Efficient metabolic engineering, of GM3 on tumor cells by N-phenylacetyl-D-mannosamine
Efficient metabolic engineering, of GM3 on tumor cells by N-phenylacetyl-D-mannosamine
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DOI：
10.1021/bi052161r
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发表时间：
2006-03-21
期刊：
影响因子：
2.9
通讯作者：
Harding, CV
中科院分区：
文献类型：
--
作者：
Chefalo, P;Pan, YB;Harding, CV
肿瘤细胞上表达的异常碳水化合物被称为肿瘤相关碳水化合物抗原（TACAs），是开发癌症疫苗的潜在靶点。然而，对TACAs的免疫耐受严重阻碍了这一领域的进展。为了克服这一问题，我们开发了一种基于合成癌症疫苗和TACAs对肿瘤细胞代谢工程的新型免疫治疗策略。这种新策略的一个关键步骤是癌症的代谢工程，即通过向肿瘤提供人工单糖前体来诱导人工形式的TACA的表达。为了确定适合此应用的前体，n -丙酰…合成了d -甘露糖胺的n -丁醇基、n -异丁醇基和n -苯乙酰基衍生物，并研究了它们作为生物合成前体修饰唾液酸和诱导修饰形式GM3抗原在肿瘤细胞上表达的效率。为此，用不同的n-酰基- d -甘露糖胺培养肿瘤细胞，用抗原特异性抗血清流式细胞术分析肿瘤细胞上表达的GM3的修饰形式。n -苯乙酰基-d -甘露糖胺以时间和剂量依赖的方式有效地掺入到几种肿瘤细胞系中，包括K562、SKMEL-28和B16-F0，生物工程GM3表达。此外，这些肿瘤细胞系对抗pacgm3免疫血清和补体介导的细胞毒性也表现出manpac依赖的敏感性。这些结果为这种新的治疗策略提供了重要的验证。由于n -苯乙酰基GM3蛋白偶联物具有特别的免疫原性，因此n -苯乙酰基GM3蛋白偶联疫苗与系统n -苯乙酰基- d -甘露胺治疗相结合是一种很有前景的免疫治疗方法，可用于黑色素瘤和其他携带GM3的肿瘤。
Abnormal carbohydrates expressed on tumor cells, which are termed tumor-associated carbohydrate antigens (TACAs), are potential targets for the 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-isobutanoyl, and N-phenylacetyl derivatives Of D-mannosamine were synthesized, and their efficiency as biosynthetic precursors in modifying sialic acid and inducing 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. N-Phenylacetyl-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 exhibited ManPAc-dependent sensitivity to cytotoxicity mediated 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.