Protein-Programmed Accumulation of Yeast Cytosine Deaminase in Cancer Cells in Response to Mock-Hypoxia
Protein-Programmed Accumulation of Yeast Cytosine Deaminase in Cancer Cells in Response to Mock-Hypoxia
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DOI：
10.1021/acssynbio.9b00036
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发表时间：
2019-05-01
影响因子：
4.7
通讯作者：
Ostermeier, Marc
中科院分区：
文献类型：
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作者：
Warren, Tiana D.;Patel, Krishna;Ostermeier, Marc
基因导向的酶前体药物治疗（GDEPT）的一个限制是难以用编码前体药物转化酶的基因选择性地和有效地转导癌细胞。为了避免这个问题，我们试图将选择性从基因递送水平转移到蛋白质水平。我们开发了前药活化酶酵母胞嘧啶脱氨酶（yCD）和HIF-1 α的氧依赖性降解结构域（ODDD）的融合蛋白，该结构域以氧依赖性方式调节HIF-1 α的积累。我们将这些HOPE融合物称为HIF 1-α氧依赖性降解结构域/前药转化酶。HOPE融合物被设计为选择性地在经历缺氧的细胞中积累，并因此选择性地引起前药5-氟胞嘧啶（5-FC）转化为化疗剂5-氟尿嘧啶（5-FU），其中氧水平低（例如，在肿瘤的中心）。与我们的假设一致，HT 1080纤维肉瘤细胞转导HOPE融合基因表现出增加的融合蛋白的积累和增加的敏感性，5-FC在模拟缺氧。
One limitation of gene-directed enzyme prodrug therapy (GDEPT) is the difficulty in selectively and efficiently transducing cancer cells with the gene encoding a prodrug-converting enzyme. To circumvent this issue, we sought to move the selectivity from the gene delivery level to the protein level. We developed fusion proteins of the prodrug-activating enzyme yeast cytosine deaminase (yCD) and the oxygen-dependent degradation domain (ODDD) of HIF-1 alpha, a domain that regulates the accumulation of HIF-1 alpha in an oxygen-dependent manner. We called these HOPE fusions for HIF1-alpha Oxygen-dependent degradation domain/Prodrug-converting Enzyme. The HOPE fusions were designed to selectively accumulate in cells experiencing hypoxia and thus selectively cause conversion of the prodrug 5-fluorocytosine (5-FC) to the chemotherapeutic 5-fluorouracil (5-FU) where oxygen levels are low (e.g., at the center of a tumor). Consistent with our hypothesis, HT1080 fibrosarcoma cells transduced with HOPE fusion genes exhibited increased fusion protein accumulation and increased sensitization to 5-FC in mock-hypoxia.