期刊
EBIOMEDICINE
卷 58, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.ebiom.2020.102897
关键词
CRISPR/Cas9; Gene therapy; Local delivery; Poly (beta-amino ester); Vaginal gel
资金
- National Science and Technology Major Project of the Ministry of science and technology of China [2018ZX10301402]
- National Natural Science Foundation of China [81761148025, 81871473, 81402158]
- Guangzhou Science and Technology Programme [201704020093]
- National Ten Thousand Plan-Young Top Talents of China
- Fundamental Research Funds for the Central Universities [17ykzd15, 19ykyjs07]
- Three Big Constructions-Supercomputing Application Cultivation Projects - National Supercomputer Center In Guangzhou
- National Research foundation (NRF) South Africa under BRICS Multilateral Joint Call for Proposals
- Russian Foundation for Basic Research [17-54-80078]
Background: Gene therapy has held promises for treating specific genetic diseases. However, the key to clinical application depends on effective gene delivery. Methods: Using a large animal model, we developed two pharmaceutical formulations for gene delivery in the pigs' vagina, which were made up of poly (beta-amino ester) (PBAE)-plasmid polyplex nanoparticles (NPs) based two gel materials, modified montmorillonite (mMMT) and hectorite (HTT). Findings: By conducting flow cytometry of the cervical cells, we found that PBAE-GFP-NPs-mMMT gel was more efficient than PBAE-GFP-NPs-HTT gel in delivering exogenous DNA intravaginally. Next, we designed specific CRISPR/SpCas9 sgRNAs targeting porcine endogenous retroviruses (PERVs) and evaluated the genome editing efficacy in vivo. We discovered that PERV copy number in vaginal epithelium could be significantly reduced by the local delivery of the PBAE-SpCas9/sgRNA NPs-mMMT gel. Comparable genome editing results were also obtained by high-fidelity version of SpCas9, SpCas9-HF1 and eSpCas9, in the mMMT gel. Further, we confirmed that the expression of topically delivered SpCas9 was limited to the vagina/cervix and did not diffuse to nearby organs, which was relatively safe with low toxicity. Interpretation: Our data suggested that the PBAE-NPs mMMT vaginal gel is an effective preparation for local gene therapy, yielding insights into novel therapeutic approaches to sexually transmitted disease in the genital tract. (c) 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license. (http://creativecommons.org/licenses/by-nc-nd/4.0/)
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