Pub Date : 2025-11-20DOI: 10.1016/j.chembiol.2025.10.006
Rachel P.M. Abrams , Rebecca G. Donahue , Jessica Ma , Ying Mao , Morgan E. Diolaiti , Alan Ashworth
Dysregulation of cysteine-dependent processes is implicated in many diseases, including cancer. Despite the importance of cysteine in crucial cellular functions, including protein synthesis, redox balance, and glutathione production, a lack of efficient assays to measure cellular cysteine has limited efforts to identify agents that affect physiological cysteine levels. We employed circular permutation to engineer a fluorescent sensor that changes conformation upon cysteine binding. Biochemical experiments showed that this sensor is selective for cysteine, operating in the 10 μM–10 mM range. To demonstrate the sensor’s applicability, we performed high-throughput screens for compounds that reduce cellular cysteine. Liquid chromatography of cell extracts validated the effect of two hit compounds, and mechanistic investigations showed that one was dependent on the anticancer target, xCT. Future application of this sensor in cell biology and drug discovery will advance understanding of cysteine metabolism and drive the development of therapeutics that restore cysteine homeostasis.
{"title":"An engineered cysteine sensor optimized for high-throughput screening identifies regulators of intracellular thiol levels","authors":"Rachel P.M. Abrams , Rebecca G. Donahue , Jessica Ma , Ying Mao , Morgan E. Diolaiti , Alan Ashworth","doi":"10.1016/j.chembiol.2025.10.006","DOIUrl":"10.1016/j.chembiol.2025.10.006","url":null,"abstract":"<div><div>Dysregulation of cysteine-dependent processes is implicated in many diseases, including cancer. Despite the importance of cysteine in crucial cellular functions, including protein synthesis, redox balance, and glutathione production, a lack of efficient assays to measure cellular cysteine has limited efforts to identify agents that affect physiological cysteine levels. We employed circular permutation to engineer a fluorescent sensor that changes conformation upon cysteine binding. Biochemical experiments showed that this sensor is selective for cysteine, operating in the 10 μM–10 mM range. To demonstrate the sensor’s applicability, we performed high-throughput screens for compounds that reduce cellular cysteine. Liquid chromatography of cell extracts validated the effect of two hit compounds, and mechanistic investigations showed that one was dependent on the anticancer target, xCT. Future application of this sensor in cell biology and drug discovery will advance understanding of cysteine metabolism and drive the development of therapeutics that restore cysteine homeostasis.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 11","pages":"Pages 1381-1396.e8"},"PeriodicalIF":7.2,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145477871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-20DOI: 10.1016/j.chembiol.2025.10.012
Dasa He , Guohong Hu
Aberrant Wnt signaling activation occurs in various cancers but has limited druggable targets. In this issue of Cell Chemical Biology, He et al.1 established a double death trap Wnt reporter system. Combined with genome-wide CRISPR screening, this approach identified STT3A as an essential Wnt signaling regulator with therapeutic potential.
{"title":"STT3A: Finding the sugar in Wnt signaling","authors":"Dasa He , Guohong Hu","doi":"10.1016/j.chembiol.2025.10.012","DOIUrl":"10.1016/j.chembiol.2025.10.012","url":null,"abstract":"<div><div>Aberrant Wnt signaling activation occurs in various cancers but has limited druggable targets. In this issue of <em>Cell Chemical Biology</em>, He et al.<span><span><sup>1</sup></span></span> established a double death trap Wnt reporter system. Combined with genome-wide CRISPR screening, this approach identified STT3A as an essential Wnt signaling regulator with therapeutic potential.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 11","pages":"Pages 1308-1309"},"PeriodicalIF":7.2,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145554789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-20DOI: 10.1016/j.chembiol.2025.10.001
Zhengjin He , Shishuang Chen , Jinlong Suo , Kai Xia , Mingxian Liu , Jingchuan Ma , Yankai Chu , Chao Wang , Yueru Xie , Wei Jiang , Hui Du , Shiyang Chen , Zhilei Zhou , Man Li , Qing Wei , Yun Zhao , Jianfeng Chen , Lin Li , Yi Zeng , Weiguo Zou , Hai Jiang
Abnormalities in the Wnt pathway are major drivers of cancer. RNF43 loss-of-function mutations are frequently detected in aggressive cancers lacking targeted therapies, underscoring the need to uncover key regulators and targets of this pathway. Using a double death trap (DDT) Wnt reporter and genome-wide CRISPR screen, we identified STT3A as an essential regulator of Wnt signaling. Genetic and pharmacological inhibition of STT3A suppressed aberrant Wnt activity caused by RNF43/ZNRF3 loss. Importantly, suppression of STT3A blocked the growth of RNF43-deficient cancer cell lines, patient-derived organoids, and spontaneous tumors. Mechanistically, STT3A regulates Wnt/β-catenin signaling via LRP6, but not LRP5. Glycosylation of LRP6 by STT3A is required for Wnt ligand binding. Notably, STT3A depletion displayed milder effects on bone homeostasis, as supported by phenotypes in STT3A-deficient patients. Together, this study established STT3A as a critical Wnt regulator through LRP6 glycosylation and a therapeutic target for RNF43-deficient cancers.
{"title":"STT3A is essential for Wnt signaling and represents a target for cancers driven by RNF43 deficiency","authors":"Zhengjin He , Shishuang Chen , Jinlong Suo , Kai Xia , Mingxian Liu , Jingchuan Ma , Yankai Chu , Chao Wang , Yueru Xie , Wei Jiang , Hui Du , Shiyang Chen , Zhilei Zhou , Man Li , Qing Wei , Yun Zhao , Jianfeng Chen , Lin Li , Yi Zeng , Weiguo Zou , Hai Jiang","doi":"10.1016/j.chembiol.2025.10.001","DOIUrl":"10.1016/j.chembiol.2025.10.001","url":null,"abstract":"<div><div>Abnormalities in the Wnt pathway are major drivers of cancer. RNF43 loss-of-function mutations are frequently detected in aggressive cancers lacking targeted therapies, underscoring the need to uncover key regulators and targets of this pathway. Using a double death trap (DDT) Wnt reporter and genome-wide CRISPR screen, we identified STT3A as an essential regulator of Wnt signaling. Genetic and pharmacological inhibition of STT3A suppressed aberrant Wnt activity caused by RNF43/ZNRF3 loss. Importantly, suppression of STT3A blocked the growth of RNF43-deficient cancer cell lines, patient-derived organoids, and spontaneous tumors. Mechanistically, STT3A regulates Wnt/β-catenin signaling via LRP6, but not LRP5. Glycosylation of LRP6 by STT3A is required for Wnt ligand binding. Notably, STT3A depletion displayed milder effects on bone homeostasis, as supported by phenotypes in STT3A-deficient patients. Together, this study established STT3A as a critical Wnt regulator through LRP6 glycosylation and a therapeutic target for RNF43-deficient cancers.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 11","pages":"Pages 1321-1335.e6"},"PeriodicalIF":7.2,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145353172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-20DOI: 10.1016/j.chembiol.2025.10.003
Yi Yuan , Zhimin Xu , Chenfang Si , Zan Gao , Xiaosong Liu , Yamei Yuan , Daichao Xu , Bing Shan , Yaoyang Zhang
Stress granules (SGs) are stress-induced membraneless organelles whose dynamics are tightly regulated by protein interactions and modifications. However, whether SUMOylation directly targets SG core proteins G3BP1/2 and which ligase is involved remains unclear, partly due to their transient and membraneless nature. To investigate this SUMOylation and its ligase, we applied our low-concentration formaldehyde crosslinking (lcFAX) method to stabilize SGs and enhance analysis. Using lcFAX-MS, we identified TRIM28 as a previously undefined SG-associated protein and showed that it SUMOylates G3BP1 at K287 and G3BP2 at K281, establishing a critical mechanism regulating SG dynamics that ultimately impacts cellular ROS and apoptosis. In addition, lcFAX-seq provides insights into SG RNA composition. Altogether, our study uncovers an essential role for TRIM28-mediated SUMOylation in modulating SG dynamics. TRIM28 may act as a versatile regulator, and with the aid of lcFAX, this mechanism could be further explored across diverse membraneless organelles and regulatory pathways.
{"title":"TRIM28-mediated SUMOylation of G3BP1/2 regulates stress granule dynamics","authors":"Yi Yuan , Zhimin Xu , Chenfang Si , Zan Gao , Xiaosong Liu , Yamei Yuan , Daichao Xu , Bing Shan , Yaoyang Zhang","doi":"10.1016/j.chembiol.2025.10.003","DOIUrl":"10.1016/j.chembiol.2025.10.003","url":null,"abstract":"<div><div>Stress granules (SGs) are stress-induced membraneless organelles whose dynamics are tightly regulated by protein interactions and modifications. However, whether SUMOylation directly targets SG core proteins G3BP1/2 and which ligase is involved remains unclear, partly due to their transient and membraneless nature. To investigate this SUMOylation and its ligase, we applied our low-concentration formaldehyde crosslinking (lcFAX) method to stabilize SGs and enhance analysis. Using lcFAX-MS, we identified TRIM28 as a previously undefined SG-associated protein and showed that it SUMOylates G3BP1 at K287 and G3BP2 at K281, establishing a critical mechanism regulating SG dynamics that ultimately impacts cellular ROS and apoptosis. In addition, lcFAX-seq provides insights into SG RNA composition. Altogether, our study uncovers an essential role for TRIM28-mediated SUMOylation in modulating SG dynamics. TRIM28 may act as a versatile regulator, and with the aid of lcFAX, this mechanism could be further explored across diverse membraneless organelles and regulatory pathways.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 11","pages":"Pages 1336-1352.e6"},"PeriodicalIF":7.2,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145442046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-20DOI: 10.1016/j.chembiol.2025.10.004
Bin Wang , Shouqing Luo , Pengkai Sun
Mutant isocitrate dehydrogenases (IDH1/IDH2) catalyze the conversion of α-ketoglutarate (αKG) to D-2-hydroxyglutarate (D2HG), a hallmark of many lower-grade gliomas. Elevated D2HG levels promote tumorigenesis through epigenetic reprogramming and immunosuppressive mechanisms, although paradoxically, D2HG can also inhibit tumor growth. To explore D2HG’s biological functions, we developed genetically encoded D2HG biosensors (DHsers) based on the prokaryotic transcriptional regulator DhdR. Structural analysis of DhdR, including its apo form, D2HG-bound complex, and DNA-bound complex, revealed that D2HG binding induces DhdR conformational changes that regulate DNA interaction. Leveraging these insights, we engineered biosensors (DHsers) that detect a wide range of concentrations of D2HG (0.3–30 mM) with high sensitivity. We also established a standardized protocol for quantifying subcellular D2HG levels in living cells. Notably, STING activation promotes D2HG production, suggesting a role of D2HG in immune modulation. Our findings reveal D2HG-induced transcriptional regulation in prokaryotes, offering a platform for studying the role of D2HG in cellular metabolism and tumorigenesis.
{"title":"Development of D2HG biosensors inspired by the molecular mechanism of D2HG regulation of DhdR","authors":"Bin Wang , Shouqing Luo , Pengkai Sun","doi":"10.1016/j.chembiol.2025.10.004","DOIUrl":"10.1016/j.chembiol.2025.10.004","url":null,"abstract":"<div><div>Mutant isocitrate dehydrogenases (IDH1/IDH2) catalyze the conversion of α-ketoglutarate (αKG) to D-2-hydroxyglutarate (D2HG), a hallmark of many lower-grade gliomas. Elevated D2HG levels promote tumorigenesis through epigenetic reprogramming and immunosuppressive mechanisms, although paradoxically, D2HG can also inhibit tumor growth. To explore D2HG’s biological functions, we developed genetically encoded D2HG biosensors (DHsers) based on the prokaryotic transcriptional regulator DhdR. Structural analysis of DhdR, including its apo form, D2HG-bound complex, and DNA-bound complex, revealed that D2HG binding induces DhdR conformational changes that regulate DNA interaction. Leveraging these insights, we engineered biosensors (DHsers) that detect a wide range of concentrations of D2HG (0.3–30 mM) with high sensitivity. We also established a standardized protocol for quantifying subcellular D2HG levels in living cells. Notably, STING activation promotes D2HG production, suggesting a role of D2HG in immune modulation. Our findings reveal D2HG-induced transcriptional regulation in prokaryotes, offering a platform for studying the role of D2HG in cellular metabolism and tumorigenesis.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 11","pages":"Pages 1397-1411.e7"},"PeriodicalIF":7.2,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145447868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-16DOI: 10.1016/j.chembiol.2025.09.007
Selina Bopp , Lọla Fagbami , Amy Deik , Claudia Taccheri , Akansha Pant , Madeline Luth , Daisy Chen , Mark A. Tye , Imran Ullah , Johannes Kreuzer , Robert Morris , Wilhelm Haas , Elizabeth A. Winzeler , Clary Clish , Amanda K. Lukens , Ralph Mazitschek , Dyann F. Wirth
Plasmodium falciparum evades the antimalarial activity of proline-competitive prolyl-tRNA synthetase (PfProRS) inhibitors, such as halofuginone (HFG), by a resistance mechanism termed the adaptive proline response (APR). The APR is characterized by a marked elevation of intracellular proline following drug exposure. Contrary to initial expectations, the APR is not mediated by alterations in canonical proline metabolic pathways involving arginase (P. falciparum arginase [PfARG]) and ornithine aminotransferase (P. falciparum ornithine aminotransferase [PfOAT]). Instead, we identified loss-of-function mutations in the apicomplexan amino acid transporter 2 (P. falciparum apicomplexan amino acid transporter 2 [PfApiAT2]) as the primary genetic driver of this resistance phenotype. Importantly, reversion of these mutations to wild type effectively suppresses the APR, establishing PfApiAT2 as the molecular determinant of this resistance mechanism.
The elucidation of the APR significantly advances our understanding of antimalarial drug resistance. By delineating the role of PfApiAT2 in this process, we establish critical insights for the development of strategies to circumvent PfProRS inhibitor resistance for future antimalarial therapies.
{"title":"Disruption of P. falciparum amino acid transporter elevates intracellular proline and induces resistance to Prolyl-tRNA synthetase inhibitors","authors":"Selina Bopp , Lọla Fagbami , Amy Deik , Claudia Taccheri , Akansha Pant , Madeline Luth , Daisy Chen , Mark A. Tye , Imran Ullah , Johannes Kreuzer , Robert Morris , Wilhelm Haas , Elizabeth A. Winzeler , Clary Clish , Amanda K. Lukens , Ralph Mazitschek , Dyann F. Wirth","doi":"10.1016/j.chembiol.2025.09.007","DOIUrl":"10.1016/j.chembiol.2025.09.007","url":null,"abstract":"<div><div><em>Plasmodium falciparum</em> evades the antimalarial activity of proline-competitive prolyl-tRNA synthetase (PfProRS) inhibitors, such as halofuginone (HFG), by a resistance mechanism termed the adaptive proline response (APR). The APR is characterized by a marked elevation of intracellular proline following drug exposure. Contrary to initial expectations, the APR is not mediated by alterations in canonical proline metabolic pathways involving arginase (<em>P. falciparum</em> arginase [PfARG]) and ornithine aminotransferase (<em>P. falciparum</em> ornithine aminotransferase [PfOAT]). Instead, we identified loss-of-function mutations in the apicomplexan amino acid transporter 2 (<em>P. falciparum</em> apicomplexan amino acid transporter 2 [PfApiAT2]) as the primary genetic driver of this resistance phenotype. Importantly, reversion of these mutations to wild type effectively suppresses the APR, establishing PfApiAT2 as the molecular determinant of this resistance mechanism.</div><div>The elucidation of the APR significantly advances our understanding of antimalarial drug resistance. By delineating the role of PfApiAT2 in this process, we establish critical insights for the development of strategies to circumvent PfProRS inhibitor resistance for future antimalarial therapies.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 10","pages":"Pages 1293-1302.e5"},"PeriodicalIF":7.2,"publicationDate":"2025-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145241395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-16DOI: 10.1016/j.chembiol.2025.09.002
Mohamed A. Anany , Daniela Siegmund , Olena Zaitseva , Isabell Lang , Bayan Mouhandes , Mervat Salah , Daniela Weisenberger , Svetlana Stepanzow , Juan Gamboa Vargas , Julia Dahlhoff , Theresa Schneider , Giel Tanghe , Frederik Stevenaert , Tugsan Tezil , Tom Van Belle , Bipasa Kar , Thomas Dandekar , Heike Margarete Hermanns , Luc van Rompaey , Andreas Beilhack , Harald Wajant
We identified several TNFR2-specific nanobodies (Nbs). When formatted as dimeric Fc fusion proteins, these nanobodies exhibited no agonistic activity. To improve activity, we genetically fused one, two, or three copies of the TNFR2-specific Nb:188 to the constant regions of an IgG1 antibody, an Fab fragment, an Fc domain or the trimerization domain of tenascin-C resulting in constructs with 2–12 Nb:188 domains. Constructs with 2 or 3 Nb:188 domains displayed no or minimal activity, while those with 4 and 5 Nb:188 domains demonstrated moderate activity. However, constructs with 6 or more Nb:188 domains exhibited potent agonism, reaching half-maximal TNFR2 activation at concentrations in the low picomolar range. Similarly, hexameric constructs generated with other TNFR2-specific Nb domains demonstrated robust agonism, too. Benchmarking against various other ligand- and antibody-based TNFR2 agonists revealed that the hexameric 3xNb:188-Fc format displays superior specific activity and efficiently expands regulatory T cells (Tregs).
{"title":"Valence and avidity determine the agonistic activity of anti-TNFR2 nanobody fusion proteins","authors":"Mohamed A. Anany , Daniela Siegmund , Olena Zaitseva , Isabell Lang , Bayan Mouhandes , Mervat Salah , Daniela Weisenberger , Svetlana Stepanzow , Juan Gamboa Vargas , Julia Dahlhoff , Theresa Schneider , Giel Tanghe , Frederik Stevenaert , Tugsan Tezil , Tom Van Belle , Bipasa Kar , Thomas Dandekar , Heike Margarete Hermanns , Luc van Rompaey , Andreas Beilhack , Harald Wajant","doi":"10.1016/j.chembiol.2025.09.002","DOIUrl":"10.1016/j.chembiol.2025.09.002","url":null,"abstract":"<div><div>We identified several TNFR2-specific nanobodies (Nbs). When formatted as dimeric Fc fusion proteins, these nanobodies exhibited no agonistic activity. To improve activity, we genetically fused one, two, or three copies of the TNFR2-specific Nb:188 to the constant regions of an IgG1 antibody, an Fab fragment, an Fc domain or the trimerization domain of tenascin-C resulting in constructs with 2–12 Nb:188 domains. Constructs with 2 or 3 Nb:188 domains displayed no or minimal activity, while those with 4 and 5 Nb:188 domains demonstrated moderate activity. However, constructs with 6 or more Nb:188 domains exhibited potent agonism, reaching half-maximal TNFR2 activation at concentrations in the low picomolar range. Similarly, hexameric constructs generated with other TNFR2-specific Nb domains demonstrated robust agonism, too. Benchmarking against various other ligand- and antibody-based TNFR2 agonists revealed that the hexameric 3xNb:188-Fc format displays superior specific activity and efficiently expands regulatory T cells (Tregs).</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 10","pages":"Pages 1279-1292.e6"},"PeriodicalIF":7.2,"publicationDate":"2025-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145183027","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-16DOI: 10.1016/j.chembiol.2025.09.006
Louis P. Conway , Michelle A. Estrada , Weichao Li , Stephen Walker , Benjamin Mielich-Süss , Anurupa Shrestha , Matthew Townsend , Jürgen Korffmann , Greg Potts , Janice Lee , Kenneth P. Robinson , Shiyao Wang , Brian Bierie , John R. Koenig , Phil Cox , Paul Richardson , Manisha Jhala , Becca McCloud , Sujatha Gopalakrishnan , Kevin Woller , Christopher G. Parker
The improper folding and aggregation of tau are linked to several neurodegenerative diseases affecting millions worldwide. However, the pathogenesis of tauopathies remains poorly understood, resulting in limited effective treatments. Here, we employ an integrated chemoproteomic phenotypic strategy to identify druggable targets and corresponding chemical probes for the treatment of tauopathies. We identified and optimized an indole-amine compound that potently and extensively clears tau aggregates in two human tauopathy models. Mechanistic and chemoproteomic studies implicate protein disulfide isomerase 1 (P4HB) as the primary target, forming covalent adducts upon metabolic activation. Knockdown of P4HB reduced tau aggregates in three tauopathy models, including an ex vivo murine neuron preclinical model. Functional characterization revealed the compound induces mild endoplasmic reticulum (ER)-stress responses as assessed by RNA sequencing and whole proteomic profiling. Our findings highlight P4HB as a potential therapeutic target for treatment of tauopathies.
{"title":"Discovery of a tau-aggregate clearing compound that covalently targets P4HB","authors":"Louis P. Conway , Michelle A. Estrada , Weichao Li , Stephen Walker , Benjamin Mielich-Süss , Anurupa Shrestha , Matthew Townsend , Jürgen Korffmann , Greg Potts , Janice Lee , Kenneth P. Robinson , Shiyao Wang , Brian Bierie , John R. Koenig , Phil Cox , Paul Richardson , Manisha Jhala , Becca McCloud , Sujatha Gopalakrishnan , Kevin Woller , Christopher G. Parker","doi":"10.1016/j.chembiol.2025.09.006","DOIUrl":"10.1016/j.chembiol.2025.09.006","url":null,"abstract":"<div><div>The improper folding and aggregation of tau are linked to several neurodegenerative diseases affecting millions worldwide. However, the pathogenesis of tauopathies remains poorly understood, resulting in limited effective treatments. Here, we employ an integrated chemoproteomic phenotypic strategy to identify druggable targets and corresponding chemical probes for the treatment of tauopathies. We identified and optimized an indole-amine compound that potently and extensively clears tau aggregates in two human tauopathy models. Mechanistic and chemoproteomic studies implicate protein disulfide isomerase 1 (P4HB) as the primary target, forming covalent adducts upon metabolic activation. Knockdown of P4HB reduced tau aggregates in three tauopathy models, including an <em>ex vivo</em> murine neuron preclinical model. Functional characterization revealed the compound induces mild endoplasmic reticulum (ER)-stress responses as assessed by RNA sequencing and whole proteomic profiling. Our findings highlight P4HB as a potential therapeutic target for treatment of tauopathies.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 10","pages":"Pages 1235-1248.e34"},"PeriodicalIF":7.2,"publicationDate":"2025-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145229399","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-16DOI: 10.1016/j.chembiol.2025.09.009
José Cubillán-Marín , Tim W. Gilberger
In this issue of Cell Chemical Biology, Bopp et al.1 discover that malaria parasite resistance to halofuginone is mediated by mutations in PfApiAT2, an amino acid transporter, rather than halofuginone’s target prolyl-tRNA synthetase. This rapid and distinctive resistance mechanism highlights amino acid transport as a promising avenue for drug discovery.
{"title":"New twists in anti-malarial drug resistance","authors":"José Cubillán-Marín , Tim W. Gilberger","doi":"10.1016/j.chembiol.2025.09.009","DOIUrl":"10.1016/j.chembiol.2025.09.009","url":null,"abstract":"<div><div>In this issue of <em>Cell Chemical Biology</em>, Bopp et al.<span><span><sup>1</sup></span></span> discover that malaria parasite resistance to halofuginone is mediated by mutations in <em>Pf</em>ApiAT2, an amino acid transporter, rather than halofuginone’s target prolyl-tRNA synthetase. This rapid and distinctive resistance mechanism highlights amino acid transport as a promising avenue for drug discovery.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 10","pages":"Pages 1199-1201"},"PeriodicalIF":7.2,"publicationDate":"2025-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145295968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-16DOI: 10.1016/j.chembiol.2025.09.004
Gregory G. Theophall , Michaele B. Manigrasso , Parastou Nazarian , Aaron Premo , Sergey Reverdatto , Gautham Yepuri , David S. Burz , Sally M. Vanegas , Kaamashri Mangar , Yanan Zhao , Huilin Li , Robert J. DeVita , Ravichandran Ramasamy , Ann Marie Schmidt , Alexander Shekhtman
RAGE and its intracellular effector molecule, the actin polymerase DIAPH1, mediate inflammation and the complications of diabetes. Using NMR spectroscopy and mass spectrometry, we built a structural model of the RAGE-DIAPH1 complex, revealing how binding of the cytoplasmic tail of RAGE (ctRAGE) to DIAPH1 stimulates its actin polymerization activity, which is inhibited by a small molecule antagonist of RAGE-DIAPH1 interaction, RAGE406R. The solution structure of the RAGE406R - ctRAGE suggests that RAGE406R prevents the formation of the RAGE-DIAPH1. FRET, actin polymerization assays, smooth muscle cell migration, and THP1 cell inflammation experiments, together with the in vivo interrogation of the effects of RAGE406R in mouse models of inflammation and diabetic wound healing, support this mode of RAGE-DIAPH1 antagonism. Finally, the treatment of macrophages differentiated from peripheral blood-derived mononuclear cells from humans with type 1 diabetes with RAGE406R reduces the mRNA expression of the chemokine CCL2, diminishing the expression of a key node in the inflammatory response.
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