Glycosylation Is a Global Target for Androgen Control in Prostate Cancer Cells

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J Munkley The AR and glycosylation in 24:3 R49–R64 Review prostate cancer

Glycosylation is a global target for androgen control in prostate cancer cells

Correspondence Jennifer Munkley should be addressed to J Munkley Institute of Genetic Medicine, Newcastle University, Newcastle-upon-Tyne, UK Email [email protected]

Abstract

Changes in glycan composition are common in cancer and can play important roles Key Words in all of the recognised hallmarks of cancer. We recently identified glycosylation as a ff androgen receptor global target for androgen control in prostate cancer cells and further defined a set ff glycosylation of 8 glycosylation enzymes (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ff prostate cancer ST6GAL1 and EDEM3), which are also significantly upregulated in prostate cancer tissue. ff glycans These 8 enzymes are under direct control of the androgen receptor (AR) and are linked to the synthesis of important cancer-associated glycans such as sialyl-Tn (sTn), sialyl LewisX (SLeX), O-GlcNAc and chondroitin sulfate. Glycosylation has a key role in many important biological processes in cancer including cell adhesion, migration, interactions with the cell matrix, immune surveillance, cell signalling and cellular metabolism. Our results suggest that alterations in patterns of glycosylation via androgen control might

Endocrine-Related Cancer Endocrine-Related modify some or all of these processes in prostate cancer. The prostate is an abundant secretor of glycoproteins of all types, and alterations in glycans are, therefore, attractive as potential biomarkers and therapeutic targets. Emerging data on these often overlooked glycan modifications have the potential to improve risk stratification and Endocrine-Related Cancer therapeutic strategies in patients with prostate cancer. (2017) 24, R49–R64

Introduction

The prostate gland is principally responsible for secreting cell viability, proliferation and invasion (Haag et al. fluid, which nourishes and protects sperm during 2005, Hara et al. 2008, Snoek et al. 2009). Although reproduction. Both the development of the prostate ADT is usually initially effective, after 2–3 years, many gland and maintenance of these secretory functions patients develop castrate-resistant prostate cancer depend on the actions of androgen steroid hormones (CRPC), which is ultimately lethal. Progression to (Livermore et al. 2016). Androgens also play a critical role CRPC is believed to involve reprogramming of the AR in the development and progression of prostate cancer, transcriptional landscape and a selective pressure for cells and androgen deprivation therapy (ADT) is usually the to maintain AR activity even in lower concentrations first-line treatment for metastatic disease. Androgens of circulating androgens (Sharma et al. 2013, Mills control gene expression via the androgen receptor (AR) 2014). AR signalling can be maintained through gene transcription factor. The AR is important for many amplification (Visakorpi et al. 1995), activating mutations signalling pathways and is essential for prostate cancer (Veldscholte et al. 1990, Steinkamp et al. 2009), AR splice

http://erc.endocrinology-journals.org © 2017 Society for Endocrinology Published by Bioscientifica Ltd. DOI: 10.1530/ERC-16-0569 Printed in Great Britain Downloaded from Bioscientifica.com at 09/25/2021 08:03:34PM via free access

10.1530/ERC-16-0569 Review J Munkley The AR and glycosylation in 24:3 R50 prostate cancer

variants (Sun et al. 2010, Cao et al. 2016) or crosstalk with –P=0.05 (signi cant enrichment) other oncogenic signalling pathways (Zhu & Kyprianou Glycosylation 2008, Liao et al. 2013). Macromolecule glycosylation The AR classically acts to control the expression of Protein glycosylation target genes. Androgen binding promotes dimerisation of AR and its translocation to the nucleus where it can Protein N-linked glycosylation act as either a transcriptional repressor or activator Glycoprotein metabolic process Protein N-linked glycosylation (Cai et al. 2011, Karantanos et al. 2013, Munkley et al. via asparagine 2014, 2015a,b). AR target genes can be classified based on Glycoprotein biosynthetic process the transcripts and proteins that respond to androgens. 0 246 Numerous genomic and transcriptomic studies have been –log10 (signi cance) carried out to identify AR-binding sites and target genes; Figure 1 however, these have been limited by the genome coverage Glycosylation is a global target for androgen control in prostate cancer on microarrays (Massie et al. 2007, 2011, Wang et al. patients. RNA sequencing analysis of prostate cancer cell lines and 2009, Rajan et al. 2011) or by the challenges of directly patients identified a set of 700 androgen-regulated genes. Gene ontology (GO) analysis of these genes identified 72 terms with associating an AR-binding site with a given gene and its significant gene enrichment (P < 0.05) and further defined glycosylation expression (Massie et al. 2011, Wu et al. 2011, Sharma et al. as an androgen-regulated process in prostate cancer cells. 2013, Barfeld et al. 2014b). To address these issues and to further understand the mechanisms by which androgens of numerous glycosylation enzymes and takes place in drive the development and growth of prostate cancer, the Golgi apparatus and the lumen of the endoplasmic RNA sequencing has been used to comprehensively reticulum. The two most common mechanisms by which profile how the prostate cancer transcriptome changes glycans can be linked to lipids and proteins are O-linked in response to androgens. We directly correlated gene and N-linked glycosylation. In O-linked glycosylation, expression data from LNCaP prostate cancer cells exposed sugars are added sequentially to the hydroxyl oxygen of to androgens (Munkley et al. 2016b) with data from 7 serine and/or threonine residues on the target protein, prostate cancer patients before and after ADT (Rajan et al. whereas in N-linked glycosylation, preassembled blocks of 2014). Our approach produced a comprehensive map 14 sugars are transferred co-translationally via the amide of 700 genes controlled by the AR in clinical prostate group of an asparagine residue on the target protein Endocrine-Related Cancer Endocrine-Related cancer and provides a new window through which to (Schwarz & Aebi 2011, Kudelka et al. 2015) (Fig. 2). How understand the signalling pathways downstream of the much a given protein or lipid is glycosylated depends on AR (Munkley et al. 2016b). Gene set enrichment analysis of the number of glycosylation sites present, as well as the these 700 genes identified known AR-regulated processes expression and activities of specific glycosylation enzymes including fatty acid metabolism (Swinnen et al. 1997a, within the cell or tissue (Marth & Grewal 2008). 2000, Liu 2006), response to endoplasmic reticulum stress Aberrant glycosylation can play an important (Segawa et al. 2002, Sheng et al. 2015) and cholesterol and role in cancer progression. Glycans can have a key lipid biosynthesis (Swinnen et al. 1997b, Suburu & Chen role in cell adhesion, migration, interactions with the 2012, Wu et al. 2014, Butler et al. 2016). In addition to cell matrix, immune surveillance, cell signalling and these well-characterised AR-regulated pathways, our data cellular metabolism (Munkley & Elliott 2016a). It is well highlighted ‘glycosylation’ as a previously unidentified documented that the development and progression of global target for androgen control in prostate cancer cells cancer results in fundamental changes to the glycome; (Munkley et al. 2016b). Gene ontology analysis identified these are associated with all the recognised hallmarks 7 androgen-regulated processes containing the term of cancer (Munkley & Elliott 2016a). Numerous studies ‘glycosylation’ (Fig. 1). have described aberrant glycosylation in prostate cancer; however, despite this, the study of glycans has lagged behind our characterisation of the genome and proteome Glycosylation (Munkley et al. 2016a). As mounting evidence links Glycosylation is an enzymatic process that links glycan changes in the glycan composition of prostate cancer sugars to other glycans, lipids or proteins. The complete cells to disease progression (Munkley et al. 2016a), we pattern of glycan modifications in a cell or tissue (known further analysed our RNA-sequencing data to identify as the glycome) is assembled by the synchronised action 25 genes with roles in the glycosylation process, which

N-glycans

Fucose Branched N-glycans Sialic acid

GlcNAc

GalNAc

Gal O-glycans Mannose Ser/Thr Asn Asn

Ser/Thr Ser/Thr

Extracellular

Cytosol

Figure 2 Representative O-linked and N-linked glycans on the surface of a mammalian cell. Proteins can be glycosylated by O-linkage to serine/threonine residues or by N-linkage to asparagine residues. O-glycans can be extended to produce various core and terminal structures that can be sialylated and/or fucosylated. N-glycans contain a common pentasaccharide core region consisting of three mannose and two N-acetylglucosamine (GlcNAc) subunits Endocrine-Related Cancer Endocrine-Related (Man3GlcNAc2Asn). This can be further modified by the addition of terminal Gal (galactose), GlcNAc and sialic acid moieties.

are regulated by androgens in prostate cancer cells structure of the Golgi and the availability and levels of sugar (Munkley et al. 2016b). Further analysis of these 25 donors (Kudelka et al. 2015). In O-linked glycosylation, genes indicated that androgens control the expression of sugars are added incrementally to the hydroxyl oxygen enzymes and lectins operating at multiple steps within of serine and threonine residues on the target protein. the glycosylation synthetic pathways. These include O-glycans become altered in the early stages of cellular enzymes regulating the synthesis and processing of transformation and are important for cancer initiation, both O- and N-glycans, the hexosamine biosynthetic invasion and metastasis (Kudelka et al. 2015). We pathway (HBP) and the synthesis of chondroitin sulfate identified five androgen-regulated glycosylation enzymes (Table 1). Importantly, and consistent with AR-dependent (GALNT7, ST6GalNAC1, GCNT1, GCNT2 and FUT1) with regulation of key glycosylation enzymes, androgens also roles in O-glycan biosynthesis (Munkley et al. 2016b). control the synthesis of several cancer-associated glycans These enzymes control key steps in the initiation of in prostate cancer cells (Fig. 3). O-glycosylation and core synthesis and have been linked to the production of several cancer-associated glycans. Initiation of O-glycosylation is carried out by a family Androgens control O-glycan biosynthesis of GALNT sialyltransferase enzymes, including GALNT7, O-glycans consist of branched and linear arrangements which catalyse the transfer of GalNAc to serine/threonine of sugar groups that are transferred to glycoproteins by residues on target proteins to produce the Tn antigen glycosylation enzymes in the Golgi apparatus. O-glycan (GalNAc-O-Ser/Thr) (Ten Hagen et al. 2003). GALNT7 has synthesis depends on many factors, including the been identified as oncogene in several types of cancer expression and localisation of glycosylation enzymes, the (Gaziel-Sovran et al. 2011, Peng et al. 2012, Li et al. 2014b,

Table 1 Overview of the glycosylation genes regulated by androgens in prostate cancer cells.

Enzyme Function Role in cancer? Previously linked to prostate cancer? O-glycan biosynthesis ST6GALNAC1 Sialylation of O-glycans to produce Associated with poor prognosis in ST6GALNAC1 shown to synthesise the cancer-associated sTn antigen numerous cancer types (Munkley sTn and reduce cell adhesion in (Munkley 2016) 2016). Promotes tumourigenicity prostate cancer cells (Munkley et al. and metastasis in breast and 2015c, Munkley & Elliott 2016b). gastric cancer cells (Julien et al. sTn antigen expressed in high 2006, Ozaki et al. 2012) grade prostate tumours (Myers et al. 1994, Genega et al. 2000). Androgen regulated in prostate cancer cells (Munkley et al. 2015c, Munkley et al. 2016b) GALNT7 Initiation of O-glycosylation Identified as oncogene in several Upregulated in malignant PCa as types of cancer and controlled by part of a glycosylation gene micro-RNAs (Gaziel-Sovran et al. signature (Barfeld et al. 2014a). 2011, Peng et al. 2012, Li et al. Androgen regulated and linked to 2014b, Nie et al. 2015, Lu et al. prostate cancer cell viability 2016) (Munkley et al. 2016b) GCNT1 Formation of core 2 branched Upregulated in endometrial Increased tumour growth in O-glycans carcinoma and associated with orthotopic mouse models poor survival (Miyamoto et al. (Hagisawa et al. 2005). Promotes 2013) resistance to NK cell immunity (Okamoto et al. 2013). Upregulated in prostate cancer and implicated in the synthesis SLeX (Hagisawa et al. 2005, Chen et al. 2014). Role in synthesising the F77 antigen (Nonaka et al. 2014) which increases prostate cancer cell growth (Zhang et al. 2010). Correlates with aggressive disease (Kojima et al. 2015) and is a predictor of recurrence after radical prostatectomy (Sato et al. 2016). Detected in patient

Endocrine-Related Cancer Endocrine-Related urine (Kojima et al. 2015). Androgen regulated and linked to prostate cancer cell viability (Munkley et al. 2016b) GCNT2 Formation of core 2 branched Suppression of GCNT2 linked to Role in synthesising the F77 antigen O-glycans metastasis in colorectal cancer (Nonaka et al. 2014) which (Nakamura et al. 2015). Increased increases prostate cancer cell expression promotes EMT growth (Zhang et al. 2010) transition and metastasis in breast cancer (Zhang et al. 2011) FUT1 O-fucosyltransferase important for Linked to prognosis in breast cancer Role in synthesising the F77 antigen the biosynthesis of H antigens, (Milde-Langosch et al. 2014). Acts (Nonaka et al. 2014) which Lewis B, and Lewis Y as an oncogene in several cancer increases prostate cancer cell types (Hao et al. 2008, Zhang et al. growth (Zhang et al. 2010) 2008, Yan et al. 2010) Hexosamine biosynthetic pathway (HBP) UAP1 Final enzyme in the hexosamine Established role in synthesising Over-expressed in prostate cancer biosynthetic pathway (HBP) O-GlcNAc which is elevated in and linked to increased synthesis of numerous cancer types and has O-GlcNAc (Itkonen et al. 2014). itself been described as a hallmark Over-expression of O-GlcNAc linked of cancer (Fardini et al. 2013, Ma to poor prognosis (Kamigaito et al. & Vosseller 2014) 2014). Promotes resistance against inhibitors of N-glycosylation (Itkonen et al. 2014). Detected in patient serum and urine (Ma et al. 2014, Albitar et al. 2016). HBP potential target in CRPC (Kaushik et al. 2016)

(Continued)

Table 1 Continued.

Enzyme Function Role in cancer? Previously linked to prostate cancer? PGM3 Penultimate enzyme in the Established role in synthesising Androgen regulated and linked to hexosamine biosynthetic pathway O-GlcNAc which is elevated in prostate cancer cell viability (HBP) numerous cancer types and has (Lee et al. 2010, Munkley et al. itself been described as a hallmark 2016b). HBP is a potential target in of cancer (Fardini et al. 2013, Ma CRPC (Kaushik et al. 2016) & Vosseller 2014) GNPNAT1 Second enzyme in the hexosamine Established role in synthesising GNPNAT1 is decreased in CRPC biosynthetic pathway (HBP) O-GlcNAc which is elevated in (Kaushik et al. 2016). Inhibition of numerous cancer types and has HBP suggested as a potential itself been described as a hallmark therapy in CRPC (Kaushik et al. of cancer (Fardini et al. 2013, Ma 2016) & Vosseller 2014) Chondroitin sulfate synthesis CSGALNACT1 Initiation of chondroitin sulfate (CS) CS is a key molecule in cancer CSGALNACT1 is androgen regulated synthesis progression (Theocharis et al. and required for prostate cancer 2006). The large CS proteoglycan cell viability (Munkley et al. Versican is linked to poor 2016b). Versican is androgen prognosis in many different cancer regulated (Read et al. 2007) and is types (Ricciardelli et al. 2009b). linked to disease progression in CSGALNACT1 is upregulated by early stage prostate cancer NEDD9 in breast cancer cells (Ricciardelli et al. 1998). CS levels (Iida et al. 2015) can predict prostate cancer progression and are increased in metastatic disease (Ricciardelli et al. 1997, 2009a). Targeting oncofoetal CS using glycosaminoglycan binding malaria protein inhibits PC3 cell growth in mouse models (Salanti et al. 2015) CHPF Chondroitin sulfate synthesis CS is a key molecule in cancer Not reported progression (Theocharis et al. 2006). The large CS proteoglycan Versican is linked to poor

Endocrine-Related Cancer Endocrine-Related prognosis in many different cancer types (Ricciardelli et al. 2009b). Differential methylation of CHPF is induced by FOXM1 (Hwang et al. 2013) Lectins MLEC Mannose binding lectin Not reported Not reported LMAN2 Lectin that binds high mannose Over-expressed in the gastric cancer Not reported glycoproteins secretome (Marimuthu et al. 2013) ERLEC1 (CIM) Lectin with role in N-glycan Critical for lung cancer metastasis recognition (Yanagisawa et al. 2010) CALR Lectin chaperone with role in Upregulated in lung cancer Suppressive role in prostate cancer glycoprotein folding (Rho et al. 2009) cell growth and metastasis (Alur et al. 2009) OST complex RPN2 Subunit of Linked to malignancy Not reported Oligosaccharyltransferase (OST) (Tominaga et al. 2014) and confers complex which catalyses central resistance to docetaxel in breast reaction in N-glycosylation cancer (Honma et al. 2008). Potential biomarker in colorectal cancer (Zhang et al. 2015). Linked to survival and invasion in lung cancer (Fujita et al. 2015) STT3A Subunit of Not reported Not reported Oligosaccharyltransferase (OST) complex which catalyses central reaction in N-glycosylation

(Continued)

Table 1 Continued.

Enzyme Function Role in cancer? Previously linked to prostate cancer? TUSC3 Subunit of oligosaccharyltransferase Tumour suppressor in ovarian cancer Downregulated in prostate cancer (OST) complex which catalyses (Kratochvilova et al. 2015), tissue – acts as a tumour suppressor central reaction in N-glycosylation pancreatic cancer (Fan et al. 2016), (Horak et al. 2012, 2014) glioblastoma (Jiang et al. 2016). Overexpressed in colorectal cancer cells – induces EMT, tumour growth and invasion (Gu et al. 2016) N-glycan processing EDEM3 Mannose trimming of N-glycans Not reported in other cancers Upregulated in malignant PCa as part of a glycosylation gene signature (Barfeld et al. 2014a). Androgen regulated and linked to prostate cancer cell viability (Munkley et al. 2016b). May play a role in creating these aberrant N-glycans (Munkley et al. 2016b) ST6GAL1 Sialylation of terminal N-glycans Over-expressed in many types of Androgen regulated and linked to cancer (Swindall et al. 2013). prostate cancer cell viability Expression reduced by epigenetic (Munkley et al. 2016b) inactivation in bladder cancer (Antony et al. 2014). Linked to EMT transition and malignancy (Lu et al. 2014) B4GALT1 Late processing of N-glycans Induced by oestrogen in breast Upregulated by TNFα in prostate (transfers a galactose residue to cancer cells (Choi et al. 2012). cancer cells – role in motility and terminal N-acetylglucosamine) Linked to adverse outcome in invasion (Radhakrishnan et al. patients with non-metastatic clear 2011) cell renal cell carcinoma (Xie et al. 2016). Associated with multi-drug resistance in leukemia (Zhou et al. 2013) UGGT1 Recognizes and reglucosylates Binds to mutation specific isoform Not reported N-glycans with minor folding of EGFR (Erdem-Eraslan et al.

Endocrine-Related Cancer Endocrine-Related defects. 2015) ALG2 Mannosyltransferase with role in Role in TRAIL-induced apoptosis Not reported N-glycan biosynthesis. (Ovcharenko et al. 2007) NEU1 Sialidase that cleaves terminal sialic Not reported Not reported acid from glycoproteins and glycolipids Other NANS Sialic acid biosynthesis (sialic acid Not reported Not reported synthase) SERP1 Stabilizes membrane proteins Involved in tumour cell survival Not reported during stress and facilitates under stress – correlates with subsequent glycosylation survival in glioblastoma patients (Mucaj et al. 2015)

The 25 androgen-regulated genes have roles in O-glycan biosynthesis, the hexosamine biosynthetic pathway (HBP), chondroitin sulfate (CS) synthesis, N-glycan processing and the oligosaccharyltransferase (OST) complex.

Nie et al. 2015, Lu et al. 2016) and is upregulated in acid to the Tn antigen to produce the cancer-associated malignant prostate cancer as part of a glycosylation gene onco-foetal sialyl-Tn (sTn) antigen, which is linked to signature (Barfeld et al. 2014a). As GALNT7 initiates poor prognosis in numerous cancer types (Munkley 2016). O-glycosylation, upregulation of this enzyme could be STn is a truncated O-glycan containing a sialic acid α-2,6 linked to a range of changes to O-glycans in prostate linked to GalNAc α-O-Ser/Thr and has been shown to cancer cells. promote tumour growth and metastasis in mouse models We also identified the sialyltransferase ST6GalNAc1 as (Julien et al. 2006, Ozaki et al. 2012). In prostate cancer, being directly regulated by androgens in prostate cancer the sTn antigen is detected in up to half of all high-grade cells (Munkley et al. 2015c, 2016b). ST6GalNAc1 adds sialic tumours (Myers et al. 1994, Genega et al. 2000), and

No androgens Androgens modification of the cell surface adhesion protein MUC1 with sTn is positively correlated with prostate-specific antigen (PSA) levels and negatively correlated with survival outcome (Arai et al. 2005). Increased expression of sialylated glycans creates negative charges and can inhibit the cell adhesion through electrostatic repulsion DNA (Munkley & Elliott 2016b). We found previously that sTn in prostate cancer cells, the androgen receptor controls the expression of the cancer-associated sTn antigen and cell adhesion through induction of ST6GalNAc1 (Munkley et al. 2015c) (Fig. 3). Induction of ST6GalNAc1 was found to promote a more mesenchymal cell phenotype in prostate cancer cells and to inhibit the formation of stable tumour masses in mouse models. DNA Further analysis of prostate cancer tissue showed that PNA although ST6GalNAc1 is increased in primary tumours, expression is dramatically downregulated in metastatic disease, indicating that ST6GalNAc1 may have a transient role in prostate cancer progression (Munkley et al. 2015c). Our analysis of the prostate cancer transcriptome also identified the core 2 branching enzyme GCNT1 as an AR target gene in prostate cancer patients (Munkley et al. DNA 2016b). GCNT1 is upregulated in prostate cancer in which sLeX its expression correlates with aggressive disease and is a predictor of recurrence after radical prostatectomy (Hagisawa et al. 2005, Kojima et al. 2015, Sato et al. 2016). GCNT1 can be detected in patient urine where in combination with PSA it is a reliable indicator for Endocrine-Related Cancer Endocrine-Related extracapsular extension of prostate cancer (Kojima et al. 2015). Upregulated GCNT1 in prostate cancer cells DNA increases cell adhesion and dramatically increases tumour CS growth in orthotopic mouse models (Hagisawa et al. 2005). GCNT1 is implicated in the synthesis of the cancer-associated Sialyl-LewisX (SLeX) antigen, which is formed by sequential addition of sialic acid and fucose to the core to branch structure (Pinho & Reis 2015). SLeX is also regulated by androgens in prostate cancer cells (Munkley et al. 2016b) (Fig. 3). Consistent with this, in DNA prostate cancer tissue, upregulated GCNT1 is associated O-GlcNAc with higher levels of SLeX in PSA, PAP and MUC1 proteins (Chen et al. 2014). SLeX is highly expressed in many VCaP malignant cancers, where it is associated with reduced

Figure 3 rates of survival (Amado et al. 1998, Baldus et al. 1998). Androgens control the synthesis of several cancer-associated glycans in In prostate cancer, upregulation of SLeX is associated with prostate cancer cells. Immunofluorescent detection of cancer-associated hormonal-resistant, aggressive disease (Jorgensen et al. glycans in VCaP prostate cancer cells. Cells were grown with or without X 10 nM R1881 (androgens) for 72 h. Glycans were detected using 1995). Increased SLe could influence prostate cancer glycan-specific antibodies or lectins as described previously (Munkley et al. progression through a number of mechanisms including 2016b). Sialyl-Tn (sTn), Sialyl Lewis X (SLeX), chondroitin sulfate (CS) and immune recognition by selectins; modification of the O-N-Acetylglucosamine (O-GlcNAc) were detected with antibodies, X whereas the Tn antigen (GalNAc linked to serine or threonine) was MUC1 protein with SLe may enable prostate cancer cells detected using peanut agglutinin (PNA) lectin. The scale bar is 10 μm. to evade destruction by NK cells (Okamoto et al. 2013).

The core 2 branching enzyme GCNT2 is also complex is essential for EGFR localisation and signalling, directly regulated by androgens in prostate cancer cells and therapeutic inhibition of OST has been shown (Munkley et al. 2016b). Although GCNT2 is not well studied to induce senescence (Lopez-Sambrooks et al. 2016). in prostate cancer, increased GCNT2 has been shown to Upregulation of the RPN2 subunit is linked to malignancy promote an epithelial-to-mesenchymal transition (EMT) and confers resistance to docetaxel in breast cancer and metastasis in breast cancer cells (Zhang et al. 2011). (Honma et al. 2008, Tominaga et al. 2014). In contrast, Similarly, the fucosyltransferase FUT1 is also androgen the TUSC3 subunit has been identified as a tumour regulated in prostate cancer cells (Munkley et al. 2016b) suppressor in several cancer types (Kratochvilova et al. and can act as an oncogene in several cancer types 2015, Fan et al. 2016, Jiang et al. 2016). Although there (Hao et al. 2008, Zhang et al. 2008, Yan et al. 2010, Milde- is no reported evidence to date linking RPN2 or STT3A to Langosch et al. 2014). GCNT1, GCNT2 and FUT1 are prostate cancer, TUSC3 has been shown to act as a tumour believed to play a role in synthesising the prostate cancer- suppressor in prostate cancer cells and is downregulated associated F77 antigen (Nonaka et al. 2014). The F77 in prostate tumours (Horak et al. 2012, 2014). antibody was isolated from mice in 2010 after the injection After initiation of N-glycosylation by the OST of PC3 tumour cells and provoked great interest based complex, N-glycans are then further processed by specific on its diagnostic and therapeutic potential (Zhang et al. glycosylation enzymes. N-glycan structures frequently 2010). The F77 antibody was found to stain 141 out of 150 change in cancer and are linked to disease progression and prostate cancer tissue samples with minimal staining of metastasis (Taniguchi & Kizuka 2015). Increased levels of non-malignant prostate tissue and to inhibit the growth branched and cryptic N-glycans are common in prostate of DU145 and PC3 tumour xenografts in mouse models cancer, have been linked to EMT and metastasis and (Zhang et al. 2010). The F77 antigen was later determined are currently being investigated as potential diagnostic using an O-glycome array and characterised as a glycolipid markers (Kyselova et al. 2007, Newsom-Davis et al. 2009, with α1,2-fucose linkages (Gao et al. 2014). Expression Pinho et al. 2012, Wang et al. 2013, Ishibashi et al. 2014, analysis of glycosyltransferase genes later identified the Li et al. 2014a, Taniguchi & Kizuka 2015). The expression branching enzymes (including GCNT1 and 2) and FUT1 as of six enzymes with roles in N-glycan processing (EDEM3, essential for F77 antigen formation (Nonaka et al. 2014). ST6GAL1, B4GALT1, UGGT1, ALG2 and NEU1) is Taken together, these findings suggest that androgen- controlled by the AR in prostate cancer cells (Munkley et al. mediated upregulation of GCNT1, GCNT2 and FUT1 may 2016b). Three of these enzymes (UGGT1, ALG2 and Endocrine-Related Cancer Endocrine-Related have a role in the production of F77. NEU1) have not been previously linked to prostate cancer (Table 1). Here, I discuss the potential roles of the other three enzymes (EDEM3, ST6GAL1 and UGGT1) in cancer N-glycan biosynthesis (OST complex, and in prostate cancer. The EDEM3 enzyme is upregulated N-glycan processing) in prostate cancer tissue as part of a glycosylation gene In addition to the O-glycosylation enzymes discussed signature (Chen et al. 2014, Munkley et al. 2016b) and previously, our study also identified a set of androgen- is believed to stimulate mannose trimming of N-glycans regulated enzymes with roles in the synthesis and from total glycoproteins (Hirao et al. 2006). Elevated processing of N-glycans (Table 1). N-glycosylation begins levels of EDEM3 may play a role in creating aberrant with a common precursor consisting of 14 sugars, which branched and cryptic N-glycans, which occur frequently is added to target proteins in the endoplasmic reticulum in malignant transformation (Munkley et al. 2016b). (ER) (Taniguchi & Kizuka 2015). Three androgen-regulated EDEM3 also enhances endoplasmic reticulum-associated glycosylation enzymes (RPN2, STT3A and TUSC3) are degradation (ERAD) of misfolded glycoproteins, which subunits of the oligosaccharyltransferase (OST) complex, is particularly important in the context of cancer where which is the central enzyme of N-linked protein increased metabolic needs lead to the accumulation of glycosylation. OST transfers the 14-sugar preassembled faulty proteins (Hirao et al. 2006, Olivari & Molinari 2007, oligosaccharide to selected asparagine residues within Olzmann et al. 2013). the consensus sequence asparagine-X-serine/threonine ST6GAL1 plays a role in terminal sialylation of (Mohorko et al. 2011). Emerging evidence links both N-glycans and is upregulated in many types of cancer the OST complex and its individual subunits to cancer (Hedlund et al. 2008, Schultz et al. 2013, Swindall et al. progression. In non-small-cell lung cancer cells, the OST 2013). Expression of ST6GAL1 is linked to EMT and

malignancy, but the mechanisms driving this transition Initiation of CS begins with the addition of xylose to serine remain to be explored (Lu et al. 2014). Increased residues of core proteins in the ER and is followed by the sialylation can influence cancer cell metastasis, invasion addition of two galactose residues in the Golgi (Silbert & and survival (Munkley 2016). In prostate cancer cells, Sugumaran 2002). Two enzymes with roles in CS synthesis increased sialylation of PSA is linked to more aggressive (CSGALNACT1 and CHPF) are regulated by androgens in disease (Llop et al. 2016, Munkley et al. 2016a). Our prostate cancer cells. CSGALNACT1 is required for the analysis suggested that the role of ST6GAL1 in prostate initiation and elongation processes (Sakai et al. 2007, cancer is likely to be influenced by cell background Watanabe et al. 2010), whereas CHPF plays a role in in context-dependent manner (Munkley et al. 2016b). polymerisation (Kitagawa et al. 2003, Mikami & Kitagawa B4GALT1 transfers a galactose residue to terminal 2013). The CSGALNACT1 enzyme is upregulated in prostate N-acetylglucosamine and is involved in the late processing cancer cells and has been shown to be essential for prostate of N-glycans. Upregulated B4GALT1 can predict adverse cancer cell viability, making it an attractive therapeutic outcome in patients with non-metastatic clear cell renal target (Munkley et al. 2016b). Recent data suggest that cell carcinoma (Xie et al. 2016) and is associated with targeting oncofoetal CS using glycosaminoglycan-binding multi-drug resistance in leukaemia (Zhou et al. 2013). malaria protein can inhibit the growth of PC3 prostate In prostate cancer cells, B4GALT1 can be upregulated by cancer cells in mouse models (Salanti et al. 2015) (as PC3 TNFα and is thought to have a role in cell motility and cells are an AR-negative prostate cancer cell line, it will invasion (Radhakrishnan et al. 2011). Taken together, be interesting to explore this finding further in a range of these findings suggest that both the initiation and AR-negative and -positive cell types). processing of N-glycosylation is under extensive control of androgens in prostate cancer cells. A wide range of Hexosamine biosynthetic pathway (HBP) changes to N-glycans have been observed in prostate cancer (Munkley et al. 2016a), and it is likely that AR The hexosamine biosynthetic pathway (HBP) is a major regulation of N-glycosylation enzymes plays a role in metabolic pathway, which produces uridine diphosphate– driving these changes during disease progression. N-acetylglucosamine (UDP–GlcNAc), an essential building block for glycan biosynthesis. The pathway is well positioned to affect glycans that accelerate cancer Chondroitin sulfate synthesis progression and is becoming of increasing importance in Endocrine-Related Cancer Endocrine-Related Our study also detected androgen regulation of the cancer biology (Vasconcelos-Dos-Santos et al. 2015). HBP is pathway producing chondroitin sulfate (CS), as well as CS sustained by metabolites produced by metabolic processes itself and the large CS proteoglycan Versican (Read et al. and is a major metabolic integration point influencing 2007, Munkley et al. 2016b) (Fig. 3). CS is a type of cell growth, metabolism, cell stress and epigenetics glycosaminoglycan (GAG) found in the extracellular (Hanover et al. 2012, Fardini et al. 2013, Bond & Hanover matrix and on the cell surface of many cell types (Gandhi & 2015). HBP produces an amino sugar conjugate, UDP- Mancera 2008) and is a key molecule in cancer progression GlcNAc, which acts as a substrate for the post-translational (Theocharis et al. 2006). CS is believed to play a role in modification of a wide range of proteins (Schwarz & Aebi cellular proliferation and differentiation (Hardingham & 2011) (Fig. 4). UDP-GlcNAc can be added to target proteins Fosang 1992), can be used to predict disease progression in in the cytoplasm, nucleus or mitochondria by the enzyme early stage prostate cancer (Ricciardelli et al. 1997), is linked OGT (O-GlcNAc transferase) (Butkinaree et al. 2010) or as to poor prognosis (Ricciardelli et al. 1999) and is increased a modification to glycoproteins in the Golgi apparatus or in metastatic prostate cancer (Ricciardelli et al. 2009a). ER (Schwarz & Aebi 2011). Both OGT and the O-GlcNAc Versican is linked to poor prognosis in many different glycan are elevated in patients with localised prostate cancer types (Ricciardelli et al. 2009b) and is associated cancer and associated with poor prognosis (Lynch et al. with disease progression in early-stage prostate cancer 2012, Kamigaito et al. 2014). OGT can modify a range of (Ricciardelli et al. 1998). CS is synthesised as GalNAc- target proteins including the transcription factors FOXM1 containing galactosaminoglycan polymers alternating and cMYC and may promote metabolic reprogramming with glucuronic acid. Chondroitin glycosaminoglycans in tumours cells (Itkonen et al. 2013). are covalently attached to core proteins and can be Our recent study showed that the HBP is under sulfated to varying degrees depending on the tissue source. extensive androgen control in prostate cancer cells

Fructose-6-P to enhance tumour growth and is a key biochemical

GFPT1 mediator of CRPC progression (Kaushik et al. 2016). The transcript levels of HBP genes are downregulated Glucosamine-6-P in CRPC tissue compared with localised hormone- GNPNAT1 sensitive tumours, and loss of GNPNAT1 in CRPC cells increases tumour growth and aggressiveness in mouse N-acetyl glucosamine-6-P models in cells containing either AR-full length of AR-V7 (Kaushik et al. 2016). In castrate-resistant disease, the PGM3 HBP can be targeted therapeutically by treatment with N-acetyl glucosamine-1-P UDP-GlcNAc, which remarkably increases the efficacy of the anti-androgen enzalutamide also (Kaushik et al. UAP1 2016). Increased glycolysis is one of the main metabolic UDP-GlcNAc adaptations found in CRPC (Biernacka et al. 2013). Hence, although HBP is increased in localised prostate cancer, UDP metabolic rewiring during disease progression is believed to promote a reduction in HBP in castrate-resistant disease. This could offer a selective adaptation to the

O-GlcNAc bioenergetics demands of the tumour and the increased N- and O-linked OGT need for glycolysis (Kaushik et al. 2016). glycosylation Protein Protein OGA Conclusions and future perspectives

The mechanisms driving the growth and spread of prostate cancer are complex and not fully understood, yet, are crucially important for disease management. GlcNAc H2O We recently identified glycosylation (the enzymatic O-GlcNAc modification of proteins attachment of glycan sugar moieties to lipids and proteins) as a novel androgen-regulated process in Endocrine-Related Cancer Endocrine-Related Figure 4 prostate cancer patients (Munkley et al. 2016b). Our The hexosamine biosynthetic pathway (HBP). The HBP is sustained by study identified 25 glycosylation enzymes and lectins, metabolites produced by metabolic processes and is a major metabolic integration point in the cell. HBP produces an amino sugar conjugate, which are regulated by androgens and respond to UDP-GlcNAc that serves as a major sugar donor for O-GlcNAcylation and androgen deprivation in prostate cancer patients. These for classical N-linked and O-linked glycosylation. O-GlcNAc transferase glycosylation enzymes operate at multiple steps within (OGT) catalyses the transfer of GlcNAc from the sugar donor UDP-GlcNAc to serine and/or threonine residues. O-GlcNAcase (OGA) carries out the the glycosylation synthetic pathways, including the reverse reaction. The enzymes GNPNAT1, PGM3 and UAP1 (shown in red) synthesis and processing of both O- and N-glycans, are directly controlled by androgens in prostate cancer cells, as is the control of the hexosamine biosynthetic pathway (HBP) O-GlcNAc modification itself. and the synthesis of chondroitin sulfate. A wide variety (Munkley et al. 2016b). The final enzyme in the HBP, of changes to the glycoproteome are frequently observed UAP1 (UDP–N-acetylglucosamine pyrophosphorylase 1), in prostate cancer. Glycans can influence cell survival, is regulated by AR activity and is highly overexpressed in proliferation and metastasis and likely play a key role in patients with prostate cancer (Itkonen et al. 2014). Our these processes in prostate cancer (Munkley et al. 2016a). data showed that in addition to UAP1, both the second Androgens control key steps in glycan synthesis and are enzyme (GNPNAT1) and the penultimate enzyme (PGM3) linked to several cancer-associated glycans (including in the pathway are also controlled by androgens in sTn, SLeX, OGlcNAc, chondroitin sulfate, branched and prostate cancer patients, as is the O-GlcNAc modification cryptic N-glycans, and potentially, the F77 antigen). itself (Munkley et al. 2016b) (Fig. 3). Prostate cancer is a unique disease characterised by Although HBP is regulated by the AR and is upregulated prognostic heterogeneity, and there is an urgent clinical in localised hormone-sensitive prostate cancer where it need to identify biomarkers to help distinguish indolent has a positive influence on disease progression, in the from aggressive disease and to develop new treatments castrate-resistant state, downregulation of HBP appears for castrate-resistant disease. The identification of

glycosylation as a global target for androgen control in Barfeld SJ, East P, Zuber V & Mills IG 2014a Meta-analysis of prostate prostate cancer cells has important clinical implications cancer gene expression data identifies a novel discriminatory signature enriched for glycosylating enzymes. BMC Medical Genomics in terms of both diagnosis and treatment. Glycoproteins 7 513. (doi:10.1186/s12920-014-0074-9) are the most commonly used serological biomarkers Barfeld SJ, Itkonen HM, Urbanucci A & Mills IG 2014b Androgen- for cancer diagnosis, and monitoring the glycan regulated metabolism and biosynthesis in prostate cancer. Endocrine- Related Cancer 21 T57–T66. (doi:10.1530/ERC-13-0515) composition of current biomarkers (such as PSA) can Biernacka JM, Geske J, Jenkins GD, Colby C, Rider DN, Karpyak VM, dramatically improve their specificity (Reis et al. 2010, Choi DS & Fridley BL 2013 Genome-wide gene-set analysis for Gilgunn et al. 2013). Glycans also likely play roles in all identification of pathways associated with alcohol dependence. International Journal of Neuropsychopharmacology 16 271–278. aspects of cancer progression and are therefore attractive (doi:10.1017/S1461145712000375) targets for therapeutic intervention (Pinho & Reis 2015, Bond MR & Hanover JA 2015 A little sugar goes a long way: the cell Munkley et al. 2016a). Glycans that hold particular biology of O-GlcNAc. Journal of Cell Biology 208 869–880. (doi:10.1083/jcb.201501101) promise as therapeutic targets in prostate cancer include Butkinaree C, Park K & Hart GW 2010 O-linked beta-N- chondroitin sulfate (Salanti et al. 2015), the F77 antigen acetylglucosamine (O-GlcNAc): extensive crosstalk with (Zhang et al. 2010), SLeX (Hagisawa et al. 2005) and STn phosphorylation to regulate signaling and transcription in response to nutrients and stress. Biochimica et Biophysica Acta 1800 96–106. (Munkley & Elliott 2016b). An increased understanding (doi:10.1016/j.bbagen.2009.07.018) of how glycosylation modulates the biological function Butler LM, Centenera MM & Swinnen JV 2016 Androgen control of lipid of prostate cancer cells will allow the development of a metabolism in prostate cancer: novel insights and future applications. Endocrine-Related Cancer 23 R219–R227. (doi:10.1530/ERC-15-0556) relatively unexploited field of drugs based on inhibitors, Cai C, He HH, Chen S, Coleman I, Wang H, Fang Z, Chen S, Nelson PS, glycan antagonists and glycan function modulators. Liu XS, Brown M, et al. 2011 Androgen receptor gene expression in prostate cancer is directly suppressed by the androgen receptor through recruitment of lysine-specific demethylase 1.Cancer Cell 20 457–471. (doi:10.1016/j.ccr.2011.09.001) Declaration of interest Cao S, Zhan Y & Dong Y 2016 Emerging data on androgen receptor The author declares that there is no conflict of interest that could be splice variants in prostate cancer. Endocrine-Related Cancer 23 perceived as prejudicing the impartiality of this review. T199–T210. (doi:10.1530/ERC-16-0298) Chen Z, Gulzar ZG, St Hill CA, Walcheck B & Brooks JD 2014 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. Prostate 74 Funding 1059–1067. (doi:10.1002/pros.22826) This work was funded by Prostate Cancer UK (PG12-34), Newcastle Choi HJ, Chung TW, Kim CH, Jeong HS, Joo M, Youn B & Ha KT 2012 University Special Trustees and The J G W Patterson Foundation. Estrogen induced beta-1,4-galactosyltransferase 1 expression Endocrine-Related Cancer Endocrine-Related regulates proliferation of human breast cancer MCF-7 cells. Biochemical and Biophysical Research Communications 426 620–625. References (doi:10.1016/j.bbrc.2012.08.140) Erdem-Eraslan L, Gao Y, Kloosterhof NK, Atlasi Y, Demmers J, Albitar M, Ma W, Lund L, Albitar F, Diep K, Fritsche HA & Shore N 2016 Sacchetti A, Kros JM, Sillevis Smitt P, Aerts J & French PJ 2015 Predicting prostate biopsy results using a panel of plasma and urine Mutation specific functions of EGFR result in a mutation-specific biomarkers combined in a scoring system. Journal of Cancer 7 downstream pathway activation. European Journal of Cancer 51 297–303. (doi:10.7150/jca.12771) 893–903. (doi:10.1016/j.ejca.2015.02.006) Alur M, Nguyen MM, Eggener SE, Jiang F, Dadras SS, Stern J, Kimm S, Fan X, Zhang X, Shen J, Zhao H, Yu X, Chen Y, Zhuang Z, Deng X, Roehl K, Kozlowski J, Pins M, et al. 2009 Suppressive roles of Feng H, Wang Y, et al. 2016 Decreased TUSC3 promotes pancreatic calreticulin in prostate cancer growth and metastasis. American cancer proliferation, invasion and metastasis. PLoS ONE 11 Journal of Pathology 175 882–890. (doi:10.2353/ajpath.2009.080417) e0149028. (doi:10.1371/journal.pone.0149028) Amado M, Carneiro F, Seixas M, Clausen H & Sobrinho-Simoes M 1998 Fardini Y, Dehennaut V, Lefebvre T & Issad T 2013 O-GlcNAcylation: a Dimeric sialyl-Le(x) expression in gastric carcinoma correlates with new cancer hallmark? Frontiers in Endocrinology 4 99. (doi:10.3389/ venous invasion and poor outcome. Gastroenterology 114 462–470. fendo.2013.00099) (doi:10.1016/S0016-5085(98)70529-3) Fujita Y, Yagishita S, Takeshita F, Yamamoto Y, Kuwano K & Ochiya T Antony P, Rose M, Heidenreich A, Knuchel R, Gaisa NT & Dahl E 2014 2015 Prognostic and therapeutic impact of RPN2-mediated tumor Epigenetic inactivation of ST6GAL1 in human bladder cancer. BMC malignancy in non-small-cell lung cancer. Oncotarget 6 3335–3345. Cancer 14 901. (doi:10.1186/1471-2407-14-901) (doi:10.18632/oncotarget.2793) Arai T, Fujita K, Fujime M & Irimura T 2005 Expression of sialylated Gandhi NS & Mancera RL 2008 The structure of glycosaminoglycans MUC1 in prostate cancer: relationship to clinical stage and and their interactions with proteins. Chemical Biology and Drug prognosis. International Journal of Urology 12 654–661. Design 72 455–482. (doi:10.1111/j.1747-0285.2008.00741.x) (doi:10.1111/j.1442-2042.2005.01112.x) Gao C, Liu Y, Zhang H, Zhang Y, Fukuda MN, Palma AS, Kozak RP, Baldus SE, Zirbes TK, Monig SP, Engel S, Monaca E, Rafiqpoor K, Childs RA, Nonaka M, Li Z, et al. 2014 Carbohydrate sequence of the Hanisch FG, Hanski C, Thiele J, Pichlmaier H, et al. 1998 prostate cancer-associated antigen F77 assigned by a mucin Histopathological subtypes and prognosis of gastric cancer are O-glycome designer array. Journal of Biological Chemistry 289 correlated with the expression of mucin-associated sialylated 16462–16477. (doi:10.1074/jbc.M114.558932) antigens: Sialosyl-Lewis(a), Sialosyl-Lewis(x) and sialosyl-Tn. Tumour Gaziel-Sovran A, Segura MF, Di Micco R, Collins MK, Hanniford D, Biology 19 445–453. (doi:10.1159/000030036) Vega-Saenz de Miera E, Rakus JF, Dankert JF, Shang S, Kerbel RS, et al.

2011 miR-30b/30d regulation of GalNAc transferases enhances breast cancer cell growth. Experimental Cell Research 330 358–370. invasion and immunosuppression during metastasis. Cancer Cell 20 (doi:10.1016/j.yexcr.2014.11.002) 104–118. (doi:10.1016/j.ccr.2011.05.027) Ishibashi Y, Tobisawa Y, Hatakeyama S, Ohashi T, Tanaka M, Narita S, Genega EM, Hutchinson B, Reuter VE & Gaudin PB 2000 Koie T, Habuchi T, Nishimura S, Ohyama C, et al. 2014 Serum tri- Immunophenotype of high-grade prostatic adenocarcinoma and and tetra-antennary N-glycan is a potential predictive biomarker for urothelial carcinoma. Modern Pathology 13 1186–1191. (doi:10.1038/ castration-resistant prostate cancer. Prostate 74 1521–1529. modpathol.3880220) (doi:10.1002/pros.22869) Gilgunn S, Conroy PJ, Saldova R, Rudd PM & O’Kennedy RJ 2013 Itkonen HM, Minner S, Guldvik IJ, Sandmann MJ, Tsourlakis MC, Aberrant PSA glycosylation – a sweet predictor of prostate cancer. Berge V, Svindland A, Schlomm T & Mills IG 2013 O-GlcNAc Nature Reviews Urology 10 99–107. (doi:10.1038/nrurol.2012.258) transferase integrates metabolic pathways to regulate the stability of Gu Y, Wang Q, Guo K, Qin W, Liao W, Wang S, Ding Y & Lin J 2016 c-MYC in human prostate cancer cells. Cancer Research 73 TUSC3 promotes colorectal cancer progression and epithelial- 5277–5287. (doi:10.1158/0008-5472.CAN-13-0549) mesenchymal transition (EMT) through WNT/beta-catenin and Itkonen HM, Engedal N, Babaie E, Luhr M, Guldvik IJ, Minner S, MAPK signalling. Journal of Pathology 239 60–71. (doi:10.1002/ Hohloch J, Tsourlakis MC, Schlomm T & Mills IG 2014 UAP1 is path.4697) overexpressed in prostate cancer and is protective against inhibitors Haag P, Bektic J, Bartsch G, Klocker H & Eder IE 2005 Androgen receptor of N-linked glycosylation. Oncogene 34 3744–3750. (doi:10.1038/ down regulation by small interference RNA induces cell growth onc.2014.307) inhibition in androgen sensitive as well as in androgen independent Jiang Z, Guo M, Zhang X, Yao L, Shen J, Ma G, Liu L, Zhao L, Xie C, prostate cancer cells. Journal of Steroid Biochemistry and Molecular Liang H, et al. 2016 TUSC3 suppresses glioblastoma development by Biology 96 251–258. (doi:10.1016/j.jsbmb.2005.04.029) inhibiting Akt signaling. Tumour Biology 37 12039–12047. Hagisawa S, Ohyama C, Takahashi T, Endoh M, Moriya T, Nakayama J, (doi:10.1007/s13277-016-5072-4) Arai Y & Fukuda M 2005 Expression of core 2 beta1,6-N- Jorgensen T, Berner A, Kaalhus O, Tveter KJ, Danielsen HE & Bryne M acetylglucosaminyltransferase facilitates prostate cancer progression. 1995 Up-regulation of the oligosaccharide sialyl LewisX: a new Glycobiology 15 1016–1024. (doi:10.1093/glycob/cwi086) prognostic parameter in metastatic prostate cancer. Cancer Research Hanover JA, Krause MW & Love DC 2012 Bittersweet memories: linking 55 1817–1819. metabolism to epigenetics through O-GlcNAcylation. Nature Reviews Julien S, Adriaenssens E, Ottenberg K, Furlan A, Courtand G, Vercoutter- Molecular Cell Biology 13 312–321. (doi:10.1038/nrm3334) Edouart AS, Hanisch FG, Delannoy P & Le Bourhis X 2006 Hao YY, Lin B, Zhao Y, Zhang YH, Li FF, Diao B, Ou YL & Zhang SL ST6GalNAc I expression in MDA-MB-231 breast cancer cells greatly 2008 Alpha1,2-fucosyltransferase gene transfection influences on modifies their O-glycosylation pattern and enhances their biological behavior of ovarian carcinoma-derived RMG-I cells. Fen Zi tumourigenicity. Glycobiology 16 54–64. (doi:10.1093/glycob/cwj033) Xi Bao Sheng Wu Xue Bao 41 435–442. Kamigaito T, Okaneya T, Kawakubo M, Shimojo H, Nishizawa O & Hara T, Miyazaki H, Lee A, Tran CP & Reiter RE 2008 Androgen receptor Nakayama J 2014 Overexpression of O-GlcNAc by prostate cancer and invasion in prostate cancer. Cancer Research 68 1128–1135. cells is significantly associated with poor prognosis of patients. (doi:10.1158/0008-5472.CAN-07-1929) Prostate Cancer and Prostatic Diseases 17 18–22. (doi:10.1038/ Hardingham TE & Fosang AJ 1992 Proteoglycans: many forms and pcan.2013.56) many functions. FASEB Journal 6 861–870. Karantanos T, Corn PG & Thompson TC 2013 Prostate cancer Hedlund M, Ng E, Varki A & Varki NM 2008 alpha 2-6-Linked sialic progression after androgen deprivation therapy: mechanisms of Endocrine-Related Cancer Endocrine-Related acids on N-glycans modulate carcinoma differentiation in vivo. castrate resistance and novel therapeutic approaches. Oncogene 32 Cancer Research 68 388–394. (doi:10.1158/0008-5472.CAN-07- 5501–5511. (doi:10.1038/onc.2013.206) 1340) Kaushik AK, Shojaie A, Panzitt K, Sonavane R, Venghatakrishnan H, Hirao K, Natsuka Y, Tamura T, Wada I, Morito D, Natsuka S, Romero P, Manikkam M, Zaslavsky A, Putluri V, Vasu VT, Zhang Y, et al. 2016 Sleno B, Tremblay LO, Herscovics A, et al. 2006 EDEM3, a soluble Inhibition of the hexosamine biosynthetic pathway promotes EDEM homolog, enhances glycoprotein endoplasmic reticulum- castration-resistant prostate cancer. Nature Communications 7 11612. associated degradation and mannose trimming. Journal of Biological (doi:10.1038/ncomms11612) Chemistry 281 9650–9658. (doi:10.1074/jbc.M512191200) Kitagawa H, Izumikawa T, Uyama T & Sugahara K 2003 Molecular Honma K, Iwao-Koizumi K, Takeshita F, Yamamoto Y, Yoshida T, cloning of a chondroitin polymerizing factor that cooperates with Nishio K, Nagahara S, Kato K & Ochiya T 2008 RPN2 gene confers chondroitin synthase for chondroitin polymerization. Journal of docetaxel resistance in breast cancer. Nature Medicine 14 939–948. Biological Chemistry 278 23666–23671. (doi:10.1074/jbc. (doi:10.1038/nm.1858) M302493200) Horak P, Tomasich E, Anees M, Marhold M, Gerschpacher M, Pils D & Kojima Y, Yoneyama T, Hatakeyama S, Mikami J, Sato T, Mori K, Krainer M 2012 Abstract 3997: TUSC3 (N33) affects protein Hashimoto Y, Koie T, Ohyama C, Fukuda M, et al. 2015 Detection of N-glycosylation and proliferation of prostate cancer cells. Cancer core2 beta-1,6-N-acetylglucosaminyltransferase in post-digital rectal Research 72 Abstract nr 3997. (doi:10.1158/1538-7445.AM2012-3997) examination urine is a reliable indicator for extracapsular extension Horak P, Tomasich E, Vanhara P, Kratochvilova K, Anees M, Marhold M, of prostate cancer. PLoS ONE 10 e0138520. (doi:10.1371/journal. Lemberger CE, Gerschpacher M, Horvat R, Sibilia M, et al. 2014 TUSC3 pone.0138520) loss alters the ER stress response and accelerates prostate cancer Kratochvilova K, Horak P, Esner M, Soucek K, Pils D, Anees M, growth in vivo. Scientific Reports 4 3739. (doi:10.1038/srep03739) Tomasich E, Drafi F, Jurtikova V, Hampl A, et al. 2015 Tumor Hwang S, Mahadevan S, Qadir F, Hutchison IL, Costea DE, suppressor candidate 3 (TUSC3) prevents the epithelial-to- Neppelberg E, Liavaag PG, Waseem A & Teh MT 2013 Identification mesenchymal transition and inhibits tumor growth by modulating of FOXM1-induced epigenetic markers for head and neck squamous the endoplasmic reticulum stress response in ovarian cancer cells. cell carcinomas. Cancer 119 4249–4258. (doi:10.1002/cncr.28354) International Journal of Cancer 137 1330–1340. (doi:10.1002/ Iida J, Dorchak J, Clancy R, Slavik J, Ellsworth R, Katagiri Y, ijc.29502) Pugacheva EN, van Kuppevelt TH, Mural RJ, Cutler ML, et al. 2015 Kudelka MR, Ju T, Heimburg-Molinaro J & Cummings RD 2015 Simple Role for chondroitin sulfate glycosaminoglycan in NEDD9-mediated sugars to complex disease – mucin-type O-glycans in cancer.

Advances in Cancer Research 126 53–135. (doi.org/10.1016/bs. Marth JD & Grewal PK 2008 Mammalian glycosylation in immunity. acr.2014.11.002) Nature Reviews Immunology 8 874–887. (doi:10.1038/nri2417) Kyselova Z, Mechref Y, Al Bataineh MM, Dobrolecki LE, Hickey RJ, Massie CE, Adryan B, Barbosa-Morais NL, Lynch AG, Tran MG, Neal DE Vinson J, Sweeney CJ & Novotny MV 2007 Alterations in the serum & Mills IG 2007 New androgen receptor genomic targets show an glycome due to metastatic prostate cancer. Journal of Proteome interaction with the ETS1 transcription factor. EMBO Reports 8 Research 6 1822–1832. (doi:10.1021/pr060664t) 871–878. (doi:10.1038/sj.embor.7401046) Lee CH, Jeong SJ, Yun SM, Kim JH, Lee HJ, Ahn KS, Won SH, Kim HS, Massie CE, Lynch A, Ramos-Montoya A, Boren J, Stark R, Fazli L, Lee HJ, Ahn KS, et al. 2010 Down-regulation of phospho Warren A, Scott H, Madhu B, Sharma N, et al. 2011 The androgen glucomutase 3 mediates sulforaphane-induced cell death in LNCaP receptor fuels prostate cancer by regulating central metabolism and prostate cancer cells. Proteome Science 8 67. (doi:10.1186/1477- biosynthesis. EMBO Journal 30 2719–2733. (doi:10.1038/ 5956-8-67) emboj.2011.158) Li N, Xu H, Fan K, Liu X, Qi J, Zhao C, Yin P, Wang L, Li Z & Zha X Mikami T & Kitagawa H 2013 Biosynthesis and function of chondroitin 2014a Altered beta1,6-GlcNAc branched N-glycans impair TGF-beta- sulfate. Biochimica et Biophysica Acta 1830 4719–4733. (doi:10.1016/j. mediated epithelial-to-mesenchymal transition through Smad bbagen.2013.06.006) signalling pathway in human lung cancer. Journal of Cellular and Milde-Langosch K, Karn T, Schmidt M, zu Eulenburg C, Oliveira-Ferrer L, Molecular Medicine 18 1975–1991. (doi:10.1111/jcmm.12331) Wirtz RM, Schumacher U, Witzel I, Schutze D & Muller V 2014 Li W, Ma H & Sun J 2014b MicroRNA34a/c function as tumor Prognostic relevance of glycosylation-associated genes in breast suppressors in Hep2 laryngeal carcinoma cells and may reduce cancer. Breast Cancer Research and Treatment 145 295–305. GALNT7 expression. Molecular Medicine Reports 9 1293–1298. (doi:10.1007/s10549-014-2949-z) Liao RS, Ma S, Miao L, Li R, Yin Y & Raj GV 2013 Androgen receptor- Mills IG 2014 Maintaining and reprogramming genomic androgen mediated non-genomic regulation of prostate cancer cell receptor activity in prostate cancer. Nature Reviews Cancer 14 proliferation. Translational Andrology and Urology 2 187–196. 187–198. (doi:10.1038/nrc3678) Liu Y 2006 Fatty acid oxidation is a dominant bioenergetic pathway in Miyamoto T, Suzuki A, Asaka R, Ishikawa K, Yamada Y, Kobara H, prostate cancer. Prostate Cancer and Prostatic Diseases 9 230–234. Nakayama J & Shiozawa T 2013 Immunohistochemical expression of (doi:10.1038/sj.pcan.4500879) core 2 beta1,6-N-acetylglucosaminyl transferase 1 (C2GnT1) in Livermore KE, Munkley J & Elliott DJ 2016 Androgen receptor and endometrioid-type endometrial carcinoma: a novel potential prostate cancer. AIMS Molecular Science 3 280–299. (doi:10.3934/ prognostic factor. Histopathology 62 986–993. (doi:10.1111/his.12107) molsci.2016.2.280) Mohorko E, Glockshuber R & Aebi M 2011 Oligosaccharyltransferase: Llop E, Ferrer-Batalle M, Barrabes S, Guerrero PE, Ramirez M, Saldova R, the central enzyme of N-linked protein glycosylation. Journal of Rudd PM, Aleixandre RN, Comet J, de Llorens R, et al. 2016 Inherited Metabolic Disease 34 869–878. (doi:10.1007/s10545-011- Improvement of prostate cancer diagnosis by detecting PSA 9337-1) glycosylation-specific changes.Theranostics 6 1190–1204. Mucaj V, Lee SS, Skuli N, Giannoukos DN, Qiu B, Eisinger-Mathason TS, (doi:10.7150/thno.15226) Nakazawa MS, Shay JE, Gopal PP, Venneti S, et al. 2015 Lopez-Sambrooks C, Shrimal S, Khodier C, Flaherty DP, Rinis N, MicroRNA-124 expression counteracts pro-survival stress responses Charest JC, Gao N, Zhao P, Wells L, Lewis TA, et al. 2016 in glioblastoma. Oncogene 34 2204–2214. (doi:10.1038/ Oligosaccharyltransferase inhibition induces senescence in RTK- onc.2014.168) driven tumor cells. Nature Chemical Biology 12 1023–1030. Munkley J 2016 The role of Sialyl-Tn in cancer. International Journal of Endocrine-Related Cancer Endocrine-Related (doi:10.1038/nchembio.2194) Molecular Sciences 17 275. (doi:10.3390/ijms17030275) Lu J, Isaji T, Im S, Fukuda T, Hashii N, Takakura D, Kawasaki N & Gu J Munkley J & Elliott DJ 2016a Hallmarks of glycosylation in cancer. 2014 beta-Galactoside alpha2,6-sialyltranferase 1 promotes Oncotarget 7 35478–35489. (doi:10.18632/oncotarget.8155) transforming growth factor-beta-mediated epithelial-mesenchymal Munkley J & Elliott DJ 2016b Sugars and cell adhesion: The role of transition. Journal of Biological Chemistry 289 34627–34641. ST6GalNAc1 in prostate cancer progression. Cancer Cell and (doi:10.1074/jbc.M114.593392) Microenvironment 3 e1174. (doi:10.14800/ccm.1174) Lu Q, Xu L, Li C, Yuan Y, Huang S & Chen H 2016 miR-214 inhibits Munkley J, Rajan P, Lafferty NP, Dalgliesh C, Jackson RM, Robson CN, invasion and migration via downregulating GALNT7 in esophageal Leung HY & Elliott DJ 2014 A novel androgen-regulated isoform of squamous cell cancer. Tumour Biology 37 14605–14614. (doi:10.1007/ the TSC2 tumour suppressor gene increases cell proliferation. s13277-016-5320-7) Oncotarget 5 131–139. (doi:10.18632/oncotarget.1405) Lynch TP, Ferrer CM, Jackson SR, Shahriari KS, Vosseller K & Munkley J, Lafferty NP, Kalna G, Robson CN, Leung HY, Rajan P & Reginato MJ 2012 Critical role of O-Linked beta-N-acetylglucosamine Elliott DJ 2015a Androgen-regulation of the protein tyrosine transferase in prostate cancer invasion, angiogenesis, and metastasis. phosphatase PTPRR activates ERK1/2 signalling in prostate cancer Journal of Biological Chemistry 287 11070–11081. (doi:10.1074/jbc. cells. BMC Cancer 15 9. (doi:10.1186/s12885-015-1012-8) M111.302547) Munkley J, Livermore KE, McClurg UL, Kalna G, Knight B, McCullagh P, Ma Z & Vosseller K 2014 Cancer metabolism and elevated O-GlcNAc in McGrath J, Crundwell M, Leung HY, Robson CN, et al. 2015b The oncogenic signaling. Journal of Biological Chemistry 289 PI3K regulatory subunit gene PIK3R1 is under direct control of 34457–34465. (doi:10.1074/jbc.R114.577718) androgens and repressed in prostate cancer cells. Oncoscience 2 Ma W, Diep K, Fritsche HA, Shore N & Albitar M 2014 Diagnostic and 755–764. (doi:10.18632/oncoscience.243) prognostic scoring system for prostate cancer using urine and plasma Munkley J, Oltean S, Vodak D, Wilson BT, Livermore KE, Zhou Y, Star E, biomarkers. Genet Test Mol Biomarkers 18 156–163. (doi:10.1089/ Floros VI, Johannessen B, Knight B, et al. 2015c The androgen gtmb.2013.0424) receptor controls expression of the cancer-associated sTn antigen Marimuthu A, Subbannayya Y, Sahasrabuddhe NA, Balakrishnan L, and cell adhesion through induction of ST6GalNAc1 in prostate Syed N, Sekhar NR, Katte TV, Pinto SM, Srikanth SM, Kumar P, et al. cancer. Oncotarget 6 34358–34374. (doi:10.18632/oncotarget.6024) 2013 SILAC-based quantitative proteomic analysis of gastric cancer Munkley J, Mills IG & Elliott DJ 2016a The role of glycans in the secretome. Proteomics Clinical Applications 7 355–366. (doi:10.1002/ development and progression of prostate cancer. Nature Reviews prca.201200069) Urology 13 324–333. (doi:10.1038/nrurol.2016.65)

Munkley J, Vodak D, Livermore KE, James K, Wilson BT, Knight B, Biochemical and Biophysical Research Communications 409 436–441. McCullagh P, McGrath J, Crundwell M, Harries LW, et al. 2016b (doi:10.1016/j.bbrc.2011.05.019) Glycosylation is an androgen-regulated process essential for prostate Rajan P, Dalgliesh C, Carling PJ, Buist T, Zhang C, Grellscheid SN, cancer cell viability. EBioMedicine 8 103–116. (doi:10.1016/j. Armstrong K, Stockley J, Simillion C, Gaughan L, et al. 2011 ebiom.2016.04.018) Identification of novel androgen-regulated pathways and mRNA Myers RB, Meredith RF, Schlom J, LoBuglio AF, Bueschen AJ, isoforms through genome-wide exon-specific profiling of the LNCaP Wheeler RH, Stockard CR & Grizzle WE 1994 Tumor associated transcriptome. PLoS ONE 6 e29088. (doi:10.1371/journal. glycoprotein-72 is highly expressed in prostatic adenocarcinomas. pone.0029088) Journal of Urology 152 243–246. Rajan P, Sudbery IM, Villasevil ME, Mui E, Fleming J, Davis M, Ahmad I, Nakamura K, Yamashita K, Sawaki H, Waraya M, Katoh H, Nakayama N, Edwards J, Sansom OJ, Sims D, et al. 2014 Next-generation Kawamata H, Nishimiya H, Ema A, Narimatsu H, et al. 2015 sequencing of advanced prostate cancer treated with androgen- Aberrant methylation of GCNT2 is tightly related to lymph node deprivation therapy. European Urology 66 32–39. (doi:10.1016/j. metastasis of primary CRC. Anticancer Research 35 1411–1421. eururo.2013.08.011) Newsom-Davis TE, Wang D, Steinman L, Chen PF, Wang LX, Simon AK Read JT, Rahmani M, Boroomand S, Allahverdian S, McManus BM & & Screaton GR 2009 Enhanced immune recognition of cryptic Rennie PS 2007 Androgen receptor regulation of the versican gene glycan markers in human tumors. Cancer Research 69 2018–2025. through an androgen response element in the proximal promoter. (doi:10.1158/0008-5472.CAN-08-3589) Journal of Biological Chemistry 282 31954–31963. (doi:10.1074/jbc. Nie GH, Luo L, Duan HF, Li XQ, Yin MJ, Li Z & Zhang W 2015 M702099200) GALNT7, a target of miR-494, participates in the oncogenesis of Reis CA, Osorio H, Silva L, Gomes C & David L 2010 Alterations in nasopharyngeal carcinoma. Tumour Biology 37 4559–4567. glycosylation as biomarkers for cancer detection. Journal of Clinical (doi:10.1007/s13277-015-4281-6) Pathology 63 322–329. (doi:10.1136/jcp.2009.071035) Nonaka M, Fukuda MN, Gao C, Li Z, Zhang H, Greene MI, Peehl DM, Rho JH, Roehrl MH & Wang JY 2009 Glycoproteomic analysis of human Feizi T & Fukuda M 2014 Determination of carbohydrate structure lung adenocarcinomas using glycoarrays and tandem mass recognized by prostate-specific F77 monoclonal antibody through spectrometry: differential expression and glycosylation patterns of expression analysis of glycosyltransferase genes. Journal of vimentin and fetuin A isoforms. Protein Journal 28 148–160. Biological Chemistry 289 16478–16486. (doi:10.1074/jbc. (doi:10.1007/s10930-009-9177-0) M114.559047) Ricciardelli C, Mayne K, Sykes PJ, Raymond WA, McCaul K, Okamoto T, Yoneyama MS, Hatakeyama S, Mori K, Yamamoto H, Koie T, Marshall VR, Tilley WD, Skinner JM & Horsfall DJ 1997 Elevated Saitoh H, Yamaya K, Funyu T, Fukuda M, et al. 2013 Core2 O-glycan- stromal chondroitin sulfate glycosaminoglycan predicts progression expressing prostate cancer cells are resistant to NK cell immunity. in early-stage prostate cancer. Clinical Cancer Research 3 983–992. Molecular Medicine Reports 7 359–364. (doi:10.3892/mmr.2012.1189) Ricciardelli C, Mayne K, Sykes PJ, Raymond WA, McCaul K, Marshall VR Olivari S & Molinari M 2007 Glycoprotein folding and the role of & Horsfall DJ 1998 Elevated levels of versican but not decorin EDEM1, EDEM2 and EDEM3 in degradation of folding-defective predict disease progression in early-stage prostate cancer. Clinical glycoproteins. FEBS Letters 581 3658–3664. (doi:10.1016/j. Cancer Research 4 963–971. febslet.2007.04.070) Ricciardelli C, Quinn DI, Raymond WA, McCaul K, Sutherland PD, Olzmann JA, Kopito RR & Christianson JC 2013 The mammalian Stricker PD, Grygiel JJ, Sutherland RL, Marshall VR, Tilley WD, et al. endoplasmic reticulum-associated degradation system. Cold Spring 1999 Elevated levels of peritumoral chondroitin sulfate are predictive Endocrine-Related Cancer Endocrine-Related Harbor Perspectives in Medicine 5 a013185. (doi:10.1101/cshperspect. of poor prognosis in patients treated by radical prostatectomy for a013185) early-stage prostate cancer. Cancer Research 59 2324–2328. Ovcharenko D, Kelnar K, Johnson C, Leng N & Brown D 2007 Genome- Ricciardelli C, Sakko AJ, Stahl J, Tilley WD, Marshall VR & Horsfall DJ scale microRNA and small interfering RNA screens identify small 2009a Prostatic chondroitin sulfate is increased in patients with RNA modulators of TRAIL-induced apoptosis pathway. Cancer metastatic disease but does not predict survival outcome. Prostate 69 Research 67 10782–10788. (doi:10.1158/0008-5472.CAN-07-1484) 761–769. (doi:10.1002/pros.20926) Ozaki H, Matsuzaki H, Ando H, Kaji H, Nakanishi H, Ikehara Y & Ricciardelli C, Sakko AJ, Ween MP, Russell DL & Horsfall DJ 2009b The Narimatsu H 2012 Enhancement of metastatic ability by ectopic biological role and regulation of versican levels in cancer. Cancer and expression of ST6GalNAcI on a gastric cancer cell line in a mouse Metastasis Reviews 28 233–245. (doi:10.1007/s10555-009-9182-y) model. Clinical and Experimental Metastasis 29 229–238. Sakai K, Kimata K, Sato T, Gotoh M, Narimatsu H, Shinomiya K & (doi:10.1007/s10585-011-9445-1) Watanabe H 2007 Chondroitin sulfate Peng RQ, Wan HY, Li HF, Liu M, Li X & Tang H 2012 MicroRNA-214 N-acetylgalactosaminyltransferase-1 plays a critical role in suppresses growth and invasiveness of cervical cancer cells by chondroitin sulfate synthesis in cartilage. Journal of Biological targeting UDP-N-acetyl-alpha-D-galactosamine:polypeptide Chemistry 282 4152–4161. (doi:10.1074/jbc.M606870200) N-acetylgalactosaminyltransferase 7. Journal of Biological Chemistry Salanti A, Clausen TM, Agerbaek MO, Al Nakouzi N, Dahlback M, 287 14301–14309. (doi:10.1074/jbc.M111.337642) Oo HZ, Lee S, Gustavsson T, Rich JR, Hedberg BJ, et al. 2015 Pinho SS & Reis CA 2015 Glycosylation in cancer: mechanisms and Targeting human cancer by a glycosaminoglycan binding malaria clinical implications. Nature Reviews Cancer 15 540–555. protein. Cancer Cell 28 500–514. (doi:10.1016/j.ccell.2015.09.003) (doi:10.1038/nrc3982) Sato T, Yoneyama T, Tobisawa Y, Hatakeyama S, Yamamoto H, Kojima Y, Pinho SS, Oliveira P, Cabral J, Carvalho S, Huntsman D, Gartner F, Mikami J, Mori K, Hashimoto Y, Koie T, et al. 2016 Core 2 beta-1, Seruca R, Reis CA & Oliveira C 2012 Loss and recovery of Mgat3 and 6-N-acetylglucosaminyltransferase-1 expression in prostate biopsy GnT-III mediated E-cadherin N-glycosylation is a mechanism specimen is an indicator of prostate cancer aggressiveness. involved in epithelial-mesenchymal-epithelial transitions. PLoS ONE Biochemical and Biophysical Research Communications 470 150–156. 7 e33191. (doi:10.1371/journal.pone.0033191) (doi:10.1016/j.bbrc.2016.01.011) Radhakrishnan P, Chachadi V, Lin MF, Singh R, Kannagi R & Cheng PW Schultz MJ, Swindall AF, Wright JW, Sztul ES, Landen CN & Bellis SL 2011 TNFalpha enhances the motility and invasiveness of prostatic 2013 ST6Gal-I sialyltransferase confers cisplatin resistance in ovarian cancer cells by stimulating the expression of selective glycosyl- and tumor cells. Journal of Ovarian Research 6 25. (doi:10.1186/1757- sulfotransferase genes involved in the synthesis of selectin ligands. 2215-6-25)

Schwarz F & Aebi M 2011 Mechanisms and principles of N-linked atherosclerosis and cancer progression. Advances in Pharmacology 53 protein glycosylation. Current Opinion in Structural Biology 21 281–295. (doi:10.1016/s1054-3589(05)53013-8) 576–582. (doi:10.1016/j.sbi.2011.08.005) Tominaga N, Hagiwara K, Kosaka N, Honma K, Nakagama H & Ochiya T Segawa T, Nau ME, Xu LL, Chilukuri RN, Makarem M, Zhang W, 2014 RPN2-mediated glycosylation of tetraspanin CD63 regulates Petrovics G, Sesterhenn IA, McLeod DG, Moul JW, et al. 2002 breast cancer cell malignancy. Molecular Cancer 13 134. Androgen-induced expression of endoplasmic reticulum (ER) stress (doi:10.1186/1476-4598-13-134) response genes in prostate cancer cells. Oncogene 21 8749–8758. Vasconcelos-Dos-Santos A, Oliveira IA, Lucena MC, Mantuano NR, (doi:10.1038/sj.onc.1205992) Whelan SA, Dias WB & Todeschini AR 2015 Biosynthetic machinery Sharma NL, Massie CE, Ramos-Montoya A, Zecchini V, Scott HE, involved in aberrant glycosylation: promising targets for developing Lamb AD, MacArthur S, Stark R, Warren AY, Mills IG, et al. 2013 The of drugs against cancer. Frontiers in Oncology 5 138. (doi:10.3389/ androgen receptor induces a distinct transcriptional program in fonc.2015.00138) castration-resistant prostate cancer in man. Cancer Cell 23 35–47. Veldscholte J, Ris-Stalpers C, Kuiper GG, Jenster G, Berrevoets C, (doi:10.1016/j.ccr.2012.11.010) Claassen E, van Rooij HC, Trapman J, Brinkmann AO & Mulder E Sheng X, Arnoldussen YJ, Storm M, Tesikova M, Nenseth HZ, Zhao S, 1990 A mutation in the ligand binding domain of the androgen Fazli L, Rennie P, Risberg B, Waehre H, et al. 2015 Divergent receptor of human LNCaP cells affects steroid binding characteristics androgen regulation of unfolded protein response pathways drives and response to anti-androgens. Biochemical and Biophysical Research prostate cancer. EMBO Molecular Medicine 7 788–801. (doi:10.15252/ Communications 173 534–540. (doi:10.1016/S0006-291X(05)80067-1) emmm.201404509) Visakorpi T, Hyytinen E, Koivisto P, Tanner M, Keinanen R, Palmberg C, Silbert JE & Sugumaran G 2002 Biosynthesis of chondroitin/dermatan Palotie A, Tammela T, Isola J & Kallioniemi OP 1995 In vivo sulfate. IUBMB Life 54 177–186. (doi:10.1080/15216540214923) amplification of the androgen receptor gene and progression of Snoek R, Cheng H, Margiotti K, Wafa LA, Wong CA, Wong EC, Fazli L, human prostate cancer. Nature Genetics 9 401–406. (doi:10.1038/ Nelson CC, Gleave ME & Rennie PS 2009 In vivo knockdown of the ng0495-401) androgen receptor results in growth inhibition and regression of Wang Q, Li W, Zhang Y, Yuan X, Xu K, Yu J, Chen Z, Beroukhim R, well-established, castration-resistant prostate tumors. Clinical Cancer Wang H, Lupien M, et al. 2009 Androgen receptor regulates a Research 15 39–47. (doi:10.1158/1078-0432.CCR-08-1726) distinct transcription program in androgen-independent prostate Steinkamp MP, O’Mahony OA, Brogley M, Rehman H, Lapensee EW, cancer. Cell 138 245–256. (doi:10.1016/j.cell.2009.04.056) Dhanasekaran S, Hofer MD, Kuefer R, Chinnaiyan A, Wang D, Dafik L, Nolley R, Huang W, Wolfinger RD, Wang LX & Rubin MA, et al. 2009 Treatment-dependent androgen receptor Peehl DM 2013 Anti-oligomannose antibodies as potential serum mutations in prostate cancer exploit multiple mechanisms to evade biomarkers of aggressive prostate cancer. Drug Development and therapy. Cancer Research 69 4434–4442. (doi:10.1158/0008-5472. Research 74 65–80. (doi:10.1002/ddr.21063) CAN-08-3605) Watanabe Y, Takeuchi K, Higa Onaga S, Sato M, Tsujita M, Abe M, Suburu J & Chen YQ 2012 Lipids and prostate cancer. Prostaglandins Natsume R, Li M, Furuichi T, Saeki M, et al. 2010 Chondroitin and Other Lipid Mediators 98 1–10. (doi:10.1016/j. sulfate N-acetylgalactosaminyltransferase-1 is required for normal prostaglandins.2012.03.003) cartilage development. Biochemical Journal 432 47–55. (doi:10.1042/ Sun S, Sprenger CC, Vessella RL, Haugk K, Soriano K, Mostaghel EA, BJ20100847) Page ST, Coleman IM, Nguyen HM, Sun H, et al. 2010 Castration Wu D, Zhang C, Shen Y, Nephew KP & Wang Q 2011 Androgen resistance in human prostate cancer is conferred by a frequently receptor-driven chromatin looping in prostate cancer. Trends in Endocrine-Related Cancer Endocrine-Related occurring androgen receptor splice variant. Journal of Clinical Endocrinology and Metabolism 22 474–480. (doi:10.1016/j. Investigation 120 2715–2730. (doi:10.1172/JCI41824) tem.2011.07.006) Swindall AF, Londono-Joshi AI, Schultz MJ, Fineberg N, Buchsbaum DJ Wu X, Daniels G, Lee P & Monaco ME 2014 Lipid metabolism in & Bellis SL 2013 ST6Gal-I protein expression is upregulated in prostate cancer. American Journal of Clinical and Experimental Urology human epithelial tumors and correlates with stem cell markers in 2 111–120. normal tissues and colon cancer cell lines. Cancer Research 73 Xie H, Zhu Y, An H, Wang H, Zhu Y, Fu H, Wang Z, Fu Q, Xu J & Ye D 2368–2378. (doi:10.1158/0008-5472.CAN-12-3424) 2016 Increased B4GALT1 expression associates with adverse outcome Swinnen JV, Esquenet M, Goossens K, Heyns W & Verhoeven G 1997a in patients with non-metastatic clear cell renal cell carcinoma. Androgens stimulate fatty acid synthase in the human prostate Oncotarget 7 32723–32730. (doi:10.18632/oncotarget.8737) cancer cell line LNCaP. Cancer Research 57 1086–1090. Yan L, Lin B, Gao L, Gao S, Liu C, Wang C, Wang Y, Zhang S & Swinnen JV, Ulrix W, Heyns W & Verhoeven G 1997b Coordinate Iwamori M 2010 Lewis (y) antigen overexpression increases the regulation of lipogenic gene expression by androgens: evidence for a expression of MMP-2 and MMP-9 and invasion of human ovarian cascade mechanism involving sterol regulatory element binding cancer cells. International Journal of Molecular Sciences 11 4441–4452. proteins. PNAS 94 12975–12980. (doi:10.1073/pnas.94.24.12975) (doi:10.3390/ijms11114441) Swinnen JV, Vanderhoydonc F, Elgamal AA, Eelen M, Vercaeren I, Yanagisawa K, Konishi H, Arima C, Tomida S, Takeuchi T, Shimada Y, Joniau S, Van Poppel H, Baert L, Goossens K, Heyns W, et al. 2000 Yatabe Y, Mitsudomi T, Osada H & Takahashi T 2010 Novel Selective activation of the fatty acid synthesis pathway in human metastasis-related gene CIM functions in the regulation of multiple prostate cancer. International Journal of Cancer 88 176–179. cellular stress-response pathways. Cancer Research 70 9949–9958. (doi:10.1002/1097-0215(20001015)88:2<176::AID- (doi:10.1158/0008-5472.CAN-10-1055) IJC5>3.0.CO;2-3) Zhang Z, Sun P, Liu J, Fu L, Yan J, Liu Y, Yu L, Wang X & Yan Q 2008 Taniguchi N & Kizuka Y 2015 Glycans and cancer: role of N-glycans in Suppression of FUT1/FUT4 expression by siRNA inhibits tumor cancer biomarker, progression and metastasis, and therapeutics. growth. Biochimica et Biophysica Acta 1783 287–296. (doi:10.1016/j. Advances in Cancer Research 126 11–51. bbamcr.2007.10.007) Ten Hagen KG, Fritz TA & Tabak LA 2003 All in the family: the UDP- Zhang G, Zhang H, Wang Q, Lal P, Carroll AM, de la Llera-Moya M, GalNAc:polypeptide N-acetylgalactosaminyltransferases. Glycobiology Xu X & Greene MI 2010 Suppression of human prostate tumor 13 1R–16R. (doi:10.1093/glycob/cwg007) growth by a unique prostate-specific monoclonal antibody F77 Theocharis AD, Tsolakis I, Tzanakakis GN & Karamanos NK 2006 targeting a glycolipid marker. PNAS 107 732–737. (doi:10.1073/ Chondroitin sulfate as a key molecule in the development of pnas.0911397107)

Zhang H, Meng F, Wu S, Kreike B, Sethi S, Chen W, Miller FR & Wu G 2011 Zhou H, Ma H, Wei W, Ji D, Song X, Sun J, Zhang J & Jia L 2013 Engagement of I-branching {beta}-1, 6-N-acetylglucosaminyltransferase B4GALT family mediates the multidrug resistance of human 2 in breast cancer metastasis and TGF-{beta} signaling. Cancer Research leukemia cells by regulating the hedgehog pathway and the 71 4846–4856. (doi:10.1158/0008-5472.CAN-11-0414) expression of p-glycoprotein and multidrug resistance-associated Zhang J, Yan B, Spath SS, Qun H, Cornelius S, Guan D, Shao J, protein 1. Cell Death and Disease 4 e654. (doi:10.1038/ Hagiwara K, Van Waes C, Chen Z, et al. 2015 Integrated cddis.2013.186) transcriptional profiling and genomic analyses reveal RPN2 and Zhu ML & Kyprianou N 2008 Androgen receptor and growth factor HMGB1 as promising biomarkers in colorectal cancer. Cell and signaling cross-talk in prostate cancer cells. Endocrine-Related Cancer Bioscience 5 53. (doi:10.1186/s13578-015-0043-9) 15 841–849. (doi:10.1677/ERC-08-0084)

Received in final form 30 January 2017 Accepted 3 February 2017 Accepted Preprint published online 3 February 2017 Endocrine-Related Cancer Endocrine-Related