B4GALNT1 induces angiogenesis, anchorage independence growth and motility, and promotes tumorigenesis in melanoma by induction of ganglioside GM2/GD2

Abstract

β-1,4-N-Acetyl-Galactosaminyltransferase 1 (B4GALNT1) encodes the key enzyme B4GALNT1 to generate gangliosides GM2/GD2. GM2/GD2 gangliosides are surface glycolipids mainly found on brain neurons as well as peripheral nerves and skin melanocytes and are reported to exacerbate the malignant potential of melanomas. In order to elucidate the mechanism, we performed functional analyses of B4GALNT1-overexpressing cells. We analyzed ganglioside pattern on four melanoma and two neuroblastoma cell lines by high performance liquid chromatography (HPLC). We overexpressed B4GALNT1 in GM2/GD2-negative human melanoma cell line (SH4) and confirmed production of GM2/GD2 by HPLC. They showed higher anchorage independence growth (AIG) in colony formation assay, and exhibited augmented motility. In vitro, cell proliferation was not affected by GM2/GD2 expression. In vivo, GM2/GD2-positive SH4 clones showed significantly higher tumorigenesis in NOD/ Scid/IL2Rγ-null mice, and immunostaining of mouse CD31 revealed that GM2/GD2 induced remarkable angiogenesis. No differences were seen in melanoma stem cell and Epithelial-Mesenchymal Transition markers between GM2/GD2-positive and -negative SH4 cells. We therefore concluded that B4GALNT1, and consequently GM2/GD2, enhanced tumorigenesis via induction of angiogenesis, AIG, and cell motility. RNA-Seq suggested periostin as a potential key factor for angiogenesis and AIG. These findings may lead to development of novel therapy for refractory melanoma.abundant in the nervous system, especially brain neurons 14 . They also exist in peripheral nerves and skin melanocytes 15,16 . These molecules are reported to have important biological functions, such as intercellular communication, cell cycling, cell growth, adhesion, differentiation, and cell motility [17][18][19] . Gangliosides are not only detected at high levels in tumors of neuroectodermal cell origin but also related to the biological and clinical behavior of many kinds of tumors 20 . Recently, some analysis revealed that patients with higher expression of B4GALNT1 and GM2/GD2 correlated with poorer prognosis in renal cell carcinoma (TCGA data set; Human Protein Atlas), neuroblastoma 21 , and melanoma 22 . Thus, B4GALNT1 gene is considered to be key tumor-associated antigens [23][24][25][26][27] , indicating that their expression is a meaningful marker for metastatic condition and are potential therapeutic targets for melanoma.Our findings indicate the involvement of B4GALNT1 and GM2/GD2 in tumor establishment and progression as well as a potential direction of therapeutic approach via controlling B4GALNT1, and consequently GM2/GD2 expression in cancers such as melanoma. Scientific RepoRtS |(2020) 10:1199 | https://doi.Generation of GM2/GD2-positive SH4 melanoma clones. The SH4 cells were transfected with expression vectors with or without B4GALNT1 gene cassette, to establish GM2/GD2-positive and -negative SH4 clones. Two GM2/GD2-high clones were selected by single cell isolation (#4 and #5, Fig. S2A). These two clones showed significa...

In Silico Analysis for Sphingolipid Metabolism-Related Genes in Human Kidney Clear Cell Carcinoma Using The Cancer Genome Atlas.

“…SPHK1 is a major enzyme of the S1P metabolism pathway, and plays an important role in tumorigenesis [20], cancer angiogenesis [21], intercellular communication of the tumor environment [22], and formation of the prometastatic environment [23]. B4GALNT1 is a key enzyme in generation of the gangliosides GM2/GD2, and has also been shown to be involved in tumorigenesis [24]. Thus, our results suggest that SPHK1 and B4GALNT1 may play crucial roles as oncogenes in ccRCC.…”

Exploiting Gangliosides for the Therapy of Ewing’s Sarcoma and H3K27M-Mutant Diffuse Midline Glioma

“…For example, gangliosides have been described as key drivers for glioblastoma stem cells and tumorigenicity [ 69 ]. Induction of ganglioside GM2/GD2 enhances tumor incidence and growth speed of melanoma in vivo [ 70 ]. Therefore, enzymes of the ganglioside synthesis could represent realistic targets for the development of antitumoral reagents.…”

Gangliosides and Neuroblastomas

“…Overexpression of MYCN allows it to form heterodimers with MYC-associated protein X (MAX) which permits it to act as a transcriptional factor and support continued tumor growth [135]. The effect of MYCN-MAX on tumor growth coupled with the observation that GD2 is able to enhance angiogenesis [93], thereby increasing the supply of blood borne nutrients needed by proliferating tumor cells, supports the hypothesis that they act synergistically to enhance/maintain cell growth. The finding that children expressing both amplified MYCN oncogene and kinase mutations in ALK were reported to have <15% survival rate after three years underscores the need to develop effective therapeutic approaches for inhibiting both.…”