Peptides derived from evolutionarily conserved domains in Beclin-1 and Beclin-2 enhance the entry of lentiviral vectors into human cells
Peptides derived from evolutionarily conserved domains in Beclin-1 and Beclin-2 enhance the entry of lentiviral vectors into human cells
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DOI：
10.1074/jbc.m117.800813
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发表时间：
2017-09
期刊：
影响因子：
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通讯作者：
S. Majdoul;J. Cosette;A. Seye;Eric Bernard;Sophie Frin;N. Holic;N. Chazal;L. Briant;L. Espert;A. Galy;D. Fenard
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文献类型：
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作者：
S. Majdoul;J. Cosette;A. Seye;Eric Bernard;Sophie Frin;N. Holic;N. Chazal;L. Briant;L. Espert;A. Galy;D. Fenard
自噬相关蛋白如Beclin-1参与了一系列复杂的过程，包括抗病毒反应，也可能调节基因治疗病毒载体的效率。Tat-Beclin-1（TB 1）肽已被报道为抑制病原体如HIV，1型（HIV-1）复制的自噬诱导因子。然而，自噬相关蛋白对于HIV-1感染的早期步骤也是必不可少的。因此，我们研究了TB 1中Beclin-1进化保守结构域对单轮HIV感染或非复制型HIV-1衍生慢病毒载体中病毒转导和自噬的影响。TB 1增强了各种假型的转导，但不诱导自噬过程。TB 1增强了人CD 34+造血干/祖细胞的转导，同时保持了它们体内移植到人源化小鼠中的能力。TB 1与其他转导添加剂一样有效，并且通过增强病毒颗粒与靶细胞的粘附和融合而不是它们的聚集来起作用。我们还发现，N-末端L1环是TB 1的转导增强活性的关键。有趣的是，Tat-Beclin-2（TB 2）肽，来源于人类Beclin-2蛋白，在促进病毒转导和感染方面甚至比TB 1更有效。综上所述，我们的研究结果表明TB 1和TB 2肽增强了病毒进入步骤。因此，Tat-Beclin肽代表了一个新的家族的病毒转导增强剂的基因治疗的潜在用途。
Autophagy-related proteins such as Beclin-1 are involved in an array of complex processes, including antiviral responses, and may also modulate the efficiency of gene therapy viral vectors. The Tat-Beclin-1 (TB1) peptide has been reported as an autophagy-inducing factor inhibiting the replication of pathogens such as HIV, type 1 (HIV-1). However, autophagy-related proteins are also essential for the early steps of HIV-1 infection. Therefore, we examined the effects of the Beclin-1 evolutionarily conserved domain in TB1 on viral transduction and autophagy in single-round HIV infection or with nonreplicative HIV-1–derived lentiviral vectors. TB1 enhanced transduction with various pseudotypes but without inducing the autophagy process. TB1 augmented the transduction of human CD34+ hematopoietic stem/progenitor cells while maintaining their capacity to engraft in vivo into humanized mice. TB1 was as effective as other transduction additives and functioned by enhancing the adhesion and fusion of viral particles with target cells but not their aggregation. We also found that the N-terminal L1 loop was critical for TB1 transduction–enhancing activity. Interestingly, the Tat-Beclin-2 (TB2) peptide, derived from the human Beclin-2 protein, was even more potent than TB1 in promoting viral transduction and infection. Taken together, our findings suggest that the TB1 and TB2 peptides enhance the viral entry step. Tat-Beclin peptides therefore represent a new family of viral transduction enhancers for potential use in gene therapy.