ObjectiveTo investigate the current status of research in gene therapy for retinitis pigmentosa (RP) from 2005 to 2024. MethodsThe literature related to gene therapy for RP included in the Web of Science Core Collection dataset from January 1, 2005 to September 15, 2024 was retrieved and screened. The bibliometrix package of R software was used to analyze the annual trend of the number of publications, citation frequency, distribution of countries/regions of the literature, and distribution of journals containing the articles. CiteSpace software was used to perform keyword clustering analysis and the keywords bursts analysis. ResultsA total of 209 articles were included. There was an overall fluctuating upward trend of annual publications from 2005 to 2024, with the highest number of publications in 2023 at 26 (12.4%, 26/209), and the lowest number of publications in 2006 at 2 (0.9%, 2/209). There was an overall increasing trend in the frequency of citations to relevant literature. Corresponding authors from the United States had the highest total number of publications with 98 (46.9%, 98/209). Among authors, Hauswirth from the University of Florida, USA, had the most with 25 (12.0%, 25/209). Among institutions, Columbia University, USA, had the most with 55 (26.3%, 55/209). Among journals, Mol Ther had the most with 25 (12.0%, 25/209), and it had the highest 2023 impact factor of 12.1. Keyword clustering analysis yielded eight valid clusters, namely #0 P23H, #1 AAV, #2 PDE6B, #3 CRB1, #4 RPGR, #5 antisense oligonucleotide, #6 NR2E3, and #7 NRL, which intersected with each other with good continuity. The keywords bursts analysis showed that the keyword with the longest emergence time was RNAi, followed by PDE and PDE6. USH2A, CRB1, CRISPR Cas9, base editing, and ORF15 were keywords that emerged in recent years and were continuously studied. ConclusionsRP gene therapy research literature has shown an increasing trend from 2005 to 2024, with the highest number of publications from research organizations and scholars in the United States. Currently, studies focus on RHO, PDE6B, CRB1, RPGR, NR2E3, and NRL gene. In recent years, there has been a gradual increase in studies on USH2A, CRB1 genes, and the RPGR ORF15 region. CRISPR Cas9 and base editing gene therapy strategies are being developed.
Objective To investigate the expression of T cell receptor (TCR) Vβ8.3 gene on CD4+ T lymphocytes in the rats with experimental autoimmune uveoretinitis (EAU). Methods Eighteen Lewis rats were divided into EAU, complete Freund′s adjuvant, and the control group. Inter photoreceptor retinoid-binding protein (IRBP) R16 peptide was synthesized using Fmoc procedure for induction of EAU. Magnetic absorption cell sorting (MACS) me thod was used to isolate the CD4+T lymphocytes from the spleen of the rats. Flow cytometry was used to monitor the efficiency of isolation. The expression of TCR Vβ8.3 gene segment on CD4+T lymphocytes was determined by fluorescent quantitative polymerase chain reaction. Results EAU was successfully induced in the Lewis rats immunized with IRBP R16 peptide. The proportion of CD4+T lymphocytes isolated by means of MACS was statistically higher than that before isolation (P＜0.001). The expression of TCR Vβ8.3 gene segment on CD4+ T lymphocytes in EAU rats was significantly higher than that in the control (P＜0.05). Conclusions There is a predominant usage of antigen-specific TCR Vβ 8.3 gene in EAU rats induced by IR BP R16 peptide, which may serve as a target for immunotherapy of EAU. (Chin J Ocul Fundus Dis，2004，20：165-167)
Objective To investigate whether mutations exist in codon 58 and codon 347 of the rhodopsin gene in patients with autosomal dominant retinitis pigmentosa(ADRP)． Methods Point mutations at codons 58 and 347 were detected by restriction endonuclease digestion of exons 1 and 5 amplified by polymerase chain reaction(PCR)．This method was applied to screen genomic DNAs from 57 patients of 38 families with ADRP and 60 normal controls． Results Four patients from one family of ADRP were confirmed to have a point mutation at the second nucleotide of codon 58,and 6 patients from two families of ADRP were found to have a mutation at codon 347．None of these mutations were found in 60 normal subjects． Conclusion It is suggested that molecular genetic heterogeneity exists within ADRP and some subtypes of ADRP are caused by points mutations of the rhodopsin gene． (Chin J Ocul Fundus Dis,1998,14:108-110)
Objective To detect and analyse the mutations in rhodopsin gene of members in a family affected by autosomal dominant retinitis pigmentosa (ADRP). Methods Using the polymerase chain reaction (PCR), we amplified exon 1-5 of rhodopsin gene in patients with ADRP,and analyzed it with direct sequence measuement. Results The Gly-182-Asp mutation in the rhodopsin gene was detected in most of affected members of this ADRP family, but no mutation was detected in two affected members and the control ones. Conclusion We cannot regard the Gly-182-Asp mutation in the rhodopsin gene as the pathagenic factor of the ADRP family. It is likely there is a new gene next to the rhodopsin gene. (Chin J Ocul Fundus Dis, 2002, 18: 256-258)
Objective To observe the interferon-gamma; (IFN-gamma;), interleukin-2 (IL-2) levels of Th1 cytokine and IL-4、IL-10 levels of Th2 cytokine in serum and culture supernatants of splenic cells of the rats in the prevention of experimental autoimmune uveoretinitis（EAU）by oral tolerance. Methods 72 Lewis rats were randomly divided into EAU group，oral tolerance group (which including 10 mu;g、100 mu;g、1 mg、10 mg of S antigen group respectively) and control group，12 rats in each group. The animal model of EAU was induced by immunization with S antigen（50 mu;g）and Freundrsquo;s complete adjuvant. Oral tolerance 10 mu;g、100 mu;g、1 mg and 10 mg group were fed with 1 ml mixture of 10 mu;g、100 mu;g、1 mg、10 mg S antigen and 1 mg trypsin inhibitor respectively by intubation，once the other day，totally 7 times，and then induced EAU according to above methods；control group was fed with 1 ml mixture of phosphate buffered saline and 1 mg trypsin inhibitor，once the other day，totally 7 times，and then induced EAU. The clinical manifestation of EAU in the eye were recorded,the eyeballs were enucleated at the peak of EAU，followed by pathological grading. Meanwhile the serum was colleced; splentic cells were separated and cultured to collect the supernatant. Cytokine levels of IFN-gamma;, IL-2, IL-4 and IL-10 in serum, cultured supernatant of splenic cells were determined by enzyme-linked immunosorbent assay (ELISA). Results Compared with EAU and control group, the levels of IFN-gamma; and IL-2 (Th1 cytokine) in the serum in 100 mu;g and 1 mg group were decreased while the levels of IL4 and IL10 (Th2 cytokine) were increased，the differences were statistically significant（F=51.9, 68.8, 35.7,7.5,P＜0.01). Compared the levels of Th1 and Th2 cytokines in the serum in 10 mu;g, 10 mg group with EAU and control group, the differences were not statistically significant. In 100 mu;g、1 mg group, the levels of IFN-gamma; and IL-2 (Th1 cytokine) in the culture supernatant of splenic cells were decreased while the levels of IL-4 and IL-10 (Th2 cytokine) were increased, compared with EAU and control group, the differences were statistically significant（F=57.1,15.6,33.1,167.7, P＜0.01）. Compared the levels of Th1 and Th2 cytokine in the culture supernatant of splenic cells in 10 mu;g、10 mg groups with EAU and control group, the difference are not statistically significant. Conclusions In the process to prevent EAU by oral intake, the levels of IFN-gamma; and IL-2 (Th1 cytokine ) were decrease while the levels of IL-4 and IL-10 (Th2 cytokine). Oral administration with too high or low dose of the antigen can not prevent EAU as well as the cytokine levels do not change obviously. Cytokines has played an important role in the prevention of EAU.
Objective To detect characteristics and the pathogenesis of rhodopsin (RHO) gene mutation in an inbreeding family with autosomal recessive retinitis pigmentosa (ARRP). Methods Peripheral venous blood 5-8 ml was abstracted from 8 members in the inbreeding ARRP family and 10 control individuals. DNA gene group was picked. Extron 1-5 of RHO gene was amplified by polymerase chain reaction (PCR)，and the mutation of RHO gene was screened by direct DNA sequence measurement. Results The Gln-344-Arg mutation in the RHO gene was detected in 3 patients with ARRP and homozygotes of the mutation in 3 patients were found. Heterozygous of the mutation was detected in the parent of patients and 1 healthy family member. No mutation of RHO gene was found in 2 healthy family members and 10 control individuals. Conclusions The Gln-344-Arg mutation in the RHO gene may be the pathogenic factor of the ARRP family; the frequency of the mutation of RHO gene may increase in the in breeding ARRP family.(Chin J Ocul Fundus Dis，2004，20：145-148)
Retinitis pigmentosa (RP) is an inherited retinal disease characterized by degeneration of retinal pigment epithelial cells. Precision medicine is a new medical model that applies modern genetic technology, combining living environment, clinical data of patients, molecular imaging technology and bio-information technology to achieve accurate diagnosis and treatment, and establish personalized disease prevention and treatment model. At present, precise diagnosis of RP is mainly based on next-generation sequencing technology and preimplantation genetic diagnosis, while precise therapy is mainly reflected in gene therapy, stem cell transplantation and gene-stem cell therapy. Although the current research on precision medicine for RP has achieved remarkable results, there are still many problems in the application process that is needed close attention. For instance, the current gene therapy cannot completely treat dominant or advanced genetic diseases, the safety of gene editing technology has not been solved, the cells after stem cell transplantation cannot be effectively integrated with the host, gene sequencing has not been fully popularized, and the big data information platform is imperfect. It is believed that with the in-depth research of gene sequencing technology, regenerative medicine and the successful development of clinical trials, the precision medicine for RP will be gradually improved and is expected to be applied to improve the vision of patients with RP in the future.
Objective To evaluate the therapeutic effects of vitrectomy on chronic and refractory Behcet′s diseases. Methods The clinical data of 8 patients (10 eyes) with Behcet′s diseases from 0.5 to 3 years after vitrectomy were retrospectively analyzed. Pre-and post-operative visual acuity, control of inflammation after the surgery, and the recurrence were observed and analyzed. Both of the eyes were involved in all of the patients, including complete Behcet′s diseases in 5 and incomplete in 3. Results Vitreous opacity and liquefaction were found in all of the involved eyes, dark retina and thin retinal vessels were seen in most of the eyes, and vascular obstruction or sheath-like changes in different degrees were detected in some eyes. Improvement of visual acuity was observed in all of the eyes which had undergone the treatment after the operation. There was significant difference between the visual acuity before the surgery and at the 1st and 2nd week and the 1st and 6th month after the surgery. During the follow-up period, recurrence of the inflammation was found in 3 eyes within 1 month and in 6 eyes within 6 months; no recurrence was found in 4 eyes after the operation. No acute recurrence of inflammation was found after operation in the involved eyes. Conclusion Vitrectomy for chronic and refractory Behcet′s diseases may improve the visual acuity of the involved eyes, and the surgery is safe and effective. (Chin J Ocul Fundus Dis, 2005, 21: 357-359)
ObjectiveTo explore the light response, retinal inflammation and apoptosis of the retinal ganglion cells (RGCs) 1 year after the new type of channelrhodopsin PsCatCh2.0 was transfected into the retina of rd1 mice. MethodsTwenty-four male rd1 mice were randomly divided into rd1 experimental group and rd1 control group, 12 mice in each group. 1.5 μl of recombinant adeno-associated virus (rAAV)2/2-cytomegalovirus (CMV)-PsCatCh2.0-enhanced green fluorescent protein (EGFP) was injected into the vitreous cavity 1 mm below the corneoscleral limbus of mice in the rd1 experimental group, and the same dose of recombinant virus was injected 2 weeks later at temporal side 1 mm below the corneoscleral limbus. One year after virus injection, the light response of RGCs expressing PsCatCh2.0 was recorded by patch clamp technique; the expression of PsCatCh2.0 in the retina was evaluated by immunofluorescence staining; the transfection efficiency of recombinant virus was evaluated by the transfection efficiency of virus and the number of RGCs. Hematoxylin-eosin staining was performed to measure the inner retinal thickness. Western blotting was used to detect the protein expression of nuclear factor (NF)-κB p65 in retina; real-time quantitative polymerase chain reaction was used to detect the relative expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and Bax mRNA. Terminal deoxynucleotidyl transferase kit was used to observe the apoptosis of retinal cells in each group of mice. ResultsOne year after the intravitreal injection of recombinant virus, PsCatCh2.0-expressing RGCs can still generate 30 pA photocurrent. The virus PsCatCh2.0-EGFP was mainly transfected into RGCs, and partly transfected into amacrine cells, almost no transfection was seen in bipolar and horizontal cells. There were no significant differences in the number of RGCs and thickness of the inner retina between the rd1 experimental group and the rd1 control group (F=14.35, 0.05; P＞0.05), while the rd1 experimental group NF-κB p65 protein expression, TNF-α and IL-6 mRNA quantification were significantly lower than those of rd1 control group (F=4.61, 5.91, 5.78; P＜0.05). The number of red fluorescent apoptotic cells in the retina of mice in the rd1 experimental group was less than that in the rd1 control group, and the Bax mRNA expression was lower than that in the rd1 control group, and the difference was statistically significant (F=7.52, P＜0.01). ConclusionOne year after intravitreal injection of recombinant virus, the PsCatCh2.0 expressing RGCs can still generate photocurrent. Long term transfection and expression of PsCatCh2.0 has no obvious cytotoxic effect on RGCs, nor it increases the inflammatory effect of the retina of rd1 mice with retinal degeneration.
Objective To observe the mutifocal electroretinogram (mfERG) characteristics of rod and cone cells in patients with retinitis pigmentosa (RP) and to evaluate the function of photosensory cell.Methods The mfERG recording technique for rod cell in eight normal subjects (eight eyes) were established and the influence of different brightness lightstimulus in P1 wave amplitude were analyzed. The cone and rod cells mfERG of 38 eyes in 19 patients were recorded and then calculated positive ratio from local signalnoise ratio. The average visual acuity and P1 wave amplitude density of cone mfERG in different types were compared and statistically analyzed. Meanwhile, the changes in P1 wave amplitude of cone and rod mfERG in four quadrants also compared and analyzed. Results Rod cell mfERG in normal subjects can be recorded stably by using blue flashes with low light intensity as 0.04 cd/m2. In patients with RP, the cone and rod cells mfERG can be detectd 65.79% and 10.51% respectively. P1 wave amplitude density in type I of cone cell mfERG was significantly higher than that in type II (t=5.21,P=0.000). There were no differences in average visual acuity (t=1.15, P=0.612). P1 wave amplitude density in type I was negatively related to logMAR visual acuity （r=-0.48，P=0.04）.The comparison of rod and cone cells mfERG in local wave characteristics showed that P1 wave amplitude densities had spatial relationship in each area. Conclusions The results suggested highly variable central responses in cone cell in RP patients, higher positive recorded ratio in cone cell than rod cell and spatial correspondence between the function of reserved cone and rod cells.