Pub Date : 2025-10-31DOI: 10.1038/s41551-025-01542-1
Elena Garreta,Daniel Moya-Rull,Alberto Centeno,Andrés Marco,Asier Ullate-Agote,Gaia Amato,Carlos J Aranda,Roger Oria,Daniel Lozano-Ojalvo,Merel B F Pool,Tim L Hamelink,Idoia Lucía Selfa,Federico González,Carolina Tarantino,Alejandro Montero Salinas,Patricia López San Martín,Priyanka Koshy,Aleix Gavaldà-Navarro,Amaia Vilas-Zornoza,Juan R Rodríguez-Madoz,Antón Fernández García,Inmaculada Marquez-Leiva,Henri G D Leuvenink,Cristobal Belda-Iniesta,Maarten Naesens,Beatriz Dominguez-Gil,Marcelino González-Martín,Javier Rodríguez-Rivera,Jordi Ochando,Felipe Prosper,Cyril Moers,Nuria Montserrat
Organoids derived from human pluripotent stem (hPS) cells hold promise for therapeutic purposes. However, technological advances to overcome their massive production while ensuring differentiation fidelity are still lacking. Here we report a procedure sustaining the derivation of kidney organoids from hPS cells (hPSC-kidney organoids) using a scalable, reproducible and affordable approach that allows hPSC-kidney organoid differentiation into different renal cell types. Using single-cell RNA sequencing, confocal image analysis, metabolic assays and CRISPR-Cas9 engineering for generation of fluorescent reporters, we show that hPSC-kidney organoids exhibit transcriptional variety and cellular composition following cell-to-cell contact. We infuse human kidney organoids into ex vivo porcine kidneys using normothermic machine perfusion, and demonstrate in vivo engraftment of hPSC-kidney organoids. We further evaluate the immune response, confirming the feasibility and viability of the procedure. We identify cells of human origin after normothermic machine perfusion and in vivo transplantation by means of in situ hybridization, immunohistochemistry, confocal microscopy, image analysis and quantification, in vivo imaging, and flow cytometry. This work provides a foundation for using hPSC-kidney organoids for ex vivo cell-based therapies in clinical trials.
{"title":"Systematic production of human kidney organoids for transplantation in porcine kidneys during ex vivo machine perfusion.","authors":"Elena Garreta,Daniel Moya-Rull,Alberto Centeno,Andrés Marco,Asier Ullate-Agote,Gaia Amato,Carlos J Aranda,Roger Oria,Daniel Lozano-Ojalvo,Merel B F Pool,Tim L Hamelink,Idoia Lucía Selfa,Federico González,Carolina Tarantino,Alejandro Montero Salinas,Patricia López San Martín,Priyanka Koshy,Aleix Gavaldà-Navarro,Amaia Vilas-Zornoza,Juan R Rodríguez-Madoz,Antón Fernández García,Inmaculada Marquez-Leiva,Henri G D Leuvenink,Cristobal Belda-Iniesta,Maarten Naesens,Beatriz Dominguez-Gil,Marcelino González-Martín,Javier Rodríguez-Rivera,Jordi Ochando,Felipe Prosper,Cyril Moers,Nuria Montserrat","doi":"10.1038/s41551-025-01542-1","DOIUrl":"https://doi.org/10.1038/s41551-025-01542-1","url":null,"abstract":"Organoids derived from human pluripotent stem (hPS) cells hold promise for therapeutic purposes. However, technological advances to overcome their massive production while ensuring differentiation fidelity are still lacking. Here we report a procedure sustaining the derivation of kidney organoids from hPS cells (hPSC-kidney organoids) using a scalable, reproducible and affordable approach that allows hPSC-kidney organoid differentiation into different renal cell types. Using single-cell RNA sequencing, confocal image analysis, metabolic assays and CRISPR-Cas9 engineering for generation of fluorescent reporters, we show that hPSC-kidney organoids exhibit transcriptional variety and cellular composition following cell-to-cell contact. We infuse human kidney organoids into ex vivo porcine kidneys using normothermic machine perfusion, and demonstrate in vivo engraftment of hPSC-kidney organoids. We further evaluate the immune response, confirming the feasibility and viability of the procedure. We identify cells of human origin after normothermic machine perfusion and in vivo transplantation by means of in situ hybridization, immunohistochemistry, confocal microscopy, image analysis and quantification, in vivo imaging, and flow cytometry. This work provides a foundation for using hPSC-kidney organoids for ex vivo cell-based therapies in clinical trials.","PeriodicalId":19063,"journal":{"name":"Nature Biomedical Engineering","volume":"12 1","pages":""},"PeriodicalIF":28.1,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145411645","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 : 2025-10-28DOI: 10.1038/s41551-025-01532-3
Jan A Rath,Lucas S P Rudden,Nazila Nouraee,Tiffany X Y Que,Christine Von Gunten,Cynthia Perez,Flora Birch,Yashashvi Bhugowon,Andreas Fueglistaler,Aisima Chatzi Souleiman,Patrick Barth,Caroline Arber
The tumour microenvironment (TME) plays a key role in tumour progression, and soluble and cellular TME components can limit CAR-T cell function and persistence. Targeting soluble TME factors to enhance anti-tumour responses of engineered T cells through chimeric receptors is not broadly explored owing to the unpredictable signalling characteristics of synthetic protein receptors. Here we develop a computational protein design platform for the de novo bottom-up assembly of allosteric receptors with programmable input-output behaviours that respond to soluble TME factors with co-stimulation and cytokine signals in T cells, called TME-sensing switch receptor for enhanced response to tumours (T-SenSER). We develop two sets of T-SenSERs targeting vascular endothelial growth factor (VEGF) or colony-stimulating factor 1 (CSF1) that are both selectively enriched in a variety of tumours. Combination of CAR and T-SenSER in human T cells enhances anti-tumour responses in models of lung cancer and multiple myeloma, in a VEGF- or CSF1-dependent manner. Our study sets the stage for the accelerated development of synthetic biosensors with custom-built sensing and responses for basic and translational cell engineering applications.
{"title":"Computational design of synthetic receptors with programmable signalling activity for enhanced cancer T cell therapy.","authors":"Jan A Rath,Lucas S P Rudden,Nazila Nouraee,Tiffany X Y Que,Christine Von Gunten,Cynthia Perez,Flora Birch,Yashashvi Bhugowon,Andreas Fueglistaler,Aisima Chatzi Souleiman,Patrick Barth,Caroline Arber","doi":"10.1038/s41551-025-01532-3","DOIUrl":"https://doi.org/10.1038/s41551-025-01532-3","url":null,"abstract":"The tumour microenvironment (TME) plays a key role in tumour progression, and soluble and cellular TME components can limit CAR-T cell function and persistence. Targeting soluble TME factors to enhance anti-tumour responses of engineered T cells through chimeric receptors is not broadly explored owing to the unpredictable signalling characteristics of synthetic protein receptors. Here we develop a computational protein design platform for the de novo bottom-up assembly of allosteric receptors with programmable input-output behaviours that respond to soluble TME factors with co-stimulation and cytokine signals in T cells, called TME-sensing switch receptor for enhanced response to tumours (T-SenSER). We develop two sets of T-SenSERs targeting vascular endothelial growth factor (VEGF) or colony-stimulating factor 1 (CSF1) that are both selectively enriched in a variety of tumours. Combination of CAR and T-SenSER in human T cells enhances anti-tumour responses in models of lung cancer and multiple myeloma, in a VEGF- or CSF1-dependent manner. Our study sets the stage for the accelerated development of synthetic biosensors with custom-built sensing and responses for basic and translational cell engineering applications.","PeriodicalId":19063,"journal":{"name":"Nature Biomedical Engineering","volume":"19 1","pages":""},"PeriodicalIF":28.1,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145380925","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 : 2025-10-28DOI: 10.1038/s41551-025-01545-y
Madelynn N. Whittaker, Lauren C. Testa, Aidan Quigley, Dominique L. Brooks, Sarah A. Grandinette, Hooda Said, Garima Dwivedi, Ishaan Jindal, Daphne Volpp, Julia L. Hacker, Ping Qu, Josh Zhiyong Wang, Michael A. Levine, Rebecca C. Ahrens-Nicklas, Qiaoli Li, Kiran Musunuru, Mohamad-Gabriel Alameh, William H. Peranteau, Xiao Wang
{"title":"Improved specificity and efficiency of in vivo adenine base editing therapies with hybrid guide RNAs","authors":"Madelynn N. Whittaker, Lauren C. Testa, Aidan Quigley, Dominique L. Brooks, Sarah A. Grandinette, Hooda Said, Garima Dwivedi, Ishaan Jindal, Daphne Volpp, Julia L. Hacker, Ping Qu, Josh Zhiyong Wang, Michael A. Levine, Rebecca C. Ahrens-Nicklas, Qiaoli Li, Kiran Musunuru, Mohamad-Gabriel Alameh, William H. Peranteau, Xiao Wang","doi":"10.1038/s41551-025-01545-y","DOIUrl":"https://doi.org/10.1038/s41551-025-01545-y","url":null,"abstract":"","PeriodicalId":19063,"journal":{"name":"Nature Biomedical Engineering","volume":"4 1","pages":""},"PeriodicalIF":28.1,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145382062","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 : 2025-10-24DOI: 10.1038/s41551-025-01537-y
Laura Ferrante,Anna Boesendorfer,Deren Y Barsakcioglu,Benedikt Baumgartner,Yazan Al-Ajam,Alexander Woollard,Norbert V Kang,Oskar C Aszmann,Dario Farina
Targeted muscle reinnervation surgery reroutes residual nerve signals into spare muscles, enabling the recovery of neural information through electromyography (EMG). However, EMG signals are often overlapping, making the interpretation of limb functions complicated. Regenerative peripheral nerve interfaces surgically partition the nerve into individual fascicles that reinnervate specific muscle grafts, isolating distinct neural sources for precise control and interpretation of EMG signals. Here we combine targeted muscle reinnervation surgery of polyvalent nerves with a high-density microelectrode array implanted at a single site within a reinnervated muscle, and via mathematical source separation methods, we separate all neural signals that are redirected into a single muscle. In participants with upper-limb amputation, the deconvolution of EMG signals from four reinnervated muscles into motor unit spike trains revealed distinct clusters of motor neurons associated with diverse functional tasks. Our method enabled the extraction of multiple neural commands within a single reinnervated muscle, eliminating the need for surgical nerve division. This approach holds promises for enhancing control over prosthetic limbs and for understanding how the central nervous system encodes movement after reinnervation.
{"title":"Implanted microelectrode arrays in reinnervated muscles allow separation of neural drives from transferred polyfunctional nerves.","authors":"Laura Ferrante,Anna Boesendorfer,Deren Y Barsakcioglu,Benedikt Baumgartner,Yazan Al-Ajam,Alexander Woollard,Norbert V Kang,Oskar C Aszmann,Dario Farina","doi":"10.1038/s41551-025-01537-y","DOIUrl":"https://doi.org/10.1038/s41551-025-01537-y","url":null,"abstract":"Targeted muscle reinnervation surgery reroutes residual nerve signals into spare muscles, enabling the recovery of neural information through electromyography (EMG). However, EMG signals are often overlapping, making the interpretation of limb functions complicated. Regenerative peripheral nerve interfaces surgically partition the nerve into individual fascicles that reinnervate specific muscle grafts, isolating distinct neural sources for precise control and interpretation of EMG signals. Here we combine targeted muscle reinnervation surgery of polyvalent nerves with a high-density microelectrode array implanted at a single site within a reinnervated muscle, and via mathematical source separation methods, we separate all neural signals that are redirected into a single muscle. In participants with upper-limb amputation, the deconvolution of EMG signals from four reinnervated muscles into motor unit spike trains revealed distinct clusters of motor neurons associated with diverse functional tasks. Our method enabled the extraction of multiple neural commands within a single reinnervated muscle, eliminating the need for surgical nerve division. This approach holds promises for enhancing control over prosthetic limbs and for understanding how the central nervous system encodes movement after reinnervation.","PeriodicalId":19063,"journal":{"name":"Nature Biomedical Engineering","volume":"108 1","pages":""},"PeriodicalIF":28.1,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145357631","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}
Stem-cell-based neural tissue engineering and spinal cord organoids show promises for spinal cord injury repair. However, the native spinal cord presents cell heterogeneity and a stereotypical spatial structure that makes difficult their recapitulation within an organoid architecture, which requires an assembly encompassing cellular composition, segmental organization and dorsoventral features. Here we engineer a thoracic vertebral segment-specific spinal cord organoid (enTsOrg) model that can precisely match the transplantation site, establish synaptic connections and enhance in vivo neuroelectric conduction. The organoids are generated from fibroblasts-derived induced pluripotent stem cells and a layered double-hydroxide matrix in a basement membrane hydrogel (Matrigel). Grafted in a spinal cord injury mouse model, enTsOrg presents advanced maturation, functionalization and organized distribution of critical neuronal subtypes with thoracic segmental heterogeneity, including various motor neuron and interneuron subtypes, that serve essentially to restore motor functions. Transplantation of enTsOrg can restructure neural circuits in paralysed animals and restore hind-limb motor function. The robust neurological function and therapeutic efficacy of enTsOrg highlight a potential avenue for organoid designing for specific anatomical regions in neurological injury treatments.
{"title":"Engineered thoracic spinal cord organoids for transplantation after spinal cord injury.","authors":"Yanjing Zhu,Ruiqi Huang,Liqun Yu,Zhibo Liu,Yuchen Liu,Wenyong Fan,Gufa Lin,Zhaojie Wang,Xiaolie He,Xu Xu,Bei Ma,Youwei Chen,Yuxin Bai,Jing Li,Bairu Chen,Liming Cheng,Rongrong Zhu","doi":"10.1038/s41551-025-01549-8","DOIUrl":"https://doi.org/10.1038/s41551-025-01549-8","url":null,"abstract":"Stem-cell-based neural tissue engineering and spinal cord organoids show promises for spinal cord injury repair. However, the native spinal cord presents cell heterogeneity and a stereotypical spatial structure that makes difficult their recapitulation within an organoid architecture, which requires an assembly encompassing cellular composition, segmental organization and dorsoventral features. Here we engineer a thoracic vertebral segment-specific spinal cord organoid (enTsOrg) model that can precisely match the transplantation site, establish synaptic connections and enhance in vivo neuroelectric conduction. The organoids are generated from fibroblasts-derived induced pluripotent stem cells and a layered double-hydroxide matrix in a basement membrane hydrogel (Matrigel). Grafted in a spinal cord injury mouse model, enTsOrg presents advanced maturation, functionalization and organized distribution of critical neuronal subtypes with thoracic segmental heterogeneity, including various motor neuron and interneuron subtypes, that serve essentially to restore motor functions. Transplantation of enTsOrg can restructure neural circuits in paralysed animals and restore hind-limb motor function. The robust neurological function and therapeutic efficacy of enTsOrg highlight a potential avenue for organoid designing for specific anatomical regions in neurological injury treatments.","PeriodicalId":19063,"journal":{"name":"Nature Biomedical Engineering","volume":"356 1","pages":""},"PeriodicalIF":28.1,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145357630","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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