Epigenetic Targeting of Adipocytes Inhibits High-Grade Serous Ovarian Cancer Cell Migration and Invasion

Ovarian cancer (OC) cells frequently metastasize to the omentum, and adipocytes play a significant role in ovarian tumor progression. Therapeutic interventions targeting aberrant DNA methylation in ovarian tumors have shown promise in the clinic, but the effects of epigenetic therapy on the tumor microenvironment are understudied. Here, we examined the effect of adipocytes on OC cell behavior in culture and impact of targeting DNA methylation in adipocytes on OC metastasis. The presence of adipocytes increased OC cell migration and invasion, and proximal and direct coculture of adipocytes increased OC proliferation alone or after treatment with carboplatin. Treatment of adipocytes with hypomethylating agent guadecitabine decreased migration and invasion of OC cells toward adipocytes. Subcellular protein fractionation of adipocytes treated with guadecitabine revealed decreased DNA methyltransferase 1 (DNMT1) levels even in the presence of DNA synthesis inhibitor, aphidicolin. Methyl-Capture- and RNA-sequencing analysis of guadecitabine-treated adipocytes revealed derepression of tumor-suppressor genes and epithelial–mesenchymal transition inhibitors. SUSD2, a secreted tumor suppressor downregulated by promoter CpG island methylation in adipocytes, was upregulated after guadecitabine treatment, and recombinant SUSD2 decreased OC cell migration and invasion. Integrated analysis of the methylomic and transcriptomic data identified pathways associated with inhibition of matrix metalloproteases and fatty acid α-oxidation, suggesting a possible mechanism of how epigenetic therapy of adipocytes decreases metastasis. In conclusion, the effect of DNMT inhibitor on fully differentiated adipocytes suggests that hypomethylating agents may affect the tumor microenvironment to decrease cancer cell metastasis. Implications: Epigenetic targeting of tumor microenvironment can affect metastatic behavior of ovarian cancer cells. Mol Cancer Res; 16(8); 1226–40. ©2018 AACR.

[1]  D. Matei,et al.  A Phase I Clinical Trial of Guadecitabine and Carboplatin in Platinum-Resistant, Recurrent Ovarian Cancer: Clinical, Pharmacokinetic, and Pharmacodynamic Analyses , 2018, Clinical Cancer Research.

[2]  Ana Lago-Sampedro,et al.  Dietary fatty acids modulate adipocyte TNFa production via regulation of its DNA promoter methylation levels. , 2017, The Journal of nutritional biochemistry.

[3]  Xia Li,et al.  The fat and the bad: Mature adipocytes, key actors in tumor progression and resistance , 2017, Oncotarget.

[4]  J. Issa,et al.  DNA Hypomethylating Drugs in Cancer Therapy. , 2017, Cold Spring Harbor perspectives in medicine.

[5]  R. Müller,et al.  Tumor-Host Cell Interactions in Ovarian Cancer: Pathways to Therapy Failure. , 2017, Trends in cancer.

[6]  K. Marycz,et al.  The effects of the DNA methyltranfserases inhibitor 5‐Azacitidine on ageing, oxidative stress and DNA methylation of adipose derived stem cells , 2016, Journal of cellular and molecular medicine.

[7]  W. Steegenga,et al.  Comprehensive DNA Methylation and Gene Expression Profiling in Differentiating Human Adipocytes , 2016, Journal of cellular biochemistry.

[8]  R. Divella,et al.  Obesity and cancer: the role of adipose tissue and adipo-cytokines-induced chronic inflammation , 2016, Journal of Cancer.

[9]  C. Ling,et al.  Human adipogenesis is associated with genome-wide DNA methylation and gene-expression changes. , 2016, Epigenomics.

[10]  X. Li,et al.  Identification of a role for TRIM29 in the control of innate immunity in the respiratory tract , 2016, Nature Immunology.

[11]  R. Drapkin,et al.  SUSD2 expression in high-grade serous ovarian cancer correlates with increased patient survival and defective mesothelial clearance , 2016, Oncogenesis.

[12]  K. Wellen,et al.  Metabolic control of epigenetics in cancer , 2016, Nature Reviews Cancer.

[13]  D. F. Chedom,et al.  Salt-Inducible Kinase 2 Couples Ovarian Cancer Cell Metabolism with Survival at the Adipocyte-Rich Metastatic Niche. , 2016, Cancer cell.

[14]  Matthew N. Metzinger,et al.  Quantitation of Intra-peritoneal Ovarian Cancer Metastasis. , 2016, Journal of Visualized Experiments.

[15]  Philipp Niethammer,et al.  The Cell Nucleus Serves as a Mechanotransducer of Tissue Damage-Induced Inflammation , 2016, Cell.

[16]  B. Xue,et al.  DNA Methylation Biphasically Regulates 3T3-L1 Preadipocyte Differentiation. , 2016, Molecular endocrinology.

[17]  Jun Zeng,et al.  Decreased expression of Sushi Domain Containing 2 correlates to progressive features in patients with hepatocellular carcinoma , 2016, Cancer Cell International.

[18]  Tingting Li,et al.  SUSD2 is frequently downregulated and functions as a tumor suppressor in RCC and lung cancer , 2016, Tumor Biology.

[19]  Jeffrey M. Bhasin,et al.  MethylAction: detecting differentially methylated regions that distinguish biological subtypes , 2015, Nucleic acids research.

[20]  Jun Zeng,et al.  Reduced expression of sushi domain containing 2 is associated with progression of non-small cell lung cancer. , 2015, Oncology letters.

[21]  Matthew N. Metzinger,et al.  Obesity Contributes to Ovarian Cancer Metastatic Success through Increased Lipogenesis, Enhanced Vascularity, and Decreased Infiltration of M1 Macrophages. , 2015, Cancer research.

[22]  M. Beckmann,et al.  Abstract B32: Inhibiting DNA methylation causes an interferon response in cancer via dsRNA including endogenous retroviruses , 2016 .

[23]  S. Kwak,et al.  Obesity-induced DNA hypermethylation of the adiponectin gene mediates insulin resistance , 2015, Nature Communications.

[24]  K. Kim,et al.  Assaying Cell Cycle Status Using Flow Cytometry , 2015, Current protocols in molecular biology.

[25]  E. Hoffman,et al.  An analysis of DNA methylation in human adipose tissue reveals differential modification of obesity genes before and after gastric bypass and weight loss , 2015, Genome Biology.

[26]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[27]  J. Ji,et al.  CSBF/C10orf99, a novel potential cytokine, inhibits colon cancer cell growth through inducing G1 arrest , 2014, Scientific Reports.

[28]  D. Matei,et al.  The Novel, Small-Molecule DNA Methylation Inhibitor SGI-110 as an Ovarian Cancer Chemosensitizer , 2014, Clinical Cancer Research.

[29]  D. Clegg,et al.  Obesity and cancer—mechanisms underlying tumour progression and recurrence , 2014, Nature Reviews Endocrinology.

[30]  W. Shen,et al.  Epigenetic Modification of the Leptin Promoter in Diet-Induced Obese Mice and the Effects of N-3 Polyunsaturated Fatty Acids , 2014, Scientific Reports.

[31]  C. Gieger,et al.  DNA methylation and body-mass index: a genome-wide analysis , 2014, The Lancet.

[32]  E. Lengyel,et al.  Adipose tissue and adipocytes support tumorigenesis and metastasis. , 2013, Biochimica et biophysica acta.

[33]  Robert V Farese,et al.  Cellular fatty acid metabolism and cancer. , 2013, Cell metabolism.

[34]  Leif Groop,et al.  A Six Months Exercise Intervention Influences the Genome-wide DNA Methylation Pattern in Human Adipose Tissue , 2013, PLoS genetics.

[35]  G. Trinchieri,et al.  Molecular Pathways Molecular Pathways : Toll-like Receptors in the Tumor Microenvironment — Poor Prognosis or New Therapeutic Opportunity , 2013 .

[36]  Rick L. Evans,et al.  Multiple Functions of Sushi Domain Containing 2 (SUSD2) in Breast Tumorigenesis , 2012, Molecular Cancer Research.

[37]  Andy H. Lee,et al.  Ovarian Cancer and Body Size: Individual Participant Meta-Analysis Including 25,157 Women with Ovarian Cancer from 47 Epidemiological Studies , 2012, PLoS medicine.

[38]  S. Baylin,et al.  Oxidative damage targets complexes containing DNA methyltransferases, SIRT1, and polycomb members to promoter CpG Islands. , 2011, Cancer cell.

[39]  G. Mills,et al.  Adipocytes promote ovarian cancer metastasis and provide energy for rapid tumor growth , 2011, Nature Medicine.

[40]  M. Lazar,et al.  Forming functional fat: a growing understanding of adipocyte differentiation , 2011, Nature Reviews Molecular Cell Biology.

[41]  W. Samlowski,et al.  The depletion of DNA methyltransferase-1 and the epigenetic effects of 5-aza-2’deoxycytidine (decitabine) are differentially regulated by cell cycle progression , 2011, Epigenetics.

[42]  Peter A. Jones,et al.  S110, a 5-Aza-2′-Deoxycytidine–Containing Dinucleotide, Is an Effective DNA Methylation Inhibitor In vivo and Can Reduce Tumor Growth , 2010, Molecular Cancer Therapeutics.

[43]  Y. Hayashizaki,et al.  Isolation of a novel mouse gene, mSVS‐1/SUSD2, reversing tumorigenic phenotypes of cancer cells in vitro , 2007, Cancer science.

[44]  H. Sul,et al.  Pref-1, a preadipocyte secreted factor that inhibits adipogenesis. , 2006, The Journal of nutrition.

[45]  Pearlly Yan,et al.  Antimitogenic and chemosensitizing effects of the methylation inhibitor zebularine in ovarian cancer , 2005, Molecular Cancer Therapeutics.

[46]  K. Ghoshal,et al.  5-Aza-Deoxycytidine Induces Selective Degradation of DNA Methyltransferase 1 by a Proteasomal Pathway That Requires the KEN Box, Bromo-Adjacent Homology Domain, and Nuclear Localization Signal , 2005, Molecular and Cellular Biology.

[47]  Peter A. Jones,et al.  Cellular differentiation, cytidine analogs and DNA methylation , 1980, Cell.

[48]  A. Jemal,et al.  Cancer statistics, 2017 , 2017, CA: a cancer journal for clinicians.

[49]  V. Zachar,et al.  Adipogenic differentiation of human mesenchymal stem cells. , 2011, Methods in molecular biology.

[50]  M. Wabitsch,et al.  Characterization of a human preadipocyte cell strain with high capacity for adipose differentiation , 2001, International Journal of Obesity.

[51]  N. Dubrawsky.  Cancer statistics , 1989, CA: a cancer journal for clinicians.