An effective cell-penetrating antibody delivery platform
An effective cell-penetrating antibody delivery platform
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DOI：
10.1172/jci.insight.127474
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发表时间：
2019-07-25
期刊：
影响因子：
8
通讯作者：
Yu, Hua
中科院分区：
文献类型：
--
作者：
Herrmann, Andreas;Nagao, Toshikage;Yu, Hua
尽管抗体在临床上取得了公认的成功，但它们通常不会穿透细胞膜来靶向细胞内分子，而其中许多分子是不治之症的基础。在这里，我们表明，将硫代磷酸化 DNA 寡核苷酸与抗体共价缀合，使其能够有效地细胞内化。抗体细胞渗透部分由膜电位改变介导。此外，在没有抗原结合的情况下，修饰抗体的细胞内水平会经历细胞清除，这涉及流出和溶酶体降解，从而能够检测到在成纤维细胞、肿瘤细胞和 T 细胞中测试的预期细胞内分子。这种修饰抗体的靶标依赖性细胞保留扩展到体内研究。局部和全身给予低剂量的修饰抗体均能有效抑制细胞内靶标，如转录因子Myc、干扰素调节因子4和酪氨酸蛋白激酶SRC，及其下游基因在肿瘤中的表达，从而导致肿瘤细胞凋亡和肿瘤生长抑制。这种简单的修饰使得能够使用抗体来检测和调节培养的活细胞和整个动物中的细胞内分子，从而为基于抗体的研究、诊断和治疗的新范例奠定了基础。
Despite their well-recognized success in the clinic, antibodies generally do not penetrate cellular membranes to target intracellular molecules, many of which underlie incurable diseases. Here we show that covalently conjugating phosphorothioated DNA oligonucleotides to antibodies enabled their efficient cellular internalization. Antibody cell penetration was partially mediated by membrane potential alteration. Moreover, without an antigen to bind, intracellular levels of the modified antibodies underwent cellular clearance, which involved efflux and lysosomal degradation, enabling detection of intended intracellular molecules as tested in fibroblasts, tumor cells, and T cells. This target-dependent cellular retention of modified antibodies extended to in vivo studies. Both local and systemic administrations of low doses of modified antibodies effectively inhibited intracellular targets, such as transcription factors Myc, interferon regulatory factor 4, and tyrosine-protein kinase SRC, and expression of their downstream genes in tumors, resulting in tumor cell apoptosis and tumor growth inhibition. This simple modification enables the use of antibodies to detect and modulate intracellular molecules in both cultured living cells and in whole animals, forming the foundation for a new paradigm for antibody-based research, diagnostics, and therapeutics.