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Maintenance and pharmacologic targeting of ROR1 protein levels via UHRF1 in t(1;19) pre-B-ALL

Oncogene volume 37, pages 5221–5232 (2018)Cite this article

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Abstract

Expression of the transmembrane pseudokinase ROR1 is required for survival of t(1;19)-pre-B-cell acute lymphoblastic leukemia (t(1;19) pre-B-ALL), chronic lymphocytic leukemia, and many solid tumors. However, targeting ROR1 with small-molecules has been challenging due to the absence of ROR1 kinase activity. To identify genes that regulate ROR1 expression and may, therefore, serve as surrogate drug targets, we employed an siRNA screening approach and determined that the epigenetic regulator and E3 ubiquitin ligase, UHRF1, is required for t(1;19) pre-B-ALL cell viability in a ROR1-dependent manner. Upon UHRF1 silencing, ROR1 protein is reduced without altering ROR1 mRNA, and ectopically expressed UHRF1 is sufficient to increase ROR1 levels. Additionally, proteasome inhibition rescues loss of ROR1 protein after UHRF1 silencing, suggesting a role for the proteasome in the UHRF1-ROR1 axis. Finally, we show that ROR1-positive cells are twice as sensitive to the UHRF1-targeting drug, naphthazarin, and undergo increased apoptosis compared to ROR1-negative cells. Naphthazarin elicits reduced expression of UHRF1 and ROR1, and combination of naphthazarin with inhibitors of pre-B cell receptor signaling results in further reduction of cell survival compared with either inhibitor alone. Therefore, our work reveals a mechanism by which UHRF1 stabilizes ROR1, suggesting a potential targeting strategy to inhibit ROR1 in t(1;19) pre-B-ALL and other malignancies.

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Acknowledgements

The pCS2-6XMYC vector was a generous gift from Dr. Monika Davare. The authors would like to thank David K. Edwards V, Samantha L. Savage, Anna M. Reister Schultz, Dr. Haijiao Zhang, and Janet Pittsenbarger for their technical expertise and Dr. Kevin M. Watanabe-Smith for editing the manuscript. MC was supported in part by the Oregon Clinical and Translational Research Institute (OCTRI) (TL1TR000129) from the National Center for Advancing Translational Sciences (NCATS) at the National Institutes of Health (NIH). BHC is supported by the Hyundai Hope on Wheels. JWT is supported by The Leukemia and Lymphoma Society, the V Foundation for Cancer Research, Gabrielle’s Angel Foundation for Cancer Research, and the National Cancer Institute (5R00CA151457-04; 1R01CA183947-01).

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Authors and Affiliations

  1. Department of Cell, Developmental, and Cancer Biology, Oregon Health and Science University, Portland, USA

    Marilynn Chow & Jeffrey W. Tyner

  2. Knight Cancer Institute, Oregon Health and Science University, Portland, USA

    Lina Gao, Jason D. MacManiman, Vincent T. Bicocca, Bill H. Chang, Joshi J. Alumkal & Jeffrey W. Tyner

  3. Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, USA

    Lina Gao & Joshi J. Alumkal

  4. Division of Hematology and Medical Oncology, Oregon Health and Science University, Portland, USA

    Jason D. MacManiman & Jeffrey W. Tyner

  5. Division of Pediatric Hematology and Oncology at Doernbecher Children’s Hospital, Oregon Health and Science University, Portland, USA

    Bill H. Chang

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Dr. JWT's work has received research support from Agios Pharmaceuticals, Array Biopharma, Aptose Biosciences, AstraZeneca, Constellation Pharmaceuticals, Genentech, Gilead, Incyte Corporation, Janssen Pharmaceutica, Seattle Genetics, Syros, Takeda Pharmaceutical Company and is a co-founder of Leap Oncology. All the remaining authors declare no conflicts of interest.

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Chow, M., Gao, L., MacManiman, J.D. et al. Maintenance and pharmacologic targeting of ROR1 protein levels via UHRF1 in t(1;19) pre-B-ALL. Oncogene 37, 5221–5232 (2018). https://doi.org/10.1038/s41388-018-0299-8

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