Magnetically guided non-invasive CRISPR-Cas9/gRNA delivery across blood-brain barrier to eradicate latent HIV-1 infection
Magnetically guided non-invasive CRISPR-Cas9/gRNA delivery across blood-brain barrier to eradicate latent HIV-1 infection
复制标题
DOI：
10.1038/s41598-019-40222-4
复制
发表时间：
2019-03-08
影响因子：
4.6
通讯作者：
Nair, Madhavan
中科院分区：
文献类型：
--
作者：
Kaushik, Ajeet;Yndart, Adriana;Nair, Madhavan
CRISPR-Cas9/gRNA对潜伏的人类免疫缺陷病毒（HIV）基因组表现出治疗功效，但将这种治疗性货物递送到大脑仍然是一个挑战。在这项研究中，我们首次证明了磁性引导的纳米制剂（NF）的非侵入性递送，该纳米制剂由与磁电纳米颗粒（MENP）结合的Cas9/gRNA组成，穿过血脑屏障（BBB）以抑制小胶质细胞（h mu glia）/HIV（HC 69）细胞中的潜伏性HIV-1感染。在NF上施加60 Oe的优化ac磁场以从MENP表面释放Cas9/gRNA并促进NF细胞摄取，导致细胞内释放和HIV的抑制。结果表明，与HIV潜伏性h μ glia/HIV（HC 69）细胞中未结合的Cas9/gRNA相比，发展的NF显著降低了HIV-LTR表达。这些发现也使用荧光显微镜定性验证，以评估NF对小胶质细胞中潜伏HIV的功效。我们相信，通过BBB的NF（CRISPR/Cas9-gRNA-MENP）的CNS递送肯定将具有临床实用性，作为未来管理神经HIV/AIDS的个性化纳米医学。
CRISPR-Cas9/gRNA exhibits therapeutic efficacy against latent human immunodeficiency virus (HIV) genome but the delivery of this therapeutic cargo to the brain remains as a challenge. In this research, for the first time, we demonstrated magnetically guided non-invasive delivery of a nano-formulation (NF), composed of Cas9/gRNA bound with magneto-electric nanoparticles (MENPs), across the blood-brain barrier (BBB) to inhibit latent HIV-1 infection in microglial (h mu glia)/HIV (HC69) cells. An optimized ac-magnetic field of 60 Oe was applied on NF to release Cas9/gRNA from MENPs surface and to facilitate NF cell uptake resulting in intracellular release and inhibition of HIV. The outcomes suggested that developed NF reduced HIV-LTR expression significantly in comparison to unbound Cas9/gRNA in HIV latent h mu glia/HIV (HC69) cells. These findings were also validated qualitatively using fluorescence microscopy to assess NF efficacy against latent HIV in the microglia cells. We believe that CNS delivery of NF (CRISPR/Cas9-gRNA-MENPs) across the BBB certainly will have clinical utility as future personalized nanomedicine to manage neuroHIV/AIDS.