Biochemical Engineering of Neural Cell Surfaces by the SyntheticN-Propanoyl-substituted Neuraminic Acid Precursor

Schmidt, Carolin; Stehling, Peer; Schnitzer, Jutta; Reutter, Werner; Horstkorte, Rüdiger

doi:10.1074/jbc.273.30.19146

Cited by 87 publications

(90 citation statements)

References 30 publications

(25 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Biochemical engineering of the acyl side chain of sialic acid, using ManNProp as an unnatural precursor, has been shown to stimulate glia cells and interfere with transmitter functions (Schmidt et al, 1998(Schmidt et al, , 2000. In the present study, we inquired whether the most important prerequisite for regeneration of neural cells, namely neurite outgrowth, is affected by this new kind of biochemical engineering.…”

Section: Resultsmentioning

confidence: 99%

“…Human diploid lung fibroblasts displayed a loss of density-dependent growth control after biochemical engineering (Wieser et al, 1996). Treatment of neural cell cultures of newborn rats with ManNProp led to proliferation of astrocytes and microglia and increased the number of oligodendrocyte progenitor cells (Schmidt et al, 1998). These oligodendrocytes show calcium spiking in response to GABA after biochemical engineering of their cell surface with ManNProp (Schmidt et al, 2000).…”

mentioning

confidence: 99%

“…In previous studies we have shown that the novel nonphysiological N-propanoylmannosamine (ManNProp) is metabolized (like the physiological ManNAc) to N-propanoylneuraminic acid (Neu5Prop) in vitro and in vivo using the same metabolic route as ManNAc (see Scheme 1). The simple addition of ManNProp to the cell culture medium leads to the expression of Neu5Prop on cell surface glycoconjugates (Kayser et al, 1992;Keppler et al, 1995;Schmidt et al, 1998Schmidt et al, , 2000. This biochemical engineering, applied to different cell systems, has so far revealed several important biological functions of the N-acyl side chain of sialic acid.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Biochemical Engineering of Cell Surface Sialic Acids Stimulates Axonal Growth

et al. 2002

Self Cite

View full text Add to dashboard Cite

Sialylation is essential for development and regeneration in mammals. Using N-propanoylmannosamine, a novel precursor of sialic acid, we were able to incorporate unnatural sialic acids with a prolonged N-acyl side chain (e.g., N-propanoylneuraminic acid) into cell surface glycoconjugates. Here we report that this biochemical engineering of sialic acid leads to a stimulation of neuronal cells. Both PC12 cells and cerebellar neurons showed a significant increase in neurite outgrowth after treatment with this novel sialic acid precursor. Furthermore, also the reestablishment of the perforant pathway was stimulated in brain slices. In addition, we surprisingly identified several cytosolic proteins with regulatory functions, which are differentially expressed after treatment with N-propanoylmannosamine. Because sialic acid is the only monosaccharide that is activated in the nucleus, we hypothesize that transcription could be modulated by the unnatural CMP-Npropanoylneuraminic acid and that sialic acid activation might be a general tool to regulate cellular functions, such as neurite outgrowth.

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Biochemical Engineering of Cell Surface Sialic Acids Stimulates Axonal Growth

et al. 2002

Self Cite

View full text Add to dashboard Cite

show abstract

“…Levels of GM3NPhAc expression on B16F0 and 3T3 A31 cells treated with ManNPhAc. After B16F0 and 3T3 A31 cells were incubated with 0, 0.02, 0.1, 0.5, and 2.0 mM of ManNPhAc for indicated time (24,48 or 72 h), they were analyzed by cellular ELISA using mAb 2H3 and goat anti-mouse IgM antibody as the primary and secondary antibodies, respectively. GM3NPhAc levels were presented as the mean OD values obtained from triplicate experiments.…”

Section: Antibody-mediated Complement-dependent Cytotoxicity (Cdc) Asmentioning

confidence: 99%

Efficient glycoengineering of GM3 on melanoma cell and monoclonal antibody-mediated selective killing of the glycoengineered cancer cell

Wang

Zhang

Guo

2007

Bioorganic & Medicinal Chemistry

View full text Add to dashboard Cite

To verify the principal of a new immunotherapeutic strategy for cancer, a monoclonal antibody 2H3 against N-phenylacetyl GM3, an unnatural form of the tumor-associated antigen GM3, was prepared and employed to demonstrate that murine melanoma cell B16F0 could be effectively glycoengineered by N-phenylacetyl-D-mannosamine to express N-phenylacetyl GM3 and that 2H3 was highly cytotoxic to the glycoengineered B16F0 cell in the presence of complements. It was further demonstrated that B16F0 cell could be glycoengineered 4-5 times more effectively than 3T3 A31 cell, a normal murine embryo fibroblast cell, and that the antibody and complement mediated cytotoxicity was at least 200 times more potent to the glycoengineered B16F0 cell than to the Nphenylacetyl-D-mannosamine-treated 3T3 A31 cell. These results show the promise for developing useful melanoma immunotherapies based on vaccination against N-phenylacetyl GM3 followed by treatment with N-phenylacetyl-D-mannosamine.

show abstract

“…2). A number of artificial sialic acids have been delivered onto cell surface by this technique (41,42,(46)(47)(48)(49)(50)(51)(52)(53)(54).…”

mentioning

confidence: 99%

Efficient Metabolic Engineering of GM3 on Tumor Cells by N-Phenylacetyl-d-mannosamine

et al. 2006

View full text Add to dashboard Cite

Abnormal carbohydrates expressed on tumor cells, which are referred to as tumor-associated carbohydrate antigens (TACAs), are potential targets for development of cancer vaccines. However, immune tolerance to TACAs has severely hindered progress in this area. To overcome this problem, we have developed a novel immunotherapeutic strategy based on synthetic cancer vaccines and metabolic engineering of TACAs on tumor cells. One critical step of this new strategy is metabolic engineering of cancer, namely to induce expression of an artificial form of a TACA by supplying tumors with an artificial monosaccharide precursor. To identify the proper precursor for this application, N-propionyl, N-butanoyl, N-iso-butanoyl and N-phenylacetyl derivatives of Dmannosamine were synthesized, and their efficiency as biosynthetic precursors to modify sialic acid and induce expression of modified forms of GM3 antigen on tumor cells was investigated. For this purpose, tumor cells were incubated with different N-acyl-D-mannosamines, and modified forms of GM3 expressed on tumor cells were analyzed by flow cytometry using antigen-specific antisera. Nphenylacetyl-D-mannosamine was efficiently incorporated in a time and dose dependent manner to bioengineer GM3 expression by several tumor cell lines including K562, SKMEL-28 and B16-F0. Moreover, these tumor cell lines also showed ManPAc-dependent sensitivity to cytotoxicity medicated by anti-PAcGM3 immune serum and complement. These results provide an important validation for this novel therapeutic strategy. Because N-phenylacetyl GM3-protein conjugates are particularly immunogenic, the combination of an N-phenylacetyl GM3 conjugate vaccine with systemic N-phenylacetyl-D-mannosamine treatment is a promising immunotherapy for future development and application to melanoma and other GM3-bearing tumors. KeywordsMetabolic engineering; cancer vaccine; GM3; D-mannosamine; sialic acid Sialic acids are unique nine-carbon acidic monosaccharides ubiquitously distributed in high animals and human beings (1). The most common form of sialic acids is N-acetyl-D-neuraminic acid (Neu5Ac, also knows as N-acetyl sialic acid). Neu5Ac is a fascinating sugar in that it usually appears at the nonreducing end of cell surface glycans and thus plays forefront roles in various biological functions such as cell proliferation, immune recognition and pathogenic invasion (2-8).

show abstract

Biochemical Engineering of Neural Cell Surfaces by the SyntheticN-Propanoyl-substituted Neuraminic Acid Precursor

Cited by 87 publications

References 30 publications

Biochemical Engineering of Cell Surface Sialic Acids Stimulates Axonal Growth

Biochemical Engineering of Cell Surface Sialic Acids Stimulates Axonal Growth

Efficient glycoengineering of GM3 on melanoma cell and monoclonal antibody-mediated selective killing of the glycoengineered cancer cell

Efficient Metabolic Engineering of GM3 on Tumor Cells by N-Phenylacetyl-d-mannosamine

Contact Info

Product

Resources

About