Investigative ophthalmology & visual science最新文献

{"title":"Liposome Encapsulation Enhances Ripasudil Therapeutic Efficacy Against Proliferative Vitreoretinal Diseases: Implications in Advanced Ocular Treatment.","authors":"Rui Ji, Keijiro Ishikawa, Wei Tan, Kenichiro Mori, Ryotaro Tsukamoto, Naoya Matsunaga, Kohei Kiyohara, Yosuke Fukuda, Iori Wada, Tomoyuki Isobe, Tomohito Tanihara, Yuya Yoshida, Kouta Mayanagi, Kosuke Oyama, Yuma Terada, Kaita Otsuki, Kengo Hamamura, Hiroshi Kikuchi, Shintaro Nakao, Shigeo Yoshida, Ram Kannan, Shigehiro Ohdo, Koh-Hei Sonoda","doi":"10.1167/iovs.66.6.56","DOIUrl":"10.1167/iovs.66.6.56","url":null,"abstract":"<p><strong>Purpose: </strong>Proliferative vitreoretinal diseases, such as proliferative vitreoretinopathy (PVR) and neovascular age-related macular degeneration (nAMD), pose substantial challenges in their advanced stages owing to the development of retinal fibrous membranes. Current therapeutic modalities, including surgical interventions for PVR and antivascular endothelial growth factor therapy for nAMD, cannot effectively manage intraocular fibrosis associated with epithelial-to-mesenchymal transition (EMT) in retinal pigment epithelium (RPE) cells. Through drug screening, we identified ripasudil, a Rho-kinase inhibitor, as a remarkable suppressor of RPE-EMT. However, the short vitreal half-lives of small-molecule drugs, coupled with the limited stability of ripasudil in the ocular environment, impede its application in vitreoretinal diseases. Considering the advances in nanotechnology-assisted improvement in drug stability and cellular uptake as well as controlled release, we aimed to enhance the efficacy of ripasudil through liposome encapsulation.</p><p><strong>Methods: </strong>After ripasudil encapsulation, we performed comprehensive in vivo and in vitro analyses and pharmacokinetic studies.</p><p><strong>Results: </strong>Liposome-encapsulated ripasudil (Lipo-Ripa) demonstrated a substantial reduction in subretinal fibrosis in an advanced AMD model and more effective inhibition of PVR progression in rabbits than that induced by ripasudil alone. Pharmacokinetic studies revealed that Lipo-Ripa exhibited improved retention capacity in the vitreous and retina, alongside reduced permeability through the RPE barrier and increased cellular uptake. These characteristics resulted in a sustained elevation of drug concentration within the ocular tissues over time.</p><p><strong>Conclusions: </strong>Our findings suggest that liposomal encapsulation of ripasudil supports enhanced bioavailability and effectiveness of the drug, presenting a promising innovative therapeutic approach for the treatment of proliferative vitreoretinopathy.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"56"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12178431/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144316930","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Machine Learning Identifies Key Gene Markers Related to Fetal Retina Development at Single-Cell Transcription Level.","authors":"Jiyu Zhang, Tong Du, Yiqing Jin, Yusheng Bao, Qinglan Ma, Yu-Dong Cai, Jian Zhang","doi":"10.1167/iovs.66.6.60","DOIUrl":"10.1167/iovs.66.6.60","url":null,"abstract":"<p><strong>Purpose: </strong>The retina is part of the central nervous system, and its function is vision. During the embryonic development of the fetus, it diversifies the seven major types of cells from a retinal progenitor cell. The purpose of this study is to investigate essential features that are necessary for the development of the fetal retina.</p><p><strong>Methods: </strong>We generated a comprehensive single-cell transcriptional atlas of the human retina by leveraging datasets from the Chan Zuckerberg Initiative Single-Cell Biology collection of human fetal retinas. The eight critical types of retinal cells were investigated, including amacrine, bipolar, cone, horizontal, Müller glia, retinal ganglion, rod, and retinal progenitor cells. We evaluated a total of 36,503 gene features across three developmental stages (early, middle, and late) for each cell type. Using seven feature ranking algorithms and incremental feature selection method, we identified key gene features, constructed efficient classifiers and classification rules.</p><p><strong>Results: </strong>For amacrine cells, RELN and DAB1 are critical; for bipolar cells, ANK3 and RIMS2; for cone cells, PDE6H and FRMPD2; for horizontal cells, NFIA; for Müller glial cells, WIF1 and TF; for retinal ganglion cells, IL1RAPL2 and PCP4; for rod cells, RIMS2 and NRG1; and for retinal progenitor cells, BTG1. These gene features bring into focus the pattern of gene regulation and developmental pathways of the retinal cells for deeper insights into retinal development.</p><p><strong>Conclusions: </strong>This study explored molecular features related to the development of the fetal retina and their potential roles in certain pathways, which may provide novel insights into retinal development and contribute to a better understanding of other retinal diseases.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"60"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12180603/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144325722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-Cell RNA Sequencing of Rabbit Sclera at Different Developmental Stages: Unveiling Scleral Cells Atlas and the Heterogeneity of Fibroblasts.","authors":"Fayuan Li, Chengqi Gu, Chengpeng Liang, Yang Li, Shuo Wang, Qingqing Tang, Huan Jiang, Shaorong Linghu, Tingting Dan, Rong Shi, Xin Luo, Taixiang Liu","doi":"10.1167/iovs.66.6.83","DOIUrl":"https://doi.org/10.1167/iovs.66.6.83","url":null,"abstract":"<p><strong>Purpose: </strong>This study aims to construct a single-cell transcriptomic atlas of the developing rabbit sclera to elucidate fibroblast heterogeneity, differentiation trajectories, matrisome expression patterns, and intercellular communication, while revealing conserved molecular features of scleral cells through cross-species analysis.</p><p><strong>Methods: </strong>Single-cell RNA sequencing (scRNA-seq) was performed on scleral tissues from New Zealand rabbits at embryonic day 25 (E25) and postnatal days 7 (P7), 21 (P21), and 180 (P180). Libraries were prepared using the DNBelab C Series Kit and sequenced on the BGISEQ-2000 platform. Sequencing reads were aligned to the OryCun2.0 genome using STAR, and unique molecular identifier (UMI) count matrices were generated with PISA. Data preprocessing was conducted using Seurat. Fibroblast lineage differentiation was analyzed via VIA, intercellular communication via CellChat, matrisome expression patterns via AUCell, and cross-species analyses via CACIMAR and hdWGCNA.</p><p><strong>Results: </strong>We identified 7 major cell types and 15 subpopulations, with fibroblasts dominating the cellular landscape. Distinct fibroblast subtypes exhibited varied expression profiles and functions: KERAlow SPARCL1⁺ fibroblasts showed stem/progenitor-like features, while KERAhigh myocilin (MYOC)⁺ fibroblasts displayed senescence-associated phenotypes. Matrisome analysis revealed dynamic alterations in collagen and extracellular matrix (ECM)-related genes, and intercellular communication analysis highlighted complex signaling networks, particularly the MDK/PTN pathway. Cross-species comparisons demonstrated high conservation of fibroblasts between rabbit and human sclera, identifying four conserved co-expression modules.</p><p><strong>Conclusions: </strong>This study presents the first single-cell atlas of rabbit scleral development, unveiling fibroblast heterogeneity, ECM remodeling mechanisms, and cross-species conserved features. These findings enhance our understanding of scleral biology and provide valuable insights for future research on ocular development and associated diseases, including myopia.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"83"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144505726","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interplay Between γ-Zone Peripapillary Atrophy and Optic Disc Parameters in Central Visual Field Impairment in Highly Myopic Eyes.","authors":"Nan Zhou, Takeshi Yoshida, Keigo Sugisawa, Sota Yoshimoto, Kyoko Ohno-Matsui","doi":"10.1167/iovs.66.6.74","DOIUrl":"10.1167/iovs.66.6.74","url":null,"abstract":"<p><strong>Purpose: </strong>The purpose of this study was to investigate the association between the development of glaucoma-like central visual field (VF) defects and γ-zone peripapillary atrophy (γPPA) in eyes with high myopia (HM).</p><p><strong>Methods: </strong>This retrospective study measured γPPA length, prelaminar tissue (PLT) thickness, and lamina cribrosa (LC) defects' width in HM eyes with central VF defects by swept-source optical coherence tomography (OCT). The central VF was assessed by using the Humphrey Field Analyzer 10-2 program, and average light sensitivity in the papillomacular bundle region (C1 region) and mean deviation (MD) values were calculated. Interactions among the central VF and structural abnormalities were analyzed using correlation tests and linear regression.</p><p><strong>Results: </strong>Forty-five eyes from 45 patients were included. PLT thickness was positively associated with light sensitivity in the C1 region and MD values, whereas γPPA was associated only with MD values. LC defects showed no association with either the C1 region or MD values. LC defects' width was negatively correlated with γPPA length. Stratified analysis showed that in the group with γPPA length ≥763 µm, PLT thickness was positively associated, and γPPA length was negatively associated with the C1 region and MD values. No such associations were found when the γPPA length was <763 µm. The association between LC defects with central VF remained nonsignificant.</p><p><strong>Conclusions: </strong>Our findings identified γPPA length and PLT thickness as risk factors of glaucoma-like central VF defects in HM, particularly when γPPA length exceeds 763 µm. We suggest that HM eyes with larger γPPA and thinner PLT should be paid special attention for early detection of central VF defects.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"74"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12204232/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144475179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adrenomedullin-RAMP2 System Modulates Inflammation and Tissue Repair in Experimental Autoimmune Uveitis Via T-Cell and M2 Macrophage Regulation.","authors":"Yorishige Matsuda, Megumu Tanaka, Yunlu Zhao, Shinji Kakihara, Ken Hoshiyama, Takayuki Sakurai, Akiko Kamiyoshi, Yuka Ichikawa-Shindo, Hisaka Kawate, Yan Zhang, Qianqian Guo, Peixuan Li, Jiake Li, Jun Duan, Marina Hayashi, Hideki Sanjo, Toshinori Murata, Takayuki Shindo","doi":"10.1167/iovs.66.6.12","DOIUrl":"10.1167/iovs.66.6.12","url":null,"abstract":"<p><strong>Purpose: </strong>Adrenomedullin (AM), a peptide produced by various cells, exerts diverse physiological effects and is regulated by receptor activity-modifying proteins (RAMP2 and 3). Experimental autoimmune uveitis (EAU) is a well-established model for studying human autoimmune uveitis. Hence, we investigated the pathophysiological roles of the AM-RAMP system in uveitis using an optimized EAU mouse model.</p><p><strong>Methods: </strong>Female wild-type (WT), AM knockout, RAMP2KO, and RAMP3KO mice were immunized with the human interphotoreceptor retinoid-binding protein. The expression of macrophage-related genes and inflammatory cytokines in the retina and spleen was analyzed using real-time polymerase chain reaction. EAU-induced WT mice received human recombinant AM; therapeutic effects were evaluated via clinical and histologic scores, quantification of T-cell and macrophage infiltration in the retina, and the number of splenic regulatory T cells (Tregs) and M2 macrophages using flow cytometry.</p><p><strong>Results: </strong>Compared with WT mice, EAU-induced AMKO and RAMP2KO mice had significantly increased retinal inflammatory cell infiltration and worsened clinical scores, whereas RAMP3KO mice did not. Proinflammatory cytokine expression was suppressed in the retina of EAU-induced WT mice that received AM. However, anti-inflammatory cytokine expression was upregulated compared with that in the vehicle group. Additionally, there was reduced retinal infiltration of T cells and macrophages, leading to improved clinical and histologic scores. AM administration also suppressed EAU-induced splenomegaly and increased the number of Tregs and M2 macrophages, possibly contributing to resolving inflammation.</p><p><strong>Conclusions: </strong>AM exerts an anti-inflammatory effect in uveitis by activating Tregs and M2 macrophages through RAMP2. Its administration is a potential adjunctive therapy for uveitis.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"12"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12147046/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144215819","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expansion of the ABCA4-Associated Retinopathy Spectrum: Severe Variants Can be Associated With Early-Onset Severe Retinal Dystrophy.","authors":"Daan M Panneman, Rebekkah J Hitti-Malin, Martin McKibbin, Suzanne E de Bruijn, Erica G M Boonen, Andrea L Vincent, Glenda Vargas, Jordi Corominas, Galuh Astuti, Christian Gilissen, Elfride De Baere, Chris F Inglehearn, Susanne Roosing, Robert K Koenekoop, Frans P M Cremers","doi":"10.1167/iovs.66.6.19","DOIUrl":"10.1167/iovs.66.6.19","url":null,"abstract":"<p><strong>Purpose: </strong>Leber congenital amaurosis (LCA) and early-onset severe retinal dystrophy (EOSRD) are inherited retinal diseases that are characterized by severe visual loss very early in life. Whereas LCA is characterized by loss of vision in the first year of life, nystagmus, and absent or abnormal electrical signals on electroretinogram, persons with EOSRD show onset of disease between 1 and 5 years of age, with better preserved visual acuity and some signals on an electroretinogram. We investigated the genetic cause of disease and clinical characteristics in three probands with EOSRD.</p><p><strong>Methods: </strong>All patients were examined by at least two ophthalmologists to reach a clinical diagnosis. APEX microarray screening, smMIP-based sequencing, and whole exome and whole genome sequencing were used to obtain a genetic diagnosis and investigate potential modifiers.</p><p><strong>Results: </strong>The EOSRD phenotype of these three probands was established through ophthalmological investigation. Biallelic severe ABCA4 variants were identified after the phenotypic diagnosis of EOSRD in these probands. We then asked whether additional gene defects may be involved and worsen the phenotype. Through whole genome sequencing we identified two NBAS variants in patient 1 and a well-known homozygous, hypomorphic missense variant in CNGB3 in patient 3.</p><p><strong>Conclusions: </strong>We propose that biallelic severe ABCA4 variants can be implicated in EOSRD. We hypothesize that the ABCA4 and CNGB3 variants could have an additive effect given the colocalization of the encoded proteins in cone photoreceptors cell membranes. Whether the CNGB3 and NBAS variants play a modifying role remains to be investigated.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"19"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12147048/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144215824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Myopia Correction, Myopia Control and Myopia Management: Definitions and Recommended Usage.","authors":"Ian Flitcroft, Mark A Bullimore, Kate L Gifford, Jost B Jonas, Deborah Jones, Lyndon W Jones, Pauline Kang, Serge Resnikoff, Jeffrey Walline, Christine F Wildsoet","doi":"10.1167/iovs.66.6.41","DOIUrl":"10.1167/iovs.66.6.41","url":null,"abstract":"","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"41"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12169461/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144274910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modeling the Ocular Pharmacokinetics and Pharmacodynamics of Ranibizumab for Improved Understanding and Data Collection Strategies in Ocular Diseases.","authors":"Jessica R Crawshaw, Eamonn A Gaffney, Michael Gertz, Philip K Maini, Antonello Caruso","doi":"10.1167/iovs.66.6.20","DOIUrl":"10.1167/iovs.66.6.20","url":null,"abstract":"<p><strong>Purpose: </strong>Improving our understanding of the ocular pharmacokinetics and pharmacodynamics of anti-vascular endothelial growth factor (VEGF) therapies, such as ranibizumab, is essential to enhance treatment strategies for a range of retinal diseases, and will help inform the development of novel anti-VEGF drug candidates.</p><p><strong>Methods: </strong>In this study, we examine a two-compartment pharmacokinetic/pharmacodynamic model of an intravitreal ranibizumab injection to understand its impact on ocular VEGF suppression. We use Bayesian inference to infer the model parameters from aqueous humor data extracted from healthy cynomolgus macaques. We leverage this approach to explore various sources of uncertainty in the data, offering practical recommendations for minimizing avoidable uncertainty.</p><p><strong>Results: </strong>The model provides a robust description of ranibizumab pharmacokinetics and pharmacodynamics, identifying the recovery region of the aqueous humor VEGF concentration-time profile as critical for the precise estimation of parameters. Our results advocate focusing on this region in future studies for optimal data collection. We consider standard data correction techniques to reduce the data uncertainty introduced by the lower limit of quantification, identifying the most preferable technique for this model and data. Using a Bayesian approach we obtain an inferred mean posterior distribution of 1459 ± 98 pM for the ranibizumab dissociation constant, a pharmacodynamic parameter with notable variability across the literature.</p><p><strong>Conclusions: </strong>This study extends our understanding of the ocular pharmacokinetics and pharmacodynamics of ranibizumab and provides theoretical insights for enhanced data collection schemes to be considered for clinical trials and in the development of novel anti-VEGF therapies.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"20"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12155695/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144234108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erratum in: A Novel AAV Capsid-Mediated RS1 Gene Therapy Restored Retinal Function to Wild-Type Levels in Rs1R213W Mouse Model.","authors":"","doi":"10.1167/iovs.66.6.29","DOIUrl":"10.1167/iovs.66.6.29","url":null,"abstract":"","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"29"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12161413/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144247915","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the Immunogenic Potential of Variations in Host Cell Protein Levels in Clinical-Grade AAV8 Products.","authors":"Immanuel P Seitz, Eduardo Rodríguez-Bocanegra, Kirsten Bucher, Felix F Reichel, Stylianos Michalakis, Dimitri Romanovsky, Martin Biel, Bernd Wissinger, Karl-Ulrich Bartz-Schmidt, Tobias Peters, Manuel D Fischer","doi":"10.1167/iovs.66.6.38","DOIUrl":"10.1167/iovs.66.6.38","url":null,"abstract":"<p><strong>Purpose: </strong>Adeno-associated virus (AAV) formulations for gene therapy contain manufacturing-associated impurities such as residual host cell protein (HCP). The aim of this study was to investigate whether high levels of HCP in AAV formulations are associated with increased inflammation and reduced ocular tolerability.</p><p><strong>Methods: </strong>Three lots of clinical-grade AAV8 vector were analyzed for the presence of manufacturing-associated impurities. The HCP component of these impurities was characterized using mass spectrometry. Lots were then compared regarding their capacity to induce a cytokine response in primary human plasmacytoid dendritic cells (pDCs) and THP-1 cells. Furthermore, the results of an ocular safety study in healthy nonhuman primates were analyzed post hoc to investigate the influence of HCP levels on clinical signs of inflammation and chorioretinal atrophy (CRA) development.</p><p><strong>Results: </strong>Vector lots displayed up to a ∼40-fold variation in HCP levels. Human galactin-3-binding protein was the only major HCP contaminant. Stimulation of human pDCs and THP-1 cells with a high HCP lot did not result in an increased cytokine response. High HCP also did not exacerbate clinical signs of inflammation. However, on retinal imaging, CRA lesions were significantly larger in high HCP-treated eyes (P = 0.001-0.048).</p><p><strong>Conclusions: </strong>HCP impurities were of low complexity, but pronounced variations in their abundance were observed between lots. High HCP levels were not overtly immunogenic in vivo and in vitro. However, despite statistical limitations, they seemed to be associated with increased CRA. Thus, a negative effect of high HCP levels on retinal tolerability could not be ruled out.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"38"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12166503/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144266236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}