CA209837 (to S. Li). The results published here are, in part, based upon data generated by The Cancer Genome Atlas project established by the NCI and NHGRI (dbGaP accession: phs000178.v6.p6). The computational infrastructure was supported by the Dan L. Duncan Comprehensive Cancer Center Biostatistics and Informatics Shared Resource and the University of Pittsburgh Center for Research Computing. We thank the Washington University HAMLET Core for providing the tissue microarrays of WHIM PDX tumors and the WHIM11J tumor cells for retransplantation. The RPPA experiment was supported, in part, by Cancer Prevention & Research Institute of Texas Proteomics & Metabolomics Core Facility Support Award (RP170005 to S.G. Hilsenbeck) and NCI Cancer Center Support Grant to Antibody-based Proteomics Core/Shared Resource (P30CA125123to S.G. Hilsenbeck). We thank Dr. Shixia Huang, Hsin-Yi Cincy Lu, and Mr. Carlos Ramos from the Antibody-based Proteomics Core/Shared Resource for their excellent technical assistant in performing RPPA experiments. We thank Drs. Kimal Rajapakshe and Cristian Coarfa and Mr. Dimuthu Perera for RPPA data processing and normalization. We thank Dr. Bert W. O'Malley at Baylor College of Medicine (Houston, TX) for providing recombinant SRC-3 protein.
Funding Information:
Informatics Shared Resource and the University of Pittsburgh Center for Research Computing. We thank the Washington University HAMLET Core for providing the tissue microarrays of WHIM PDX tumors and the WHIM11J tumor cells for retransplantation. The RPPA experiment was supported, in part, by Cancer Prevention & Research Institute of Texas Proteomics & Metabolomics Core Facility Support Award (RP170005 to S.G. Hilsenbeck) and NCI Cancer Center Support Grant to Antibody-based Proteomics Core/Shared Resource (P30CA125123to S.G. Hilsenbeck). We thank Dr. Shixia Huang, Hsin-Yi Cincy Lu, and Mr. Carlos Ramos from the Antibody-based Proteomics Core/Shared Resource for their excellent technical assistant in performing RPPA experiments. We thank Drs.
Funding Information:
This study was supported by NIH grant R01CA181368 (to X.-S. Wang), R01CA183976 (to X.-S. Wang), Susan G. Komen Foundation PDF15333523 (to X. Wang), Congressionally Directed Medical Research Program W81XWH-12-1-0166 (to X.-S. Wang), W81XWH-12-1-0167 (to R. Schiff), W81XWH-13-1-0201 (to X.-X. Cao), and Nancy Owens Foundation. This study was also supported, in part, by NIH grant R21CA237964 (to X.-S. Wang); CDMRP W81XWH-13-1-0431 (to J. Veeraraghavan); Susan G. Komen Foundation PDF12231561 (to J.-A. Kim); the Breast Cancer Research Foundation grants BCRF-16-142, 17-143, 18-145 (to R. Schiff), Stand Up To Cancer-American Association for Cancer Research Dream Team Translational Research Grant, grant number SU2C-AACR-DT0409 (to R. Schiff); Commonwealth of PA Tobacco Phase 15 Formula Fund (to X.-S. Wang), the Shear Family Foundation, and the Hillman Foundation (to X.-S. Wang); and U24CA209837 (to S. Li). The results published here are, in part, based upon data generated by The Cancer Genome Atlas project established by the NCI and NHGRI (dbGaP accession: phs000178.v6.p6). The computational infrastructure was supported by the Dan L. Duncan Comprehensive Cancer Center Biostatistics and
Funding Information:
S. Li reported grants from NIH 5U24CA209837 during the conduct of the study and personal fees from Envigo outside the submitted work. R. Schiff reported grants from Breast Cancer Research Foundation and DoD during the conduct of the study, grants from PUMA Biotechnology, AstraZeneca, Gilead Sciences, and CPRIT, and personal fees from MacroGenics outside the submitted work. X.-S. Wang reported grants from NIH/NCI, Congressionally Directed Medical Research Program, Susan G. Komen Foundation, Commonwealth of PA, and Nancy Owens Foundation and other from Vertex Pharmaceuticals Incorporated during the conduct of the study. No disclosures were reported by the other authors.
Publisher Copyright:
© 2021 American Association for Cancer Research.
PY - 2021/5/1
Y1 - 2021/5/1
N2 - Purpose: Endocrine resistance remains a major clinical challenge in estrogen receptor (ER)-positive breast cancer. Despite the encouraging results from clinical trials for the drugs targeting known survival signaling, relapse is still inevitable. There is an unmet need to discover new drug targets in the unknown escape pathways. Here, we report Nemo-like kinase (NLK) as a new actionable kinase target that endows previously uncharacterized survival signaling in endocrine-resistant breast cancer. Experimental Design: The effects of NLK inhibition on the viability of endocrine-resistant breast cancer cell lines were examined by MTS assay. The effect of VX-702 on NLK activity was verified by kinase assay. The modulation of ER and its coactivator, SRC-3, by NLK was examined by immunoprecipitation, kinase assay, luciferase assay, and RNAsequencing. The therapeutic effects of VX-702 and everolimus were tested on cell line- and patientderived xenograft (PDX) tumor models. Results: NLK overexpression endows reduced endocrine responsiveness and is associated with worse outcome of patients treated with tamoxifen. Mechanistically, NLK may function, at least in part, via enhancing the phosphorylation of ERa and its key coactivator, SRC-3, to modulate ERa transcriptional activity. Through interrogation of a kinase profiling database, we uncovered and verified a highly selective dual p38/NLK inhibitor, VX-702. Coadministration of VX-702 with the mTOR inhibitor, everolimus, demonstrated a significant therapeutic effect in cell line-derived xenograft and PDX tumor models of acquired or de novo endocrine resistance. Conclusions: Together, this study reveals the potential of therapeutic modulation of NLK for the management of the endocrineresistant breast cancers with active NLK signaling.
AB - Purpose: Endocrine resistance remains a major clinical challenge in estrogen receptor (ER)-positive breast cancer. Despite the encouraging results from clinical trials for the drugs targeting known survival signaling, relapse is still inevitable. There is an unmet need to discover new drug targets in the unknown escape pathways. Here, we report Nemo-like kinase (NLK) as a new actionable kinase target that endows previously uncharacterized survival signaling in endocrine-resistant breast cancer. Experimental Design: The effects of NLK inhibition on the viability of endocrine-resistant breast cancer cell lines were examined by MTS assay. The effect of VX-702 on NLK activity was verified by kinase assay. The modulation of ER and its coactivator, SRC-3, by NLK was examined by immunoprecipitation, kinase assay, luciferase assay, and RNAsequencing. The therapeutic effects of VX-702 and everolimus were tested on cell line- and patientderived xenograft (PDX) tumor models. Results: NLK overexpression endows reduced endocrine responsiveness and is associated with worse outcome of patients treated with tamoxifen. Mechanistically, NLK may function, at least in part, via enhancing the phosphorylation of ERa and its key coactivator, SRC-3, to modulate ERa transcriptional activity. Through interrogation of a kinase profiling database, we uncovered and verified a highly selective dual p38/NLK inhibitor, VX-702. Coadministration of VX-702 with the mTOR inhibitor, everolimus, demonstrated a significant therapeutic effect in cell line-derived xenograft and PDX tumor models of acquired or de novo endocrine resistance. Conclusions: Together, this study reveals the potential of therapeutic modulation of NLK for the management of the endocrineresistant breast cancers with active NLK signaling.
UR - http://www.scopus.com/inward/record.url?scp=85105289374&partnerID=8YFLogxK
U2 - 10.1158/1078-0432.CCR-20-2961
DO - 10.1158/1078-0432.CCR-20-2961
M3 - Article
C2 - 33542078
AN - SCOPUS:85105289374
SN - 1078-0432
VL - 27
SP - 2648
EP - 2662
JO - Clinical Cancer Research
JF - Clinical Cancer Research
IS - 9
ER -