High-throughput screening identifies idasanutlin as a resensitizing drug for venetoclax-resistant neuroblastoma cells

Lindy Vernooij; Laurel T. Bate-Eya; Lindy K. Alles; Jasmine Y. Lee; Bianca Koopmans; Hunter C. Jonus; Nil A. Schubert; Linda Schild; Daphne Lelieveld; David A. Egan; Mark Kerstjens; Ronald W. Stam; Jan Koster; Kelly C. Goldsmith; Jan J. Molenaar; M. M. Emmy Dolman

doi:https://doi.org/10.1158/1535-7163.MCT-20-0666

High-throughput screening identifies idasanutlin as a resensitizing drug for venetoclax-resistant neuroblastoma cells

Lindy Vernooij, Laurel T. Bate-Eya, Lindy K. Alles, Jasmine Y. Lee, Bianca Koopmans, Hunter C. Jonus, Nil A. Schubert, Linda Schild, Daphne Lelieveld, David A. Egan, Mark Kerstjens, Ronald W. Stam, Jan Koster, Kelly C. Goldsmith, Jan J. Molenaar, M. M. Emmy Dolman

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

8 Citations (Scopus)

Abstract

Neuroblastoma tumors frequently overexpress the anti-apoptotic protein B-cell lymphoma/leukemia 2 (BCL-2). We previously showed that treating BCL-2-dependent neuroblastoma cells with the BCL-2 inhibitor venetoclax results in apoptosis, but unfortunately partial therapy resistance is observed. The current study describes the identification of drugs capable of resensitizing venetoclax-resistant neuroblastoma cells to venetoclax. To examine these effects, venetoclax resistance was induced in BCL-2-dependent neuroblastoma cell lines KCNR and SJNB12 by continuous exposure to high venetoclax concentrations. Nonresistant and venetoclax-resistant neuroblastoma cell lines were exposed to a 209-compound library in the absence and presence of venetoclax to identify compounds that were more effective in the venetoclax-resistant cell lines under venetoclax pressure. Top hits were further validated in combination with venetoclax using BCL-2-dependent neuroblastoma model systems. Overall, high-throughput drug screening identified the MDM2 inhibitor idasanutlin as a promising resensitizing agent for venetoclax-resistant neuroblastoma cell lines. Idasanutlin treatment induced BAX-mediated apoptosis in venetoclax-resistant neuroblastoma cells in the presence of venetoclax, whereas it caused p21-mediated growth arrest in control cells. In vivo combination treatment showed tumor regression and superior efficacy over single-agent therapies in a BCL-2-dependent neuroblastoma cell line xenograft and a patient-derived xenograft. However, xenografts less dependent on BCL-2 were not sensitive to venetoclax-idasanutlin combination therapy. This study demonstrates that idasanutlin can overcome resistance to the BCL-2 inhibitor venetoclax in preclinical neuroblastoma model systems, which supports clinical development of a treatment strategy combining the two therapies.

Original language	English
Pages (from-to)	1161-1172
Number of pages	12
Journal	Molecular Cancer Therapeutics
Volume	20
Issue number	6
DOIs	https://doi.org/10.1158/1535-7163.MCT-20-0666
Publication status	Published - 1 Jun 2021

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https://doi.org/10.1158/1535-7163.MCT-20-0666

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Vernooij, L., Bate-Eya, L. T., Alles, L. K., Lee, J. Y., Koopmans, B., Jonus, H. C., Schubert, N. A., Schild, L., Lelieveld, D., Egan, D. A., Kerstjens, M., Stam, R. W., Koster, J., Goldsmith, K. C., Molenaar, J. J., & Dolman, M. M. E. (2021). High-throughput screening identifies idasanutlin as a resensitizing drug for venetoclax-resistant neuroblastoma cells. Molecular Cancer Therapeutics, 20(6), 1161-1172. https://doi.org/10.1158/1535-7163.MCT-20-0666

@article{ce471bc6acf74c88bbec536229bb33d6,

title = "High-throughput screening identifies idasanutlin as a resensitizing drug for venetoclax-resistant neuroblastoma cells",

abstract = "Neuroblastoma tumors frequently overexpress the anti-apoptotic protein B-cell lymphoma/leukemia 2 (BCL-2). We previously showed that treating BCL-2-dependent neuroblastoma cells with the BCL-2 inhibitor venetoclax results in apoptosis, but unfortunately partial therapy resistance is observed. The current study describes the identification of drugs capable of resensitizing venetoclax-resistant neuroblastoma cells to venetoclax. To examine these effects, venetoclax resistance was induced in BCL-2-dependent neuroblastoma cell lines KCNR and SJNB12 by continuous exposure to high venetoclax concentrations. Nonresistant and venetoclax-resistant neuroblastoma cell lines were exposed to a 209-compound library in the absence and presence of venetoclax to identify compounds that were more effective in the venetoclax-resistant cell lines under venetoclax pressure. Top hits were further validated in combination with venetoclax using BCL-2-dependent neuroblastoma model systems. Overall, high-throughput drug screening identified the MDM2 inhibitor idasanutlin as a promising resensitizing agent for venetoclax-resistant neuroblastoma cell lines. Idasanutlin treatment induced BAX-mediated apoptosis in venetoclax-resistant neuroblastoma cells in the presence of venetoclax, whereas it caused p21-mediated growth arrest in control cells. In vivo combination treatment showed tumor regression and superior efficacy over single-agent therapies in a BCL-2-dependent neuroblastoma cell line xenograft and a patient-derived xenograft. However, xenografts less dependent on BCL-2 were not sensitive to venetoclax-idasanutlin combination therapy. This study demonstrates that idasanutlin can overcome resistance to the BCL-2 inhibitor venetoclax in preclinical neuroblastoma model systems, which supports clinical development of a treatment strategy combining the two therapies.",

author = "Lindy Vernooij and Bate-Eya, {Laurel T.} and Alles, {Lindy K.} and Lee, {Jasmine Y.} and Bianca Koopmans and Jonus, {Hunter C.} and Schubert, {Nil A.} and Linda Schild and Daphne Lelieveld and Egan, {David A.} and Mark Kerstjens and Stam, {Ronald W.} and Jan Koster and Goldsmith, {Kelly C.} and Molenaar, {Jan J.} and Dolman, {M. M. Emmy}",

note = "Funding Information: The research in this article was supported by grants from the Villa Joep Foundation (grant BCL-2 in neuroblastoma), the Kinderen Kankervrij Foundation (KiKa; grant 189), ERC-START (grant PREDICT-716079), NWO-Vidi (grant 91716482) and Aflac Pilot grant (to K.C. Goldsmith). We would like to thank Dr. C. Patrick Reynolds and Cogcell.org for providing us with the COG-N-424x PDX model and the CHOA Pediatric Biorepository for the CHOA-NBX-4 PDX model. Publisher Copyright: {\textcopyright} 2021 American Association for Cancer Research.",

year = "2021",

month = jun,

day = "1",

doi = "https://doi.org/10.1158/1535-7163.MCT-20-0666",

language = "English",

volume = "20",

pages = "1161--1172",

journal = "Molecular Cancer Therapeutics",

issn = "1535-7163",

publisher = "American Association for Cancer Research Inc.",

number = "6",

}

Vernooij, L, Bate-Eya, LT, Alles, LK, Lee, JY, Koopmans, B, Jonus, HC, Schubert, NA, Schild, L, Lelieveld, D, Egan, DA, Kerstjens, M, Stam, RW, Koster, J, Goldsmith, KC, Molenaar, JJ & Dolman, MME 2021, 'High-throughput screening identifies idasanutlin as a resensitizing drug for venetoclax-resistant neuroblastoma cells', Molecular Cancer Therapeutics, vol. 20, no. 6, pp. 1161-1172. https://doi.org/10.1158/1535-7163.MCT-20-0666

TY - JOUR

T1 - High-throughput screening identifies idasanutlin as a resensitizing drug for venetoclax-resistant neuroblastoma cells

AU - Vernooij, Lindy

AU - Bate-Eya, Laurel T.

AU - Alles, Lindy K.

AU - Lee, Jasmine Y.

AU - Koopmans, Bianca

AU - Jonus, Hunter C.

AU - Schubert, Nil A.

AU - Schild, Linda

AU - Lelieveld, Daphne

AU - Egan, David A.

AU - Kerstjens, Mark

AU - Stam, Ronald W.

AU - Koster, Jan

AU - Goldsmith, Kelly C.

AU - Molenaar, Jan J.

AU - Dolman, M. M. Emmy

N1 - Funding Information: The research in this article was supported by grants from the Villa Joep Foundation (grant BCL-2 in neuroblastoma), the Kinderen Kankervrij Foundation (KiKa; grant 189), ERC-START (grant PREDICT-716079), NWO-Vidi (grant 91716482) and Aflac Pilot grant (to K.C. Goldsmith). We would like to thank Dr. C. Patrick Reynolds and Cogcell.org for providing us with the COG-N-424x PDX model and the CHOA Pediatric Biorepository for the CHOA-NBX-4 PDX model. Publisher Copyright: © 2021 American Association for Cancer Research.

PY - 2021/6/1

Y1 - 2021/6/1

N2 - Neuroblastoma tumors frequently overexpress the anti-apoptotic protein B-cell lymphoma/leukemia 2 (BCL-2). We previously showed that treating BCL-2-dependent neuroblastoma cells with the BCL-2 inhibitor venetoclax results in apoptosis, but unfortunately partial therapy resistance is observed. The current study describes the identification of drugs capable of resensitizing venetoclax-resistant neuroblastoma cells to venetoclax. To examine these effects, venetoclax resistance was induced in BCL-2-dependent neuroblastoma cell lines KCNR and SJNB12 by continuous exposure to high venetoclax concentrations. Nonresistant and venetoclax-resistant neuroblastoma cell lines were exposed to a 209-compound library in the absence and presence of venetoclax to identify compounds that were more effective in the venetoclax-resistant cell lines under venetoclax pressure. Top hits were further validated in combination with venetoclax using BCL-2-dependent neuroblastoma model systems. Overall, high-throughput drug screening identified the MDM2 inhibitor idasanutlin as a promising resensitizing agent for venetoclax-resistant neuroblastoma cell lines. Idasanutlin treatment induced BAX-mediated apoptosis in venetoclax-resistant neuroblastoma cells in the presence of venetoclax, whereas it caused p21-mediated growth arrest in control cells. In vivo combination treatment showed tumor regression and superior efficacy over single-agent therapies in a BCL-2-dependent neuroblastoma cell line xenograft and a patient-derived xenograft. However, xenografts less dependent on BCL-2 were not sensitive to venetoclax-idasanutlin combination therapy. This study demonstrates that idasanutlin can overcome resistance to the BCL-2 inhibitor venetoclax in preclinical neuroblastoma model systems, which supports clinical development of a treatment strategy combining the two therapies.

AB - Neuroblastoma tumors frequently overexpress the anti-apoptotic protein B-cell lymphoma/leukemia 2 (BCL-2). We previously showed that treating BCL-2-dependent neuroblastoma cells with the BCL-2 inhibitor venetoclax results in apoptosis, but unfortunately partial therapy resistance is observed. The current study describes the identification of drugs capable of resensitizing venetoclax-resistant neuroblastoma cells to venetoclax. To examine these effects, venetoclax resistance was induced in BCL-2-dependent neuroblastoma cell lines KCNR and SJNB12 by continuous exposure to high venetoclax concentrations. Nonresistant and venetoclax-resistant neuroblastoma cell lines were exposed to a 209-compound library in the absence and presence of venetoclax to identify compounds that were more effective in the venetoclax-resistant cell lines under venetoclax pressure. Top hits were further validated in combination with venetoclax using BCL-2-dependent neuroblastoma model systems. Overall, high-throughput drug screening identified the MDM2 inhibitor idasanutlin as a promising resensitizing agent for venetoclax-resistant neuroblastoma cell lines. Idasanutlin treatment induced BAX-mediated apoptosis in venetoclax-resistant neuroblastoma cells in the presence of venetoclax, whereas it caused p21-mediated growth arrest in control cells. In vivo combination treatment showed tumor regression and superior efficacy over single-agent therapies in a BCL-2-dependent neuroblastoma cell line xenograft and a patient-derived xenograft. However, xenografts less dependent on BCL-2 were not sensitive to venetoclax-idasanutlin combination therapy. This study demonstrates that idasanutlin can overcome resistance to the BCL-2 inhibitor venetoclax in preclinical neuroblastoma model systems, which supports clinical development of a treatment strategy combining the two therapies.

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U2 - https://doi.org/10.1158/1535-7163.MCT-20-0666

DO - https://doi.org/10.1158/1535-7163.MCT-20-0666

M3 - Article

C2 - 33850004

SN - 1535-7163

VL - 20

SP - 1161

EP - 1172

JO - Molecular Cancer Therapeutics

JF - Molecular Cancer Therapeutics

IS - 6

ER -

High-throughput screening identifies idasanutlin as a resensitizing drug for venetoclax-resistant neuroblastoma cells

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