这项研究评估了基于微针的脉络膜上(SC)给药后非病毒DNA纳米颗粒(DNP)的眼部耐受性和可转染性,与视网膜下(SR)给药相比。
DNP由单拷贝的质粒DNA和用10kDaPEG取代的赖氨酸30聚体肽(CK30PEG10k)压紧的聚泛素C/荧光素酶转录盒组成。新西兰白兔(每组n=4)接受单侧SC注射(通过微针技术0.1mL)椭圆形DNP,杆状DNP,或盐水(阴性对照)。一组兔(n=4)还接受了杆状DNP的单次单侧SR注射(0.05mL,通过跨玻璃体方法)。在第7天,通过生物发光测定在视网膜和视网膜色素上皮(RPE)-脉络膜中测量荧光素酶活性。一组兔子接受了类似DNP的SC注射,以通过光学相干断层扫描和组织学评估DNP注射液在脉络膜上腔(SCS)中的扩散。
脉络膜上注射DNP导致SCS周向和向后可逆打开,并且通常具有良好的耐受性。眼部检查评分无明显变化,眼内压异常,或第7天与基线相比视网膜电图幅度的变化。与对照组相比,在接受SCDNP(杆状和椭圆形)和SRDNP(杆状)的眼睛的视网膜和RPE脉络膜中观察到高荧光素酶活性。RPE脉络膜和视网膜中的平均荧光素酶活性在SC和SR给药之间相当。在SC或SR施用DNP后,RPE脉络膜中的转染比视网膜中的转染高约10倍。
脉络膜上和SR给予DNP导致视网膜和RPE脉络膜的相当转染。
脉络膜上递送DNP提供了精确靶向脉络膜视网膜组织的潜力,同时避免了与SR注射相关的手术风险。它可能为视网膜疾病的治疗提供基于办公室的非手术基因治疗选择。
This study evaluated ocular tolerability and transfectability of nonviral DNA nanoparticles (DNPs) after microneedle-based suprachoroidal (SC) administration, in comparison to subretinal (SR) administration.
The DNPs consisted of a single copy of plasmid DNA with a polyubiquitin C/luciferase transcriptional cassette compacted with 10 kDa PEG-substituted lysine 30-mer peptides (CK30PEG10k). New Zealand White rabbits (n = 4 per group) received a unilateral SC injection (0.1 mL via a microneedle technique) of ellipsoid-shaped DNPs, rod-shaped DNPs, or saline (negative control). A cohort of rabbits (n = 4) also received a single unilateral SR injection (0.05 mL via a transvitreal approach) of rod-shaped DNPs. At day 7, luciferase activity was measured in the retina and retinal pigment epithelium (RPE)-choroid via bioluminescence assay. A cohort of rabbits received a SC injection of analogous DNPs to assess spread of DNP injectate in the suprachoroidal space (SCS) via optical coherent tomography and histology.
Suprachoroidal injection of DNPs resulted in reversible opening of the SCS circumferentially and posteriorly and was generally well tolerated, with no significant ocular examination score changes, intraocular pressure abnormalities, or changes in electroretinography amplitudes on day 7 compared to the baseline. High luciferase activity was observed in the retina and RPE-choroid of eyes that received SC DNPs (rod and ellipsoid shape) and SR DNPs (rod shape) compared to controls. The mean luciferase activity in RPE-choroid and retina was comparable between SC and SR administrations. Transfection in the RPE-choroid was approximately 10-fold higher than in the retina after either SC or SR administration of DNPs.
Suprachoroidal and SR administration of DNPs resulted in comparable transfection of retina and RPE-choroid.
Suprachoroidal delivery of DNPs offers the potential to precisely target chorioretinal tissues while avoiding surgical risks associated with SR injection, and it may offer an office-based nonsurgical gene therapy option for the treatment of retinal diseases.