Cellular and Molecular Life Sciences最新文献

{"title":"Tumor-associated macrophages contribute to cisplatin resistance via regulating Pol η-mediated translesion DNA synthesis in ovarian cancer.","authors":"Bilash Chatterjee, Mrinmoy Sarkar, Debanjana Ghosh, Sangita Mishra, Subhankar Bose, Md Maqsood Ahamad Khan, Senthil Kumar Ganesan, Nabanita Chatterjee, Amit Kumar Srivastava","doi":"10.1007/s00018-025-05731-8","DOIUrl":"https://doi.org/10.1007/s00018-025-05731-8","url":null,"abstract":"<p><p>Tumor-associated macrophages (TAMs) are known to be involved in the manifestation of aggressive and therapy-resistant phenotypes in solid tumors. Nevertheless, the effects of dynamic intervention by TAMs on the DNA damage response of cancer cells are largely unexplored. Herein, we report that TAMs modulate the DNA damage repair pathways of ovarian cancer cells in response to platinum-(Pt) based therapeutic regimen. We demonstrate that coculture of TAMs with cancer cells directly upregulate Pol η, along with RAD18 and REV1 of the Translesion DNA synthesis (TLS) pathway, while concurrently downregulating components of the high-fidelity nucleotide excision repair (NER) mechanism. Consequently, we observed a better survival probability, DNA repair capacity, and enrichment of stemness properties in ovarian cancer cells. DNA bulky adducts produced by cisplatin are resolved through differential activation NER and TLS pathways. However, we elucidated that TAMs provide favorable conditions for activating the error-prone TLS pathway for lesion bypass over damage resolution. Furthermore, cellular crosstalk in cocultured cancer cells stimulates the nuclear translocation and expression of RelA, which recruits Pol η by acting as a potent transcription factor. In fact, with pristimerin-mediated disruption of p65 (RelA) translocation, the cancer cells become more prone to DNA damage-induced cell death and compromised regenerative potential. In both in vitro cell cultures and in vivo mouse xenograft models, cocultured macrophages exhibited predominantly M2-like phenotype with prevalence in the invasive zone of xenograft tumor margins. Taken together, our investigation revealed multifaceted crosstalk-mediated regulation of DNA damage repair between TAMs and ovarian cancer cells.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"220"},"PeriodicalIF":6.2,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144172854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"GPR35 prevents drug-induced liver injury via the Gαs-cAMP-PKA axis in macrophages.","authors":"Xueqin Zhao, Yuanhao Li, Liu Yang, Xi Chen, Jialong Zhang, Tong Chen, Haoqi Wang, Fei Li, Chen Cheng, Jingjing Wu, Jingjing Cong, Wenwei Yin, Jing Li, Xuefu Wang","doi":"10.1007/s00018-025-05751-4","DOIUrl":"https://doi.org/10.1007/s00018-025-05751-4","url":null,"abstract":"<p><p>Acetaminophen (APAP) overdose induces acute liver injury and represents the most frequent cause of drug-induced liver injury worldwide. Macrophage-mediated inflammation plays detrimental roles during the early stage of liver injury. However, the potential targets regulating inflammation to improve drug-induced liver injury remains undefined. In this study, we reported that G protein-coupled receptor 35 (GPR35) improves drug-induced liver injury by blocking macrophage-mediated inflammation via the Gαs-cyclic AMP-protein kinase A (Gαs-cAMP-PKA) pathway. The ablation of GPR35 exacerbates APAP-induced liver injury, characterized by higher levels of alanine aminotransferase and aspartate aminotransferase in sera, larger damaged areas, and increased levels of pro-inflammatory cytokines. More hepatic macrophages appeared in the inflamed liver of mice with GPR35 deficiency. In contrast, the agonists of GPR35 alleviated APAP-induced liver injury. The depletion of macrophages abolished GPR35-mediated protection. Mechanistically, GPR35 ablation facilitated the activation of pro-inflammatory AKT, MAPK, and NF-κB signaling pathways at the downstream of Toll-like receptors in macrophages. GPR35 agonists activated Gαs-cAMP-PKA signaling to inhibit the activation of these pro-inflammatory signaling pathways and then suppress the inflammatory response in macrophages. Thus, our findings demonstrate that GPR35 prevents drug-induced liver injury by blocking macrophage-mediated inflammation via the Gαs-cAMP-PKA pathway, indicating that GPR35 is a potential target for the development of novel medicines that control drug-induced liver injury.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"219"},"PeriodicalIF":6.2,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144172851","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual phosphorylation of glycogen synthase kinase 3β differentially integrates metabolic programs to determine T cell immunity across vertebrates.","authors":"Wei Liang, Ming Geng, Wenzhuo Rao, Kang Li, Yating Zhu, Yuying Zheng, Xiumei Wei, Jialong Yang","doi":"10.1007/s00018-025-05746-1","DOIUrl":"https://doi.org/10.1007/s00018-025-05746-1","url":null,"abstract":"<p><p>The integration of metabolic programs with T cell signaling establishes a molecular foundation for immune metabolism. As a key metabolic regulator, GSK3β's activity is dynamically modulated by phosphorylation at Ser9 and Tyr216. However, the contribution of these phosphorylation sites on metabolism-driven T cell response remains unclear. Using tilapia and mouse models, we investigated the regulation of GSK3β on T cell metabolism and its evolutionary variation. In tilapia, T cell activation induces GSK3β signaling, linking to both glycolysis and oxidative phosphorylation (OXPHOS). Tyr216 phosphorylation preferentially promotes glycolysis, facilitating T cell activation, proliferation, and antibacterial immunity; while inhibition of Ser9 phosphorylation specifically enhances OXPHOS to sustain T cell responses. Differently, Tyr216 phosphorylation supports both glycolysis and OXPHOS in mouse, ensuring CD4⁺ T and CD8⁺ T cell activation, proliferation, and cytokine production. Although Ser9 phosphorylation controls OXPHOS, its inhibition impairs rather than enhances OXPHOS and CD4⁺ T cell responses in mouse. We thus revealed a previously unknown mechanism underlying T cell metabolism and proposed that, through evolution, GSK3β has restructured the regulatory strategy, enabling bidirectional control of T cell metabolism and immunity in mammals and enhancing the flexibility of the adaptive immune system.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"218"},"PeriodicalIF":6.2,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144157102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sulfasalazine combined with anti-IL-1β mAb induces ferroptosis and immune modulation in oral squamous cell carcinoma.","authors":"Rui Zhou, Jiaying Zhou, Yuwen Xiong, Kai Su, Changlin Liu, Bin Cheng, Tong Wu","doi":"10.1007/s00018-025-05742-5","DOIUrl":"10.1007/s00018-025-05742-5","url":null,"abstract":"<p><p>Oral squamous cell carcinoma (OSCC), one of the most prevalent and aggressive forms of head and neck squamous cell carcinoma, has a five-year survival rate of about 50% ~ 60%, emphasizing the urgent need for more effective therapeutic strategies. Solute carrier family 7 member 11 (SLC7A11) is overexpressed in various cancers and represents a potential therapeutic target. Sulfasalazine (SAS), a Food and Drug Administration-approved drug, is a potent inhibiter of SLC7A11. However, SAS can also increase the levels of pro-inflammatory cytokines such as IL-1β, which may suppress the immune response. Here, we investigate the effect of SAS combined with anti-IL-1β monoclonal antibody (anti-IL-1β mAb) as a novel treatment strategy for OSCC. In this study, SLC7A11 was markedly increased in OSCC tissues, and high SLC7A11 expression predicted poor prognosis. SAS treatment was shown to suppress OSCC cell proliferation and trigger ferroptosis, as evidenced by elevated reactive oxygen species, reduced glutathione and enhanced lipid peroxidation. SAS also elevated IL-1β levels, leading to T cell exhaustion. Combining SAS with anti-IL-1β mAb reversed T cell exhaustion and amplified the anti-tumor effects in vitro. In the 4-nitroquinoline-1-oxide-induced oral cancergenisis model, the combination treatment significantly inhibited oral carcinogenesis compared to monotherapy. Our results suggest that combining SAS with anti-IL-1β mAb enhances the anti-tumor efficacy against OSCC through tumor growth inhibition and immune modulation, offering a promising therapeutic strategy.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"216"},"PeriodicalIF":6.2,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144157197","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"α-Ketoglutarate alleviates the pathogenesis of lupus and inhibits the activation and differentiation of B cells by promoting the expression of CD39.","authors":"Yangzhe Gao, Yucai Xiao, Yuxin Hu, Lu Yu, Jiakun Liu, Zhengyi Zhang, Tianqi Zhao, Shuo Zhao, Lili Zhang, Yonghong Yang, Huabao Xiong, Guanjun Dong","doi":"10.1007/s00018-025-05734-5","DOIUrl":"https://doi.org/10.1007/s00018-025-05734-5","url":null,"abstract":"<p><p>The abnormal activation and differentiation of B cells play an important role in the pathogenesis of autoimmune diseases, including systemic lupus erythematosus (SLE). Alpha-ketoglutarate (α-KG), a key metabolite in the tricarboxylic acid cycle, has been shown to be involved in the pathogenesis of many diseases by regulating the immune response. However, the role of α-KG in the pathogenesis of SLE, as well as the activation and differentiation of B cells, remains unclear. In this study, we used organic acid-targeted metabolomics to analyze the changes in the levels of 100 organic acids in the serum of SLE patients and healthy controls, and found a significant increase in the α-KG level in SLE patients compared to that in healthy controls. Notably, α-KG significantly could inhibit the activation and differentiation of B cells and alleviate disease progression in lupus-prone mice. Mechanistically, RNA-seq revealed that α-KG upregulated the expression of ENTPD1, which encodes an important immune checkpoint molecule CD39; B-cell-specific loss of ENTPD1 could significantly promote the Toll-like receptors-mediated activation and differentiation of B cells and aggravate the disease conditions of lupus-prone mice. The findings of our study demonstrate that α-KG alleviates the pathogenesis of lupus and inhibits the activation and differentiation of B cells by increasing the expression of CD39. Our findings laid a theoretical foundation for understanding the pathogenesis of SLE. Based on our study, α-KG might be further examined as a drug for the effective treatment of SLE.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"217"},"PeriodicalIF":6.2,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144157198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"IGFBP5 promotes EndoMT and renal fibrosis through H3K18 lactylation in diabetic nephropathy.","authors":"Xiaofang Hu, Wei Chen, Ming Yang, Mengwei Li, Xiangyi Li, Shaxi Ouyang","doi":"10.1007/s00018-025-05718-5","DOIUrl":"10.1007/s00018-025-05718-5","url":null,"abstract":"<p><strong>Objective: </strong>Diabetic nephropathy (DN) is an important complication in diabetic patients that severely impacts their quality of life and life expectancy. Although metabolic and inflammatory responses induced by hyperglycemia are considered the primary pathogenic factors of DN, the specific molecular mechanisms involved remain unclear. Here, we investigated the role of insulin-like growth factor-binding protein 5 (IGFBP5) in DN using in vitro cell experiments and mouse models.</p><p><strong>Methods: </strong>We assessed the effects of high-glucose conditions on IGFBP5 expression in glomerular endothelial cells and evaluated its regulatory effects on glycolysis, NLRP3 inflammasome activation, endothelial‒mesenchymal transition (EndoMT), and histone lactylation via the suppression of IGFBP5. Furthermore, we evaluated the effects of IGFBP5 on renal fibrosis and confirmed its regulatory mechanisms in DN model mice.</p><p><strong>Results: </strong>Knockdown of IGFBP5 inhibited high glucose-induced EndoMT in glomerular endothelial cells, which could also be suppressed by the NLRP3 inflammasome inhibitor MCC950. In addition, silencing of IGFBP5 decreased glycolytic activity and histone lactylation, thereby inhibiting the activation of the NLRP3 inflammasome and EndoMT. Furthermore, in mouse models of DN, IGFBP5 knockdown alleviated renal fibrosis and reduced glycolysis, histone lactylation, NLRP3 inflammasome activation and EndoMT.</p><p><strong>Conclusions: </strong>IGFBP5 promotes NLRP3 inflammasome-induced EndoMT and renal fibrosis by regulating glycolysis-mediated histone lactylation, accelerating the progression of DN. These findings provide a new potential therapeutic target for DN.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"215"},"PeriodicalIF":6.2,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LncRNA SYISL promotes fibroblast myofibroblast transition via miR-23a-mediated TRIOBP regulation.","authors":"Cong Xia, Lianhui Cheng, Wenyu Zhao, Airu Chang, Zhixia Wang, Huibing Liu, Xin Pan, Wenwen Li, Sakamoto Koji, Zhongzheng Li, Bin Li, Lan Wang, Guoying Yu","doi":"10.1007/s00018-025-05729-2","DOIUrl":"10.1007/s00018-025-05729-2","url":null,"abstract":"<p><p>Long non-coding RNAs (lncRNAs) play critical roles in the process of lung tissue injury and repair which abnormal repair leads to disease including fibrosis, yet the physiopathology remains elusive. Here, we identified the lncRNA SYISL as a key regulator that is markedly upregulated in idiopathic pulmonary fibrosis (IPF) patients and bleomycin (BLM)-induced murine fibrotic lungs. Inhibition of SYISL significantly attenuates TGF-β1-driven fibroblast myofibroblast transition (FMT), a process confers to tissue injury repair and regeneration. Which demonstrates SYISL interaction with miR-23a function as a potent suppressor of fibrotic activation. Mechanistically, SYISL acts as a competing endogenous RNA (ceRNA) that directly binds miR-23a, thereby derepressing TRIO and F-actin binding protein (TRIOBP) via targeting its 3' untranslated region (UTR). Knockdown of TRIOBP amplifies the anti-fibrotic effects of miR-23a mimics while abolishing the pro-fibrotic activity of miR-23a inhibitors, establishing TRIOBP as a downstream effector of the SYISL/miR-23a axis. In vivo, intratracheal delivery of SYISL-targeting shRNA via adeno-associated virus (AAV) robustly reduces collagen deposition, hydroxyproline content, and expression of fibrotic markers in BLM-induced mice. Our findings elucidate a lncRNA-driven regulatory circuit in which SYISL promotes pulmonary fibrosis by sequestering miR-23a to elevate TRIOBP expression, nominating this axis as a novel therapeutic target for IPF.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"214"},"PeriodicalIF":6.2,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12106260/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanisms of drug induced liver injury.","authors":"J Skat-Rørdam, J Lykkesfeldt, L L Gluud, P Tveden-Nyborg","doi":"10.1007/s00018-025-05744-3","DOIUrl":"10.1007/s00018-025-05744-3","url":null,"abstract":"<p><p>Drug induced liver injury (DILI) is a serious and potentially life-threatening condition resulting from an adverse drug reaction. Both the clinical manifestations and pathological mechanisms of DILI vary depending on drug characteristics, dose, duration of exposure as well as host specific factors. Disease onset can occur within days or months after the introduction of a drug. This has challenged identification of disease specific biomarkers and resulted in delayed and even erroneous diagnosis of patients. Apart from discontinuation of current pharmacotherapy, options for DILI patients are scarce and the condition can sometimes continue or worsen after drugs are discontinued or result in irreversible liver damage such as cirrhosis. This illustrates the need to uncover relevant pathological pathways that will pave the road for targeted interventions. In an effort to accommodate these needs, novel insights from preclinical and cellular disease modeling have allowed coupling of specific drugs to potential mechanisms of toxicity. This review outlines three signaling pathways of DILI: organelle stress, cholestasis, and immune responses, discusses their interplay with oxidative stress, and provides examples of drugs specifically targeting one or more steps in these pathways. A systematic approach identifying specific mechanisms of DILI could allow for the assembly of large databases, in turn enabling advanced computational modelling to provide accurate predictions of the DILI potential of both known drugs and future drug candidates.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"213"},"PeriodicalIF":6.2,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12106265/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144141636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LncRNA SNHG15 promotes angiogenesis and improves cardiac repair after myocardial infarction through MiR-665-mediated KDR expression.","authors":"Xiaoyun Liang, Shangyu Liu, Gang Liu, Qiankun Fan, Fangfang Ma, Yajuan Yin, ZhaoMing Li, Yuming Wu, Mingqi Zheng","doi":"10.1007/s00018-025-05737-2","DOIUrl":"10.1007/s00018-025-05737-2","url":null,"abstract":"<p><p>Angiogenesis is crucial for prolonging survival of the injured myocardium following myocardial infarction (MI). Long non-coding RNAs (lncRNAs), recognized as a novel class of regulatory RNAs, play significant roles in various biological processes. However, their role in cardiac angiogenesis is not well elucidated. This study aimed to identify angiogenic lncRNAs and investigate their roles and mechanisms following MI. In our study utilizing lncRNA sequencing within a mouse model of MI, systematic lncRNA profiling identified differentially expressed transcripts in the MI border zone at 7 days post-MI, with SNHG15 being notably upregulated in cardiac tissue and endothelial cells (ECs) of the peri-infarct area. Overexpression of SNHG15 in human coronary artery endothelial cells (HCAECs) led to an increase in kinase insert domain receptor (KDR) expression and enhanced angiogenic activity. Furthermore, adeno-associated virus 9 (AAV9)-mediated overexpression of SNHG15, under the control of an endothelial-specific promoter, resulted in improved cardiac function, reduced infarct size, and increased angiogenesis in the infarcted myocardium in vivo. However, after endothelial-specific knockdown of SNHG15, cardiac function in mice with MI deteriorated. Localization studies revealed that SNHG15 is primarily found in the cytoplasm of HCAECs and mechanistic investigations indicated that SNHG15 acts as a competing endogenous RNA for miR-665, thereby regulating KDR signaling and expression. And KDR overexpression rescues both MI exacerbation and EC dysfunction induced by SNHG15 silencing in MI hearts. Collectively, our study has uncovered lncRNA SNHG15 as a novel regulator of angiogenesis that enhances the endogenous repair mechanisms of ECs in response to pathophysiological remodeling post-MI. These findings position SNHG15 as a promising therapeutic target for inhibiting infarct expansion and promoting cardiac repair and regeneration following MI.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"211"},"PeriodicalIF":6.2,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12106210/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144141634","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Good manufacturing practice production of an off-the-shelf CD34⁺ progenitor-derived NK cell product with preserved anti-tumor functionality post-infusion in NOD/SCID/IL2Rg^null mice.","authors":"Laura Hooijmaijers, Marcos Vidal-Manrique, Bart Spils, Diede van Ens, Verónica Castaño Rodriguez, Willemijn Hobo, Anna L de Goede, Suzanne van Dorp, Joop H Jansen, Anniek B van der Waart, Paul K J D de Jonge, Harry Dolstra","doi":"10.1007/s00018-025-05727-4","DOIUrl":"10.1007/s00018-025-05727-4","url":null,"abstract":"<p><p>The adoptive transfer of natural killer (NK) cells represents a promising cancer therapy due to their intrinsic ability to distinguish between malignant and healthy cells in an allogeneic context, enabling off-the-shelf manufacturing possibilities. On demand availability of cryopreserved advanced therapy medicinal products (ATMPs) could promote enrolment in clinical trials and eventually commercialization with repeated dosing possibilities. However, NK cells are considered highly sensitive to cryopreservation-induced defects, including impaired viability, anti-tumor cytotoxicity, and in vivo expansion capacity. Here, we present the GMP-compliant manufacturing of an off-the-shelf NK cell product (RNK001), derived ex vivo from CD34⁺ hematopoietic stem and progenitor cells (HSPCs). To facilitate scalability and reduce hands-on time, the production process was adapted to a G-rex bioreactor, yielding high numbers of a pure CD56⁺CD3^- NK cell product. Cryopreservation of HSPC-NK cells using an optimized freeze-thaw protocol resulted in a consistently high post-thawing viability of mature and differentiated cells. Surviving HSPC-NK cells post-thawing exhibited enhanced proliferative capacity compared to fresh cells in vitro and their persistence in vivo was similar to that of fresh cells when administrated intravenously or intraperitoneally in NOD/SCID/IL2Rg^null mice. Moreover, cryopreserved HSPC-NK cells had robust anti-tumor functionality, efficiently killing tumor spheroids embedded in a 3D collagen matrix and maintaining degranulation and interferon-γ production capacity comparable to fresh cells following in vivo infusion. Together, these findings show the potential of cryopreserved HSPC-NK cells with potent effector functions, allowing the manufacturing of an off-the-shelf therapeutical NK cell product for hematological and solid malignancies.</p>","PeriodicalId":10007,"journal":{"name":"Cellular and Molecular Life Sciences","volume":"82 1","pages":"210"},"PeriodicalIF":6.2,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12104124/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144141609","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}