Hypothesis: Hypoxia induces de novo synthesis of NeuGc gangliosides in humans through CMAH domain substitute

Bousquet, Paula A.; Sandvik, Joe Alexander; Edin, Nina Frederike Jeppesen; Krengel, Ute

doi:10.1016/j.bbrc.2017.11.183

Cited by 25 publications

(32 citation statements)

References 56 publications

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“…In contrast, Neu5Gc-sialoconjugates are often highly expressed in human tumors 6–8 , a phenomenon that has been attributed to the altered metabolic properties of cancer cells with enhanced uptake and incorporation of nonhuman Neu5Gc from Neu5Gc-containing foods, such as red meat 9–11 . To which extent proposed CMAH-independent, alternate biosynthetic pathways also contribute to Neu5Gc expression in cancer cells in patients 12–14 , remains to be explored.…”

Section: Introductionmentioning

confidence: 99%

Antitumor effects of the GM3(Neu5Gc) ganglioside-specific humanized antibody 14F7hT against Cmah-transfected cancer cells

Dorvignit

Boligan

Relova-Hernández

et al. 2019

Sci Rep

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The GM3(Neu5Gc) ganglioside represents a tumor-specific antigen that is considered a promising target for cancer immunotherapy. We previously demonstrated that the humanized antibody 14F7hT, specific for this ganglioside, exhibited significant antitumor effects in preclinical hematological tumor models. As this antibody recognizes human tumor tissues from several origins, we addressed its potential effect on different tumor types. The use of cell lines for testing GM3(Neu5Gc)-targeting strategies, in particular for human malignancies, is complicated by the absence in humans of functional cytidine monophospho- N -acetyl-neuraminic acid hydroxylase (CMAH), the enzyme required for Neu5Gc sialic acid biosynthesis. Quantitative flow cytometry revealed the absence of surface GM3(Neu5Gc) in several human but also mouse cell lines, in the last case due to low expression of the enzyme. Hypoxia-induced expression of this ganglioside on human SKOV3 cells was observed upon culture in Neu5Gc-containing medium without evidence for CMAH-independent biosynthesis. However, only transfection of the mouse Cmah gene into human SKOV3 and mouse 3LL cells induced a stable expression of GM3(Neu5Gc) on the cancer cell surface, resulting in effective models to evaluate the antitumor responses by 14F7hT in vitro and in vivo . This antibody exerted antibody-dependent cell-mediated cytotoxicity (ADCC) and in vivo antitumor effects on these Cmah -transfected non-hematological tumors from both mouse and human origin. These results contribute to validate GM3(Neu5Gc) as a relevant target for cancer immunotherapy and reinforces the value of 14F7hT as a novel anti-cancer drug.

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Section: Introductionmentioning

confidence: 99%

Antitumor effects of the GM3(Neu5Gc) ganglioside-specific humanized antibody 14F7hT against Cmah-transfected cancer cells

Dorvignit

Boligan

Relova-Hernández

et al. 2019

Sci Rep

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“…NeuGc GM3 is structurally highly similar to the ganglioside N -acetyl GM3 (NeuAc GM3), with the only difference being an additional oxygen atom found in NeuGc GM3. The presence of NeuGc GM3 in certain human cancer types remains enigmatic 18 – 20 . NeuAc GM3 can be converted to NeuGc GM3 by cytidine monophosphate- N -acetylneuraminic acid hydroxylase (cmah) 21 .…”

Section: Introductionmentioning

confidence: 99%

Crystal structure of an L chain optimised 14F7 anti-ganglioside Fv suggests a unique tumour-specificity through an unusual H-chain CDR3 architecture

Bjerregaard-Andersen

Johannesen

Abdel-Rahman

et al. 2018

Sci Rep

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Targeted cancer immunotherapy offers increased efficacy concomitantly with reduced side effects. One antibody with promising clinical potential is 14F7, which specifically recognises the NeuGc GM3 ganglioside. This antigen is found in the plasma membrane of a range of tumours, but is essentially absent from healthy human cells. 14F7 can discriminate NeuGc GM3 from the very similar NeuAc GM3, a common component of cell membranes. The molecular basis for this unique specificity is poorly understood. Here we designed and expressed 14F7-derived single-chain Fvs (scFvs), which retained the specificity of the parent antibody. Detailed expression and purification protocols are described as well as the synthesis of the NeuGc GM3 trisaccharide. The most successful scFv construct, which comprises an alternative variable light chain (VLA), allowed structure determination to 2.2 Å resolution. The structure gives insights into the conformation of the important CDR H3 loop and the suspected antigen binding site. Furthermore, the presence of VLA instead of the original VL elucidates how this subdomain indirectly stabilises the CDR H3 loop. The current work may serve as a guideline for the efficient production of scFvs for structure determination.

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“…Along these lines, the expression of NeuGc-containing gangliosides has been shown to be triggered by hypoxic conditions [17]. In particular, Bousquet et al confirmed the capacity of hypoxia to induce NeuGcGM3 in HeLa cells [18].…”

Section: Discussionmentioning

confidence: 99%

Aberrant expression of N-glycolyl GM3 ganglioside is associated with the aggressive biological behavior of human sarcomas

et al. 2019

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Background The aberrant expression of N-glycolyl GM3 ganglioside (NeuGcGM3) in patients with sarcomas was reevaluated by assessing the relation of this molecule with some clinicopathological features and overall survival (OS) of patients. Methods Fifty formalin-fixed and paraffin-embedded specimens from patients diagnosed with sarcomas were included. For the evaluation of NeuGcGM3, the 14F7 monoclonal antibody followed by a peroxidase avidin-biotin system was used. Clinicopathological features were obtained from patient records. Survival rates were estimated by the Kaplan-Meier method and compared with the log-rank test. For multivariate analyses, the Cox regression model was used to identify independent prognostic factors for OS. Results The majority of samples had high levels of NeuGcGM3 expression (66.0%) that showed statistical correlation with age ( p = 0.014), TNM stage ( p = 0.022), histological grade ( p = 0.013) and proliferation rates ( p = 0.012). In addition, a tendency for association with tumor depth ( p = 0.070) was evidenced. In univariate survival analysis, TNM stage ( p = 0.000), occurrence of metastasis (p = 0.000) and expression of NeuGcGM3 ( p = 0.034) were significant prognostic factors for OS, while a tendency for association was evidenced for histological grade ( p = 0.091). Among these variables, only the presence of metastasis ( p = 0.001) was an independent prognostic factor on multivariate analysis. Conclusions The present research suggests the evaluation of NeuGcGM3 expression as a complementary prognostic factor in sarcoma, although our results need to be validated in a larger series and prospective studies. Moreover, our results could support the use of this molecule as a target for immunotherapy.

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Hypothesis: Hypoxia induces de novo synthesis of NeuGc gangliosides in humans through CMAH domain substitute

Cited by 25 publications

References 56 publications

Antitumor effects of the GM3(Neu5Gc) ganglioside-specific humanized antibody 14F7hT against Cmah-transfected cancer cells

Antitumor effects of the GM3(Neu5Gc) ganglioside-specific humanized antibody 14F7hT against Cmah-transfected cancer cells

Crystal structure of an L chain optimised 14F7 anti-ganglioside Fv suggests a unique tumour-specificity through an unusual H-chain CDR3 architecture

Aberrant expression of N-glycolyl GM3 ganglioside is associated with the aggressive biological behavior of human sarcomas

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