Abstract
Tumor organoids recapitulate pathological properties and would serve as an excellent ex vivo model for drug discovery. Here, we performed an unbiased drug screening on drivers-defined tumor organoids from mouse endometrial cancer, the most prevalent gynecological malignancy in human, with a small molecule library targeting epigenetic factors. Among them, menin-MLL inhibitors MI-136 and MI-463 scored. The therapeutic capacity of MI-136 was further validated in tumor organoids in vitro and an orthotopic model in vivo. CRISPR/cas9-mediated mutations of major components of the menin-MLL complex, Men1, Kmt2a and Ash2l, inhibited the growth of tumor organoids, suggesting that the complex was the target of MI-136. Transcriptome analysis showed that the hypoxia-inducible factor (HIF) pathway was the most significantly downregulated pathway by MI-136 treatment. Consistently, Men1, Kmt2a, and Ash2l knockout also repressed the expressions of the HIF target genes. Loss of Hif1a or Hif1b partially phenocopied the inhibition of the menin-MLL complex by MI-136 or mutations in term of tumor organoid growth. Further, we found that MEN1 was upregulated in human endometrial cancers, which were tightly correlated with the expression levels of HIF1A, and associated with poor prognosis. Importantly, MI-136 also significantly inhibited the growth of endometrial cancer organoids derived from patients. Thus, our study identified MI-136 as a potential inhibitor for endometrial cancer through regulating the HIF pathway, a novel molecular mechanism distinguished from those in AML and prostate cancer.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Roy A, Matzuk MM. Reproductive tract function and dysfunction in women. Nat Rev Endocrinol. 2011;7:517–25.
Jemal A, Bray F, Cente MM, Ferlay JJ, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90.
Morice P, Leary A, Creutzberg C, Abu-Rustum N, Darai E, Morice P, et al. Endometrial cancer. Lancet 2016;387:1094–108.
Grywalska E, Sobstyl M, Putowski L, Rolinski J. Current possibilities of gynecologic cancer treatment with the use of immune checkpoint inhibitors. Int J Mol Sci. 2019;20:4705.
Lheureux S, Oza AM. Endometrial cancer-targeted therapies myth or reality? Review of current targeted treatments. Eur J Cancer 2016;59:99–108.
Cherniack AD, Shen H, Walter V, Stewart C, Murray BA, Bowlby R, et al. Integrated molecular characterization of uterine carcinosarcoma. Cancer Cell 2017;31:411–23.
Getz G, Gabriel SB, Cibulskis K, Lander E, Sivachenko A, Sougnez C, et al. Integrated genomic characterization of endometrial carcinoma. Nature 2013;497:67–73.
Kuhn E, Wu RC, Guan B, Wu G, Zhang JH, Wang Y. Identification of molecular pathway aberrations in uterine serous carcinoma by genome-wide analyses. J Natl Cancer Inst. 2012;104:1503–13.
Cheung LWT, Hennessy BT, Li J, Yu S, Myers AP, Djordjevic B, et al. High frequency of PIK3R1 and PIK3R2 mutations in endometrial cancer elucidates a novel mechanism for regulation of PTEN protein stability. Cancer Disco. 2011;1:170–85.
Levine RL, Cargile CB, Blazes MS, van Rees B, Kurman RJ, Ellenson LH. PTEN mutations and microsatellite instability in complex atypical hyperplasia, a precursor lesion to uterine endometrioid carcinoma. Cancer Res. 1998;58:3254–8.
Petrella BL, Brinckerhoff CE. PTEN suppression of YY1 induces HIF-2 activity in von-Hippel-Lindau-null renal-cell carcinoma. Cancer Biol Ther. 2009;8:1389–401.
Bartosch C, Lopes JM, Jerónimo C. Epigenetics in endometrial carcinogenesis - part 1: DNA methylation. Epigenomics 2017;9:737–55.
Salvesen HB, Macdonald N, Ryan A, Jacobs IJ, Lynch ED, Akslen LA, et al. PTEN methylation is associated with advanced stage and microsatellite instability in endometrial carcinoma. Int J Cancer 2001;91:22–6.
Dewdney SB, Rimel B, Thaker H, Thompson DM, Schmidt A. Huettner, et al. Aberrant methylation of the X-linked ribosomal S6 kinase RPS6KA6 (RSK4) in endometrial cancers. Clin Cancer Res. 2011;17:2120–9.
Seeber LMS, Zweemer R, Marchionni L, Massuger LFAG, Smit VTHBM, Van Baal WM, et al. Methylation profiles of endometrioid and serous endometrial cancers. Endocr Relat Cancer 2010;17:663–73.
Suehiro Y, Okada T, Okada T, Anno K, Okayama N, Ueno K, et al. Aneuploidy predicts outcome in patients with endometrial carcinoma and is related to lack of CDH13 hypermethylation. Clin Cancer Res. 2008;14:3354–61.
Guida M, Sanguedolce F, Bufo, Sardo ADS, Pannone G. Aberrant DNA hypermethylation of hMLH-1 and CDKN2A/p16 genes in benign, premalignant and malignant endometrial lesions. Eur J Gynaecol Oncol. 2009;30:267–70.
Daniela F, Ileana C, Barbara M, Roberta C, Emanuele D, Carlo C, et al. The high frequency of de novo promoter methylation in synchronous primary endometrial and ovarian carcinomas. Clin Cancer Res. 2006;12:3329–36.
Xu S, Ren J, Bin Chen H, Wang Y, Liu Q, Zhang R, et al. Cytostatic and apoptotic effects of DNMT and HDAC inhibitors in endometrial cancer cells. Curr Pharm Des. 2014;20:1881–7.
Satoshi I, Jyoji I, Keiko S. Protein expression, mRNA expression and gene amplification of DNA methyltransferase 1 in endometrial tumor tissues. Mol Clin Oncol. 2013;1:423–9.
Claudia AK, Anne JV, Irmgard CL, Ulrike von R, Henning MB, Joachim A. Class I histone deacetylase expression in the human cyclic endometrium and endometrial adenocarcinomas. Hum Reprod. 2007;22:2956–66.
Wilko W, Carsten D, Aurelia N, Silvia DE, Manfred D, Steve EK, et al. Expression of class I histone deacetylases indicates poor prognosis in endometrioid subtypes of ovarian and endometrial carcinomas. Neoplasia 2008;10:1021–7.
Asaka R, Miyamoto T, Yamada Y, Ando H, Mvunta DH, Kobara H, et al. Sirtuin 1 promotes the growth and cisplatin resistance of endometrial carcinoma cells: a novel therapeutic target. Lab Invest. 2015;95:1363–73.
Theisen ER, Gajiwala S, Bearss J, Sorna V, Sharma S, Janat-Amsbury M. Reversible inhibition of lysine specific demethylase 1 is a novel anti-tumor strategy for poorly differentiated endometrial carcinoma. BMC Cancer 2014;14:752.
Bartosch C, Lopes JM, jeronimo C. Epigenetics in endometrial carcinogenesis - part 2: histone modifications, chromatin remodeling and noncoding RNAs. Epigenomics 2017;9:873–92.
Tom VN, Moiola CP, Eva C, Daniela A. Modeling endometrial cancer: past, present, and future. Int J Mol Sci. 2018;19:2348.
Fatehullah A, Si Hui T, Barker N. Organoids as an in vitro model of human development and disease. Nat Cell Biol. 2016;18:246–54.
Huang L, Holtzinger A, Jagan I, Nostro C, Wang RN, Muthuswamy L, et al. Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell- and patient-derived tumor organoids. Nat Med. 2015;21:1364–71.
Drost J, Clevers H. Organoids in cancer research. Nat Rev Cancer 2018;18:407–18.
Sachs N, de Ligt J, Kopper O, Gogola E, Bounova G, Weeber F, et al. A living biobank of breast cancer organoids captures disease heterogeneity. Cell 2018;172:373–86.
Turco MY, Gardner L, Hughes J, Cindrova-Davies T, Gomez MJ, Farrell L, et al. Long-term, hormone-responsive organoid cultures of human endometrium in a chemically defined medium. Nat Cell Biol. 2017;19:568–77.
Boretto M, Maenhoudt N, Luo XL, Hennes A, Boeckx B, Bui B, et al. Patient-derived organoids from endometrial disease capture clinical heterogeneity and are amenable to drug screening. Nat Cell Biol. 2019;21:1041–51.
Takai N. Histone deacetylase inhibitors have a profound antigrowth activity in endometrial cancer cells. Clin Cancer Res. 2004;10:1141–9.
Kiyoko U, Umene K, Yanokura M, Banno K, Irie H, Adachi M, et al. Aurora kinase A has a significant role as a therapeutic target and clinical biomarker in endometrial cancer. Int J Oncol. 2015;46:1498–506.
Bestvina CM, Fleming GF. Chemotherapy for endometrial cancer in adjuvant and advanced disease settings. Oncologist 2016;21:1250–9.
Shi A, Murai MJ, He S, Lund G, Hartley T, Purohit T, et al. Structural insights into inhibition of the bivalent menin-MLL interaction by small molecules in leukemia. Blood 2012;120:4461–9.
Grembecka J, He S, Shi A, Purohit T, Muntean AG, Sorenson RJ, et al. Menin-MLL inhibitors reverse oncogenic activity of MLL fusion proteins in leukemia. Nat Chem Biol. 2012;8:277–84.
Malik R, Khan AP, Asangani IA, Cielik M, Chinnaiyan AM. Targeting the MLL complex in castration-resistant prostate cancer. Nat Med. 2015;21:344–52.
Salimian Rizi B, Caneba C, Nowicka A, Nabiyar AW, Liu X, Chen K, et al. Nitric oxide mediates metabolic coupling of omentum-derived adipose stroma to ovarian and endometrial cancer cells. Cancer Res. 2015;75:456–71.
Basudhar D, Glynn SA, Greer M, Somasundaram V, No JH, Scheiblin DA, et al. Coexpression of NOS2 and COX2 accelerates tumor growth and reduces survival in estrogen receptor-negative breast cancer. Proc Natl Acad Sci USA. 2017;114:13030–5.
Shi L, Pan H, Liu Z, Xie J, Han W. Roles of PFKFB3 in cancer. Signal Transduct Target Ther. 2017;2:17044.
Gustafsson NMS, Färnegårdh K, Bonagas N, Ninou AH, Groth P, Wiita E, et al. Targeting PFKFB3 radiosensitizes cancer cells and suppresses homologous recombination. Nat Commun. 2018;9:3872.
Nwosu ZC, Ebert MP, Dooley S, Meyer C. Caveolin-1 in the regulation of cell metabolism: a cancer perspective. Mol Cancer 2016;15:71.
Krivtsov AV, Evans K, Gadrey JY, Eschle BK, Hatton C, Uckelmann HJ, et al. A menin-MLL inhibitor induces specific chromatin changes and eradicates disease in models of MLL-rearranged leukemia. Cancer Cell 2019;36:660–73.
Ning Z, Jia NF, Ting CC. Synergistic combination of microtubule targeting anticancer fludelone with cytoprotective panaxytriol derived from panax ginseng against MX-1 cells in vitro: experimental design and data analysis using the combination index method. Am J Cancer Res. 2016;6:97–104.
Alexander D, Carrie AD, Felix S, Jorg D, Chris Z, Sonali J, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics 2013;29:15–21.
Guang CY, Li GW, Yan YH, Qing YH. clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS 2012;16:284–7.
Aravind S, Pablo T, Vamsi KM, Sayan M, Benjamin LE, Michael AG, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA. 2005;102:15545–50.
Hanbo C, Paul CB. VennDiagram: a package for the generation of highly-customizable Venn and Euler diagrams in R. BMC Bioinforma 2011;12:35.
Heinz S, Benner C, Spann N, Bertolino E, Lin YC, Laslo P, et al. Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities. Mol Cell 2010;38:576–89.
Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15:550.
H Wickham. ggplot2: Elegant Graphics for Data Analysis. 2nd edn. (Springer Nature, New York, 2016).
Robinson JT, Thorvaldsdóttir H, Winckler W, Guttman M, Lander ES, Getz G, et al. Integrative genomics viewer. Nat Biotechnol. 2011;29:24–6.
Acknowledgements
We thank all Chen-Liu laboratory members for their invaluable discussions and technical support.
Funding
This work was supported by the National Key R&D Program of China (2017YFA0505600).
Author information
Authors and Affiliations
Contributions
JC, LZ, YL, CC, and FN conceived the project, designed experiments, and wrote the paper. JC, HP, SD, MW, JW, YQ, ZB, YZ, and SZ performed experiments and analyzed data. LZ and JC performed RNA-seq and ChIP-seq analysis.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval and consent to participate
All the collection of specimens and animal handling in this work was reviewed and approved by the Medical Ethics Committee of the Sichuan University.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Rights and permissions
About this article
Cite this article
Chen, J., Zhao, L., Peng, H. et al. An organoid-based drug screening identified a menin-MLL inhibitor for endometrial cancer through regulating the HIF pathway. Cancer Gene Ther 28, 112–125 (2021). https://doi.org/10.1038/s41417-020-0190-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41417-020-0190-y
This article is cited by
-
Collagen-based biomaterials in organoid technology for reproductive medicine: composition, characteristics, and applications
Collagen and Leather (2023)
-
Combinatorial targeting of menin and the histone methyltransferase DOT1L as a novel therapeutic strategy for treatment of chemotherapy-resistant ovarian cancer
Cancer Cell International (2022)
-
Effect of chronic intermittent hypoxia-induced HIF-1α/ATAD2 expression on lung cancer stemness
Cellular & Molecular Biology Letters (2022)
-
Strategies for modelling endometrial diseases
Nature Reviews Endocrinology (2022)
-
Organoids
Nature Reviews Methods Primers (2022)