Downregulation of dystroglycan glycosyltransferases LARGE2 and ISPD associate with increased mortality in clear cell renal cell carcinoma

BackgroundDystroglycan (DG) is a cell-surface laminin receptor that links the cytoskeleton to the extracellular matrix in a variety of epithelial tissues. Its function as a matrix receptor requires extensive glycosylation of its extracellular subunit αDG, which involves at least 13 distinct genes. Prior work has shown loss of αDG glycosylation in an assortment of carcinomas, including clear cell renal cell carcinoma (ccRCC) though the cause (s) and functional consequences of this loss are still unclear.MethodsUsing The Cancer Genome Atlas (TCGA) database, we analyzed the DG glycosylation pathway to identify changes in mRNA expression and correlation with clinical outcomes. We validated our findings with a cohort of 65 patients treated with radical nephrectomy by analyzing DG glycosylation via immunohistochemistry and gene expression via qRT-PCR.ResultsAnalysis of TCGA database revealed frequent dysregulation of a subset of DG glycosyltransferases. Most notably, there was a frequent, significant downregulation of GYLTL1B (LARGE2) and ISPD. DG glycosylation is frequently impaired in ccRCC patient samples and most strongly associates with downregulation of GYLTL1B.ConclusionsReduced levels of GYLTL1B and ISPD mRNA associated with increased patient mortality and are the likely cause of αDG hypoglycosylation in ccRCC.

[1]  H. Cedar,et al.  Role of DNA methylation in the regulation of transcription. , 1994, Current opinion in genetics & development.

[2]  Obi L Griffith,et al.  Loss of cell-surface laminin anchoring promotes tumor growth and is associated with poor clinical outcomes. , 2012, Cancer research.

[3]  R. U. Margolis,et al.  Demonstration of mammalian protein O-mannosyltransferase activity: coexpression of POMT1 and POMT2 required for enzymatic activity. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[4]  G. Rossi,et al.  Loss of nuclear p27kip1 and α‐dystroglycan is a frequent event and is a strong predictor of poor outcome in renal cell carcinoma , 2010, Cancer science.

[5]  A. Jemal,et al.  Cancer statistics, 2014 , 2014, CA: a cancer journal for clinicians.

[6]  M. Fukuda,et al.  Tumor suppressor function of laminin-binding α-dystroglycan requires a distinct β3-N-acetylglucosaminyltransferase , 2009, Proceedings of the National Academy of Sciences.

[7]  A. Postigo,et al.  ZEB1 represses E-cadherin and induces an EMT by recruiting the SWI/SNF chromatin-remodeling protein BRG1 , 2010, Oncogene.

[8]  I. Kanazawa,et al.  Structures of sialylated O-linked oligosaccharides of bovine peripheral nerve alpha-dystroglycan. The role of a novel O-mannosyl-type oligosaccharide in the binding of alpha-dystroglycan with laminin. , 1997, The Journal of biological chemistry.

[9]  Francesco Muntoni,et al.  ISPD loss-of-function mutations disrupt dystroglycan O-mannosylation and cause Walker-Warburg syndrome , 2012, Nature Genetics.

[10]  O. Ibraghimov-Beskrovnaya,et al.  Primary structure of dystrophin-associated glycoproteins linking dystrophin to the extracellular matrix , 1992, Nature.

[11]  G. Genovese,et al.  Increased expression of CD133 and reduced dystroglycan expression are strong predictors of poor outcome in colon cancer patients , 2012, Journal of experimental & clinical cancer research : CR.

[12]  H. Friess,et al.  Reduced α-dystroglycan expression correlates with shortened patient survival in pancreatic cancer. , 2011, The Journal of surgical research.

[13]  W. Yang,et al.  Increments of α‐dystroglycan expression in liver metastasis correlate with poor survival in gastric cancer , 2009, Journal of surgical oncology.

[14]  S. Cross,et al.  Loss of dystroglycan function in oesophageal cancer , 2011, Histopathology.

[15]  M. Kattan,et al.  A postoperative prognostic nomogram predicting recurrence for patients with conventional clear cell renal cell carcinoma. , 2005, The Journal of urology.

[16]  J. Cheville,et al.  Comparisons of Outcome and Prognostic Features Among Histologic Subtypes of Renal Cell Carcinoma , 2003, The American journal of surgical pathology.

[17]  Steven J. M. Jones,et al.  Comprehensive molecular characterization of clear cell renal cell carcinoma , 2013, Nature.

[18]  Liping Yu,et al.  O-Mannosyl Phosphorylation of Alpha-Dystroglycan Is Required for Laminin Binding , 2010, Science.

[19]  K. Campbell,et al.  Proteolytic Enzymes and Altered Glycosylation Modulate Dystroglycan Function in Carcinoma Cells , 2004, Cancer Research.

[20]  G. Einarsson,et al.  Histological subtyping and nuclear grading of renal cell carcinoma and their implications for survival: a retrospective nation-wide study of 629 patients. , 2005, European urology.

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

[22]  J. Herman,et al.  Hypermethylation-associated inactivation of p14(ARF) is independent of p16(INK4a) methylation and p53 mutational status. , 2000, Cancer research.

[23]  J. Brooks,et al.  Mutations of the VHL tumour suppressor gene in renal carcinoma , 1994, Nature Genetics.

[24]  K. Campbell,et al.  Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection , 2011, Proceedings of the National Academy of Sciences.

[25]  D. Faller,et al.  SIRT1 induces EMT by cooperating with EMT transcription factors and enhances prostate cancer cell migration and metastasis , 2012, Oncogene.

[26]  K. Campbell,et al.  Xylosyl- and glucuronyltransferase functions of LARGE in α-dystroglycan modification are conserved in LARGE2 , 2012, Glycobiology.

[27]  Michael R. Miller,et al.  The glycosyltransferase LARGE2 is repressed by Snail and ZEB1 in prostate cancer , 2015, Cancer biology & therapy.

[28]  S. Nelson,et al.  SGK196 Is a Glycosylation-Specific O-Mannose Kinase Required for Dystroglycan Function , 2013, Science.

[29]  Liping Yu,et al.  Dystroglycan Function Requires Xylosyl- and Glucuronyltransferase Activities of LARGE , 2012, Science.

[30]  J. Hewitt,et al.  Characterization of the LARGE family of putative glycosyltransferases associated with dystroglycanopathies. , 2005, Glycobiology.

[31]  A. Cittadini,et al.  Aberrant expression of α-dystroglycan in cervical and vulvar cancer , 2006 .

[32]  K. Campbell,et al.  Reduced expression of dystroglycan in breast and prostate cancer. , 2001, Human pathology.

[33]  Benjamin E. Gross,et al.  The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. , 2012, Cancer discovery.

[34]  G. Rossi,et al.  Dystroglycan expression is frequently reduced in human breast and colon cancers and is associated with tumor progression. , 2003, The American journal of pathology.

[35]  Susan C. Brown,et al.  Mutations in the fukutin-related protein gene (FKRP) cause a form of congenital muscular dystrophy with secondary laminin alpha2 deficiency and abnormal glycosylation of alpha-dystroglycan. , 2001, American journal of human genetics.

[36]  L. Wells.  The O-Mannosylation Pathway: Glycosyltransferases and Proteins Implicated in Congenital Muscular Dystrophy* , 2013, The Journal of Biological Chemistry.

[37]  Gyan Bhanot,et al.  Abstract 1996: Molecular stratification of clear cell renal cell carcinoma by consensus clustering reveals distinct subtypes and survival patterns , 2010 .

[38]  I. Kanazawa,et al.  An ancient retrotransposal insertion causes Fukuyama-type congenital muscular dystrophy , 1998, Nature.

[39]  Jian-guo Shen,et al.  Dystroglycan is Associated with Tumor Progression and Patient Survival in Gastric Cancer , 2011, Pathology & Oncology Research.

[40]  Benjamin E. Gross,et al.  Integrative Analysis of Complex Cancer Genomics and Clinical Profiles Using the cBioPortal , 2013, Science Signaling.

[41]  The Cancer Genome Atlas Research Network.  COMPREHENSIVE MOLECULAR CHARACTERIZATION OF CLEAR CELL RENAL CELL CARCINOMA , 2013, Nature.

[42]  J. Ervasti,et al.  A role for the dystrophin-glycoprotein complex as a transmembrane linker between laminin and actin , 1993, The Journal of cell biology.

[43]  G. Rossi,et al.  Expression of dystroglycan correlates with tumor grade and predicts survival in renal cell carcinoma , 2007, Cancer biology & therapy.

[44]  Shun–ichi Kobayashi.  Tokyo campus rising , 1998, Nature.

[45]  D. Dean,et al.  ZEB represses transcription through interaction with the corepressor CtBP. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[46]  Brian J. Smith,et al.  Loss of LARGE2 Disrupts Functional Glycosylation of α-Dystroglycan in Prostate Cancer* , 2012, The Journal of Biological Chemistry.

[47]  A. Cittadini,et al.  Aberrant expression of alpha-dystroglycan in cervical and vulvar cancer. , 2006, Gynecologic oncology.

[48]  J. Ervasti,et al.  Membrane organization of the dystrophin-glycoprotein complex , 1991, Cell.

[49]  Michael D. Henry,et al.  Loss of α-Dystroglycan Laminin Binding in Epithelium-derived Cancers Is Caused by Silencing of LARGE*S⃞♦ , 2009, Journal of Biological Chemistry.