Clinical and Translational Medicine最新文献

{"title":"Clinical translation potential of self-inspired live-cell super-resolution microscopy","authors":"Liying Qu, Jingyang Zhu, Xiangyan Ding, Haoyu Li, Weisong Zhao","doi":"10.1002/ctm2.70390","DOIUrl":"https://doi.org/10.1002/ctm2.70390","url":null,"abstract":"<p>Super-resolution microscopy (SRM) has transformed our capacity to visualise subcellular structures,<span><sup>1</sup></span> offering unparalleled detail for biomedical research and clinical diagnostics.<span><sup>2-5</sup></span> However, the inherent photon budget constraints in live-cell imaging have long impeded the full realisation of these techniques, particularly when high spatiotemporal resolution is crucial for tracking dynamic biological processes.<span><sup>6</sup></span> To address this fundamental challenge, in our recent work published in <i>Nature Methods</i>, we introduced Self-inspired Noise2Noise (SN2N),<span><sup>7</sup></span> a deep learning framework that substantially enhances photon efficiency in live-cell SRM by one to two orders of magnitude. Here, we discuss the profound clinical translational potential we believe this innovation unlocks.</p><p>A key innovation of SN2N, as we designed it, is its capacity for robust denoising without the need for clean reference images or paired noisy training data. Crucially, its ability to train effectively from a single noisy frame, by leveraging inherent spatial redundancy in super-resolution images, offers a paradigm shift. For clinical research, this translates directly to markedly improved signal-to-noise ratios under low-illumination conditions. This, in turn, enables the adoption of gentler imaging protocols, thereby minimising phototoxicity and photobleaching—persistent challenges in live-cell SRM that we aimed to overcome.</p><p>This advance is particularly pertinent for studies involving patient-derived primary cells or biopsy tissues. Here, long-term observation of disease progression, drug efficacy and cellular dynamics can be achieved with minimal sample perturbation. Traditional SRM approaches often necessitate intense illumination, risking cellular stress, altered physiological states or even cell death.<span><sup>8, 9</sup></span> Such effects can compromise observational validity, a critical concern for sensitive samples such as cancer biopsies or stem cells. Our development of SN2N seeks to mitigate these limitations.</p><p>SN2N's optimisation for 5D imaging (xyz-colour-time) represents an achievement of our framework. We demonstrated its application in facilitating the first complete observation of mitosis in live cells over a 3-h period, maintaining <100 nm spatial resolution. Given that mitosis is highly susceptible to phototoxicity, where conventional imaging often induces cell cycle arrest, chromosomal missegregation or apoptosis,<span><sup>10</sup></span> this capability to observe the entire process with such fidelity is noteworthy. The reduced phototoxicity afforded by SN2N offers a window into cell division mechanisms and the development of anti-mitotic therapies, pertinent for understanding chromosomal instability disorders and refining targeted cell cycle interventions (Figure 1).</p><p>SN2N exhibits broad compatibility with diverse imaging modalities","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 7","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70390","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144524649","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}

{"title":"EchoBack-CAR T cells: Tuning immunity with sound","authors":"Longwei Liu, Yingxiao Wang","doi":"10.1002/ctm2.70391","DOIUrl":"https://doi.org/10.1002/ctm2.70391","url":null,"abstract":"<p>Despite the transformative success of chimeric antigen receptor T cell (CAR T) therapy in hematologic malignancies, extending this paradigm to solid tumours has proven difficult. Major barriers such as on-target off-tumour toxicity, antigen heterogeneity, T cell exhaustion, and poor cell persistence have limited clinical translation in solid tumours.<span><sup>1, 2</sup></span> In our recent study, we present EchoBack-CAR T cells, a novel synthetic biology platform that integrates genetic circuit engineering with focused ultrasound (FUS) control, to address these core limitations with a non-invasive, spatiotemporally tunable strategy.<span><sup>3</sup></span></p><p>This work builds on our team's efforts at the interface of synthetic immunology, T cell biology and physical modulation based on sound. Previously, we developed the first-generation FUS-CAR, an ultrasound-controllable CAR T platform that significantly improved the safety profile of CAR T therapies by spatially confining activation to the tumour site.<span><sup>4</sup></span> However, we also recognized a limitation: the CAR expression in FUS-CAR T cells was transient and decayed quickly after stimulation which necessary the incorporation of Cre recombinase to enable permeant CAR expression after stimulation. At the same time, our group developed a mammalian cell screening platform for fluorescent biosensor screening and live CAR T cell imaging, which enabled us to dissect endogenous CAR T cell signalling networks with high spatiotemporal resolution.<span><sup>5</sup></span> This insight proved crucial: by mapping the activation dynamics of pathways in CAR T cells engaging tumour antigens, we gained a systems-level understanding of how to harness natural signalling feedback loops to sustain CAR expression.</p><p>The EchoBack CAR design lies the synergy between biophysical control and cellular logic. We first repurposed the high-throughput evolutionary screening platform for promoter screening to identify a highly heat-inducible promoter with minimal basal activity. This promoter is activated specifically when cells are exposed to a short pulse of ultrasound and its generated localized heat (e.g., 43°C for 15 min). The resulting EchoBack-CAR T cells exhibit tightly regulated CAR expression and robustly activated upon ultrasound stimulation at the tumour site. However, achieving durable anti-tumour activity requires more than a transient pulse of expression. We, therefore, incorporated a positive feedback loop into the genetic design. This circuit senses endogenous T cell signalling pathways activated by tumour engagement, specifically NFAT, NF-κB, and cAMP/MAPK pathways, and converts those signals into sustained CAR expression. The result is a synthetic immune cell that amplifies its own activation signal, maintaining cytotoxic function long after the initial FUS trigger without continuous external input.</p><p>This system overcomes a major drawback of traditional inducible CAR design","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 7","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70391","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144524650","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}

{"title":"tRF-59:76-Arg-ACG-1-M2 is upregulated during colorectal carcinogenesis and promotes cell proliferation, migration, and invasion","authors":"Xiangrong Gao, Hao Bai, Yiming Wang, Zhaohui Zhang, Tingting Lin, Hao Fu, Jianhao Xu, Xinglin Fei, Jinhua Yang, Jinghao Sheng, Xiaojiang Ying, Lihua Zhang, Mengling Tang, Jianbing Wang, Kun Chen, Mingjuan Jin","doi":"10.1002/ctm2.70378","DOIUrl":"https://doi.org/10.1002/ctm2.70378","url":null,"abstract":"<p>Dear Editor</p><p>Colorectal cancer (CRC) ranks third in incidence and second in mortality worldwide as of 2022.<span><sup>1</sup></span> Although CRC is driven by the accumulation of diverse molecular alterations,<span><sup>2, 3</sup></span> its underlying mechanisms have not been fully understood. Transfer RNA-derived small RNAs (tsRNAs) participate in diverse physiological and pathological processes by modulating gene expression at both the transcription and post-transcription levels.<span><sup>4, 5</sup></span> Evidence indicates that dysregulation of tsRNAs is widespread across various human diseases and plays a critical role in cancer development.<span><sup>6-8</sup></span> Here, we aimed to profile dynamic changes across all stages of colorectal carcinogenesis in human and to elucidate the biological functions of tRF-59:76-Arg-ACG-1-M2 through experiments.</p><p>A multi-stage study was conducted, encompassing a discovery stage, two validation stages (I and II), in vitro and in vivo experiments, and bioinformatics analyses (Figure S1). Human plasma samples were obtained from the Jiashan cohort, a population-based cohort of CRC in southeast China.<span><sup>9</sup></span> We included healthy controls (HC), non-advanced adenoma (NAA), advanced adenoma (AA), and CRC patients between June 2016 and September 2021, matched by age and sex. Follow-up was continued until death or December 31, 2024.</p><p>In the discovery stage, small RNA sequencing was performed to profile plasma tsRNA expression and identify differentially expressed tsRNAs (fold change ≥1.50, <i>p</i> < .05) (<b>Table</b> <b>S1</b>). The proportion distribution of subtypes and the number of subtypes mapped to tRNA isodecoders across HC, adenoma and cancer groups are presented in <b>Figure</b> <b>S2</b>. tRF-5c was the most abundant subtype in all samples (Figure 1A). We observed significant differences in the expression of 24 tsRNAs between CRC and adenoma, and of 25 tsRNAs between CRC and HC (Figure 1B). Among them, 10 tsRNAs exhibiting higher levels in the CRC group overlapped (Figure 1C,D; <b>Table</b> <b>S2</b>). These tsRNAs were selected as candidates for further validation. Nevertheless, primers were only available for 8 of them (<b>Table</b> <b>S3</b>).</p><p>In addition to the examination of candidate tsRNAs expression across different stages of colorectal carcinogenesis, a two-stage qRT-PCR was conducted. The baseline characteristics did not differ significantly across the four groups in both validation stage I and II (<i>p</i> > .05) (Table 1). In validation stage I, the expression levels of the 8 candidate tsRNAs were quantified among 70 HC, 70 NAA, 70 AA and 70 CRC subjects. tRF-59:76-Arg-ACG-1-M2 and tRF-59:76-Pro-AGG-1-M8 showed significant differences in expression levels across the four groups (<i>p </i>< .05), while the remaining six did not (Figure 2A). The expression levels of these two tsRNAs were further measured in the validation stage II using a ","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 7","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70378","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144519779","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}

{"title":"T-cell differentiation stage block bias confers hypermethylation and mediastinal preference in T-cell lymphoblastic lymphoma","authors":"Jiali Wang,&nbsp;Bo Qian,&nbsp;Xiaowen Yu,&nbsp;Yidan Zhang,&nbsp;Chunlei Zhou,&nbsp;Tingting Yang,&nbsp;Le Xia,&nbsp;Gang Zhang,&nbsp;Yi-Xuan Zhang,&nbsp;Yaping Wang,&nbsp;Yongjun Fang","doi":"10.1002/ctm2.70380","DOIUrl":"https://doi.org/10.1002/ctm2.70380","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>The clinical guideline classifies T-LBL and T-ALL jointly, differentiating them merely by the bone marrow blast cell proportion. However, their distinct clinical manifestations, genetic profiles, and specific pathogenic requirements have prompted us to reevaluate the differences between them.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods and Results</h3>\u0000 \u0000 <p>We established the NCH-TALL-LBL cohort, which includes flow cytometry data and somatic mutation data from our center. Additionally, we collected T-LBL samples and implemented single-cell RNA sequencing and single-cell T-cell receptor sequencing. Combining the single-cell RNA sequencing data of T-ALL, expression array data, flow cytometry data, we discovered that malignant T cells in T-LBL are predominantly in the DN- and DP-stage blocking modes (DP cells dominate). This block mode in T-LBL generates signals that drive the development of an immunosuppressive microenvironment and the mediastinum preference. Additionally, E2F2, an active transcription factor in the DP and DN stages, upregulates the expression of UHRF1, resulting in hypermethylation of tumor suppressor genes. Findings from in vivo and in vitro research clearly show that demethylation therapy targeting this mechanism effectively inhibits tumor proliferation in T-LBL.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>From the perspective of differentiation blockage, T-LBL and T-ALL represent different stages of the same disease, and the stage block bias of T-cell contributes to their heterogeneity.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Key points</h3>\u0000 \u0000 <div>\u0000 <ul>\u0000 \u0000 <li>Malignant T cells in T-LBL are primarily blocked in the DN and DP stages, which contributes to the immunosuppressive TME and mediastinum preference of T-LBL.</li>\u0000 \u0000 <li>The active transcription factor E2F2 in the DP and DN stages upregulates UHRF1 expression, leading to the hypermethylation of tumor suppressor genes in T-LBL.</li>\u0000 \u0000 <li>Demethylation therapy targeting the hypermethylation of tumor suppressor genes mediated by UHRF1 effectively inhibits tumor proliferation in T-LBL.</li>\u0000 </ul>\u0000 </div>\u0000 </section>\u0000 </div>","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 7","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70380","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503047","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}

{"title":"Decoding the pancreatic cancer microenvironment: The multifaceted regulation of microRNAs","authors":"Jie Ji,&nbsp;Dandan Jin,&nbsp;Junpeng Zhao,&nbsp;Xudong Xie,&nbsp;Yujie Jiao,&nbsp;Xuyang He,&nbsp;Yuxuan Huang,&nbsp;Lirong Zhou,&nbsp;Mingbing Xiao,&nbsp;Xiaolei Cao","doi":"10.1002/ctm2.70354","DOIUrl":"https://doi.org/10.1002/ctm2.70354","url":null,"abstract":"<p>Pancreatic cancer (PC) is an extremely deadly type of cancer, and the 5-year survival rate remains less than 10%. The tumour microenvironment (TME) affects the occurrence, progression and treatment outcomes of PC. MicroRNAs (miRNAs) are essential to regulate PC TME. This review delves into the different roles of miRNAs in the PC TME, including exosome communication, angiogenesis, interactions with cancer-associated fibroblasts, the immunological and neuronal microenvironments and metabolic reprogramming. However, research on the complex regulatory networks and synergistic effects of miRNAs in the TME is still insufficient, and their clinical translation and application face challenges. This review summarised the activities of miRNAs in the PC TME, guiding future research and therapeutic strategies involving miRNAs in PC. Future studies should integrate advanced technologies to decode the spatiotemporal dynamics of miRNA regulation within the TME and develop optimised nanodelivery systems for stable and targeted miRNA delivery, advancing clinical applications in PC treatment.</p>","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 7","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70354","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503046","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}

{"title":"A sex- and gender-informed future for Parkinson's disease care","authors":"Mariapaola Barbato,&nbsp;Beatriz Guzman,&nbsp;Santosh Dixit,&nbsp;Antonella Santuccione Chadha,&nbsp;Roberta Marongiu","doi":"10.1002/ctm2.70382","DOIUrl":"https://doi.org/10.1002/ctm2.70382","url":null,"abstract":"&lt;p&gt;Biological sex and its accompanying hormonal milieu profoundly shape Parkinson's disease (PD) risk and trajectory.&lt;span&gt;&lt;sup&gt;1, 2&lt;/sup&gt;&lt;/span&gt; Higher lifetime estrogen exposure appears neuroprotective, while menstrual-cycle fluctuations and the transition through menopause can significantly modulate PD motor and non-motor symptoms. Conversely, men frequently experience earlier motor onset and more rapid cognitive decline, indicating a distinct clinical course.&lt;span&gt;&lt;sup&gt;1, 3&lt;/sup&gt;&lt;/span&gt; Beyond biology, sociocultural gender norms influence who seeks care, how symptoms are reported, and which resources are accessible. Women with PD often face the longest diagnostic delays, suboptimal treatment and are less likely to access neurologist care or advanced therapies, underscoring the need to confront both biological and social determinants of health.&lt;span&gt;&lt;sup&gt;4&lt;/sup&gt;&lt;/span&gt; Precision medicine mandates that we honor biological diversity and lived experience. Yet, PD females remain underrepresented in preclinical research and clinical trials, and critical hormonal influences are sidelined in research study design and clinical care.&lt;span&gt;&lt;sup&gt;1, 5&lt;/sup&gt;&lt;/span&gt; Castro-Aldrete et al. call for a recalibration of research priorities and clinical frameworks, embedding sex- and gender-informed principles from discovery through policy to achieve equitable, personalized PD care (Figure 1).&lt;/p&gt;&lt;p&gt;Although awareness of sex and gender influences in PD is growing,&lt;span&gt;&lt;sup&gt;6&lt;/sup&gt;&lt;/span&gt; clinical practice remains compartmentalized and continues to lack a sex- and gender-informed approach. Neurological assessments still emphasize on motor symptoms, often overlooking broader health factors, such as hormonal transitions (menstruation, pregnancy, postpartum changes, use of contraceptives and menopause) that critically shape women's symptom profiles and treatment responses.&lt;span&gt;&lt;sup&gt;7&lt;/sup&gt;&lt;/span&gt; Indeed, hormonal states can alter dopaminergic neurotransmission,&lt;span&gt;&lt;sup&gt;3&lt;/sup&gt;&lt;/span&gt; while pharmacokinetic studies reveal that women exhibit greater levodopa bioavailability and a higher risk of dyskinesia, reinforcing the necessity for sex-specific dosing strategies.&lt;span&gt;&lt;sup&gt;8, 9&lt;/sup&gt;&lt;/span&gt; Importantly, these variables are interdependent: while hormones influence symptoms and drug response, drugs may, in turn, interact with hormonal treatments; yet current care models treat them in isolation.&lt;/p&gt;&lt;p&gt;Structural barriers further compound the problem. Many neurologists lack the tools, time, and clinical guidance required to gather and act upon sex-specific information. This represents both an equity issue and an opportunity to advance precision neurology. To overcome these limitations, we envision a truly integrated care model in which neurologists, gynecologists, endocrinologists, and mental-health specialists collaborate to develop holistic treatment plans. This multidisciplinary, collaborative environment will facilitate comprehensive care plans that address both","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 6","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70382","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144367362","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}

{"title":"OXA1L deficiency causes mitochondrial myopathy via reactive oxygen species regulated nuclear factor kappa B signalling pathway","authors":"Yongkun Zhan,&nbsp;Qian Wang,&nbsp;Ya Wang,&nbsp;Yanjie Fan,&nbsp;Dan Yan,&nbsp;Xianlong Lin,&nbsp;Yaoting Chen,&nbsp;Tingting Hu,&nbsp;Nan Li,&nbsp;Weiqian Dai,&nbsp;Hezhi Fang,&nbsp;Yongguo Yu","doi":"10.1002/ctm2.70385","DOIUrl":"https://doi.org/10.1002/ctm2.70385","url":null,"abstract":"&lt;div&gt;\u0000 \u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Background&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;OXA1L is crucial for mitochondrial protein insertion and assembly into the inner mitochondrial membrane, and its variants have been recently linked to mitochondrial encephalopathy. However, the definitive pathogenic link between &lt;i&gt;OXA1L&lt;/i&gt; variants and mitochondrial diseases as well as the underlying pathogenesis remains elusive.&lt;/p&gt;\u0000 &lt;/section&gt;\u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Methods&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;In this study, we identified bi-allelic variants of c.620G&gt;T, p.(Cys207Phe) and c.1163_1164del, p.(Val388Alafs*15) in &lt;i&gt;OXA1L&lt;/i&gt; gene in a mitochondrial myopathy patient using whole exome sequencing. To unravel the genotype–phenotype relationship and underlying pathogenic mechanism between &lt;i&gt;OXA1L&lt;/i&gt; variants and mitochondrial diseases, patient-specific human-induced pluripotent stem cells (hiPSC) were reprogrammed and differentiated into myotubes, while &lt;i&gt;OXA1L&lt;/i&gt; knockout human immortalised skeletal muscle cells (IHSMC) and a conditional skeletal muscle knockout mouse model was generated using clustered regularly interspaced short palindromic repeats/Cas9 genomic editing technology.&lt;/p&gt;\u0000 &lt;/section&gt;\u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Results&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;Both patient-specific hiPSC differentiated myotubes and &lt;i&gt;OXA1L&lt;/i&gt; knockout IHSMC showed combined mitochondrial respiratory chain defects and oxidative phosphorylation (OXPHOS) impairments. Notably, in &lt;i&gt;OXA1L&lt;/i&gt;-knockout IHSMC, transfection of wild-type human OXA1L but not truncated mutant form rescued the respiratory chain defects. Moreover, skeletal muscle conditional &lt;i&gt;Oxa1l&lt;/i&gt; knockout mice exhibited OXPHOS deficiencies and skeletal muscle morphofunctional abnormalities, recapitulating the phenotypes of mitochondrial myopathy. Further functional investigations revealed that impaired OXPHOS resulting of OXA1L deficiency led to elevated reactive oxygen species production, which possibly activated the nuclear factor kappa B signalling pathway, triggering cell apoptosis.&lt;/p&gt;\u0000 &lt;/section&gt;\u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Conclusions&lt;/h3&gt;\u0000 \u0000 &lt;p&gt;Together, our findings reinforce the genotype–phenotype association between &lt;i&gt;OXA1L&lt;/i&gt; variations and mitochondrial diseases and further delineate the potential molecular mechanisms of how OXA1L deficiency causes skeletal muscle deficits in mitochondrial myopathy.&lt;/p&gt;\u0000 &lt;/section&gt;\u0000 \u0000 &lt;section&gt;\u0000 \u0000 &lt;h3&gt; Keypoints&lt;/h3&gt;\u0000 \u0000 &lt;div&gt;\u0000 &lt;ol&gt;\u0000 \u0000 &lt;li&gt;\u0000 &lt;p&gt;&lt;i&gt;OXA1L&lt;/i&gt; gene bi-allelic variants cause mitochondrial myopathy.&lt;","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 6","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70385","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144367360","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}

{"title":"In vitro reconstitution of SPO11-mediated DNA cleavage reveals mechanistic and clinical insights into meiotic DNA double-strand break formation","authors":"Zetao Hu,&nbsp;Xinzhe Tang,&nbsp;Ming-Han Tong,&nbsp;Ying Huang","doi":"10.1002/ctm2.70383","DOIUrl":"https://doi.org/10.1002/ctm2.70383","url":null,"abstract":"&lt;p&gt;Infertility affects 8–12% of couples worldwide, with male factors contributing to up to 50% of cases. Notably, approximately 30% of couples are classified with unexplained infertility despite having normal karyotyping, hormone levels, and semen parameters.&lt;span&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/span&gt; One overlooked but fundamental cause is defective meiosis—-the specialized cell division that halves the chromosome number to produce haploid gametes.&lt;span&gt;&lt;sup&gt;2, 3&lt;/sup&gt;&lt;/span&gt; A critical step in meiosis is the formation of programmed DNA double-strand breaks (DSBs), catalyzed by the topoisomerase-like nuclease SPO11, which licenses homologous recombination and chromosome synapsis.&lt;span&gt;&lt;sup&gt;4&lt;/sup&gt;&lt;/span&gt; SPO11 is essential for meiosis: in mice, for instance, &lt;i&gt;Spo11&lt;/i&gt;-&lt;i&gt;null&lt;/i&gt; males are arrested before the pachytene stage, and females lose nearly all oocytes shortly after birth.&lt;span&gt;&lt;sup&gt;5&lt;/sup&gt;&lt;/span&gt; However, the catalytic architecture of human SPO11, its tolerance to activity loss, and the clinical significance of rare alleles remain poorly defined.&lt;/p&gt;&lt;p&gt;To address this, we established an in vitro reconstitution system for mouse SPO11-mediated DSB formation, confirming its catalytic activity outside a cellular context.&lt;span&gt;&lt;sup&gt;6&lt;/sup&gt;&lt;/span&gt; Notably, two other groups independently reported similar findings in parallel studies.&lt;span&gt;&lt;sup&gt;7, 8&lt;/sup&gt;&lt;/span&gt; Biochemical characterization revealed that although the purified SPO11 and its partner TOP6BL predominantly exist as heterodimers, cross-linking mass spectrometry, pull-down assays, and co-immunoprecipitation (Co-IP) identified a minor population of heterotetrameric complexes.&lt;span&gt;&lt;sup&gt;6&lt;/sup&gt;&lt;/span&gt; This tetrameric assembly is widely recognized as the core catalytic framework required for DNA cleavage.&lt;/p&gt;&lt;p&gt;Incubation of the purified SPO11-TOP6BL complex with both supercoiled and linear DNA substrates resulted in detectable DSBs via gel electrophoresis. Treatment of the reaction products with proteinase K or human TDP2, a tyrosyl-DNA phosphodiesterase, demonstrated that SPO11 forms a covalent phosphotyrosyl linkage with the 5′ end of the cleaved DNA, consistent with its topoisomerase-like activity. Structural alignment between the AlphaFold 3 predicted SPO11 complex and African swine fever virus topoisomerase II (AsfvTop2) revealed two adjacent conserved residues, Y137 and Y138.&lt;span&gt;&lt;sup&gt;9&lt;/sup&gt;&lt;/span&gt; Site-directed mutagenesis demonstrated that only Y138—-not Y137—-is essential for forming the covalent linkage with DNA, identifying Y138 as the catalytic nucleophile. This was further supported by biochemical assays using 46 bp Cy3-labeled DNA substrates. These findings provide a direct molecular visualization of SPO11's catalytic mechanism and significantly advance the understanding of meiotic recombination.&lt;/p&gt;&lt;p&gt;Further structural alignment highlighted three conserved acidic residues (E224, D277 and D279) likely involved in Mg&lt;sup&gt;2+&lt;/sup&gt; coordination essential for catalysis. Mutational analys","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 6","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70383","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144339453","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}

{"title":"Programmed death-ligand 1 expression and prognostic significance in bevacizumab treated ovarian cancer patients: Results from the phase IV MITO16A/MaNGO OV-2 translational study","authors":"Francesca Basso-Valentina,&nbsp;Vincenzo Canzonieri,&nbsp;Rossella De Cecio,&nbsp;Laura Arenare,&nbsp;Domenica Lorusso,&nbsp;Sabrina Chiara Cecere,&nbsp;Eliana Pivetta,&nbsp;Daniela Russo,&nbsp;Annabella Di Mauro,&nbsp;Grazia Artioli,&nbsp;Anna Spina,&nbsp;Carmine De Angelis,&nbsp;Daniela Califano,&nbsp;Saverio Cinieri,&nbsp;Giosuè Scognamiglio,&nbsp;Vanda Salutari,&nbsp;Paolo Chiodini,&nbsp;Francesco Perrone,&nbsp;Sandro Pignata,&nbsp;Gustavo Baldassarre","doi":"10.1002/ctm2.70373","DOIUrl":"https://doi.org/10.1002/ctm2.70373","url":null,"abstract":"&lt;p&gt;Dear Editor,&lt;/p&gt;&lt;p&gt;We are pleased to present our latest analyses, showing that multiple immunofluorescence (MIF) can be used in large multicenter clinical trials to define the prognostic/predictive value of programmed death-ligand 1 (PD-L1) by concomitantly and precisely assessing its expression and spatial distribution in the tumour tissue, outperforming classical immunohistochemistry (IHC).&lt;/p&gt;&lt;p&gt;Immunotherapy with Immune Checkpoint Inhibitors (ICIs) has revolutionized cancer treatment, becoming the primary therapeutic option for several types of tumours.&lt;span&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/span&gt; One of the main ICIs targets is the PD-1/PD-L1 axis, and the assessment of PD-L1 expression by IHC is used as a predictive biomarker of response. Although ICIs have not proven successful for Epithelial Ovarian Cancer (EOC) patients and PD-L1 expression did not show a predictive value,&lt;span&gt;&lt;sup&gt;2-4&lt;/sup&gt;&lt;/span&gt; new results of the Keynote-B96 trial suggest otherwise and support the research for better biomarkers of ICIs activity. Accordingly, while new studies have explored the combination of ICIs with targeted agents in EOC—such as PARP inhibitors and the anti-angiogenic agent bevacizumab (BEV)—and are supported by growing evidence that BEV can modulate the tumour microenvironment (TME) and enhance ICIs efficacy through synergistic effects, these efforts have largely failed due to the lack of reliable predictive biomarkers to identify patients most likely to benefit.&lt;span&gt;&lt;sup&gt;5, 6&lt;/sup&gt;&lt;/span&gt; Here, we analyzed by Multiple Immuno-Fluorescence (MIF) the expression and spatial localization of immune cells and PD-L1 in 292 samples from patients enrolled in the prospective phase IV MITO16A-MaNGO OV-2 clinical trial, which aimed to explore the prognostic role of selected clinical and biological factors in EOC patients treated in first line with standard chemotherapy plus BEV (Table 1 and Figure S1).&lt;span&gt;&lt;sup&gt;7&lt;/sup&gt;&lt;/span&gt; The choice of using MIF relies on the possibility of precisely assessing biomarkers’ spatial distribution in the tissue and of defining which cell types express PD-L1 in the tumour or the surrounding stroma.&lt;/p&gt;&lt;p&gt;We first compared two different anti-PD-L1 antibodies, and assessed their concordance in scoring PD-L1 on 100 selected samples, using IHC which is the gold standard for PD-L1 evaluation in the clinic.&lt;span&gt;&lt;sup&gt;8&lt;/sup&gt;&lt;/span&gt; Statistical analyses showed high correlation and concordance between two independent pathologists and the two antibodies (Figure 1A–G). The anti-PD-L1 E1L3N antibody demonstrated slightly better performance in scoring PD-L1 positivity and was selected for MIF analyses (Figure 1H and Figure S2). The correlation and the agreement between MIF and IHC using the E1L3N antibody were very high, demonstrating that the evaluation of PD-L1 by MIF is comparable to classical IHC (Figure 1I-L) although, as expected, the percentage of positive cells decreased approximately 40-fold in MIF computer-assisted count compared to the human","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 6","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70373","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144339456","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}

{"title":"The discovery of Cas9's trans-cleavage activity: Unlocking new molecular diagnostic tools","authors":"Ying Chen,&nbsp;Jiyun Chen,&nbsp;Liang Liu","doi":"10.1002/ctm2.70384","DOIUrl":"https://doi.org/10.1002/ctm2.70384","url":null,"abstract":"&lt;p&gt;Rapid and accurate nucleic acid testing is of vital importance in clinical treatment for disease prevention, diagnosis and prognosis judgment.&lt;span&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/span&gt; Since the type I,&lt;span&gt;&lt;sup&gt;2&lt;/sup&gt;&lt;/span&gt; V&lt;span&gt;&lt;sup&gt;3&lt;/sup&gt;&lt;/span&gt; and VI&lt;span&gt;&lt;sup&gt;4&lt;/sup&gt;&lt;/span&gt; CRISPR-Cas effector nucleases showed &lt;i&gt;trans&lt;/i&gt;-cleavage activity for non-specific nucleic acids after target binding, researchers have developed a series of efficient nucleic acid detection tools, which can achieve high sensitivity and specificity in pathogen detection.&lt;span&gt;&lt;sup&gt;3, 5, 6&lt;/sup&gt;&lt;/span&gt; Recently, our research innovatively revealed that the type II CRISPR-Cas9 system possesses &lt;i&gt;trans&lt;/i&gt;-cleavage activity with multiple target activation and multiple cutting substrates.&lt;span&gt;&lt;sup&gt;7&lt;/sup&gt;&lt;/span&gt; By combining the &lt;i&gt;trans&lt;/i&gt;-cleavage activity of Cas9 with nucleic acid amplification technology, we successfully developed a new nucleic acid detection platform, expanding the application of the CRISPR–Cas9 system from gene editing to nucleic acid-based diagnostics.&lt;/p&gt;&lt;p&gt;In recent years, molecular diagnostic techniques based on the CRISPR–Cas system have made breakthrough progress due to their unique nucleic acid targeting and cutting activity. Research reports that the effector protein Cas13a of the type VI CRISPR–Cas system, activates the &lt;i&gt;trans&lt;/i&gt;-cleavage activity towards non-specific single-stranded RNA (ssRNA) after specifically recognizing the target RNA.&lt;span&gt;&lt;sup&gt;5&lt;/sup&gt;&lt;/span&gt; Similarly, the effector proteins of the type V CRISPR–Cas system also exhibit the &lt;i&gt;trans&lt;/i&gt;-cleavage ability towards non-specific single-stranded DNA (ssDNA) upon target DNA binding.&lt;span&gt;&lt;sup&gt;3&lt;/sup&gt;&lt;/span&gt;&lt;/p&gt;&lt;p&gt;By combining this characteristic with isothermal amplification technology, researchers have successfully developed a variety of nucleic acid detection platforms, including SHERLOCK and DETECTR.&lt;span&gt;&lt;sup&gt;5, 8, 3&lt;/sup&gt;&lt;/span&gt; These breakthrough technologies effectively overcomethe dependence of traditional PCR methods on profssional equipment and shorten detection time, demonstrating significant advantages in portability and reaction speed. However, existing systems still have room for improvement in detection sensitivity, multiplex detection capabilities and clinical validation. To further promote the development of molecular diagnostic technologies and achieve true point-of-care (POC) testing, it is particularly important to develop more CRISPR–Cas systems with application potential.&lt;span&gt;&lt;sup&gt;5&lt;/sup&gt;&lt;/span&gt;&lt;/p&gt;&lt;p&gt;However, developing technologies that can achieve POC detection faces multiple key challenges. First, the existing amplification-free detection techniques are difficult to achieve clinical-grade detection sensitivity while ensuring rapid response. Second, mainstream CRISPR detection technologies (such as the Cas12 and Cas13 systems) have significant barriers to technological transformation, which directly leads to high costs of diagnostic reagents and limited","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"15 6","pages":""},"PeriodicalIF":7.9,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70384","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144323482","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}