A miR-192-EGR1-HOXB9 regulatory network controls the angiogenic switch in cancer

A deeper mechanistic understanding of tumour angiogenesis regulation is needed to improve current anti-angiogenic therapies. Here we present evidence from systems-based miRNA analyses of large-scale patient data sets along with in vitro and in vivo experiments that miR-192 is a key regulator of angiogenesis. The potent anti-angiogenic effect of miR-192 stems from its ability to globally downregulate angiogenic pathways in cancer cells through regulation of EGR1 and HOXB9. Low miR-192 expression in human tumours is predictive of poor clinical outcome in several cancer types. Using 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) nanoliposomes, we show that miR-192 delivery leads to inhibition of tumour angiogenesis in multiple ovarian and renal tumour models, resulting in tumour regression and growth inhibition. This anti-angiogenic and anti-tumour effect is more robust than that observed with an anti-VEGF antibody. Collectively, these data identify miR-192 as a central node in tumour angiogenesis and support the use of miR-192 in an anti-angiogenesis therapy.

[1]  George A. Calin,et al.  RNAi Therapies: Drugging the Undruggable , 2014, Science Translational Medicine.

[2]  D. Tindall,et al.  Expression of early growth response genes in human prostate cancer. , 1998, Cancer research.

[3]  G. Yousef,et al.  miR-192, miR-194 and miR-215: a convergent microRNA network suppressing tumor progression in renal cell carcinoma. , 2013, Carcinogenesis.

[4]  A. Sood,et al.  Resistance and escape from antiangiogenesis therapy: clinical implications and future strategies. , 2012, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[5]  Gerald Hoefler,et al.  miR-192, miR-194, miR-215, miR-200c and miR-141 are downregulated and their common target ACVR2B is strongly expressed in renal childhood neoplasms. , 2012, Carcinogenesis.

[6]  Axel Benner,et al.  A Systematic Approach to Defining the microRNA Landscape in Metastasis. , 2015, Cancer research.

[7]  M. Masuda,et al.  Cardiovascular , Pulmonary and Renal Pathology Early Growth Response-1 Induces and Enhances Vascular Endothelial Growth Factor-A Expression in Lung Cancer Cells , 2010 .

[8]  G. Calin,et al.  Hypoxia Mediated Downregulation of miRNA Biogenesis Promotes Tumor Progression , 2014, Nature Communications.

[9]  Prahlad T. Ram,et al.  NetWalker: a contextual network analysis tool for functional genomics , 2012, BMC Genomics.

[10]  M. Fabbri,et al.  MicroRNAs and other non-coding RNAs as targets for anticancer drug development , 2013, Nature Reviews Drug Discovery.

[11]  Oriol Casanovas,et al.  Drug resistance by evasion of antiangiogenic targeting of VEGF signaling in late-stage pancreatic islet tumors. , 2005, Cancer cell.

[12]  Jacobus Pfisterer,et al.  A phase 3 trial of bevacizumab in ovarian cancer. , 2011, The New England journal of medicine.

[13]  Jeffrey M. Fowler,et al.  Incorporation of bevacizumab in the primary treatment of ovarian cancer , 2012 .

[14]  Liz Y. Han,et al.  Anti-angiogenic properties of metronomic topotecan in ovarian carcinoma , 2009, Cancer biology & therapy.

[15]  Michael A. White,et al.  Use of Data-Biased Random Walks on Graphs for the Retrieval of Context-Specific Networks from Genomic Data , 2010, PLoS Comput. Biol..

[16]  M. Okano,et al.  Cohort Study , 2020, Definitions.

[17]  A. Sood,et al.  Silencing Survivin Splice Variant 2B Leads to Antitumor Activity in Taxane-Resistant Ovarian Cancer , 2011, Clinical Cancer Research.

[18]  Huey-Jen Lin,et al.  Downregulation of p53-inducible microRNAs 192, 194, and 215 impairs the p53/MDM2 autoregulatory loop in multiple myeloma development. , 2010, Cancer cell.

[19]  Chengzhong Xing,et al.  microRNA-192, -194 and -215 are frequently downregulated in colorectal cancer. , 2012, Experimental and therapeutic medicine.

[20]  C. Bucana,et al.  Expression of platelet-derived growth factor and activated receptor in clinical specimens of epithelial ovarian cancer and ovarian carcinoma cell lines. , 2004, Gynecologic oncology.

[21]  Arek Kasprzyk,et al.  The prognostic value of temporal in vitro and in vivo derived hypoxia gene-expression signatures in breast cancer. , 2012, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[22]  Jeffrey S. Morris,et al.  Phase II trial of infusional fluorouracil, irinotecan, and bevacizumab for metastatic colorectal cancer: efficacy and circulating angiogenic biomarkers associated with therapeutic resistance. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[23]  Jianxing He,et al.  MicroRNA-192 targeting retinoblastoma 1 inhibits cell proliferation and induces cell apoptosis in lung cancer cells , 2011, Nucleic acids research.

[24]  L. Khachigian,et al.  Early Growth Response-1 Regulates Angiopoietin-1–Induced Endothelial Cell Proliferation, Migration, and Differentiation , 2009, Arteriosclerosis, thrombosis, and vascular biology.

[25]  John J Rossi,et al.  MicroRNA-192 in diabetic kidney glomeruli and its function in TGF-β-induced collagen expression via inhibition of E-box repressors , 2007, Proceedings of the National Academy of Sciences.

[26]  A. Bhan,et al.  Homeodomain‐containing protein HOXB9 regulates expression of growth and angiogenic factors, facilitates tumor growth in vitro and is overexpressed in breast cancer tissue , 2012, The FEBS journal.

[27]  G. Rao,et al.  15(S)‐Hydroxyeicosatetraenoic acid‐induced angiogenesis requires Src‐mediated Egr‐1‐dependent rapid induction of FGF‐2 expression , 2010, Blood.

[28]  David B Jackson,et al.  Src activation by b-adrenoreceptors is a key switch for tumour metastasis , 2013 .

[29]  S. Maheswaran,et al.  HOXB9 Expression Promoting Tumor Cell Proliferation and Angiogenesis Is Associated with Clinical Outcomes in Breast Cancer Patients , 2012, Annals of Surgical Oncology.

[30]  A. Sood,et al.  Assessment of In Vivo siRNA Delivery in Cancer Mouse Models. , 2016, Methods in molecular biology.

[31]  Xin Zhang,et al.  p53-Responsive micrornas 192 and 215 are capable of inducing cell cycle arrest. , 2008, Cancer research.

[32]  Tae Jin Lee,et al.  p53 regulates epithelial–mesenchymal transition through microRNAs targeting ZEB1 and ZEB2 , 2011, The Journal of experimental medicine.

[33]  Sheila M. Reynolds,et al.  Integrated analyses identify a master microRNA regulatory network for the mesenchymal subtype in serous ovarian cancer. , 2013, Cancer cell.

[34]  George A Calin,et al.  Tumour angiogenesis regulation by the miR-200 family , 2013, Nature Communications.

[35]  Anil K Sood,et al.  Therapeutic EphA2 gene targeting in vivo using neutral liposomal small interfering RNA delivery. , 2005, Cancer research.

[36]  V. Poltoratsky,et al.  Proteasome Inhibition Increases Recruitment of IκB Kinase β (IKKβ), S536P-p65, and Transcription Factor EGR1 to Interleukin-8 (IL-8) Promoter, Resulting in Increased IL-8 Production in Ovarian Cancer Cells* , 2013, The Journal of Biological Chemistry.

[37]  M. Koch,et al.  Tumor Escape from Endogenous, Extracellular Matrix–Associated Angiogenesis Inhibitors by Up-Regulation of Multiple Proangiogenic Factors , 2008, Clinical Cancer Research.

[38]  Ming-Xi Li,et al.  VEGF suppresses epithelial-mesenchymal transition by inhibiting the expression of Smad3 and miR‑192, a Smad3-dependent microRNA. , 2013, International journal of molecular medicine.

[39]  Prahlad T. Ram,et al.  Hematogenous metastasis of ovarian cancer: rethinking mode of spread. , 2014, Cancer cell.

[40]  William C Reinhold,et al.  CellMiner: a relational database and query tool for the NCI-60 cancer cell lines , 2009, BMC Genomics.

[41]  Y. Joo,et al.  Expression of early growth response-1 in colorectal cancer and its relation to tumor cell proliferation and apoptosis. , 2014, Oncology reports.

[42]  E. Bandrés,et al.  miRNA cargo within exosome‐like vesicle transfer influences metastatic bone colonization , 2014, Molecular oncology.

[43]  G. Verdine.  Drugging the "undruggable". , 2006, Harvey lectures.

[44]  Kyung-Hee Lee,et al.  Hepatocyte growth factor induced up-regulations of VEGF through Egr-1 in hepatocellular carcinoma cells , 2009, Clinical & Experimental Metastasis.

[45]  David B Jackson,et al.  Src activation by β-adrenoreceptors is a key switch for tumor metastasis , 2012, Nature Communications.

[46]  G. Calin,et al.  Plasma circulating-microRNA profiles are useful for assessing prognosis in patients with cytogenetically normal myelodysplastic syndromes , 2015, Modern Pathology.

[47]  Prahlad T. Ram,et al.  2′-OMe-phosphorodithioate-modified siRNAs show increased loading into the RISC complex and enhanced anti-tumour activity , 2014, Nature Communications.

[48]  L. Klotz,et al.  Identification and Validation of a Five MicroRNA Signature Predictive of Prostate Cancer Recurrence and Metastasis: A Cohort Study , 2015, Journal of Cancer.

[49]  Tracy T Batchelor,et al.  AZD2171, a pan-VEGF receptor tyrosine kinase inhibitor, normalizes tumor vasculature and alleviates edema in glioblastoma patients. , 2007, Cancer cell.

[50]  G. Goodall,et al.  E-Cadherin Expression Is Regulated by miR-192/215 by a Mechanism That Is Independent of the Profibrotic Effects of Transforming Growth Factor-β , 2010, Diabetes.

[51]  T. H. van der Kwast,et al.  Expression and prognostic value of Wilms' tumor 1 and early growth response 1 proteins in nephroblastoma. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[52]  C. Creighton,et al.  The microRNA-200/Zeb1 axis regulates ECM-dependent β1-integrin/FAK signaling, cancer cell invasion and metastasis through CRKL , 2016, Scientific Reports.

[53]  Hongquan Zhang,et al.  High expression of transcriptional factor HoxB9 predicts poor prognosis in patients with lung adenocarcinoma , 2015, Histopathology.

[54]  A. Bamias,et al.  Targeting angiogenesis in renal cell carcinoma. , 2008, Current cancer drug targets.

[55]  M. Brattain,et al.  MicroRNA-192 Suppresses Liver Metastasis of Colon Cancer , 2013, Oncogene.

[56]  J. Christensen,et al.  Multiple circulating proangiogenic factors induced by sunitinib malate are tumor-independent and correlate with antitumor efficacy , 2007, Proceedings of the National Academy of Sciences.

[57]  Jan-Fang Cheng,et al.  Dicer, Drosha, and outcomes in patients with ovarian cancer. , 2008, The New England journal of medicine.

[58]  Eugene S. Kim,et al.  Effect of MDM2 and vascular endothelial growth factor inhibition on tumor angiogenesis and metastasis in neuroblastoma , 2011, Angiogenesis.

[59]  Norbert Gretz,et al.  miRWalk - Database: Prediction of possible miRNA binding sites by "walking" the genes of three genomes , 2011, J. Biomed. Informatics.

[60]  A. Schetter,et al.  Identification of a metastasis-specific MicroRNA signature in human colorectal cancer. , 2015, Journal of the National Cancer Institute.