Molecular Vision最新文献

{"title":"Whole-exome sequencing identified genes known to be responsible for retinitis pigmentosa in 28 Chinese families.","authors":"Chang Shen,&nbsp;Bing You,&nbsp;Yu-Ning Chen,&nbsp;Yang Li,&nbsp;Wei Li,&nbsp;Wen-Bin Wei","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Retinitis pigmentosa (RP) is a group of highly heterogenetic inherited retinal degeneration diseases. Molecular genetic diagnosis of RP is quite challenging because of the complicated disease-causing mutation spectrum. The aim of this study was to explore the mutation spectrum in Chinese RP patients using next-generation sequencing technology and to explore the genotype-phenotype relationship.</p><p><strong>Method: </strong>In this study, a cost-effective strategy using whole-exome sequencing (WES) was employed to address the genetic diagnosis of 28 RP families in China. One to two patients and zero to two healthy relatives were sequenced in each family. All mutations in WES data that passed through the filtering procedure were searched in relation to 662 gene defects that can cause vision-associated phenotypes (including 89 RP genes in the RetNet Database). All patients visiting the outpatient department received comprehensive ophthalmic examinations.</p><p><strong>Result: </strong>Twenty-five putative pathogenic mutations of 12 genes were detected by WES and were all confirmed by Sanger sequencing in 20 (20/28, 71.4%) families, including the 12 following genes: <i>USH2A</i>, <i>CYP4V2</i>, <i>PRPF31</i>, <i>RHO</i>, <i>RP1</i>, <i>CNGA1</i>, <i>CNGB1</i>, <i>EYS</i>, <i>PRPF3</i>, <i>RP2</i>, <i>RPGR</i>, and <i>TOPORS</i>. Three families were rediagnosed as having Bietti crystalline dystrophy (BCD). <i>USH2A</i> (4/20, 20%) and <i>CYP4V2</i> (3/20, 15%) were found to be the most frequent mutated genes. Seven novel mutations were identified in this research, including mutations in <i>USH2A1, USH2A2</i>, <i>PRPF31</i>, <i>RP2</i>, <i>TOPORS</i>, <i>CNGB1</i>, and <i>RPGR.</i> Phenotype and genotype relationships in the 12 RP genes were analyzed, which revealed later disease onset and more severe visual function defects in <i>CYP4V2</i>.</p><p><strong>Conclusion: </strong>Twenty-five putative pathogenic mutations of 12 genes were detected by WES, and these were all confirmed by Sanger sequencing in 20 (20/28, 71.4%) families, including seven novel mutations. <i>USH2A</i> and <i>CYP4V2</i> were found to be the most frequent genes in this research. Phenotype and genotype relationships were revealed, and the mutation spectrum of RP in Chinese populations was expanded in this research, which may benefit future cutting-edge therapies.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":" ","pages":"96-113"},"PeriodicalIF":2.2,"publicationDate":"2022-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ef/da/mv-v28-96.PMC9239900.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40491456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comprehensive circular RNA expression profiling with associated ceRNA network in orbital venous malformation.","authors":"Jie Yu,&nbsp;Peiwei Chai,&nbsp;Yixiong Zhou,&nbsp;Renbing Jia,&nbsp;Yefei Wang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Orbital venous malformation (OVM), the most common type of vascular malformation in adults, has a great impact on both visual and cosmetic factors. Circular RNAs (circRNAs) play important roles in various ophthalmological diseases; however, little is known about their function in the pathogenesis of OVM.</p><p><strong>Methods: </strong>We obtained differentially expressed circRNAs, mRNAs, and miRNAs based on RNA sequencing of four OVM tissues and four normal orbital vascular tissues. The circRNA-mRNA coexpression network and circRNA-miRNA-mRNA and competing endogenous RNA (ceRNA) networks were constructed using miRanda software. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to identify the up- and downregulated mRNAs in the circRNA-miRNA-mRNA ceRNA network.</p><p><strong>Results: </strong>Overall, we identified 45 upregulated and 144 downregulated circRNAs, as well as 2,175 upregulated and 1,274 downregulated mRNAs and 156 upregulated and 168 downregulated miRNAs in OVM samples compared with normal orbital vascular tissues. The expression changes of mRNAs and circRNAs detected by quantitative real-time PCR (qRT-PCR) were in line with the RNA-seq results. Then, a ceRNA regulatory network was constructed with these differentially expressed circRNAs, mRNAs, and miRNAs. GO functional analysis revealed that most related biological processes involved extracellular matrix organization, positive regulation of actin nucleation, and so on, which were thought to be involved in the evolution of OVM. KEGG pathway analysis of upregulated mRNAs showed that mucin-type O-glycan biosynthesis, glycosaminoglycan degradation, and the <i>PI3K</i> (Gene ID: 5290; OMIM: 613089)-<i>AKT</i> (Gene ID: 207; OMIM: 114500) signaling pathway were all enriched in OVM samples.</p><p><strong>Conclusions: </strong>Our study provides novel insight into the regulatory mechanism of circRNAs, miRNAs, and mRNAs in the pathogenesis of OVM.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":" ","pages":"83-95"},"PeriodicalIF":2.2,"publicationDate":"2022-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/72/83/mv-v28-83.PMC9239899.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40491455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel homozygous mutations in the transcription factor <i>NRL</i> cause non-syndromic retinitis pigmentosa.","authors":"Mohammed E El-Asrag, Marta Corton, Martin McKibbin, Almudena Avila-Fernandez, Moin D Mohamed, Fiona Blanco-Kelly, Carmel Toomes, Chris F Inglehearn, Carmen Ayuso, Manir Ali","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>To describe the clinical phenotype and genetic basis of non-syndromic retinitis pigmentosa (RP) in one family and two sporadic cases with biallelic mutations in the transcription factor neural retina leucine zipper (<i>NRL)</i>.</p><p><strong>Methods: </strong>Exome sequencing was performed in one affected family member. Microsatellite genotyping was used for haplotype analysis. PCR and Sanger sequencing were used to confirm mutations in and screen other family members where they were available. The SMART tool for domain prediction helped us build the protein schematic diagram.</p><p><strong>Results: </strong>For family MM1 of Pakistani origin, whole-exome sequencing and microsatellite genotyping revealed homozygosity on chromosome 14 and identified a homozygous stop-loss mutation in <i>NRL</i>, NM_006177.5: c.713G>T, p.*238Lext57, which is predicted to add an extra 57 amino acids to the normal protein chain. The variant segregated with disease symptoms in the family. For case RP-3051 of Spanish ancestry, clinical exome sequencing focusing on the morbid genome highlighted a homozygous nonsense mutation in <i>NRL</i>, c.238C>T, p.Gln80*, as the most likely disease candidate. For case RP-1553 of Romanian ethnicity, targeted-exome sequencing of 73 RP/LCA genes identified a homozygous nonsense mutation in <i>NRL</i>, c.544C>T, p.Gln182*. The variants were either rare or absent in the gnomAD database.</p><p><strong>Conclusions: </strong><i>NRL</i> mutations predominantly cause dominant retinal disease, but there have been five published reports of mutations causing recessive disease. Here, we present three further examples of recessive RP due to <i>NRL</i> mutations. The phenotypes observed are consistent with those in the previous reports, and the observed mutation types and distribution further confirm distinct patterns for variants in <i>NRL</i> causing recessive and dominant diseases.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"48-56"},"PeriodicalIF":1.8,"publicationDate":"2022-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9122474/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142080965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PET imaging of retinal inflammation in mice exposed to blue light using [¹⁸F]-DPA-714.","authors":"Yuan Chen,&nbsp;Yixiang Zhou,&nbsp;Xue Zhu,&nbsp;Ge Yan,&nbsp;Donghui Pan,&nbsp;Lizhen Wang,&nbsp;Min Yang,&nbsp;Ke Wang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Positron emission tomography (PET) is widely used in high-precision imaging, which may provide a simple and noninvasive method for the detection of pathology and therapeutic effects. [¹⁸F]-DPA-714 is a second-generation translocator protein (TSPO) positron emission tomography radiotracer that shows great promise in a model of neuroinflammation. In this study, [¹⁸F]-DPA-714 micro-PET imaging was used to evaluate retinal inflammation in mice exposed to blue light, a well-established model of age-related macular degeneration (AMD) for molecular mechanism research and drug screening.</p><p><strong>Methods: </strong>C57BL/6J melanized mice were subjected to 10,000, 15,000, and 20,000 lux blue light for 5 days (8 h/day) to develop the retinal injury model, and the structure and function of the retina were assessed using hematoxylin-eosin (HE) staining, electroretinography (ERG), and terminal-deoxynucleotidyl transferase (TdT)-mediated nick-end labeling (TUNEL) immunostaining. Then, [¹⁸F]-DPA-714 was injected approximately 100 μCi through each tail vein, and static imaging was performed 1 h after injection. Finally, the mice eyeballs were collected for biodistribution and immune analysis.</p><p><strong>Results: </strong>The blue light exposure significantly destroyed the structure and function of the retina, and the uptake of [¹⁸F]-DPA-714 in the retinas of the mice exposed to blue light were the most significantly upregulated, which was consistent with the biodistribution data. In addition, the immunohistochemical, western blot, and immunofluorescence data showed an increase in microglial TSPO expression.</p><p><strong>Conclusions: </strong>[¹⁸F]-DPA-714 micro-PET imaging might be a good method for evaluating early inflammatory status during retinal pathology.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"507-515"},"PeriodicalIF":2.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/43/d4/mv-v28-507.PMC10115360.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9380332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The long noncoding RNA MALAT1/microRNA-598-3p axis regulates the proliferation and apoptosis of retinoblastoma cells through the PI3K/AKT pathway.","authors":"Xiaoli Lin,&nbsp;Xionggao Huang,&nbsp;Ling Wang,&nbsp;Weixian Liu","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>This study was designed to dissect the role of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in retinoblastoma (RB) and its underlying mechanism.</p><p><strong>Methods: </strong>Gain- and loss-of-function experiments were adopted to explore the effects of MALAT1 and microRNA (miR)-598-3p on the biologic behaviors of RB cells. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to assess the expression of MALAT1 and miR-598-3p in Y79 and HXO-RB44 cells. The proliferation of RB cells was determined with the cell counting kit-8 (CCK-8) assay and 5-ethynyl-2'-deoxyuridine (EdU) staining. Flow cytometry was employed for the measurement of the apoptotic rate, western blotting for examination of the expression of apoptosis-related proteins (Bax and Bcl-2) and phosphoinositide 3-kinase/protein kinase-B (PI3K/AKT) pathway-related factors (PI3K, AKT, p-PI3K, and p-AKT), and the luciferase reporter assay for assessment of the interaction between MALAT1 and miR-598-3p.</p><p><strong>Results: </strong>High expression of MALAT1 and low expression of miR-598-3p were noticed in Y79 and HXO-RB44 cells. MALAT1 upregulation or miR-598-3p downregulation facilitated RB cell proliferation and inhibited cell apoptosis, as evidenced by the increased proliferation rate and Bcl-2 expression, as well as diminished Bax expression and apoptotic rate, in the RB cells after transfection with pcDNA3.1-MALAT1 or miR-598-3p inhibitor. MALAT1 bound to and negatively regulated miR-598-3p. The PI3K/AKT pathway activation occurred with MALAT1 overexpression. MALAT1 promoted RB cell proliferation and repressed cell apoptosis by repressing miR-598-3p to activate the PI3K/AKT pathway.</p><p><strong>Conclusions: </strong>MALAT1 repressed miR-598-3p to activate the PI3K/AKT pathway, thus facilitating cell proliferation and inhibiting cell apoptosis in RB.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"269-279"},"PeriodicalIF":2.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/4e/80/mv-v28-269.PMC9514550.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10161853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural and functional analysis of SNP rs76740365 G>A in exon-3 of the alpha A-crystallin gene in lens epithelial cells.","authors":"Zhennan Zhao,&nbsp;Yang Sun,&nbsp;Qi Fan,&nbsp;Yongxiang Jiang,&nbsp;Yi Lu","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>To clarify the effect of a previously identified single nucleotide polymorphism (SNP; rs76740365 G>A) in the exon-3 of the <i>alpha A-crystallin (CRYAA)</i> gene on the properties of <i>CRYAA</i> and to investigate its function in human lens epithelial cells (HLECs).</p><p><strong>Methods: </strong>The human recombinant wild-type and mutant <i>CRYAA</i> (E156K) were constructed, and the molecular weight was measured by mass spectrometry. The structural changes induced by E156K mutation were analyzed by UV circular dichroism spectra and intrinsic tryptophan fluorescence and were predicted using Schrödinger software. The chaperone-like ability of wild-type and E156K mutant <i>CRYAA</i> was invested against the heat-induced aggregation of βL-crystallin and the DTT-induced aggregation of insulin. HLECs expressing wild-type and mutated <i>CRYAA</i> were subjected to quantitative PCR (qPCR) and western blot. Cell apoptosis was determined using flow cytometry analysis, and the expression of apoptosis-related proteins were determined using western blot.</p><p><strong>Results: </strong>The mass spectrometric detection revealed that E156K mutation had no significant effect on the apparent molecular mass of the <i>CRYAA</i> oligomeric complex. Evaluation of the structures of the <i>CRYAA</i> indicated that E156K mutation did not significantly affect the secondary structures, while causing perturbations of the tertiary structure. The mutant <i>CRYAA</i> displayed an increase in chaperone-like activity, which might be related to the increase of the surface hydrophobicity. We also predicted that E156K mutation would induce a change from negatively charged surface to positively charged, which was the possible reason for the disturbance to the surface hydrophobicity. Transfection studies of HLECs revealed that the E156K mutant induced anti-apoptotic function in HLECs, which was possibly associated with the activation of the p-AKT signal pathway and downregulation of Casepase3.</p><p><strong>Conclusions: </strong>Taken together, our results for the first time showed that E156K mutation in <i>CRYAA</i> associated with ARC resulted in enhanced chaperone-like function by inducing its surface hydrophobicity, which was directly related to the activation of its anti-apoptotic function.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"317-330"},"PeriodicalIF":2.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/85/70/mv-v28-317.PMC9603911.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10404310","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epac1 and PKA agonists inhibit ROS to reduce NLRP3 inflammasome proteins in retinal endothelial cells.","authors":"Li Liu,&nbsp;Youde Jiang,&nbsp;Jena J Steinle","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Reactive oxygen species (ROS) activate inflammatory pathways in several organs, including the retina. More recent work has shown that ROS activate the NOD-like receptor protein 3 (NLRP3) inflammasome pathway proteins. We recently showed that the exchange protein activated by cAMP 1 (Epac1) and protein kinase A (PKA) regulates NLRP3 proteins in the retina. Our goal was to determine whether Epac1 and PKA reduce ROS and NLRP3 inflammasome proteins.</p><p><strong>Methods: </strong>We used human primary retinal endothelial cells (RECs) grown in normal glucose (5 mM) and stimulated in normal glucose with hydrogen peroxide (H₂O₂) to induce ROS and measured NLRP3 pathway proteins. In some groups, we treated cells with an Epac1 or a PKA agonist in addition to H₂O₂ treatment to determine whether Epac1 and PKA reduced ROS and induced NLRP3 pathway proteins.</p><p><strong>Results: </strong>The data showed that 500 µM H₂O₂ was the optimal dose to increase ROS in RECs. In RECs treated with H₂O₂, NLRP3 pathway proteins were increased, which were significantly reduced by cotreatment with PKA or Epac1 agonists. H₂O₂ significantly increased NIMA-related kinase 7 (Nek7) and purinergic 2X7 receptor 7 (P2X7) levels, which were blocked by Epac1 and PKA agonists.</p><p><strong>Conclusions: </strong>Taken together, these data suggest that Epac1 and PKA reduce retinal inflammation through the reduced ROS-induced activation of NLRP3 pathway proteins.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"500-506"},"PeriodicalIF":2.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/3f/55/mv-v28-500.PMC10115359.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9380327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A five-year follow-up of <i>ABCA4</i> carriers showing deterioration of retinal function and increased structural changes.","authors":"Ulrika Kjellström,&nbsp;Sten Andréasson","doi":"","DOIUrl":"","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Purpose: &lt;/strong&gt;To investigate whether the reduced retinal function and morphological retinal changes previously demonstrated in &lt;i&gt;ABCA4&lt;/i&gt; carriers had remained stationary or had deteriorated over time at 5-year follow-up to further explore if carriers of an autosomal recessive trait also express a weak phenotype, although this is not expected for an autosomal recessive disorder.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Thirteen &lt;i&gt;ABCA4&lt;/i&gt; carriers from a previous study that included parents to patients with well known genetically verified &lt;i&gt;ABCA4&lt;/i&gt;-associated retinal degenerations were reexamined 5 years after the initial examination. As novel genes and new variants in already established genes are continuously reported, all subjects underwent renewed genetic testing with a next-generation sequencing (NGS) panel that included 288 genes associated with retinal dystrophies and an analysis of deep intronic mutations and copy number variations in the &lt;i&gt;ABCA4&lt;/i&gt; gene. Moreover, to evaluate any changes in retinal function and/or structure over time, clinical reassessment with Goldmann perimetry, visual acuity testing, fundus photography, fundus autofluorescence (FAF) imaging, optical coherence tomography (OCT), full-field electroretinography (ffERG), and multifocal ERG (mfERG) were performed 5 years after the initial investigation. The values of the ffERG parameters were compared between the two time points (the measurements obtained in the initial study versus the measurements at 5-year follow-up) and with the controls. The mfERG results of the carriers were compared with those of the controls.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The renewed genetic testing confirmed the previously established &lt;i&gt;ABCA4&lt;/i&gt; mutations but also revealed the hypomorph &lt;i&gt;ABCA4&lt;/i&gt; variant c.5603A&gt;T in five &lt;i&gt;ABCA4&lt;/i&gt; carriers. In three of them, the variant was found to be associated with known disease-causing alleles that always carry the c.5603A&gt;T in cis. According to recent publications, the subjects could still be considered &lt;i&gt;ABCA4&lt;/i&gt; carriers because both variants are on the same allele. In the remaining two subjects, c.5603A&gt;T could be in trans with the previously known &lt;i&gt;ABCA4&lt;/i&gt; variant, and the subjects were therefore excluded from the study since they could no longer be considered as carriers only. Statistical comparison of ffERG parameters showed significant reduction of the isolated rod, -as well as the combined rod-cone amplitudes over the five years of follow-up, but not compared with the controls. Concerning macular function, mfERG amplitudes were reduced for all rings in the carriers compared with the controls. Fundus photographs demonstrated morphological changes in 64% of the carriers, and 36% of them had further changes at follow-up. FAF images showed alterations in 55% of the carriers, with increased changes in 36% of them. Abnormalities on OCT were observed in 82% of the carriers, of whom 9% had newly found abnormalities at follow-up.","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"300-316"},"PeriodicalIF":2.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/06/b2/mv-v28-300.PMC9603925.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10412964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TNFAIP3 is anti-inflammatory in the retinal vasculature.","authors":"Li Liu,&nbsp;Youde Jiang,&nbsp;Jena J Steinle","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>To determine whether tumor necrosis factor alpha-induced protein 3 (TNFAIP3) regulates inflammatory and permeability proteins in the retinal vasculature.</p><p><strong>Methods: </strong>We used retinal lysates from type 1 diabetic mice and endothelial cell-specific exchange protein for cAMP 1 (Epac1) knockout mice to determine the protein levels of TNFAIP3. We also treated retinal endothelial cells (RECs) in normal (5 mM) and high (25 mM) glucose with an Epac1 agonist or with TNFAIP3 siRNA. We performed western blotting for TNFAIP3 and inflammatory and permeability proteins after treatment. TNFAIP3 siRNA was used only in cells grown in high glucose. Immunostaining was performed for localization of ZO-1 and tight junction protein 1.</p><p><strong>Results: </strong>TNFAIP3 was reduced in the diabetic retinas and the retinas of the Epac1 conditional knockout mice. The Epac1 agonist increased TNFAIP3 levels in RECs grown in high glucose. Reduction of TNFAIP3 with siRNA led to increased levels of tumor necrosis factor alpha (TNFα) and phosphorylation of nuclear factor kappa beta (NF-kB), while decreasing occludin and zonula occludens 1 (ZO-1) protein levels and inhibitory kappa beta kinase (IkB) phosphorylation. Tumor receptor-associated factor 6 (TRAF6) levels were increased above high glucose levels.</p><p><strong>Conclusions: </strong>TNFAIP3 serves as an anti-inflammatory factor in the retinal vasculature. Epac1 regulates TNFAIP3. TNFAIP3 may offer a new mechanism for regulating inflammation and permeability in the retinal vasculature.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"124-129"},"PeriodicalIF":2.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/62/f6/mv-v28-124.PMC9352365.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9703811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oxidative stress alters transcript localization of disease-associated genes in the retinal pigment epithelium.","authors":"Tadeusz J Kaczynski,&nbsp;Elizabeth D Au,&nbsp;Michael H Farkas","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Nuclear retention is a mechanism whereby excess RNA transcripts are stored in the event that a cell needs to quickly respond to a stimulus; maintaining proper nuclear-to-cytoplasmic balance is important for cellular homeostasis and cell function. There are many mechanisms that are employed to determine whether to retain a transcript or export it to the cytoplasm, although the extent to which tissue or cell type, internal and external stressors, and disease pathogenesis affect this process is not yet clear. As the most biochemically active tissue in the body, the retina must mitigate endogenous and exogenous stressors to maintain cell health and tissue function. Oxidative stress, believed to contribute to the pathogenesis or progression of age-related macular degeneration (AMD) and inherited retinal dystrophies (IRDs), is produced both internally from biochemical processes as well as externally from environmental insult. Here, we evaluate the effect of oxidative stress on transcript localization in the retinal pigment epithelium (RPE), with specific focus on transcripts related to RPE function and disease.</p><p><strong>Methods: </strong>We performed poly(A) RNA sequencing on nuclear and cytoplasmic fractions from human induced pluripotent stem cell-derived retinal pigment epithelium (iPSC-RPE) cells exposed to hydrogen peroxide (H₂O₂), as well as on untreated controls.</p><p><strong>Results: </strong>Under normal conditions, the number of mRNA transcripts retained in the nucleus exceeded that found in studies on other tissues. Further, the nuclear-to-cytoplasmic ratio of transcripts was altered following oxidative stress, as was the retention of genes associated with AMD and IRDs, as well as those that are important for RPE physiology.</p><p><strong>Conclusions: </strong>These results provide a localization catalog of all expressed mRNA in iPSC-RPE under normal conditions and after exposure to H₂O₂, shedding light on the extent to which H₂O₂ alters transcript localization and potentially offering insight into one mechanism through which oxidative stress may contribute to the progression of visual disorders.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"28 ","pages":"340-351"},"PeriodicalIF":2.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e2/fe/mv-v28-340.PMC9603899.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10404308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}