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In vivo genome-wide CRISPR screening identifies CITED2 as a driver of prostate cancer bone metastasis

Oncogene volume 43, pages 1303–1315 (2024)Cite this article

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A Correction to this article was published on 16 April 2024

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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.

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Fig. 1: In vivo genome-wide screening for activation and inhibition of gene expression results in metastasis of prostate cancer cells.
Fig. 2: Identification and analysis of metastasis hits in all genome-wide screens.
Fig. 3: Functional validation of CITED2 as a driver of prostate cancer bone metastasis.
Fig. 4: Transcriptional outcome of CITED2 overexpression and relevance to human prostate cancer patients.

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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.

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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.

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  1. Juan M. Arriaga

    Present address: Department of Oncological Sciences, Department of Urology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA

Authors and Affiliations

  1. Department of Molecular Pharmacology and Therapeutics, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, NY, 10032, USA

    Juan M. Arriaga, Florencia Picech, Francisca Nunes de Almeida & Stephanie Afari

  2. Department of Biomedical Engineering, Columbia University, New York, NY, 10032, USA

    Kacey Ronaldson-Bouchard & Gordana Vunjak-Novakovic

  3. Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA

    Houssein Chhouri

  4. Department of Medicine, Vagelos College of Physicians and Surgeons, Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY, 10032, USA

    Gordana Vunjak-Novakovic

  5. Departments of Molecular Pharmacology and Therapeutics, Urology, Medicine, Pathology & Cell Biology, and Systems Biology, Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, NY, 10032, USA

    Cory Abate-Shen

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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.

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Correspondence to Juan M. Arriaga or Cory Abate-Shen.

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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

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