[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-431651-105":59,"doc-detail-431651-en":130},{"code":4,"msg":5,"data":6},0,"success",[7,13,18,23,28,33,38,43,48,51,55],{"id":8,"doc_module":4,"doc_module_name":9,"category_name":10,"show_sort_weight":11,"slug":12},1,"Document","Story & Novel",90,"story-novel",{"id":14,"doc_module":4,"doc_module_name":9,"category_name":15,"show_sort_weight":16,"slug":17},2,"Literature",80,"literature",{"id":19,"doc_module":4,"doc_module_name":9,"category_name":20,"show_sort_weight":21,"slug":22},4,"Exam",70,"exam",{"id":24,"doc_module":4,"doc_module_name":9,"category_name":25,"show_sort_weight":26,"slug":27},5,"Comic",60,"comic",{"id":29,"doc_module":4,"doc_module_name":9,"category_name":30,"show_sort_weight":31,"slug":32},6,"Technology",50,"technology",{"id":34,"doc_module":4,"doc_module_name":9,"category_name":35,"show_sort_weight":36,"slug":37},7,"Healthcare",40,"healthcare",{"id":39,"doc_module":4,"doc_module_name":9,"category_name":40,"show_sort_weight":41,"slug":42},8,"Research & Report",30,"research-report",{"id":44,"doc_module":4,"doc_module_name":9,"category_name":45,"show_sort_weight":46,"slug":47},9,"Religion & Spirituality",20,"religion-spirituality",{"id":46,"doc_module":4,"doc_module_name":9,"category_name":49,"show_sort_weight":46,"slug":50},"World Cup","world-cup",{"id":52,"doc_module":4,"doc_module_name":9,"category_name":53,"show_sort_weight":52,"slug":54},10,"Lifestyle","lifestyle",{"id":56,"doc_module":4,"doc_module_name":9,"category_name":57,"show_sort_weight":24,"slug":58},19,"General","general",{"code":4,"msg":60,"data":61},"ok",{"site_id":62,"language":63,"slug":64,"title":65,"keywords":66,"description":67,"schema_data":68,"social_meta":123,"head_meta":125,"extra_data":127,"updated_unix":129},105,"en","large-scale-high-throughput-screen-for-cardiac-ryanodine-receptor-targeted-therapeutics","Large-scale high-throughput screen for cardiac ryanodine receptor targeted therapeutics","","Large-scale high-throughput screening (HTS) assays using fluorescence lifetime (FLT)-detected FRET identified small molecules that allosterically modulate the pathology-linked leaky ryanodine receptor (RyR) calcium release channel. By monitoring binding of calmodulin and the DPc10 peptide under pathological conditions, a 50,000-compound library yielded 603 reproducible FRET-altering candidates. Dose-response FRET validation narrowed this to 83 compounds, from which 10 scaffold-focused hits reduced RyR2 activity via [3H]ryanodine binding and highlighted isoxazole-group chemotypes as therapeutic by targeting the pathological RyR2 leak state in cellular and cardiomyocyte models.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/large-scale-high-throughput-screen-for-cardiac-ryanodine-receptor-targeted-therapeutics/431651/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/large-scale-high-throughput-screen-for-cardiac-ryanodine-receptor-targeted-therapeutics/431651.png","ImageObject",300,407,{"name":92,"@type":93},"Nguyễn Văn Học","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-09-30","2026-09-29",true,{"@type":102,"interactionType":103,"userInteractionCount":8},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"How does the study detect the leaky RyR2 state using FRET?","Question",{"text":112,"@type":113},"The assay monitors FRET changes tied to binding of calmodulin (CaM) and the DPc10 peptide to RyR2 under conditions mimicking pathology. The CaM–DPc10 binding relationship is used to report the functional leak state.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"What scale was screened and how were hits validated?",{"text":117,"@type":113},"A 50,000-compound chemical library was screened, producing 603 reproducible FRET-altering compounds. The most promising 83 were then purchased and validated using FRET dose-response evaluation.",{"name":119,"@type":110,"acceptedAnswer":120},"What experimental evidence supports that selected hits reduce RyR2 activity?",{"text":121,"@type":113},"Six representative compounds from scaffold-focused hits reduced RyR2 activity as measured by [3H]ryanodine binding. Cellular Ca2+ dynamics in HEK293 cells expressing human RyR2 and in cardiomyocytes further supported the therapeutic relevance of the isoxazole-group chemotypes.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},431651,1790740694,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":8,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145},1374402739827,"https://ap-avatar.wpscdn.com/avatar/14000c97e7351f1a627?x-image-process=image/resize,m_fixed,w_180,h_180&k=1787885694763230660","RESEARCH ARTICLE  \nLarge-scale high-throughput screen for cardiac ryanodine receptor targeted therapeutics  \nReceived for publication, May 29, 2025, and in revised form, October 29, 2025 Published, Papers in Press, November 17, 2025 [https://doi.org/10.1016/j.jbc.2025.1](https://doi.org/10.1016/j.jbc.2025.1)10951  \nRoman Nikolaienko 1,‡, Elisa Bovo 1,‡, Jonathan C. Solberg2,‡, Marzena Brinkmann3, Levy M. Treinen2,  \nAndrew R. Thompson2, Kaja Berg3, David D. Thomas2,4, Jennifer J. Thomas4, Donald M. Bers5,  \nCourtney C. Aldrich3, Aleksey V. Zima 1, Razvan L. Cornea2, and Robyn T. Rebbeck2, *  \nFrom the 1Department of Cell and Molecular Physiology, Loyola University Chicago, Stritch School of Medicine, Maywood, Illinois, USA; 2Department of Biochemistry, Molecular Biology, and Biophysics, and 3Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota, USA; 4Photonic Pharma LLC, Minneapolis, Minnesota, USA; 5Department of Pharmacology, University of California at Davis, Davis, California, USA  \nReviewed by members of the JBC Editorial Board. Edited by Roger Colbran  \nIn high-throughput screening (HTS) assays using fluorescence lifetime (FLT)-detected FRET, we have identified compounds that allosterically modulate the pathologically leakyryanodine receptor (RyR) calcium release channels. These compounds may prevent or reduce the elevated Ca2+ that fuels arrhythmia, heart failure, and age-related neurodegeneration. RyRs are responsible for intracellular Ca2+ release from endoplasmic/sarcoplasmic reticulum (ER/SR). The resulting [Ca2+] pulse is a signal for many cellular processes, whereas sustained elevated [Ca2+] is pathologic. Our FRET-based HTS detects the pathology-linked RyR leaky state by monitoring binding of the accessory protein calmodulin and the DPc10 peptide (corresponding to RyR2 residues 2460–2495) known to perturb interdomain interactions within RyR2. Under conditions mimicking a pathological state, we have screened a 50,000-compound chemical library to identify small-molecule modulators of RyR2 in cardiac SR membranes. This screen yielded 603 compounds that reproducibly altered FRET. Based on FRET response profiles that align with therapeutic potential, 83 of those most promising compounds were purchased and validated by FRET dose response evaluation. Focusing on ten chemical scaffolds that desirably increase A-CaM binding, six representative compounds reduced RyR2 activity as measured by [3H]ryanodine binding. Ca2+ dynamics in HEK293 cells expressing human RyR2 or in cardiomyocytes highlighted the isoxazole group of hits as potentially therapeutic by targeting the pathological RyR2 leak state.  \nControl of Ca2+ signaling is essential to healthy muscle and neuronal function. Important players in this signaling are theryanodine receptor (RyR) intracellular channels, which are the Ca2+ gatekeepers of the endoplasmic reticulum (ER) and its muscle-specialized variant, the sarcoplasmic reticulum (SR). Of the three mammalian isoforms, RyR2 is the dominant isoform expressed in the heart, and the main isoform  \n‡ These authors contributed equally to this work.  \n* For correspondence: Robyn T. Rebbeck, [rrebbeck@umn.edu](rrebbeck@umn.edu).  \nexpressed in the brain. Brain also expresses RyR1 (dominant isoform in skeletal muscle) and RyR3 (1, 2).  \nPharmaceutically, RyRs are increasingly attractive for therapeutic discovery for treating dysfunctional ER/SR Ca2+ leak, which is prevalent in age-related pathologies, including arrhythmia, heart failure, sarcopenia, Alzheimer’s disease and Huntington’s disease (3–8). Proof of principle for targeting RyR has been demonstrated by studies investigating RyR inhibitors that mitigate RyR Ca2+ leak in these pathologies (dantrolene (9–17), rycals (6, 18–20), carvedilol (21–24)). Thus, there is a strong scientific basis for using systematic screening approaches to find new agents that inhibit ER/SR Ca2+ leak. Indeed, this has proven effective for identifying new com","cbCaivspn6UB8pSp","https://ap.wps.com/l/cbCaivspn6UB8pSp","pdf",5441443,14,"English","# Introduction\n## Pathophysiology of RyR Ca2+ leak\n## Rationale for systematic RyR targeting screens\n# Methods and HTS platform\n## FLT-detected FRET assay design\n## Target engagement through CaM and DPc10 binding readouts\n# Results\n## Screening of a 50,000-compound library\n## Validation and scaffold-focused hit selection\n# Discussion\n## Therapeutic potential of identified modulators","[{\"question\":\"How does the study detect the leaky RyR2 state using FRET?\",\"answer\":\"The assay monitors FRET changes tied to binding of calmodulin (CaM) and the DPc10 peptide to RyR2 under conditions mimicking pathology. The CaM–DPc10 binding relationship is used to report the functional leak state.\"},{\"question\":\"What scale was screened and how were hits validated?\",\"answer\":\"A 50,000-compound chemical library was screened, producing 603 reproducible FRET-altering compounds. The most promising 83 were then purchased and validated using FRET dose-response evaluation.\"},{\"question\":\"What experimental evidence supports that selected hits reduce RyR2 activity?\",\"answer\":\"Six representative compounds from scaffold-focused hits reduced RyR2 activity as measured by [3H]ryanodine binding. Cellular Ca2+ dynamics in HEK293 cells expressing human RyR2 and in cardiomyocytes further supported the therapeutic relevance of the isoxazole-group chemotypes.\"}]","Large-scale high-throughput screen for cardiac ryanodine receptor targeted therapeutics | PDF",1790656211,35]