Gang Niu1, Zhuang Jin1, Chong Zhang2, Dan He3, Xueqin Gao4, Chenming Zou2, Wei Zhang2, Jiahui Ding2, Bhudev C Das5, Konstantin Severinov6, Inga Isabel Hitzeroth7, Priya Ranjan Debata8, Xin Ma9, Xun Tian10, Qinglei Gao11, Jun Wu12, Zeshan You1, Rui Tian1, Zifeng Cui1, Weiwen Fan1, Weiling Xie1, Zhaoyue Huang1, Chen Cao10, Wei Xu1, Hongxian Xie13, Hongyan Xu14, Xiongzhi Tang15, Yan Wang16, Zhiying Yu17, Hui Han18, Songwei Tan19, Shuqin Chen20, Zheng Hu21. 1. Department of Obstetrics and Gynecology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China. 2. School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. 3. Department of Neurology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong, China. 4. Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. 5. Amity Institute of Molecular Medicine & Stem Cell Research, Amity University, Uttar Pradesh, Noida 201313, India. 6. Skolkovo Institute of Science and Technology, Skolkovo, Moscow Region 143025, Russian Federation. 7. Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Cape Town 7701, South Africa. 8. Department of Zoology, North Orissa University, Takatpur, Baripada, Odisha 757003, India. 9. Department of Urology, General Hospital of People's Liberation Army, Beijing 100039, China. 10. Department of Obstetrics and Gynecology, Academician expert workstation, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, Hubei, China. 11. Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei, China. 12. School of Biomedical Engineering, Sun Yat-sen University, Guangzhou 510006, Guangdong, China. 13. Generulor Company Bio-X Lab, Guangzhou 510006, Guangdong, China. 14. Department of Obstetrics and Gynecology, Yuebei People's Hospital, Medical College of Shantou University, Shaoguan 512026, Guangdong, China. 15. Department of Obstetrics and Gynecology, Guilin People's Hospital, Guilin, The Guangxi Zhuang Autonomous Region, 541002, China. 16. Key Laboratory of Molecular Biophysics of the Ministry of Education, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China. 17. Department of Obstetrics & Gynecology, First Affiliated Hospital of Shenzhen University, Shenzhen 518000, Guangdong, China. 18. State Key Laboratory of Oncology in Southern China, Collaborative Innovation Center for Cancer Medicine & Department of Urology, Yat-sen University Cancer Center, Guangzhou 510080, Guangdong Province, China. 19. School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. Electronic address: tansongwei@gmail.com. 20. Department of Obstetrics and Gynecology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China. Electronic address: chenshuqin1021@163.com. 21. Department of Obstetrics and Gynecology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China; Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei, China; Precision Medicine Institute, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China. Electronic address: huzheng1998@163.com.
Abstract
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 (β-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. FUNDING: This work was supported by the National Science and Technology Major Project of the Ministry of science and technology of China (No. 2018ZX10301402); the National Natural Science Foundation of China (81761148025, 81871473 and 81402158); Guangzhou Science and Technology Programme (No. 201704020093); National Ten Thousand Plan-Young Top Talents of China, Fundamental Research Funds for the Central Universities (17ykzd15 and 19ykyjs07); Three Big Constructions-Supercomputing Application Cultivation Projects sponsored by National Supercomputer Center In Guangzhou; the National Research FFoundation (NRF) South Africa under BRICS Multilateral Joint Call for Proposals; grant 17-54-80078 from the Russian Foundation for Basic Research.
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 (β-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. FUNDING: This work was supported by the National Science and Technology Major Project of the Ministry of science and technology of China (No. 2018ZX10301402); the National Natural Science Foundation of China (81761148025, 81871473 and 81402158); Guangzhou Science and Technology Programme (No. 201704020093); National Ten Thousand Plan-Young Top Talents of China, Fundamental Research Funds for the Central Universities (17ykzd15 and 19ykyjs07); Three Big Constructions-Supercomputing Application Cultivation Projects sponsored by National Supercomputer Center In Guangzhou; the National Research FFoundation (NRF) South Africa under BRICS Multilateral Joint Call for Proposals; grant 17-54-80078 from the Russian Foundation for Basic Research.
Authors: Alicia Rodríguez-Gascón; Ana Del Pozo-Rodríguez; Arantxazu Isla; María Angeles Solinís Journal: Adv Drug Deliv Rev Date: 2015-07-17 Impact factor: 15.470
Authors: Peter Cameron; Chris K Fuller; Paul D Donohoue; Brittnee N Jones; Matthew S Thompson; Matthew M Carter; Scott Gradia; Bastien Vidal; Elizabeth Garner; Euan M Slorach; Elaine Lau; Lynda M Banh; Alexandra M Lied; Leslie S Edwards; Alexander H Settle; Daniel Capurso; Victor Llaca; Stéphane Deschamps; Mark Cigan; Joshua K Young; Andrew P May Journal: Nat Methods Date: 2017-05-01 Impact factor: 28.547