BET inhibition increases βIII-tubulin expression and sensitizes metastatic breast cancer in the brain to vinorelbine

Deepak Kanojia; Wojciech K. Panek; Alex Cordero; Jawad Fares; Annie Xiao; Solomiia Savchuk; Krishan Kumar; Ting Xiao; Katarzyna C. Pituch; Jason Miska; Peng Zhang; Kwok Ling Kam; Craig Horbinski; Irina V. Balyasnikova; Atique U. Ahmed; Maciej S. Lesniak

doi:10.1126/SCITRANSLMED.AAX2879

BET inhibition increases βIII-tubulin expression and sensitizes metastatic breast cancer in the brain to vinorelbine

Deepak Kanojia, Wojciech K. Panek, Alex Cordero, Jawad Fares, Annie Xiao, Solomiia Savchuk, Krishan Kumar, Ting Xiao, Katarzyna C. Pituch, Jason Miska, Peng Zhang, Kwok Ling Kam, Craig Horbinski, Irina V. Balyasnikova, Atique U. Ahmed, Maciej S. Lesniak^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

Metastases from primary breast cancer result in poor survival. βIII-tubulin (TUBB3) has been established as a therapeutic target for breast cancer metastases specifically to the brain. In this study, we conducted a systematic analysis to determine the regulation of TUBB3 expression in breast cancer metastases to the brain and strategically target these metastases using vinorelbine (VRB), a drug approved by the U.S. Food and Drug Administration (FDA). We found that human epidermal growth factor receptor 2 (HER2) signaling regulates TUBB3 expression in both trastuzumab-sensitive and trastuzumab-resistant neoplastic cells. We further discovered that bromodomain and extra-terminal domain (BET) inhibition increases TUBB3 expression, rendering neoplastic cells more susceptible to apoptosis by VRB. Orthotopic xenograft assays using two different breast cancer cell models revealed a reduction in tumor volume with BET inhibition and VRB treatment. In addition, in vivo studies using a model of multiple brain metastasis (BM) showed improved survival with the combination of radiation + BET inhibitor (iBET-762) + VRB (75% long-term survivors, P < 0.05). Using in silico analysis and BET inhibition, we found that the transcription factor myeloid zinc finger-1 (MZF-1) protein binds to the TUBB3 promoter. BET inhibition decreases MZF-1 expression and subsequently increases TUBB3 expression. Overexpression of MZF-1 decreases TUBB3 expression and reduces BM in vivo, whereas its knockdown increases TUBB3 expression in breast cancer cells. In summary, this study demonstrates a regulatory mechanism of TUBB3 and provides support for an application of BET inhibition to sensitize breast cancer metastases to VRB-mediated therapy.

Original language	English (US)
Article number	eaax2879
Journal	Science translational medicine
Volume	12
Issue number	558
DOIs	https://doi.org/10.1126/SCITRANSLMED.AAX2879
State	Published - Aug 2020

Funding

We would like to thank A. Lopez-Rosas for technical assistance. We thank D. Horiuchi for cell lines and V5-tagged antibody. We thank E. Gonzalez-Buendia for assistance with ChIP analysis and J. R. Kane for critically reading the manuscript. We would like to thank J. Massague (MSKCC) for providing the MDA-MB-231Br, CN34, and CN34Br cell lines. We would like to thank D. Yu (MD Anderson Cancer Centre) for providing the BT-474 and BT-474Br cell lines. This work was supported by NIH grants R35CA197725, R01NS093903, and P50CA221747 to M.S.L., R01NS87990 to M.S.L. and I.V.B., R01NS106379 01A1 to I.V.B., 1R01NS096376-01A1 to A.U.A., 5R21CA220625 to K.K., and R01NS102669 to C.H. Author

ASJC Scopus subject areas

General Medicine

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1126/SCITRANSLMED.AAX2879

Cite this

Kanojia, D., Panek, W. K., Cordero, A., Fares, J., Xiao, A., Savchuk, S., Kumar, K., Xiao, T., Pituch, K. C., Miska, J., Zhang, P., Kam, K. L., Horbinski, C., Balyasnikova, I. V., Ahmed, A. U., & Lesniak, M. S. (2020). BET inhibition increases βIII-tubulin expression and sensitizes metastatic breast cancer in the brain to vinorelbine. Science translational medicine, 12(558), Article eaax2879. https://doi.org/10.1126/SCITRANSLMED.AAX2879

@article{63cdb5de7fde44a3b1f412ddf3a03329,

title = "BET inhibition increases βIII-tubulin expression and sensitizes metastatic breast cancer in the brain to vinorelbine",

abstract = "Metastases from primary breast cancer result in poor survival. βIII-tubulin (TUBB3) has been established as a therapeutic target for breast cancer metastases specifically to the brain. In this study, we conducted a systematic analysis to determine the regulation of TUBB3 expression in breast cancer metastases to the brain and strategically target these metastases using vinorelbine (VRB), a drug approved by the U.S. Food and Drug Administration (FDA). We found that human epidermal growth factor receptor 2 (HER2) signaling regulates TUBB3 expression in both trastuzumab-sensitive and trastuzumab-resistant neoplastic cells. We further discovered that bromodomain and extra-terminal domain (BET) inhibition increases TUBB3 expression, rendering neoplastic cells more susceptible to apoptosis by VRB. Orthotopic xenograft assays using two different breast cancer cell models revealed a reduction in tumor volume with BET inhibition and VRB treatment. In addition, in vivo studies using a model of multiple brain metastasis (BM) showed improved survival with the combination of radiation + BET inhibitor (iBET-762) + VRB (75% long-term survivors, P < 0.05). Using in silico analysis and BET inhibition, we found that the transcription factor myeloid zinc finger-1 (MZF-1) protein binds to the TUBB3 promoter. BET inhibition decreases MZF-1 expression and subsequently increases TUBB3 expression. Overexpression of MZF-1 decreases TUBB3 expression and reduces BM in vivo, whereas its knockdown increases TUBB3 expression in breast cancer cells. In summary, this study demonstrates a regulatory mechanism of TUBB3 and provides support for an application of BET inhibition to sensitize breast cancer metastases to VRB-mediated therapy.",

author = "Deepak Kanojia and Panek, {Wojciech K.} and Alex Cordero and Jawad Fares and Annie Xiao and Solomiia Savchuk and Krishan Kumar and Ting Xiao and Pituch, {Katarzyna C.} and Jason Miska and Peng Zhang and Kam, {Kwok Ling} and Craig Horbinski and Balyasnikova, {Irina V.} and Ahmed, {Atique U.} and Lesniak, {Maciej S.}",

note = "Funding Information: We would like to thank A. Lopez-Rosas for technical assistance. We thank D. Horiuchi for cell lines and V5-tagged antibody. We thank E. Gonzalez-Buendia for assistance with ChIP analysis and J. R. Kane for critically reading the manuscript. We would like to thank J. Massague (MSKCC) for providing the MDA-MB-231Br, CN34, and CN34Br cell lines. We would like to thank D. Yu (MD Anderson Cancer Centre) for providing the BT-474 and BT-474Br cell lines. This work was supported by NIH grants R35CA197725, R01NS093903, and P50CA221747 to M.S.L., R01NS87990 to M.S.L. and I.V.B., R01NS106379 01A1 to I.V.B., 1R01NS096376-01A1 to A.U.A., 5R21CA220625 to K.K., and R01NS102669 to C.H. Author Publisher Copyright: Copyright {\textcopyright} 2020 The Authors",

year = "2020",

month = aug,

doi = "10.1126/SCITRANSLMED.AAX2879",

language = "English (US)",

volume = "12",

journal = "Science translational medicine",

issn = "1946-6234",

publisher = "American Association for the Advancement of Science",

number = "558",

}

Kanojia, D, Panek, WK, Cordero, A, Fares, J, Xiao, A, Savchuk, S, Kumar, K, Xiao, T, Pituch, KC , Miska, J , Zhang, P, Kam, KL, Horbinski, C , Balyasnikova, IV , Ahmed, AU & Lesniak, MS 2020, 'BET inhibition increases βIII-tubulin expression and sensitizes metastatic breast cancer in the brain to vinorelbine', Science translational medicine, vol. 12, no. 558, eaax2879. https://doi.org/10.1126/SCITRANSLMED.AAX2879

TY - JOUR

T1 - BET inhibition increases βIII-tubulin expression and sensitizes metastatic breast cancer in the brain to vinorelbine

AU - Kanojia, Deepak

AU - Panek, Wojciech K.

AU - Cordero, Alex

AU - Fares, Jawad

AU - Xiao, Annie

AU - Savchuk, Solomiia

AU - Kumar, Krishan

AU - Xiao, Ting

AU - Pituch, Katarzyna C.

AU - Miska, Jason

AU - Zhang, Peng

AU - Kam, Kwok Ling

AU - Horbinski, Craig

AU - Balyasnikova, Irina V.

AU - Ahmed, Atique U.

AU - Lesniak, Maciej S.

N1 - Funding Information: We would like to thank A. Lopez-Rosas for technical assistance. We thank D. Horiuchi for cell lines and V5-tagged antibody. We thank E. Gonzalez-Buendia for assistance with ChIP analysis and J. R. Kane for critically reading the manuscript. We would like to thank J. Massague (MSKCC) for providing the MDA-MB-231Br, CN34, and CN34Br cell lines. We would like to thank D. Yu (MD Anderson Cancer Centre) for providing the BT-474 and BT-474Br cell lines. This work was supported by NIH grants R35CA197725, R01NS093903, and P50CA221747 to M.S.L., R01NS87990 to M.S.L. and I.V.B., R01NS106379 01A1 to I.V.B., 1R01NS096376-01A1 to A.U.A., 5R21CA220625 to K.K., and R01NS102669 to C.H. Author Publisher Copyright: Copyright © 2020 The Authors

PY - 2020/8

Y1 - 2020/8

N2 - Metastases from primary breast cancer result in poor survival. βIII-tubulin (TUBB3) has been established as a therapeutic target for breast cancer metastases specifically to the brain. In this study, we conducted a systematic analysis to determine the regulation of TUBB3 expression in breast cancer metastases to the brain and strategically target these metastases using vinorelbine (VRB), a drug approved by the U.S. Food and Drug Administration (FDA). We found that human epidermal growth factor receptor 2 (HER2) signaling regulates TUBB3 expression in both trastuzumab-sensitive and trastuzumab-resistant neoplastic cells. We further discovered that bromodomain and extra-terminal domain (BET) inhibition increases TUBB3 expression, rendering neoplastic cells more susceptible to apoptosis by VRB. Orthotopic xenograft assays using two different breast cancer cell models revealed a reduction in tumor volume with BET inhibition and VRB treatment. In addition, in vivo studies using a model of multiple brain metastasis (BM) showed improved survival with the combination of radiation + BET inhibitor (iBET-762) + VRB (75% long-term survivors, P < 0.05). Using in silico analysis and BET inhibition, we found that the transcription factor myeloid zinc finger-1 (MZF-1) protein binds to the TUBB3 promoter. BET inhibition decreases MZF-1 expression and subsequently increases TUBB3 expression. Overexpression of MZF-1 decreases TUBB3 expression and reduces BM in vivo, whereas its knockdown increases TUBB3 expression in breast cancer cells. In summary, this study demonstrates a regulatory mechanism of TUBB3 and provides support for an application of BET inhibition to sensitize breast cancer metastases to VRB-mediated therapy.

AB - Metastases from primary breast cancer result in poor survival. βIII-tubulin (TUBB3) has been established as a therapeutic target for breast cancer metastases specifically to the brain. In this study, we conducted a systematic analysis to determine the regulation of TUBB3 expression in breast cancer metastases to the brain and strategically target these metastases using vinorelbine (VRB), a drug approved by the U.S. Food and Drug Administration (FDA). We found that human epidermal growth factor receptor 2 (HER2) signaling regulates TUBB3 expression in both trastuzumab-sensitive and trastuzumab-resistant neoplastic cells. We further discovered that bromodomain and extra-terminal domain (BET) inhibition increases TUBB3 expression, rendering neoplastic cells more susceptible to apoptosis by VRB. Orthotopic xenograft assays using two different breast cancer cell models revealed a reduction in tumor volume with BET inhibition and VRB treatment. In addition, in vivo studies using a model of multiple brain metastasis (BM) showed improved survival with the combination of radiation + BET inhibitor (iBET-762) + VRB (75% long-term survivors, P < 0.05). Using in silico analysis and BET inhibition, we found that the transcription factor myeloid zinc finger-1 (MZF-1) protein binds to the TUBB3 promoter. BET inhibition decreases MZF-1 expression and subsequently increases TUBB3 expression. Overexpression of MZF-1 decreases TUBB3 expression and reduces BM in vivo, whereas its knockdown increases TUBB3 expression in breast cancer cells. In summary, this study demonstrates a regulatory mechanism of TUBB3 and provides support for an application of BET inhibition to sensitize breast cancer metastases to VRB-mediated therapy.

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BET inhibition increases βIII-tubulin expression and sensitizes metastatic breast cancer in the brain to vinorelbine

Abstract

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UN SDGs

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