Abstract
Most cancer deaths are due to metastatic dissemination to distant organs. Bone is the most frequently affected organ in metastatic prostate cancer and a major cause of prostate cancer deaths. Yet, our partial understanding of the molecular factors that drive bone metastasis has been a limiting factor for developing preventative and therapeutic strategies to improve patient survival and well-being. Although recent studies have uncovered molecular alterations that occur in prostate cancer metastasis, their functional relevance for bone metastasis is not well understood. Using genome-wide CRISPR activation and inhibition screens we have identified multiple drivers and suppressors of prostate cancer metastasis. Through functional validation, including an innovative organ-on-a-chip invasion platform for studying bone tropism, our study identifies the transcriptional modulator CITED2 as a novel driver of prostate cancer bone metastasis and uncovers multiple new potential molecular targets for bone metastatic disease.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Data availability
Sequencing raw counts and differentially expressed sgRNAs are included in the manuscript tables. Plasmids and sgRNA libraries are available from Addgene. Code used for analysis is available at https://sourceforge.net/p/mageck/wiki/Home/. RNAseq data was deposited in GEO, accession number GSE253815.
Change history
16 April 2024
A Correction to this paper has been published: https://doi.org/10.1038/s41388-024-03031-2
References
Hernandez RK, Wade SW, Reich A, Pirolli M, Liede A, Lyman GH. Incidence of bone metastases in patients with solid tumors: analysis of oncology electronic medical records in the United States. BMC Cancer. 2018;18:44.
Gandaglia G, Abdollah F, Schiffmann J, Trudeau V, Shariat SF, Kim SP, et al. Distribution of metastatic sites in patients with prostate cancer: a population-based analysis. Prostate. 2014;74:210–6.
Arriaga JM, Abate-Shen C. Genetically engineered mouse models of prostate cancer in the postgenomic era. Cold Spring Harb Perspect Med. 2019;9:a030528.
Grabowska MM, DeGraff DJ, Yu X, Jin RJ, Chen Z, Borowsky AD, et al. Mouse models of prostate cancer: picking the best model for the question. Cancer Metastasis Rev. 2014;33:377–97.
Cancer Genome Atlas Research N. The molecular taxonomy of primary prostate cancer. Cell. 2015;163:1011–25.
Grasso CS, Wu YM, Robinson DR, Cao X, Dhanasekaran SM, Khan AP, et al. The mutational landscape of lethal castration-resistant prostate cancer. Nature. 2012;487:239–43.
Kumar A, Coleman I, Morrissey C, Zhang X, True LD, Gulati R, et al. Substantial interindividual and limited intraindividual genomic diversity among tumors from men with metastatic prostate cancer. Nat Med. 2016;22:369–78.
Taylor BS, Schultz N, Hieronymus H, Gopalan A, Xiao Y, Carver BS, et al. Integrative genomic profiling of human prostate cancer. Cancer Cell. 2010;18:11–22.
Abida W, Cyrta J, Heller G, Prandi D, Armenia J, Coleman I, et al. Genomic correlates of clinical outcome in advanced prostate cancer. Proc Natl Acad Sci USA. 2019;116:11428–36.
Armenia J, Wankowicz SAM, Liu D, Gao J, Kundra R, Reznik E, et al. The long tail of oncogenic drivers in prostate cancer. Nat Genet. 2018;50:645–51.
Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, et al. Genome-scale CRISPR-mediated control of gene repression and activation. Cell. 2014;159:647–61.
Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE, et al. CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes. Cell. 2013;154:442–51.
Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, et al. Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis. Cell. 2015;160:1246–60.
Bajaj J, Hamilton M, Shima Y, Chambers K, Spinler K, Van Nostrand EL, et al. An in vivo genome-wide CRISPR screen identifies the RNA-binding protein Staufen2 as a key regulator of myeloid leukemia. Nat Cancer. 2020;1:410–22.
Dong MB, Wang G, Chow RD, Ye L, Zhu L, Dai X, et al. Systematic immunotherapy target discovery using genome-scale in vivo CRISPR screens in CD8 T cells. Cell. 2019;178:1189–204.e1123.
Dai M, Yan G, Wang N, Daliah G, Edick AM, Poulet S, et al. In vivo genome-wide CRISPR screen reveals breast cancer vulnerabilities and synergistic mTOR/Hippo targeted combination therapy. Nat Commun. 2021;12:3055.
Yau EH, Kummetha IR, Lichinchi G, Tang R, Zhang Y, Rana TM. Genome-wide CRISPR screen for essential cell growth mediators in mutant KRAS colorectal cancers. Cancer Res. 2017;77:6330–9.
Cheng L, Sun J, Pretlow TG, Culp J, Yang NS. CWR22 xenograft as an ex vivo human tumor model for prostate cancer gene therapy. J Natl Cancer Inst. 1996;88:607–11.
Sramkoski RM, Pretlow TG 2nd, Giaconia JM, Pretlow TP, Schwartz S, Sy MS, et al. A new human prostate carcinoma cell line, 22Rv1. Vitr Cell Dev Biol Anim. 1999;35:403–9.
Kovar JL, Johnson MA, Volcheck WM, Chen J, Simpson MA. Hyaluronidase expression induces prostate tumor metastasis in an orthotopic mouse model. Am J Pathol. 2006;169:1415–26.
Preston Campbell J, Mulcrone P, Masood SK, Karolak M, Merkel A, Hebron K, et al. TRIzol and Alu qPCR-based quantification of metastatic seeding within the skeleton. Sci Rep. 2015;5:12635.
Horlbeck MA, Gilbert LA, Villalta JE, Adamson B, Pak RA, Chen Y, et al. Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation. Elife. 2016;5:e19760.
Li W, Xu H, Xiao T, Cong L, Love MI, Zhang F, et al. MAGeCK enables robust identification of essential genes from genome-scale CRISPR/Cas9 knockout screens. Genome Biol. 2014;15:554.
Ronaldson-Bouchard K, Teles D, Yeager K, Tavakol DN, Zhao Y, Chramiec A, et al. A multi-organ chip with matured tissue niches linked by vascular flow. Nat Biomed Eng. 2022;6:351–71.
Zou M, Toivanen R, Mitrofanova A, Floch N, Hayati S, Sun Y, et al. Transdifferentiation as a mechanism of treatment resistance in a mouse model of castration-resistant prostate cancer. Cancer Discov. 2017;7:736–49.
Arriaga JM, Panja S, Alshalalfa M, Zhao J, Zou M, Giacobbe A, et al. A MYC and RAS co-activation signature in localized prostate cancer drives bone metastasis and castration resistance. Nat Cancer. 2020;1:1082–96.
Shin SH, Lee GY, Lee M, Kang J, Shin HW, Chun YS, et al. Aberrant expression of CITED2 promotes prostate cancer metastasis by activating the nucleolin-AKT pathway. Nat Commun. 2018;9:4113.
Bhattacharya S, Michels CL, Leung MK, Arany ZP, Kung AL, Livingston DM. Functional role of p35srj, a novel p300/CBP binding protein, during transactivation by HIF-1. Genes Dev. 1999;13:64–75.
Chou YT, Hsieh CH, Chiou SH, Hsu CF, Kao YR, Lee CC, et al. CITED2 functions as a molecular switch of cytokine-induced proliferation and quiescence. Cell Death Differ. 2012;19:2015–28.
Hu C, Zhang Y, Tang K, Luo Y, Liu Y, Chen W. Downregulation of CITED2 contributes to TGFbeta-mediated senescence of tendon-derived stem cells. Cell Tissue Res. 2017;368:93–104.
Mattes K, Berger G, Geugien M, Vellenga E, Schepers H. CITED2 affects leukemic cell survival by interfering with p53 activation. Cell Death Dis. 2017;8:e3132.
Gerhauser C, Favero F, Risch T, Simon R, Feuerbach L, Assenov Y, et al. Molecular evolution of early-onset prostate cancer identifies molecular risk markers and clinical trajectories. Cancer Cell. 2018;34:996–1011.e1018.
Pettaway CA, Pathak S, Greene G, Ramirez E, Wilson MR, Killion JJ, et al. Selection of highly metastatic variants of different human prostatic carcinomas using orthotopic implantation in nude mice. Clin Cancer Res. 1996;2:1627–36.
Kim RS, Avivar-Valderas A, Estrada Y, Bragado P, Sosa MS, Aguirre-Ghiso JA, et al. Dormancy signatures and metastasis in estrogen receptor positive and negative breast cancer. PLoS ONE. 2012;7:e35569.
Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. 2020;70:7–30.
Hubbard GK, Mutton LN, Khalili M, McMullin RP, Hicks JL, Bianchi-Frias D, et al. Combined MYC activation and pten loss are sufficient to create genomic instability and lethal metastatic prostate cancer. Cancer Res. 2016;76:283–92.
Joyce JA, Pollard JW. Microenvironmental regulation of metastasis. Nat Rev Cancer. 2009;9:239–52.
Borriello L, Karagiannis GS, Duran CL, Coste A, Oktay MH, Entenberg D, et al. The role of the tumor microenvironment in tumor cell intravasation and dissemination. Eur J Cell Biol. 2020;99:151098.
Jayaraman S, Doucet M, Lau WM, Kominsky SL. CITED2 modulates breast cancer metastatic ability through effects on IKKalpha. Mol Cancer Res. 2016;14:730–9.
Lau WM, Weber KL, Doucet M, Chou YT, Brady K, Kowalski J, et al. Identification of prospective factors promoting osteotropism in breast cancer: a potential role for CITED2. Int J Cancer. 2010;126:876–84.
Valastyan S, Weinberg RA. Tumor metastasis: molecular insights and evolving paradigms. Cell. 2011;147:275–92.
Lawson H, van de Lagemaat LN, Barile M, Tavosanis A, Durko J, Villacreces A, et al. CITED2 coordinates key hematopoietic regulatory pathways to maintain the HSC pool in both steady-state hematopoiesis and transplantation. Stem Cell Rep. 2021;16:2784–97.
Wang H, Zhang W, Bado I, Zhang XH. Bone tropism in cancer metastases. Cold Spring Harb Perspect Med. 2020;10:a036848.
Easterly ME, Foltz CJ, Paulus MJ. Body condition scoring: comparing newly trained scorers and micro-computed tomography imaging. Lab Anim. 2001;30:46–49.
Villasante A, Marturano-Kruik A, Robinson ST, Liu Z, Guo XE, Vunjak-Novakovic G. Tissue-engineered model of human osteolytic bone tumor. Tissue Eng Part C Methods. 2017;23:98–107.
Reich M, Liefeld T, Gould J, Lerner J, Tamayo P, Mesirov JP. GenePattern 2.0. Nat Genet. 2006;38:500–1.
Acknowledgements
The research was supported by NIH grants R01 CA193442, R01 CA173481, R01 CA183929 and P01 CA265768 to CAS, and UH3 EB025765, P41 EB027062 and R01 CA249799 to GVN. JMA was supported by a postdoctoral training grant from the Department of Defense (DOD) Prostate Cancer Research Program (W81XWH-15-1-0185), an Irving Institute/Clinical Trials Office Pilot Award funded by the National Center for Advancing Translational Sciences, NIH (UL1TR001873), the Dean’s Precision Medicine Research Fellowship from the Irving Institute for Clinical and Translational Research at CUIMC (UL1TR001873) and a Prostate Cancer Foundation Young Investigator Award (20YOUN25). CAS is supported by the TJ Martell Foundation for Leukemia, Cancer and AIDS Research and The Prostate Cancer Foundation and is an American Cancer Society Research Professor. FP was supported by a DOD Early Investigator Research Award (W81XWH-22-1-0054). This research was funded in part through the National Institutes of Health (NIH)/NCI Cancer Center Support Grant P30CA013696 awarded to the Herbert Irving Comprehensive Cancer Center (HICCC), which supported the Molecular Pathology, Flow Cytometry, Genomics and High Throughput Screening and Oncology Precision Therapeutics and Imaging Cores at HICCC. We are grateful to Jonathan Weissman for kindly providing the genome-wide libraries used in this study. Some figure panels (as indicated) were created with BioRender.com using an institutional license sponsored by Columbia University’s VP&S Office for Research.
Author information
Authors and Affiliations
Contributions
JMA and CAS conceived and designed the study. JMA collected, assembled and analyzed data. KRB performed and analyzed organ-on-a-chip studies. FP performed and analyzed select mouse studies with assistance from SA. FNA and HC performed select in vitro studies and RNAseq. HC also performed western blot studies. GVN supervised organ-on-a-chip studies and writing of the manuscript. JMA and CAS wrote the manuscript. CAS supervised all studies. All authors approved the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Arriaga, J.M., Ronaldson-Bouchard, K., Picech, F. et al. In vivo genome-wide CRISPR screening identifies CITED2 as a driver of prostate cancer bone metastasis. Oncogene 43, 1303–1315 (2024). https://doi.org/10.1038/s41388-024-02995-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41388-024-02995-5