Advances in experimental medicine and biology最新文献

{"title":"Identification of Unexpected Pathomechanisms Underlying the Human Usher Syndrome.","authors":"Uwe Wolfrum, Joshua Linnert, Baran E Güler, Joshua Klein, Jacques S Fritze, Nicole Wenck, Kerstin Nagel-Wolfrum","doi":"10.1007/978-3-031-76550-6_28","DOIUrl":"https://doi.org/10.1007/978-3-031-76550-6_28","url":null,"abstract":"<p><p>Human Usher syndrome (USH) is the most common form of hereditary deaf-blindness, characterized by inner ear defects and late-onset vision loss. USH is a complex genetic disorder, clinically and genetically heterogeneous. To date, there is no treatment for the ocular phenotype of any USH subtype, as the underlying pathomechanisms of the disease in the eye are far from being understood. We aim to elucidate the function of USH proteins to gain insight into the pathomechanisms leading to the retinal phenotype in USH. Here, we focus on the USH1 proteins SANS (USH1G) and harmonin (USH1C), and the USH2C protein ADGRV1. Results from affinity capture approaches revealed putative interacting proteins to these USH proteins, indicative of diverse various unexpected molecular pathways and modules. Functional studies in both cellular and animal models confirmed the roles of SANS in the pre-mRNA splicing of other retinal genes, especially USH genes and harmonin as a suppressor of the canonical Wnt signaling. Additionally, ADGRV1 showed characteristics of a metabotropic mechanoreceptor regulating cell adhesions, Ca²⁺ homeostasis of the cell, and autophagy. The dysfunction of these pathways and processes may contribute to the development of USH and are novel potential targets for future therapies.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1468 ","pages":"171-175"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photoreceptor Disc Morphogenesis: Who Are the Conductors of This Highly Metronomic Process?","authors":"Roly Megaw","doi":"10.1007/978-3-031-76550-6_50","DOIUrl":"https://doi.org/10.1007/978-3-031-76550-6_50","url":null,"abstract":"<p><p>To ensure that normal vision is maintained, the photoreceptor must continually renew its outer segment, a massive expanse of ciliary membrane extending from the tip of its connecting cilium. The outer segment is organised into hundreds of flattened discs, the formation of which is highly regulated. Disc morphogenesis requires the metronomic assembly of an actin cytoskeletal network to initiate the necessary membrane deformation and subsequent network disassembly to allow disc completion. Disruption of disc turnover, due to human mutations, results in an inherited retinal dystrophy (IRD), a leading cause of visual loss in children and working adults. This chapter will describe the structural evidence that disc formation is actin-driven and discuss what is known of the molecular mechanisms that govern the process.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1468 ","pages":"305-308"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389821","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Precise Gene Editing Technologies in Retinal Applications.","authors":"Mehri Ahmadian, Iskalen Cansu Topcu Okan, Gokce Uyanik, Markus Tschopp, Cavit Agca","doi":"10.1007/978-3-031-76550-6_20","DOIUrl":"https://doi.org/10.1007/978-3-031-76550-6_20","url":null,"abstract":"<p><p>Gene therapy is emerging as a promising treatment for inherited retinal diseases (IRDs). One of the first successful applications of gene therapy for IRDs was the gene replacement therapy for the RPE65 mutation. This therapy delivers a functional copy of the RPE65 gene to patients via AAV vectors, rather than targeting the mutation itself. Gene editing technologies have advanced significantly in recent years, allowing it to make precise in vivo modifications to the genetic code. After the discovery of CRISPR-Cas9, other gene editing technologies such as base editing and prime editing have been developed by modifying and combining the original CRISPR-Cas9 technology with other methods. Moreover, recently discovered CRISPR-Cas systems allow RNA editing to correct mutations at the posttranscriptional level. These technologies have potential applications in various fields, including inherited retinal diseases. This mini-review evaluates and summarizes the most current advancements in genome editing methods, including prime editing, base editing, and RNA editing, and their applications on retinal diseases.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1468 ","pages":"119-123"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The PRPH2 D2 Loop: Biochemical Insights and Implications in Disease.","authors":"Larissa Ikelle, Muayyad R Al-Ubaidi, Muna I Naash","doi":"10.1007/978-3-031-76550-6_52","DOIUrl":"https://doi.org/10.1007/978-3-031-76550-6_52","url":null,"abstract":"<p><p>PRPH2 is a glycosylated tetraspanin protein that plays a crucial role in outer segment morphogenesis. The protein induces membrane curvature at the rim and orchestrates proper disc elaboration by forming a complex network of homo- and hetero-oligomers, primarily mediated through its EC2/D2 loop. This loop, a functionally and structurally essential domain shared by all tetraspanins, is divided into two subdomains responsible for diverse interactions. Pathological mutations in the EC2/D2 loop account for nearly 70% of all PRPH2 mutations, resulting in a spectrum of heterogenous disease phenotypes. In this chapter, we provide an overview of the structural elements of the EC2/D2 loop of PRPH2 and explore the cellular and biochemical consequences of known mutations.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1468 ","pages":"313-317"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Monocytes in Retinal Degeneration: Little Cells with a Big Impact.","authors":"Kaitryn E Ronning, Marie E Burns, Florian Sennlaub","doi":"10.1007/978-3-031-76550-6_22","DOIUrl":"https://doi.org/10.1007/978-3-031-76550-6_22","url":null,"abstract":"<p><p>Inflammation has been strongly implicated in retinal degenerative disorders, including inherited retinal degenerations (IRDs) and age-related macular degeneration (AMD). Microglia are the only immune cells in the retina during normal function, but during damage and disease, monocytes are able to invade the retina. Despite similarities to microglia, monocyte-derived cells (MdCs) may play a distinct and often pathogenic role in disease. Recent technological advances are rapidly improving our ability to investigate monocytic cells, yet many questions remain. Still, it is clear monocytes play an important role during retinal degenerative disorders and they are an exciting target for the development of therapeutic interventions.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1468 ","pages":"133-137"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Megabase Deletion of the Human EYS Locus Using CRISPR/Cas9.","authors":"Bruna Lopes da Costa, Anders Steen Knudsen, C Henrique Alves, Stephen H Tsang, Peter M J Quinn","doi":"10.1007/978-3-031-76550-6_18","DOIUrl":"https://doi.org/10.1007/978-3-031-76550-6_18","url":null,"abstract":"<p><p>Mutations in the Eyes Shut Homolog (EYS) gene are associated with autosomal recessive retinitis pigmentosa (arRP). To date, four retinal isoforms of EYS have been identified. However, the precise retinal function of EYS is not fully understood, but it has apparent roles in retinal morphogenesis, architecture, and ciliary transport. Clustered-regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated (Cas) nuclease-mediated approaches are powerful tools for genome engineering in mammalian cells. The use of paired CRISPR/Cas9-induced double-strand breaks (DSBs) using dual single guide RNAs (sgRNA) can lead to precise genomic deletions. In this study, we developed a dual sgRNA strategy to facilitate CRISPR/Cas9-mediated deletion of 1,988,210 bp of the EYS locus, removing the four currently identified human retinal EYS isoforms. This approach can be used to produce EYSdel induced pluripotent stem cell (iPSC) lines to explore the function of EYS in human iPSC-derived retinal organoids.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1468 ","pages":"107-111"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Desmosome and Hemidesmosome Disassembly in Retinal Pigmented Epithelium: Intersection with the Exosome Pathway.","authors":"Mikael Klingeborn, Emily D Reese","doi":"10.1007/978-3-031-76550-6_56","DOIUrl":"https://doi.org/10.1007/978-3-031-76550-6_56","url":null,"abstract":"<p><p>The retinal pigmented epithelium (RPE) forms the outer blood-retinal barrier, and like other epithelia it has several different types of cell-cell junctions, such as desmosomes. The RPE provides key metabolic and nutrient support to photoreceptors and the function of normal vision. The RPE is a principal location of disease-associated changes in age-related macular degeneration (AMD), due to its essential role in visual homeostasis. There are no robust early indicators of AMD or disease progression, a need that could be filled by the development of early AMD biomarkers. Exosomes are lipid bilayer membrane vesicles of nanometer sizes that are released via a dedicated machinery by all cells and carry out a multitude of functions related to cellular signaling and waste management. In the RPE, they are released from both the apical and basal sides, and the cargo composition reflects this polarization. We have recently shown that exosomes released from the basolateral side of RPE cells under chronic oxidative stress conditions contain desmosome and hemidesmosome proteins. Here we discuss the composition of desmosomes and hemidesmosomes in the RPE, and the potential of these exosome-associated components as biomarkers of early RPE dysfunction preceding AMD symptoms detectable in the current clinical setting. How cargo loading into basolateral exosomes is controlled in polarized epithelia such as RPE, is also discussed.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1468 ","pages":"339-343"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Autoimmune Regulator (AIRE) Gene, The Master Activator of Self-Antigen Expression in the Thymus.","authors":"Matthieu Giraud, Pärt Peterson","doi":"10.1007/978-3-031-77921-3_7","DOIUrl":"https://doi.org/10.1007/978-3-031-77921-3_7","url":null,"abstract":"<p><p>It has been more than 20 years since the AIRE gene was discovered. The mutations in the AIRE gene cause a rare and life-threatening autoimmune disease with severe manifestations against a variety of organs. Since the identification of the AIRE gene in 1997, more than two decades of investigations have revealed key insights into the role of AIRE and its mode of action. These studies have shown that AIRE uniquely induces the expression of thousands of tissue-restricted self-antigens in the thymus. These self-antigens are presented to developing T cells, resulting in the deletion of the self-reactive T cells and the generation of regulatory T cells. Thus, AIRE is a master guardian in establishing and maintaining central immune tolerance.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1471 ","pages":"199-221"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The CRISPR-Cas9 System Is Used to Edit the Autoimmune Regulator Gene in Vitro and in Vivo.","authors":"Pedro P Tanaka, Cíntia J Monteiro, Max J Duarte, Ernna D Oliveira, Ana C Monteleone-Cassiano, Romário S Mascarenhas, Mayara C Vieira Machado, Adriana A Matos, Letícia A Brito, Alina O Oliveira, Thiago M Cunha, Eduardo A Donadi, Geraldo A Passos","doi":"10.1007/978-3-031-77921-3_10","DOIUrl":"https://doi.org/10.1007/978-3-031-77921-3_10","url":null,"abstract":"<p><p>Although mutations in the AIRE gene in patients with autoimmune polyendocrine syndrome type 1 (APS-1) syndrome are associated with the onset of this autoimmune disease, much of what is known about its mechanisms has been obtained through studies with Aire mutant Mus musculus mouse model or with Aire mutant medullary thymic epithelial cells (mTEC) cultured in vitro. The in vivo murine model was soon established, and ten mutant strains are currently described. Most Aire mutant mice were obtained through homologous recombination, which generated Aire knockout (KO) animals. Nevertheless, long-term cultures of mTECs from APS-1 patients or Aire mutant mice are difficult to establish. The CRISPR-Cas9 system to edit Aire in a murine mTEC line in vitro and mouse embryo has been successfully used to overcome this. The ribonucleoprotein (RNP) complexes composed of the guide RNA (gRNA), the Cas9 enzyme, and single-stranded oligonucleotides (ssODN) were designed to target Aire exons 6 and 8 separately. The CRISPR-Cas9 makes it possible to produce NHEJ-derived indels or HDR-derived mutations. Efforts are being concentrated on using RNP complex rather than plasmid vectors, as RNP makes recurrent NHEJ-derived mutations among in vitro and in vivo editions. One recurrent mutation was described in the Aire exon 6 (del 3554G) and the other in the exon 8 (del 5676_5677TG), i.e., the exon 6 mutation was kept in an mTEC clone edited in vitro and in vivo in a mouse, and the exon 8 mutation was kept in several mTEC clones in vitro. In contrast, none of the mutations obtained with the nickase system (plasmid expression vector) were recurrent, indicating the participation of the RNP complex in recurring mutation, which offers advantages, as it does not involve recombinant plasmids and does not generate a genetically modified organism but rather a mutant animal or cell.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1471 ","pages":"269-283"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interaction of the Systemic Inflammatory State, Inflammatory Mediators, and the Oral Microbiome.","authors":"Mariana Caldas Oliveira Mattos, Amanda Vivacqua, Valeria Martins Araújo Carneiro, Daniela Correa Grisi, Maria do Carmo Machado Guimarães","doi":"10.1007/978-3-031-79146-8_8","DOIUrl":"https://doi.org/10.1007/978-3-031-79146-8_8","url":null,"abstract":"<p><p>Humans are biological units that host numerous microbial symbionts and their genomes, which together form a superorganism or holobiont. Changes in the balance of the oral ecosystem can have consequences for both general and oral health, such as cavities, gingivitis, and periodontitis. Periodontitis is initiated by a synergistic and dysbiotic microbial community that causes local inflammation and destruction of the tooth's supporting tissues, potentially leading to systemic inflammation. This inflammation caused by periodontal disease has been associated with various systemic alterations, and the immune system is largely responsible for the body's exacerbated response, which can induce and exacerbate chronic conditions. Studies indicate that subgingival microorganisms found in periodontitis reach the bloodstream and are distributed throughout the body and, therefore, can be found in distant tissues and organs. Among all diseases associated with periodontal disease, diabetes mellitus presents the strongest and most elucidated link, and its bidirectional relationship has already been demonstrated. Chronic hyperglycemia favors the worsening of periodontal parameters, while the aggravation of periodontal parameters can promote an increase in glycemic indexes. Other systemic diseases have been related to periodontitis, such as Alzheimer's, chronic kidney disease, atherosclerosis, and respiratory diseases. The importance of periodontal control may suggest a reduction in the chances of developing chronic inflammatory diseases because these two alterations often share inflammatory pathways and, for this reason, may influence each other.</p>","PeriodicalId":7270,"journal":{"name":"Advances in experimental medicine and biology","volume":"1472 ","pages":"121-132"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143668832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}