VHL-regulated MiR-204 suppresses tumor growth through inhibition of LC3B-mediated autophagy in renal clear cell carcinoma.

The von Hippel-Lindau tumor-suppressor gene (VHL) is lost in most clear cell renal cell carcinomas (ccRCC). Here, using human ccRCC specimens, VHL-deficient cells, and xenograft models, we show that miR-204 is a VHL-regulated tumor suppressor acting by inhibiting macroautophagy, with MAP1LC3B (LC3B) as a direct and functional target. Of note, higher tumor grade of human ccRCC was correlated with a concomitant decrease in miR-204 and increase in LC3B levels, indicating that LC3B-mediated macroautophagy is necessary for RCC progression. VHL, in addition to inducing endogenous miR-204, triggered the expression of LC3C, an HIF-regulated LC3B paralog, that suppressed tumor growth. These data reveal a function of VHL as a tumor-suppressing regulator of autophagic programs.

[1]  H. Horvitz,et al.  MicroRNA expression profiles classify human cancers , 2005, Nature.

[2]  Gareth Griffiths,et al.  Autophagosome formation from membrane compartments enriched in phosphatidylinositol 3-phosphate and dynamically connected to the endoplasmic reticulum , 2008, The Journal of cell biology.

[3]  M. Czech.  MicroRNAs as therapeutic targets. , 2006, The New England journal of medicine.

[4]  H. Tournaye,et al.  Orthotopic grafting of cryopreserved prepubertal testicular tissue: in search of a simple yet effective cryopreservation protocol. , 2012, Fertility and sterility.

[5]  J. Nemunaitis,et al.  Modulation of miRNA activity in human cancer: a new paradigm for cancer gene therapy? , 2008, Cancer Gene Therapy.

[6]  R. Bukowski,et al.  Anti-angiogenic targets in the treatment of advanced renal cell carcinoma. , 2008, Current cancer drug targets.

[7]  Kathryn A. O’Donnell,et al.  Therapeutic microRNA Delivery Suppresses Tumorigenesis in a Murine Liver Cancer Model , 2009, Cell.

[8]  C. Thompson,et al.  Ammonia-induced autophagy is independent of ULK1/ULK2 kinases , 2011, Proceedings of the National Academy of Sciences.

[9]  Hans Lehrach,et al.  MicroRNA profiling of clear cell renal cell cancer identifies a robust signature to define renal malignancy , 2009, Journal of cellular and molecular medicine.

[10]  P. Sutphin,et al.  A molecule targeting VHL-deficient renal cell carcinoma that induces autophagy. , 2008, Cancer cell.

[11]  C. Croce,et al.  Oncogenic role of miR-483-3p at the IGF2/483 locus. , 2010, Cancer research.

[12]  M. Seto,et al.  Genome‐wide microRNA expression profiling in renal cell carcinoma: significant down‐regulation of miR‐141 and miR‐200c , 2008, The Journal of pathology.

[13]  E. White,et al.  The Double-Edged Sword of Autophagy Modulation in Cancer , 2009, Clinical Cancer Research.

[14]  N. Rajewsky,et al.  Widespread changes in protein synthesis induced by microRNAs , 2008, Nature.

[15]  R. Raval,et al.  Regulation of E-cadherin expression by VHL and hypoxia-inducible factor. , 2006, Cancer research.

[16]  K. Otsu,et al.  Discovery of Atg5/Atg7-independent alternative macroautophagy , 2009, Nature.

[17]  Svetlana V. Harbaugh,et al.  The von Hippel-Lindau Tumor Suppressor Protein and Egl-9-Type Proline Hydroxylases Regulate the Large Subunit of RNA Polymerase II in Response to Oxidative Stress , 2008, Molecular and Cellular Biology.

[18]  G. Berx,et al.  VHL Promotes E2 Box-Dependent E-Cadherin Transcription by HIF-Mediated Regulation of SIP1 and Snail , 2006, Molecular and Cellular Biology.

[19]  Robert Clarke,et al.  Guidelines for the use and interpretation of assays for monitoring autophagy , 2012 .

[20]  S. Gygi,et al.  Network organization of the human autophagy system , 2010, Nature.

[21]  George A. Calin,et al.  A MicroRNA Signature of Hypoxia , 2006, Molecular and Cellular Biology.

[22]  K. Naito,et al.  Renal cell carcinoma: allelic loss at chromosome 9 using the fluorescent multiplex-polymerase chain reaction technique. , 2002, Human pathology.

[23]  Long Yu,et al.  Molecular cloning and characterization of rat LC3A and LC3B--two novel markers of autophagosome. , 2006, Biochemical and biophysical research communications.

[24]  Kevin Bray,et al.  Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis. , 2006, Cancer cell.

[25]  Franghiz Ali-Zadeh,et al.  In search of , 2005 .

[26]  Daniel J Klionsky,et al.  In search of an “autophagomometer” , 2009, Autophagy.

[27]  J. Meller,et al.  von Hippel-Lindau–Dependent Patterns of RNA Polymerase II Hydroxylation in Human Renal Clear Cell Carcinomas , 2010, Clinical Cancer Research.

[28]  D. Aust,et al.  Loss of heterozygosity and copy number abnormality in clear cell renal cell carcinoma discovered by high-density affymetrix 10K single nucleotide polymorphism mapping array. , 2008, Neoplasia.

[29]  H. Tagawa,et al.  High‐resolution analysis of DNA copy number alterations and gene expression in renal clear cell carcinoma , 2007, The Journal of pathology.

[30]  D. Klionsky,et al.  Regulation mechanisms and signaling pathways of autophagy. , 2009, Annual review of genetics.

[31]  Peter K. Kim,et al.  Mitochondria Supply Membranes for Autophagosome Biogenesis during Starvation , 2010, Cell.

[32]  Robin Mathew,et al.  Role of autophagy in cancer , 2007, Nature Reviews Cancer.

[33]  G. Bhanot,et al.  Identification of a microRNA panel for clear-cell kidney cancer. , 2010, Urology.

[34]  L. Gossage,et al.  Alterations in VHL as potential biomarkers in renal-cell carcinoma , 2010, Nature Reviews Clinical Oncology.

[35]  Z. Elazar,et al.  LC3 and GATE‐16/GABARAP subfamilies are both essential yet act differently in autophagosome biogenesis , 2010, The EMBO journal.

[36]  Marc Liesa,et al.  Pancreatic cancers require autophagy for tumor growth. , 2011, Genes & development.

[37]  D. Rubinsztein,et al.  Plasma membrane contributes to the formation of pre-autophagosomal structures , 2010, Nature Cell Biology.

[38]  Patrick J. Paddison,et al.  Genome-wide RNA-mediated interference screen identifies miR-19 targets in Notch-induced T-cell acute lymphoblastic leukaemia , 2010, Nature Cell Biology.

[39]  T. Ueno,et al.  LC3 conjugation system in mammalian autophagy , 2004, The International Journal of Biochemistry & Cell Biology.

[40]  J. Lovén,et al.  MYCN-regulated microRNAs repress estrogen receptor-α (ESR1) expression and neuronal differentiation in human neuroblastoma , 2010, Proceedings of the National Academy of Sciences.

[41]  E. White,et al.  Metabolic catastrophe as a means to cancer cell death , 2007, Journal of Cell Science.

[42]  C. Croce,et al.  MicroRNA signatures in human cancers , 2006, Nature Reviews Cancer.

[43]  Massimo Negrini,et al.  MicroRNAs in human cancer: from research to therapy , 2007, Journal of Cell Science.

[44]  John L Cleveland,et al.  Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes , 2008, Autophagy.

[45]  N. Rajewsky,et al.  Silencing of microRNAs in vivo with ‘antagomirs’ , 2005, Nature.