Pub Date : 2025-12-17DOI: 10.1126/scitranslmed.adw9446
Ankit Uniyal, Niyada Hin, Chi Zhang, Qian Xu, Qian Huang, Ilyas Berhane, Ajit G. Thomas, John Maragakis, Sadakatali Gori, Jing Liu, Qin Zheng, Xiang Cui, Qi Peng, Jieru Wan, Rana Rais, Barbara S. Slusher, Srinivasa. N. Raja, Xinzhong Dong, Takashi Tsukamoto, Yun Guan
The human Mas-related G protein–coupled receptor X1 (MrgprX1) represents a promising nonopioid analgesic target because of its selective expression in primary nociceptive sensory neurons. Positive allosteric modulators (PAMs) promote receptor signaling, depending on the availability of endogenous ligands, offering physiological selectivity over orthosteric agonists. We developed an orally bioavailable MrgprX1 PAM, 6-tert-butyl-5-(4-chlorophenyl)-4-(2-fluoro-6-(trifluoromethoxy)phenoxy)thieno[2,3-d]pyrimidine (BCFTP). BCFTP selectively potentiated the functional response of MrgprX1 in HEK293 cells, was metabolically stable, and demonstrated a favorable in vitro safety profile. BCFTP was orally bioavailable and distributed into the spinal cords of wild-type mice. BAM22, an endogenous ligand for MrgprX1, was up-regulated in the spinal cord after nerve injury in both wild-type and humanized MrgprX1 mice and was expressed in peptidergic and nonpeptidergic dorsal root ganglion neurons. Oral administration of BCFTP dose-dependently inhibited heat hyperalgesia and spontaneous pain-like behavior but not mechanical hypersensitivity after sciatic chronic constrictive injury (CCI) in MrgprX1 mice. BCFTP did not have analgesic effects in Mrgpr cluster knockout (Mrgpr−/−) mice, indicating that the analgesic effects in MrgprX1 mice were MrgprX1 dependent. BCFTP enhanced BAM8-22–induced, MrgprX1-mediated reduction of C-fiber eEPSC amplitudes in spinal lamina II neurons, indicating inhibition of spinal nociceptive synaptic transmission. BCFTP did not induce tolerance or side effects, such as itch, sedation, and motor incoordination, and had no rewarding properties. The mRNAs encoding MrgprX1 and μ-opioid receptors were colocalized in human DRG neurons, and BCFTP synergistically enhanced morphine analgesia in CCI MrgprX1 mice. Our research suggests an approach for developing safer, orally bioavailable MrgprX1 PAM as a nonopioid therapy for neuropathic pain.
{"title":"An orally bioavailable MrgprX1-positive allosteric modulator alleviates certain neuropathic pain–related behaviors in humanized mice","authors":"Ankit Uniyal, Niyada Hin, Chi Zhang, Qian Xu, Qian Huang, Ilyas Berhane, Ajit G. Thomas, John Maragakis, Sadakatali Gori, Jing Liu, Qin Zheng, Xiang Cui, Qi Peng, Jieru Wan, Rana Rais, Barbara S. Slusher, Srinivasa. N. Raja, Xinzhong Dong, Takashi Tsukamoto, Yun Guan","doi":"10.1126/scitranslmed.adw9446","DOIUrl":"10.1126/scitranslmed.adw9446","url":null,"abstract":"<div >The human <i>Mas</i>-related G protein–coupled receptor X1 (MrgprX1) represents a promising nonopioid analgesic target because of its selective expression in primary nociceptive sensory neurons. Positive allosteric modulators (PAMs) promote receptor signaling, depending on the availability of endogenous ligands, offering physiological selectivity over orthosteric agonists. We developed an orally bioavailable MrgprX1 PAM, 6-tert-butyl-5-(4-chlorophenyl)-4-(2-fluoro-6-(trifluoromethoxy)phenoxy)thieno[2,3-d]pyrimidine (BCFTP). BCFTP selectively potentiated the functional response of MrgprX1 in HEK293 cells, was metabolically stable, and demonstrated a favorable in vitro safety profile. BCFTP was orally bioavailable and distributed into the spinal cords of wild-type mice. BAM22, an endogenous ligand for MrgprX1, was up-regulated in the spinal cord after nerve injury in both wild-type and humanized MrgprX1 mice and was expressed in peptidergic and nonpeptidergic dorsal root ganglion neurons. Oral administration of BCFTP dose-dependently inhibited heat hyperalgesia and spontaneous pain-like behavior but not mechanical hypersensitivity after sciatic chronic constrictive injury (CCI) in MrgprX1 mice. BCFTP did not have analgesic effects in Mrgpr cluster knockout (Mrgpr<sup>−/−</sup>) mice, indicating that the analgesic effects in MrgprX1 mice were MrgprX1 dependent. BCFTP enhanced BAM8-22–induced, MrgprX1-mediated reduction of C-fiber eEPSC amplitudes in spinal lamina II neurons, indicating inhibition of spinal nociceptive synaptic transmission. BCFTP did not induce tolerance or side effects, such as itch, sedation, and motor incoordination, and had no rewarding properties. The mRNAs encoding MrgprX1 and μ-opioid receptors were colocalized in human DRG neurons, and BCFTP synergistically enhanced morphine analgesia in CCI MrgprX1 mice. Our research suggests an approach for developing safer, orally bioavailable MrgprX1 PAM as a nonopioid therapy for neuropathic pain.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 829","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145765548","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-17DOI: 10.1126/scitranslmed.adq4060
Yahya Mohammadzadeh, Vojislav Gligorovski, Olga Egorova, Gabriele Casagrande Raffi, Jort J. van der Schans, Ali Ghasemi, Katharina Jonas, Bruno Torchia, Alan Guichard, Rachel Marcone, Amaia Martinez-Usatorre, Anna Köck, Raphael Genolet, Nadine Fournier, Tatiana V. Petrova, Daniel E. Speiser, Sahand Jamal Rahi, Nahal Mansouri, Michele De Palma
Effective antitumor immunity requires dendritic cells (DCs) to internalize, process, and present tumor antigens to T cells. Adoptive transfer of DCs that were loaded ex vivo with tumor antigens has been shown to stimulate antitumor immunity in patients with cancer, but clinical responses have been mixed. To address the limitations of traditional DC-based therapies, we constructed and functionally screened a panel of chimeric antigen receptors (CARs) optimized for expression and activity in DCs. Through this screening, we identified key functional components that guided the development of an inducible platform centered on an instructive chimeric antigen receptor (iCAR). This iCAR enabled DCs to (i) recognize a surface molecule present on cancer cells or their extracellular vesicles (EVs), such as disialoganglioside GD2 (expressed in melanoma and other tumors of neuroectodermal origin) or HER2 (expressed in some epithelial cancers), thereby promoting the acquisition of tumor-derived material containing putative tumor antigens; (ii) undergo immunostimulatory activation to prime antigen-specific T cells via both cross-dressing and cross-presentation; and (iii) transactivate the expression of the therapeutic cytokine interleukin-12 (IL-12) in response to antigen uptake. The iCAR converted melanoma-derived EVs from immune-suppressive to stimulatory cues for DCs in cell culture assays. Moreover, systemic administration of iCAR-DCs enhanced antigen-specific T cells, expanded low-frequency T cell clonotypes, and delayed tumor growth in immunotherapy-resistant melanoma models without the need for ex vivo antigen loading or cell maturation. iCAR-DCs may therefore provide a platform for antigen-agnostic cancer immunotherapy that integrates antigen uptake with programmable DC activation.
{"title":"Coordinate tumor-antigen uptake and dendritic cell activation by chimeric antigen receptors","authors":"Yahya Mohammadzadeh, Vojislav Gligorovski, Olga Egorova, Gabriele Casagrande Raffi, Jort J. van der Schans, Ali Ghasemi, Katharina Jonas, Bruno Torchia, Alan Guichard, Rachel Marcone, Amaia Martinez-Usatorre, Anna Köck, Raphael Genolet, Nadine Fournier, Tatiana V. Petrova, Daniel E. Speiser, Sahand Jamal Rahi, Nahal Mansouri, Michele De Palma","doi":"10.1126/scitranslmed.adq4060","DOIUrl":"10.1126/scitranslmed.adq4060","url":null,"abstract":"<div >Effective antitumor immunity requires dendritic cells (DCs) to internalize, process, and present tumor antigens to T cells. Adoptive transfer of DCs that were loaded ex vivo with tumor antigens has been shown to stimulate antitumor immunity in patients with cancer, but clinical responses have been mixed. To address the limitations of traditional DC-based therapies, we constructed and functionally screened a panel of chimeric antigen receptors (CARs) optimized for expression and activity in DCs. Through this screening, we identified key functional components that guided the development of an inducible platform centered on an instructive chimeric antigen receptor (iCAR). This iCAR enabled DCs to (i) recognize a surface molecule present on cancer cells or their extracellular vesicles (EVs), such as disialoganglioside GD2 (expressed in melanoma and other tumors of neuroectodermal origin) or HER2 (expressed in some epithelial cancers), thereby promoting the acquisition of tumor-derived material containing putative tumor antigens; (ii) undergo immunostimulatory activation to prime antigen-specific T cells via both cross-dressing and cross-presentation; and (iii) transactivate the expression of the therapeutic cytokine interleukin-12 (IL-12) in response to antigen uptake. The iCAR converted melanoma-derived EVs from immune-suppressive to stimulatory cues for DCs in cell culture assays. Moreover, systemic administration of iCAR-DCs enhanced antigen-specific T cells, expanded low-frequency T cell clonotypes, and delayed tumor growth in immunotherapy-resistant melanoma models without the need for ex vivo antigen loading or cell maturation. iCAR-DCs may therefore provide a platform for antigen-agnostic cancer immunotherapy that integrates antigen uptake with programmable DC activation.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 829","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145765535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-17DOI: 10.1126/scitranslmed.adv9403
Amit Manhas, Yu Liu, Chikage Noishiki, David Wu, Dipti Tripathi, Sarah Mirza, Dilip Thomas, Lu Liu, Avirup Guha, Patricia K. Nguyen, Ian Y. Chen, Vipul Chitalia, Paul Cheng, Danish Sayed, Melinda L. Telli, Karim Sallam, Joseph C. Wu, Nazish Sayed
Tyrosine kinase inhibitors (TKIs) have improved cancer outcomes but are limited by cardiovascular toxicity, most notably hypertension and heart failure. The underlying mechanisms remain poorly understood, hindering the development of protective strategies. Here, we investigated the role of endothelial mechanotransduction in mediating vascular and cardiac injury caused by the vascular endothelial growth factor receptor–targeting TKI sunitinib. Using patient-specific induced pluripotent stem cell–derived endothelial cells (iPSC-ECs) and a mouse model of TKI-induced hypertension, we identified down-regulation of piezo-type mechanosensitive ion channel component 1 (PIEZO1), a mechanically activated ion channel, as a driver of endothelial dysfunction. Restoring PIEZO1 expression, either pharmacologically with Yoda1, a selective agonist, or through inducible overexpression in iPSC-ECs, reversed sunitinib-induced endothelial dysfunction and mitigated its hypertensive effects, providing both mechanistic and genetic validation of PIEZO1’s protective role against vascular toxicity. In mice, cotreatment with sunitinib and Yoda1 prevented the long-term cardiac dysfunction observed after sunitinib exposure and normalized elevations in circulating cardiac stress biomarkers. Single-nucleus multiomic profiling of mouse hearts revealed that sunitinib exposure activated chromatin remodeling and fibrogenic programs, which were reversed with PIEZO1 activation. Human engineered cardiac organoids further demonstrated that sunitinib impaired cardiomyocyte function only in the presence of endothelial cells, confirming a role for disrupted endothelial-cardiomyocyte cross-talk in TKI cardiotoxicity. Together, these findings identify endothelial PIEZO1 as a mediator of TKI-induced hypertension and cardiac dysfunction and highlight PIEZO1 activation as a potential therapeutic strategy for protecting cardiovascular health during cancer therapy.
{"title":"Multiscale profiling of tyrosine kinase inhibitor cardiotoxicity reveals mechanosensitive ion channel PIEZO1 as cardioprotective","authors":"Amit Manhas, Yu Liu, Chikage Noishiki, David Wu, Dipti Tripathi, Sarah Mirza, Dilip Thomas, Lu Liu, Avirup Guha, Patricia K. Nguyen, Ian Y. Chen, Vipul Chitalia, Paul Cheng, Danish Sayed, Melinda L. Telli, Karim Sallam, Joseph C. Wu, Nazish Sayed","doi":"10.1126/scitranslmed.adv9403","DOIUrl":"10.1126/scitranslmed.adv9403","url":null,"abstract":"<div >Tyrosine kinase inhibitors (TKIs) have improved cancer outcomes but are limited by cardiovascular toxicity, most notably hypertension and heart failure. The underlying mechanisms remain poorly understood, hindering the development of protective strategies. Here, we investigated the role of endothelial mechanotransduction in mediating vascular and cardiac injury caused by the vascular endothelial growth factor receptor–targeting TKI sunitinib. Using patient-specific induced pluripotent stem cell–derived endothelial cells (iPSC-ECs) and a mouse model of TKI-induced hypertension, we identified down-regulation of <i>piezo-type mechanosensitive ion channel component 1</i> (<i>PIEZO1</i>), a mechanically activated ion channel, as a driver of endothelial dysfunction. Restoring <i>PIEZO1</i> expression, either pharmacologically with Yoda1, a selective agonist, or through inducible overexpression in iPSC-ECs, reversed sunitinib-induced endothelial dysfunction and mitigated its hypertensive effects, providing both mechanistic and genetic validation of PIEZO1’s protective role against vascular toxicity. In mice, cotreatment with sunitinib and Yoda1 prevented the long-term cardiac dysfunction observed after sunitinib exposure and normalized elevations in circulating cardiac stress biomarkers. Single-nucleus multiomic profiling of mouse hearts revealed that sunitinib exposure activated chromatin remodeling and fibrogenic programs, which were reversed with PIEZO1 activation. Human engineered cardiac organoids further demonstrated that sunitinib impaired cardiomyocyte function only in the presence of endothelial cells, confirming a role for disrupted endothelial-cardiomyocyte cross-talk in TKI cardiotoxicity. Together, these findings identify endothelial PIEZO1 as a mediator of TKI-induced hypertension and cardiac dysfunction and highlight PIEZO1 activation as a potential therapeutic strategy for protecting cardiovascular health during cancer therapy.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 829","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145765533","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1126/scitranslmed.adq0143
Xu Cao, Amit Manhas, Yi-Ing Chen, Arianne Caudal, Gema Mondejar-Parreño, Wenjuan Zhu, Wenqiang Liu, Xiaohui Kong, Wenshu Zeng, Lichao Liu, Shane R. Zhao, James W. S. Jahng, Paul J. Utz, Kari C. Nadeau, Masataka Nishiga, Joseph C. Wu
Messenger RNA (mRNA) vaccines against SARS-CoV-2 are highly effective and were instrumental in curbing the COVID-19 pandemic. However, rare cases of noninfective myocarditis, particularly in young males and typically after the second dose, have been observed. Here, we explore the mediators of this myocarditis to better understand and to enhance the safety of future mRNA vaccines. Through analysis of human plasma data and in vitro experiments with human macrophages and T cells, we identified increased C-X-C motif chemokine ligand 10 (CXCL10) and interferon-γ (IFN-γ) after exposure to BNT162b2 (Pfizer) or mRNA-1273 (Moderna). Neutralization of CXCL10 and IFN-γ during the second dose (21 days after the first dose) reduced vaccine-induced cardiac injury in mice. Neutralization also reduced cardiac stress markers such as the release of N-terminal pro-B-type natriuretic peptide (NT-proBNP) and expression of inflammatory genes in human induced pluripotent stem cell (iPSC)–derived cardiac spheroids. When exposed to these cytokines in vitro, human iPSC-derived cardiomyocytes (iPSC-CMs) exhibited impaired contractility, arrhythmogenicity, and proinflammatory gene expression patterns. Genistein, a phytoestrogen implicated in reducing cardiovascular inflammation, mitigated these effects in iPSC-CMs. In mice exposed to these cytokines or receiving BNT162b2 vaccination, genistein treatment reduced cardiac injury markers and attenuated infiltration of neutrophils and macrophages into the heart. These findings implicate CXCL10–IFN-γ signaling as a contributor to myocardial injury in experimental models of mRNA vaccination and indicate that pharmacologic modulation, such as with genistein, may mitigate cytokine-driven injury.
{"title":"Inhibition of CXCL10 and IFN-γ ameliorates myocarditis in preclinical models of SARS-CoV-2 mRNA vaccination","authors":"Xu Cao, Amit Manhas, Yi-Ing Chen, Arianne Caudal, Gema Mondejar-Parreño, Wenjuan Zhu, Wenqiang Liu, Xiaohui Kong, Wenshu Zeng, Lichao Liu, Shane R. Zhao, James W. S. Jahng, Paul J. Utz, Kari C. Nadeau, Masataka Nishiga, Joseph C. Wu","doi":"10.1126/scitranslmed.adq0143","DOIUrl":"10.1126/scitranslmed.adq0143","url":null,"abstract":"<div >Messenger RNA (mRNA) vaccines against SARS-CoV-2 are highly effective and were instrumental in curbing the COVID-19 pandemic. However, rare cases of noninfective myocarditis, particularly in young males and typically after the second dose, have been observed. Here, we explore the mediators of this myocarditis to better understand and to enhance the safety of future mRNA vaccines. Through analysis of human plasma data and in vitro experiments with human macrophages and T cells, we identified increased C-X-C motif chemokine ligand 10 (CXCL10) and interferon-γ (IFN-γ) after exposure to BNT162b2 (Pfizer) or mRNA-1273 (Moderna). Neutralization of CXCL10 and IFN-γ during the second dose (21 days after the first dose) reduced vaccine-induced cardiac injury in mice. Neutralization also reduced cardiac stress markers such as the release of N-terminal pro-B-type natriuretic peptide (NT-proBNP) and expression of inflammatory genes in human induced pluripotent stem cell (iPSC)–derived cardiac spheroids. When exposed to these cytokines in vitro, human iPSC-derived cardiomyocytes (iPSC-CMs) exhibited impaired contractility, arrhythmogenicity, and proinflammatory gene expression patterns. Genistein, a phytoestrogen implicated in reducing cardiovascular inflammation, mitigated these effects in iPSC-CMs. In mice exposed to these cytokines or receiving BNT162b2 vaccination, genistein treatment reduced cardiac injury markers and attenuated infiltration of neutrophils and macrophages into the heart. These findings implicate CXCL10–IFN-γ signaling as a contributor to myocardial injury in experimental models of mRNA vaccination and indicate that pharmacologic modulation, such as with genistein, may mitigate cytokine-driven injury.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 828","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145711322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1126/scitranslmed.ado6948
Tao Zhang, Tingyun Lei, Jie Han, Yangwu Chen, Yuepeng Nie, Ru Zhang, Honglu Cai, Zijin Guo, Ruojin Yan, Zan Li, Yongqiang Xu, Xianzhu Zhang, Hongxia Xu, Jiahe Chen, Youguo Liao, Qiulin He, Hong Zhang, Jianquan Chen, Shouan Zhu, Jiansheng Guo, Yi Liu, Haihua Pan, Jingyao Chen, Wei Yin, Hongwei Ouyang, Weiliang Shen, Zi Yin, Ruikang Tang, Chen Xiao
The enthesis, a fibrocartilaginous tissue connecting tendon or ligament to bone, is critical for joint movement but lacks regenerative capacity after injury. Current clinical treatments for enthesis healing remain limited. Here, with a resolution of 2 to 3 nanometers, we found that mineral particles form a continuous cross-fibrillar phase with a discontinuous distribution in the fibrocartilage layer. Building on this finding, we developed a series of bioinspired mineralized collagen matrices, characterized by both intra- and extrafibrillar localization of crystallites, with a tunable mass percentage of inorganic content as scaffolds for enthesis repair. Our results revealed that mineralized collagen with controlled inorganic content (33% mineral content) facilitated fibrocartilage healing across multiple animal enthesis injury models, including mice, rats, rabbits, and goats. In direct comparisons with other biomaterials in a rabbit model, the bioinspired mineralized collagen resulted in 82% fibrocartilage width recovery, more than two times the healing observed with other materials. Treatment with the bioinspired mineralized collagen scaffold produced joint healing with an ability to sustain a higher maximum load in both rat and rabbit models, with the animals able to walk normally. The goat model exhibited an improvement in jumping ability. Mechanistically, we found that the bioinspired mineralized collagen modulated Hedgehog signaling intensity in a mineralization-dependent manner, which in turn up-regulated Gli1 expression. This modulation regulated the differentiation of mesenchymal progenitor cells and promoted fibrocartilage healing. Overall, we demonstrate that a bioinspired mineralized collagen scaffold effectively promotes enthesis injury repair, demonstrating potential for clinical translation.
{"title":"A bioinspired mineralized collagen scaffold promotes enthesis healing and activates Gli1 expression in preclinical models","authors":"Tao Zhang, Tingyun Lei, Jie Han, Yangwu Chen, Yuepeng Nie, Ru Zhang, Honglu Cai, Zijin Guo, Ruojin Yan, Zan Li, Yongqiang Xu, Xianzhu Zhang, Hongxia Xu, Jiahe Chen, Youguo Liao, Qiulin He, Hong Zhang, Jianquan Chen, Shouan Zhu, Jiansheng Guo, Yi Liu, Haihua Pan, Jingyao Chen, Wei Yin, Hongwei Ouyang, Weiliang Shen, Zi Yin, Ruikang Tang, Chen Xiao","doi":"10.1126/scitranslmed.ado6948","DOIUrl":"10.1126/scitranslmed.ado6948","url":null,"abstract":"<div >The enthesis, a fibrocartilaginous tissue connecting tendon or ligament to bone, is critical for joint movement but lacks regenerative capacity after injury. Current clinical treatments for enthesis healing remain limited. Here, with a resolution of 2 to 3 nanometers, we found that mineral particles form a continuous cross-fibrillar phase with a discontinuous distribution in the fibrocartilage layer. Building on this finding, we developed a series of bioinspired mineralized collagen matrices, characterized by both intra- and extrafibrillar localization of crystallites, with a tunable mass percentage of inorganic content as scaffolds for enthesis repair. Our results revealed that mineralized collagen with controlled inorganic content (33% mineral content) facilitated fibrocartilage healing across multiple animal enthesis injury models, including mice, rats, rabbits, and goats. In direct comparisons with other biomaterials in a rabbit model, the bioinspired mineralized collagen resulted in 82% fibrocartilage width recovery, more than two times the healing observed with other materials. Treatment with the bioinspired mineralized collagen scaffold produced joint healing with an ability to sustain a higher maximum load in both rat and rabbit models, with the animals able to walk normally. The goat model exhibited an improvement in jumping ability. Mechanistically, we found that the bioinspired mineralized collagen modulated Hedgehog signaling intensity in a mineralization-dependent manner, which in turn up-regulated <i>Gli1</i> expression. This modulation regulated the differentiation of mesenchymal progenitor cells and promoted fibrocartilage healing. Overall, we demonstrate that a bioinspired mineralized collagen scaffold effectively promotes enthesis injury repair, demonstrating potential for clinical translation.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 828","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145711357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1126/scitranslmed.aed9347
{"title":"Erratum for the Research Article “Impairment of stromal-epithelial regenerative cross-talk in Hirschsprung disease primes for the progression to enterocolitis” by Z. Zhang et al.","authors":"","doi":"10.1126/scitranslmed.aed9347","DOIUrl":"10.1126/scitranslmed.aed9347","url":null,"abstract":"","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 828","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145717556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1126/scitranslmed.adr0024
Erkin Kurganov, Lei Cui, Nikita Budnik, Siwei Chen, Erick Olivares, David Baez-Nieto, Ahmet S. Asan, Laina Lusk, Lacey Smith, Sooyeon Jo, Diogo Marques, Ralda Nehme, Sureni V. Mullegama, Amanda Lindy, Alfred L. George Jr., Annapurna Poduri, Ingo Helbig, Mark Daly, Jen Q. Pan
CACNA1A encodes the α1A subunit of the CaV2.1 voltage–gated calcium channel essential for neuronal excitability, neurotransmitter release, and calcium signaling in neurons. Pathogenic CACNA1A variants are linked to diverse neurological and developmental disorders, including hemiplegic migraine, epilepsy, developmental delay, and ataxia. However, the contribution of CACNA1A variants to neurodevelopmental disorders has been less well established. Clinical heterogeneity and the complex impact of missense variants on channel function have made genotype-phenotype relationships difficult to resolve. Here, we characterized the biophysical properties of 42 de novo CACNA1A missense variants identified in a neurodevelopmental disorder cohort of over 31,000 individuals, alongside eight common variants from the general population. All but one de novo variant altered biophysical properties of the CaV2.1 channel, with 50% abolishing measurable currents. Among variants with detectable currents, nearly half disrupted voltage-dependent gating, whereas common variants had no measurable effect. Similar results were obtained using representative variants in the presence of β3 or β4 auxiliary subunits. The expression of two selected variants in CACNA1A-knockdown hiPSC-derived neurons confirmed similar functional alterations in human neurons. Simulations using the NEURON model of a Purkinje neuron demonstrated that these biophysical changes profoundly affected excitability. Coupling functional data with AlphaMissense predictions, we found that CaV2.1 missense variants contribute to the risk for developmental epileptic encephalopathy. Finally, correlating clinical features with molecular consequences of each variant revealed meaningful associations between specific channel dysfunctions and distinct clinical outcomes. These findings provide insights into CACNA1A molecular pathology and highlight the potential for precision medicine treatment of CACNA1A-associated disorders.
CACNA1A编码Ca V 2.1电压门控钙通道的α1A亚基，该通道对神经元的兴奋性、神经递质释放和钙信号传导至关重要。致病性CACNA1A变异与多种神经和发育障碍有关，包括偏瘫性偏头痛、癫痫、发育迟缓和共济失调。然而，CACNA1A变异对神经发育障碍的贡献尚未得到很好的证实。临床异质性和错义变异对通道功能的复杂影响使得基因型-表型关系难以解决。在这里，我们描述了在一个超过31000人的神经发育障碍队列中发现的42个新生CACNA1A错义变异的生物物理特性，以及来自普通人群的8个常见变异。除了一个全新的变体外，其他所有变体都改变了Ca V 2.1通道的生物物理特性，其中50%的变体消除了可测量电流。在具有可检测电流的变体中，近一半破坏了电压依赖性门控，而普通变体没有可测量的影响。在β3或β4辅助亚基存在的情况下，使用具有代表性的变体获得了类似的结果。在CACNA1A敲低hipsc衍生的神经元中，两种选择的变体的表达证实了人类神经元中类似的功能改变。使用浦肯野神经元模型的模拟表明，这些生物物理变化深刻地影响了兴奋性。将功能数据与AlphaMissense预测相结合，我们发现Ca V 2.1错义变异增加了发育性癫痫性脑病的风险。最后，将临床特征与每种变异的分子后果相关联，揭示了特定通道功能障碍与不同临床结果之间有意义的关联。这些发现提供了对CACNA1A分子病理学的见解，并强调了CACNA1A相关疾病的精准医学治疗的潜力。
{"title":"Characterization of the functional and clinical impacts of CACNA1A missense variants found in neurodevelopmental disorders","authors":"Erkin Kurganov, Lei Cui, Nikita Budnik, Siwei Chen, Erick Olivares, David Baez-Nieto, Ahmet S. Asan, Laina Lusk, Lacey Smith, Sooyeon Jo, Diogo Marques, Ralda Nehme, Sureni V. Mullegama, Amanda Lindy, Alfred L. George Jr., Annapurna Poduri, Ingo Helbig, Mark Daly, Jen Q. Pan","doi":"10.1126/scitranslmed.adr0024","DOIUrl":"10.1126/scitranslmed.adr0024","url":null,"abstract":"<div ><i>CACNA1A</i> encodes the α1A subunit of the Ca<sub>V</sub>2.1 voltage–gated calcium channel essential for neuronal excitability, neurotransmitter release, and calcium signaling in neurons. Pathogenic <i>CACNA1A</i> variants are linked to diverse neurological and developmental disorders, including hemiplegic migraine, epilepsy, developmental delay, and ataxia. However, the contribution of <i>CACNA1A</i> variants to neurodevelopmental disorders has been less well established. Clinical heterogeneity and the complex impact of missense variants on channel function have made genotype-phenotype relationships difficult to resolve. Here, we characterized the biophysical properties of 42 de novo <i>CACNA1A</i> missense variants identified in a neurodevelopmental disorder cohort of over 31,000 individuals, alongside eight common variants from the general population. All but one de novo variant altered biophysical properties of the Ca<sub>V</sub>2.1 channel, with 50% abolishing measurable currents. Among variants with detectable currents, nearly half disrupted voltage-dependent gating, whereas common variants had no measurable effect. Similar results were obtained using representative variants in the presence of β3 or β4 auxiliary subunits. The expression of two selected variants in <i>CACNA1A</i>-knockdown hiPSC-derived neurons confirmed similar functional alterations in human neurons. Simulations using the <i>NEURON</i> model of a Purkinje neuron demonstrated that these biophysical changes profoundly affected excitability. Coupling functional data with AlphaMissense predictions, we found that Ca<sub>V</sub>2.1 missense variants contribute to the risk for developmental epileptic encephalopathy. Finally, correlating clinical features with molecular consequences of each variant revealed meaningful associations between specific channel dysfunctions and distinct clinical outcomes. These findings provide insights into <i>CACNA1A</i> molecular pathology and highlight the potential for precision medicine treatment of <i>CACNA1A</i>-associated disorders.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 828","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145711356","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1126/scitranslmed.adz0529
Xin Liu, Jiayi Zhang, Junjun Chu, Yi-Jou Chen, Chen Qian, Helen Y. Wang, Rong-Fu Wang
Chimeric antigen receptor (CAR) T cell therapy has shown impressive clinical responses in the treatment of blood cancers, but high percentages of disease relapse 1 year after T cell infusion and severe toxicities associated with CAR T cell therapy remain major issues. Here, we report the construction of CARs with a ZAP70-derived signaling domain (ZAP327) that enhances therapeutic antitumor activity with increasing in vivo T cell persistence. ZAP327-driven CAR T cells reduced cytokine release and expression of T cell exhaustion markers but maintained similar or better cytolytic activity against tumor cells compared with conventional CAR T cells. The costimulatory domains, such as CD28 and 4-1BB, in the ZAP327 CAR constructs are required for providing a spacer between the transmembrane domain and ZAP327 kinase domain for optimal kinase structure folding and function, as well as costimulatory signaling. Furthermore, ZAP327-driven CAR T cells outperform conventional and several recently improved CAR T cells in therapeutic antitumor immunity, particularly in an antigen-low expression tumor model, which is clinically relevant and important for immune escape and disease relapse. Mechanistically, we show that the ZAP327 domain tuned down TCR signaling, increased the pools of stem-like memory T cells, and exhibited metabolic features associated with memory T cells by using the oxidative phosphorylation pathway. These results highlight the therapeutic potential of ZAP327-driven CAR T cells to overcome the limitations of the current CAR T cell therapies and enhance the potency and persistence of antitumor T cell responses in solid tumors as well.
{"title":"ZAP327 signaling domain–driven chimeric antigen receptor generates robust and long-term antitumor immunity in mouse models","authors":"Xin Liu, Jiayi Zhang, Junjun Chu, Yi-Jou Chen, Chen Qian, Helen Y. Wang, Rong-Fu Wang","doi":"10.1126/scitranslmed.adz0529","DOIUrl":"10.1126/scitranslmed.adz0529","url":null,"abstract":"<div >Chimeric antigen receptor (CAR) T cell therapy has shown impressive clinical responses in the treatment of blood cancers, but high percentages of disease relapse 1 year after T cell infusion and severe toxicities associated with CAR T cell therapy remain major issues. Here, we report the construction of CARs with a ZAP70-derived signaling domain (ZAP327) that enhances therapeutic antitumor activity with increasing in vivo T cell persistence. ZAP327-driven CAR T cells reduced cytokine release and expression of T cell exhaustion markers but maintained similar or better cytolytic activity against tumor cells compared with conventional CAR T cells. The costimulatory domains, such as CD28 and 4-1BB, in the ZAP327 CAR constructs are required for providing a spacer between the transmembrane domain and ZAP327 kinase domain for optimal kinase structure folding and function, as well as costimulatory signaling. Furthermore, ZAP327-driven CAR T cells outperform conventional and several recently improved CAR T cells in therapeutic antitumor immunity, particularly in an antigen-low expression tumor model, which is clinically relevant and important for immune escape and disease relapse. Mechanistically, we show that the ZAP327 domain tuned down TCR signaling, increased the pools of stem-like memory T cells, and exhibited metabolic features associated with memory T cells by using the oxidative phosphorylation pathway. These results highlight the therapeutic potential of ZAP327-driven CAR T cells to overcome the limitations of the current CAR T cell therapies and enhance the potency and persistence of antitumor T cell responses in solid tumors as well.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 828","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/scitranslmed.adz0529","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145711320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1126/scitranslmed.ads1892
Courtney M. Jackson, Elizabeth Ponko, Natalie M. Vance, Erin C. Davis, Mustafa M. Mahmood, Carolina Valderrama-Hincapié, Nichole S. Diaz, Lauren Vanasse, R. John Looney, Juilee Thakar, Antti E. Seppo, Kirsi M. Järvinen
The role of antibodies in protection against food allergy is debated. Here, we designed a longitudinal birth cohort study recruiting participants from an Old Order Mennonite (OOM) community, a traditional farm lifestyle considered to be at low risk for allergic diseases, and participants from urban and suburban Rochester, NY, USA, that were at higher risk for developing allergic diseases. We collected cord and peripheral blood, stool, saliva, and maternal milk samples longitudinally during pregnancy and during the first year of the infant’s life to evaluate B cell and antibody responses. Farm exposure led to an accrual of memory and immunoglobulin G–positive (IgG+) B cells, higher titers of plasma IgG and IgA and salivary and fecal IgA in infants, and higher IgA titers in maternal milk. Moreover, OOM infants and their mothers had higher titers of plasma IgG4 and IgA antibodies to egg ovalbumin in maternal milk, respectively, a feature that we found to be associated with lower incidence of egg allergy. Infant systemic IgG4 and IgA were associated with early introduction or more frequent ingestion of lightly cooked egg, respectively, but there was also an effect independent of egg exposure. Several food antigens were detected in cord blood, with in utero exposure potentially explaining the unexpected presence of antigen-specific IgA at birth. These analyses suggest earlier maturation of B cell immunity in farming lifestyle communities.
{"title":"Farm exposure in infancy is associated with elevated systemic IgG4, mucosal IgA responses, and lower incidence of food allergy","authors":"Courtney M. Jackson, Elizabeth Ponko, Natalie M. Vance, Erin C. Davis, Mustafa M. Mahmood, Carolina Valderrama-Hincapié, Nichole S. Diaz, Lauren Vanasse, R. John Looney, Juilee Thakar, Antti E. Seppo, Kirsi M. Järvinen","doi":"10.1126/scitranslmed.ads1892","DOIUrl":"10.1126/scitranslmed.ads1892","url":null,"abstract":"<div >The role of antibodies in protection against food allergy is debated. Here, we designed a longitudinal birth cohort study recruiting participants from an Old Order Mennonite (OOM) community, a traditional farm lifestyle considered to be at low risk for allergic diseases, and participants from urban and suburban Rochester, NY, USA, that were at higher risk for developing allergic diseases. We collected cord and peripheral blood, stool, saliva, and maternal milk samples longitudinally during pregnancy and during the first year of the infant’s life to evaluate B cell and antibody responses. Farm exposure led to an accrual of memory and immunoglobulin G–positive (IgG<sup>+</sup>) B cells, higher titers of plasma IgG and IgA and salivary and fecal IgA in infants, and higher IgA titers in maternal milk. Moreover, OOM infants and their mothers had higher titers of plasma IgG<sub>4</sub> and IgA antibodies to egg ovalbumin in maternal milk, respectively, a feature that we found to be associated with lower incidence of egg allergy. Infant systemic IgG<sub>4</sub> and IgA were associated with early introduction or more frequent ingestion of lightly cooked egg, respectively, but there was also an effect independent of egg exposure. Several food antigens were detected in cord blood, with in utero exposure potentially explaining the unexpected presence of antigen-specific IgA at birth. These analyses suggest earlier maturation of B cell immunity in farming lifestyle communities.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 828","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145711321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1126/scitranslmed.adu8484
Meng Xu, Dingju Wei, Shu Zhong, Weicheng Liu, Mei Ye, Jie Luo, Shuangdie Zhu, Zhenzhen Lai, Zhiping Zheng, Rui Hu, Qin Chen, Ningning Guo, Jian Lv, Wenmin Tian, Yang Chen, Guangfu Yang, Zhihua Wang, Jingjing Tong
Signal transducer and activator of transcription 3 (STAT3) is essential for cell signaling in response to extracellular stimuli, and its overactivation is a hallmark of inflammation and tumorigenesis. The differential mechanisms underlying the physiological and pathological regulation of STAT3 remain elusive. Here, we demonstrated that cryptic splice sites in STAT3 generated heterogeneous isoforms with or without a single amino acid Ser701 (wS701/ΔS701), with the latter being more abundant in colon cancers. Intrinsic S701 underwent reversible phosphorylation catalyzed by mechanistic target of rapamycin complex 1 (mTORC1) and protein phosphatase 2A (PP2A). Upon inflammatory stimulation, phosphorylation at S701 (p-S701) sequestered Y705 phosphorylation (p-Y705) by interfering with the access of Janus kinase 1/2 and restricting STAT3 overactivation. In contrast, the STAT3_ΔS701 isoform was hyperactive because of the absence of this self-restricting mechanism. Deletion of S701 in mice increased susceptibility to colonic inflammation and tumorigenesis. Pharmacological inhibition of PP2A sustained p-S701 and alleviated colon inflammation in wild-type but not in ΔS701 mice. Our findings highlight the importance of STAT3 heterogeneity in colonic inflammation and colorectal cancer.
{"title":"A hyperactive splice variant of STAT3 promotes colonic inflammation-associated tumorigenesis in mice","authors":"Meng Xu, Dingju Wei, Shu Zhong, Weicheng Liu, Mei Ye, Jie Luo, Shuangdie Zhu, Zhenzhen Lai, Zhiping Zheng, Rui Hu, Qin Chen, Ningning Guo, Jian Lv, Wenmin Tian, Yang Chen, Guangfu Yang, Zhihua Wang, Jingjing Tong","doi":"10.1126/scitranslmed.adu8484","DOIUrl":"10.1126/scitranslmed.adu8484","url":null,"abstract":"<div >Signal transducer and activator of transcription 3 (STAT3) is essential for cell signaling in response to extracellular stimuli, and its overactivation is a hallmark of inflammation and tumorigenesis. The differential mechanisms underlying the physiological and pathological regulation of STAT3 remain elusive. Here, we demonstrated that cryptic splice sites in <i>STAT3</i> generated heterogeneous isoforms with or without a single amino acid Ser701 (wS701/ΔS701), with the latter being more abundant in colon cancers. Intrinsic S701 underwent reversible phosphorylation catalyzed by mechanistic target of rapamycin complex 1 (mTORC1) and protein phosphatase 2A (PP2A). Upon inflammatory stimulation, phosphorylation at S701 (p-S701) sequestered Y705 phosphorylation (p-Y705) by interfering with the access of Janus kinase 1/2 and restricting STAT3 overactivation. In contrast, the STAT3_ΔS701 isoform was hyperactive because of the absence of this self-restricting mechanism. Deletion of S701 in mice increased susceptibility to colonic inflammation and tumorigenesis. Pharmacological inhibition of PP2A sustained p-S701 and alleviated colon inflammation in wild-type but not in ΔS701 mice. Our findings highlight the importance of STAT3 heterogeneity in colonic inflammation and colorectal cancer.</div>","PeriodicalId":21580,"journal":{"name":"Science Translational Medicine","volume":"17 828","pages":""},"PeriodicalIF":14.6,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145711323","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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