Clinical and Translational Medicine最新文献

{"title":"TCRvβ8 chimeric antigen receptor natural killer cells exhibit potent preclinical activity against T-cell malignancies","authors":"Lianjun He, Yinmei He, Ye He, Xing Bao, Yuqiong Yang, Xueyi Qian, Ziyun Lin, Weijie He, Yao Wu, Huimin Shao, Lingjie Zhou, Lin Wan, Zhenyu Xu","doi":"10.1002/ctm2.70004","DOIUrl":"10.1002/ctm2.70004","url":null,"abstract":"<p>Dear Editor</p><p>T-cell malignancies, such as mainly T-cell lymphomas and T-cell acute lymphoblastic leukaemia (T-ALL),<span><sup>1</sup></span> are often associated with poor prognosis.<span><sup>2</sup></span> The effectiveness of immunotherapy for treating T cell leukaemias was not promising.<span><sup>3</sup></span> Recent research on therapeutic targets against T-cell malignancies has primarily focused on CD5 or CD7.<span><sup>4, 5</sup></span> However, targeting these T-cell antigens has led to the occurrence of T-cell disorders due to immune impairment. To address the above problems, we developed a chimeric antigen receptor-natural killer (CAR-NK) platform specifically eliminating malignant TCRvβ8 T-cells while preserving the majority of normal T-cells to avoid immune dysfunction. Both normal and malignant T-cells express a unique TCR β chain,<span><sup>6</sup></span> and clonal expansions of one or more TCRs are often observed in cases of T-cell malignancies,<span><sup>7</sup></span> making the TCR β chain an effective target for CAR therapy. While, there have been multiple reports about CAR-T therapy for targeting TCRvβ,<span><sup>8, 9</sup></span> using NK cells instead of autologous T-cells for CAR-T preparation, not only avoids the risk of contamination by malignant cells in the final product but also prevents fratricide during CAR-T preparation. Additionally, NK cells from healthy individuals have higher vitality and safety.</p><p>We utilized a lentiviral system to construct four TCRvβ8 CAR-NKs (Figure 1A). Among them, 4-1BB-CD3ζ exhibited higher transduction efficiency (Figure 1B and Figure S1A) and greater levels of cytotoxicity (Figure 1E), while showing no significant difference in terms of NK proportion (Figure 1C and Figure S1B) and expansion fold (Figure 1D). Therefore, we selected 4-1BB-CD3ζ CAR for further study.</p><p>The phenotype of CAR-NKs and Mock-NKs is similar (Figure S2A), while an increase in CD107a and interferon-gamma (IFN-γ) expression on CAR-NKs was induced after co-culture with Jurkat cells (Figure S2B). CAR-NKs exhibited promising cytotoxicity against TCRvβ8 positive cells (Figure 1F,G and Figure S3A,B) while having no killing effect on TCRvβ8<sup>−</sup> cells (Figure 1H,J and Figure S3C–E). Furthermore, to assess the activity of CAR-NKs against malignant T cells from a lymphoma patient, peripheral blood was collected and it was observed that CD3(+)/TCRvβ8(+) positive cells accounted for up to 80% by flow cytometry (Figure 1K). CAR-NKs exhibited an enhanced ability to eliminate malignant T cells compared to Mock-NKs (Figure 1L,M). Taken together, Vβ8-CAR-NKs may specifically target Vβ8<sup>+</sup> T leukaemia cells in vitro.</p><p>To monitor CAR-NK expansion and persistence, a repeated antigen stimulation protocol using Jurkat cells was developed (Figure S4A). TCRvβ8 CAR-NKs got an enrichment of CAR<sup>+</sup> cells and a continuous amplification after antigen stimulation (Figure S4B,C). Furthermore, CAR","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142046444","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":"PUS7-dependent pseudouridylation of ALKBH3 mRNA inhibits gastric cancer progression","authors":"Yongxia Chang, Hao Jin, Yun Cui, Feng Yang, Kanghua Chen, Wenjun Kuang, Chunxiao Huo, Zhangqi Xu, Ya Li, Aifu Lin, Bo Yang, Wei Liu, Shanshan Xie, Tianhua Zhou","doi":"10.1002/ctm2.1811","DOIUrl":"10.1002/ctm2.1811","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>RNA pseudouridylation is a critical post-transcriptional modification that influences gene expression and impacts various biological functions. Despite its significance, the role of mRNA pseudouridylation in cancer remains poorly understood. This study investigates the impact of pseudouridine synthase 7 (PUS7)-mediated pseudouridylation of Alpha-ketoglutarate-dependent Dioxygenase alkB Homolog 3 (<i>ALKBH3</i>) mRNA in gastric cancer.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Immunohistochemistry and Western blotting were used to assess PUS7 protein levels in human gastric cancer tissues. The relationship between PUS7 and gastric cancer progression was examined using 3D colony formation assays and subcutaneous xenograft models. Real-time quantitative PCR (RT-qPCR), Western blotting, and polysome profiling assays were conducted to investigate how PUS7 regulates ALKBH3. A locus-specific pseudouridine (Ψ) detection assay was used to identify Ψ sites on <i>ALKBH3</i> mRNA.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Our findings indicate a significant reduction of PUS7 in gastric cancer tissues compared to adjacent non-tumour tissues. Functional analyses reveal that PUS7 inhibits gastric cancer cell proliferation and tumour growth via its catalytic activity. Additionally, PUS7 enhances the translation efficiency of <i>ALKBH3</i> mRNA by modifying the U696 site with pseudouridine, thereby attenuating tumour growth. Importantly, ALKBH3 functions as a tumour suppressor in gastric cancer, with its expression closely correlated with PUS7 levels in tumour tissues.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>PUS7-dependent pseudouridylation of <i>ALKBH3</i> mRNA enhances its translation, thereby suppressing gastric cancer progression. These findings highlight the potential significance of mRNA pseudouridylation in cancer biology and suggest a therapeutic target for gastric cancer.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Highlights</h3>\u0000 \u0000 <div>\u0000 <ul>\u0000 \u0000 <li>PUS7 enhances the translation efficiency of ALKBH3 through its pseudouridylation activity on ALKBH3 mRNA, thereby inhibiting gastric tumourigenesis.</li>\u0000 \u0000 <li>The expression levels of PUS7 and ALKBH3 are significantly correlated in gastric tumours, which may be potential prognostic predictors and therapeutic targets for patients with gastric cancer.</li>\u0000 ","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.1811","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142035424","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":"Single-nucleus and spatial transcriptome reveal adrenal homeostasis in normal and tumoural adrenal glands","authors":"Barbara Altieri,&nbsp;A. Kerim Secener,&nbsp;Somesh Sai,&nbsp;Cornelius Fischer,&nbsp;Silviu Sbiera,&nbsp;Panagiota Arampatzi,&nbsp;Stefan Kircher,&nbsp;Sabine Herterich,&nbsp;Laura-Sophie Landwehr,&nbsp;Sarah N. Vitcetz,&nbsp;Caroline Braeuning,&nbsp;Martin Fassnacht,&nbsp;Cristina L. Ronchi,&nbsp;Sascha Sauer","doi":"10.1002/ctm2.1798","DOIUrl":"10.1002/ctm2.1798","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <p>The human adrenal gland is a complex endocrine tissue. Studies on adrenal renewal have been limited to animal models or human foetuses. Enhancing our understanding of adult human adrenal homeostasis is crucial for gaining insights into the pathogenesis of adrenal diseases, such as adrenocortical tumours.</p>\u0000 \u0000 <p>Here, we present a comprehensive cellular genomics analysis of the adult human normal adrenal gland, combining single-nuclei RNA sequencing and spatial transcriptome data to reconstruct adrenal gland homeostasis. As expected, we identified primary cells of the various zones of the adrenal cortex and medulla, but we also uncovered additional cell types. They constitute the adrenal microenvironment, including immune cells, mostly composed of a large population of M2 macrophages, and new cell populations, including different subpopulations of vascular-endothelial cells and cortical-neuroendocrine cells. Utilizing spatial transcriptome and pseudotime trajectory analysis, we support evidence of the centripetal dynamics of adrenocortical cell maintenance and the essential role played by Wnt/β-catenin, sonic hedgehog, and fibroblast growth factor pathways in the adult adrenocortical homeostasis. Furthermore, we compared single-nuclei transcriptional profiles obtained from six healthy adrenal glands and twelve adrenocortical adenomas. This analysis unveiled a notable heterogeneity in cell populations within the adenoma samples. In addition, we identified six distinct adenoma-specific clusters, each with varying distributions based on steroid profiles and tumour mutational status.</p>\u0000 \u0000 <p>Overall, our results provide novel insights into adrenal homeostasis and molecular mechanisms potentially underlying early adrenocortical tumorigenesis and/or autonomous steroid secretion. Our cell atlas represents a powerful resource to investigate other adrenal-related pathologies.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11338279/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142016526","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":"Restoring expression of tumour suppressor PTEN by engineered circular RNA-enhanced Osimertinib sensitivity in non-small cell lung cancer","authors":"Haoran Li,&nbsp;Zheng Liu,&nbsp;Shaoyi Chen,&nbsp;Jingsheng Cai,&nbsp;Peiyu Wang,&nbsp;Kezhong Chen,&nbsp;Mantang Qiu","doi":"10.1002/ctm2.1792","DOIUrl":"https://doi.org/10.1002/ctm2.1792","url":null,"abstract":"&lt;p&gt;Dear Editor,&lt;/p&gt;&lt;p&gt;This study provides a new strategy to construct circular RNA (circRNA) in vitro named NeoAna, with splicing sites concealed in CVB3_IRES. Re-storing phosphatase and tensin homologue deleted on chromosome 10 (PTEN) expression by engineered circRNA enhances sensitivity to Osimertinib in non-small lung cancer (NSCLC).&lt;/p&gt;&lt;p&gt;Previous Anabaena permuted intron-exon system could permit the circularisation of sequences up to 5 kb in length, significantly longer than previously reported; however, it is important to acknowledge the presence of ‘scar sequences’ in the final products (Figure 1A).&lt;span&gt;&lt;sup&gt;1-3&lt;/sup&gt;&lt;/span&gt; We designed the NeoAna systems to synthesise circRNAs (Figure 1B) without scar sequences (Figure S1). As shown in Figure 1C, the circRNA (enhanced green fluorescent protein [EGFP], as an example) is clearly observed and resistance to RNase R treatment. Indeed, the formation of circRNA was further confirmed by PCR and the exact splicing site was determined by Sanger sequencing (Figure 1D,E). Successful protein translation was confirmed in cells (Figure 1F,H). Then, in vitro transcription (IVT) products of NeoAna system were subjected to high-performance liquid chromatography and each fraction was transfected into 293T cells, and the main peak showed strongest protein expression (Figure 1G,I‒K). Then, we synthesised pseudo-uridine-modified linear EGFP (m1ψ-EGFP), cEGFP_Ana and cEGFP_NeoAna (Figure S2A) and transfected three RNAs into 293T and H1299 cells and green fluorescence and protein expression were observed in cells (Figure S2B,C). Compared with cEGFP_Ana, cEGFP_NeoAna induced weaker innate immunity response in 293T cells (Figure 1L). Besides, we found that the stability of cEGFP_NeoAna is comparable to that of cEGFP_Ana (Figure S3A).&lt;/p&gt;&lt;p&gt;The well-known tumour suppressor, PTEN is a negative regulator of epidermal growth factor receptor (EGFR) signalling pathway,&lt;span&gt;&lt;sup&gt;4&lt;/sup&gt;&lt;/span&gt; and PTEN protein expression is often lost in lung cancer.&lt;span&gt;&lt;sup&gt;5&lt;/sup&gt;&lt;/span&gt; Thus, restoring PTEN expression might reverse EGFR-TKI resistance.&lt;span&gt;&lt;sup&gt;6&lt;/sup&gt;&lt;/span&gt; We synthesised PTEN protein template with NeoAna system (Figure S3B‒E) and long-lasting PTEN protein expression was observed. Osimertinib-resistant cells were established in HCC827 and PC9 cells, since they harbour EGFR exon 19 deletion (Figure S4A‒E).&lt;/p&gt;&lt;p&gt;Cell Counting Kit-8 (CCK-8), colony formation and 5-ethynyl-2'-deoxyuridine (EdU) assays showed that cPTEN_NeoAna increased sensitivity to Osimertinib compared with control group (Figures 2A‒D, S4A and S3D). Annexin V-FITC and TUNEL assay both confirmed that apoptosis rate was increased by elevating the concentration of Osimertinib and the transfection of cPTEN_NeoAna (Figures 2E,F and S5) in PC9 Osimertinib-resistance (PC9OR) and HCC827 Osimertinib-resistance (HCC827OR) cells.&lt;/p&gt;&lt;p&gt;The cEGFP_NeoAna was encapsulated by lipid nanoparticles (LNP) to form the stable complex and observed under electron micr","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.1792","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142013671","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":"Extracellular vesicles as the next-generation modulators of pharmacokinetics and pharmacodynamics of medications and their potential as adjuvant therapeutics","authors":"Jiaqi Liu,&nbsp;Joel Z. Nordin,&nbsp;Andrew J. McLachlan,&nbsp;Wojciech Chrzanowski","doi":"10.1002/ctm2.70002","DOIUrl":"https://doi.org/10.1002/ctm2.70002","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background and main body</h3>\u0000 \u0000 <p>Pharmacokinetics (PK) and pharmacodynamics (PD) are central concepts to guide the dosage and administration of drug therapies and are essential to consider for both healthcare professionals and researchers in therapeutic planning and drug discovery. PK/PD properties of a drug significantly influence variability in response to treatment, including therapeutic failure or excessive medication-related harm. Furthermore, suboptimal PK properties constitute a significant barrier to further development for some candidate treatments in drug discovery. This article describes how extracellular vesicles (EVs) affect different aspects of PK and PD of medications and their potential to modulate PK and PD properties to address problematic PK/PD profiles of drugs. We reviewed EVs' intrinsic effects on cell behaviours and medication responses. We also described how surface and cargo modifications can enhance EV functionalities and enable them as adjuvants to optimise the PK/PD profile of conventional medications. Furthermore, we demonstrated that various bioengineering strategies can be used to modify the properties of EVs, hence enhancing their potential to modulate PK and PD profile of medications.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>This review uncovers the critical role of EVs in PK and PD modulation and motivates further research and the development of assays to unfold EVs’ full potential in solving PK and PD-related problems. However, while we have shown that EVs play a vital role in modulating PK and PD properties of medications, we postulated that it is essential to define the context of use when designing and utilising EVs in pharmaceutical and medical applications.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Highlights</h3>\u0000 \u0000 <div>\u0000 <ul>\u0000 \u0000 <li>Existing solutions for pharmacokinetics and pharmacodynamics modulation are limited.</li>\u0000 \u0000 <li>Extracellular vesicles can optimise pharmacokinetics as a drug delivery vehicle.</li>\u0000 \u0000 <li>Biogenesis and administration of extracellular vesicles can signal cell response.</li>\u0000 \u0000 <li>The pharmaceutical potential of extracellular vesicles can be enhanced by surface and cargo bioengineering.</li>\u0000 \u0000 <li>When using extracellular vesicles as modulators of pharmacokinetics and pharmacodynamics, the ‘context of use’ must be considered.</li>\u0000 </ul>\u0000 </div>\u0000 </section>\u0000 </div>","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142013673","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 healing power of sensory neurons: New horizons for diabetic and neuropathic tissue repair","authors":"Yen-Zhen Lu,&nbsp;Sanjay Ramakrishnan,&nbsp;Mikaël M. Martino","doi":"10.1002/ctm2.1813","DOIUrl":"https://doi.org/10.1002/ctm2.1813","url":null,"abstract":"&lt;p&gt;Tissue repair and regeneration after injury is a highly complex process involving the coordination of multiple biological systems. Therefore, successful regenerative medicine strategies should harness the key mechanisms that control the tissue healing process, particularly when these mechanisms are disrupted by pathological conditions that impede normal healing. Nociceptive sensory neurons, or nociceptors, are specialized primary sensory neurons with nerve endings in tissues such as skin, muscles, and joints that detect and respond to noxious stimuli, including inflammatory mediators.&lt;span&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/span&gt; Although nociceptors have been shown to have pro-inflammatory activities in some contexts, they generally mediate anti-inflammatory processes&lt;span&gt;&lt;sup&gt;1, 2&lt;/sup&gt;&lt;/span&gt; and their activation has been shown to be involved in skin wound healing.&lt;span&gt;&lt;sup&gt;3, 4&lt;/sup&gt;&lt;/span&gt;&lt;/p&gt;&lt;p&gt;Lu et al. investigated the role of peptidergic nociceptive sensory neurons in tissue repair and regeneration following acute injury in mice, exploring whether neuro-immune interactions could be harnessed to promote tissue healing.&lt;span&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/span&gt; They found that nociceptors extend their nerve endings into injured skin and muscle after acute injury and release calcitonin gene-related peptide (CGRP). CGRP from sensory neurons modulates neutrophils and monocytes/macrophages—the majority of immune cells accumulating in injured tissues—to create an anti-inflammatory and pro-healing environment. Mechanistically, the immunomodulatory and pro-healing effects of CGRP were mediated by the release of the extracellular matrix protein thrombospondin-1 (TSP-1) from neutrophils and macrophages, although CGRP may also exert direct effects. TSP-1 was shown to act in an autocrine/paracrine manner, promoting neutrophil efferocytosis by macrophages (clearance by engulfment) and polarizing macrophages to a pro-repair phenotype. Both processes are crucial to proper and timely tissue healing. In addition, TSP-1 was found to accelerate the death of neutrophil and pro-inflammatory macrophage , which could further contribute to reducing inflammation (Figure 1A). Despite these findings, a few questions still remain. For example, the precise mechanisms by which nociceptors are activated after acute injury and how nerve endings grow into injured tissue are not fully understood. Additionally, there may be critical feedback interactions between immune cells and nociceptors both at the injury site and in the dorsal root ganglia, where the cell bodies of sensory neurons are located. Furthermore, the neuro-immune-regenerative axis may also be significant in other tissues or contexts that involve similar mechanisms to tissue healing, such as cancer&lt;span&gt;&lt;sup&gt;2&lt;/sup&gt;&lt;/span&gt; and fibrotic tissue formation, both of which rely heavily on immunoregulation.&lt;/p&gt;&lt;p&gt;The discovery of a neuro-immune-regenerative axis mediated by CGRP is particularly relevant for patients with diabetes, as more than half","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.1813","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142013672","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":"Potential mechanisms of cancer stem-like progenitor T-cell bio-behaviours","authors":"Ling Ni","doi":"10.1002/ctm2.1817","DOIUrl":"10.1002/ctm2.1817","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <p>In situations involving continuous exposure to antigens, such as chronic infections or cancer, antigen-specific CD8⁺ T cells can become dysfunctional or exhausted. This change is marked by increased expression levels of inhibitory receptors (PD-1 and Tim-3). Stem-like progenitor exhausted (Tpex) cells, a subset of exhausted cells that express TCF-1 and are mainly found in the lymph nodes, demonstrate the ability to self-renew and exhibit a high rate of proliferation. Tpex cells can further differentiate into transitional intermediate exhausted (Tex-int) cells and terminally exhausted (Tex-term) cells. Alternatively, they can directly differentiate into Tex-term cells. Tpex cells are the predominant subset that respond to immune checkpoint inhibitors (ICI), making them a prime candidate for improving the efficacy of ICI therapy. This review article aimed to present the latest developments in the field of Tpex formation, expansion, and differentiation in the context of cancer, as well as their responses to ICIs in cancer immunotherapy. Consequently, it may be possible to develop novel treatments that exclusively target Tpex cells, thus improving overall treatment outcomes.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Key points</h3>\u0000 \u0000 <div>\u0000 <ul>\u0000 \u0000 <li>\u0000 <p>Tpex cells are located in lymph nodes and TLS.</p>\u0000 </li>\u0000 \u0000 <li>\u0000 <p>Several pathways control the differentiation trajectories of Tpex cells, including epigenetic factors, transcription factors, cytokines, age, sex, etc.</p>\u0000 </li>\u0000 </ul>\u0000 </div>\u0000 </section>\u0000 </div>","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.1817","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142016525","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":"Cyclic-di-GMP induces inflammation and acute lung injury through direct binding to MD2","authors":"Chenchen Qian,&nbsp;Weiwei Zhu,&nbsp;Jiong Wang,&nbsp;Zhe Wang,&nbsp;Weiyang Tang,&nbsp;Xin Liu,&nbsp;Bo Jin,&nbsp;Yong Xu,&nbsp;Yuyang Zhang,&nbsp;Guang Liang,&nbsp;Yi Wang","doi":"10.1002/ctm2.1744","DOIUrl":"https://doi.org/10.1002/ctm2.1744","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Severe bacterial infections can trigger acute lung injury (ALI) and acute respiratory distress syndrome, with bacterial pathogen-associated molecular patterns (PAMPs) exacerbating the inflammatory response, particularly in COVID-19 patients. Cyclic-di-GMP (CDG), one of the PAMPs, is synthesized by various Gram-positve and Gram-negative bacteria. Previous studies mainly focused on the inflammatory responses triggered by intracellular bacteria-released CDG. However, how extracellular CDG, which is released by bacterial autolysis or rupture, activates the inflammatory response remains unclear.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>The interaction between extracellular CDG and myeloid differentiation protein 2 (MD2) was investigated using in vivo and in vitro models. MD2 blockade was achieved using specific inhibitor and genetic knockout mice. Site-directed mutagenesis, co-immunoprecipitation, SPR and Bis-ANS displacement assays were used to identify the potential binding sites of MD2 on CDG.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Our data show that extracellular CDG directly interacts with MD2, leading to activation of the TLR4 signalling pathway and lung injury. Specific inhibitors or genetic knockout of MD2 in mice significantly alleviated CDG-induced lung injury. Moreover, isoleucine residues at positions 80 and 94, along with phenylalanine at position 121, are essential for the binding of MD2 to CDG.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>These results reveal that extracellular CDG induces lung injury through direct interaction with MD2 and activation of the TLR4 signalling pathway, providing valuable insights into bacteria-induced ALI mechanisms and new therapeutic approaches for the treatment of bacterial co-infection in COVID-19 patients.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.1744","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142013670","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":"Circular RNA LIPH promotes pancreatic cancer glycolysis and progression through sponge miR-769-3p and interaction with GOLM1","authors":"Yan Ma,&nbsp;Xiaomeng He,&nbsp;Yang Di,&nbsp;Wenyang Li,&nbsp;Lixiang Sun,&nbsp;Xin Zhang,&nbsp;Li Xu,&nbsp;Zhihui Bai,&nbsp;Zehuan Li,&nbsp;Lijun Cai,&nbsp;Huaqin Sun,&nbsp;Christopher Corpe,&nbsp;Jin Wang","doi":"10.1002/ctm2.70003","DOIUrl":"https://doi.org/10.1002/ctm2.70003","url":null,"abstract":"&lt;p&gt;Dear Editor,&lt;/p&gt;&lt;p&gt;Pancreatic cancer (PaCa) is a highly malignant tumour of the digestive system and is one of the major causes of cancer-related death worldwide,&lt;span&gt;&lt;sup&gt;1-3&lt;/sup&gt;&lt;/span&gt; and only approximately 10% of PaCa patients survive for 1 year after diagnosis.&lt;span&gt;&lt;sup&gt;4, 5&lt;/sup&gt;&lt;/span&gt; Thus, investigations into sensitive and specific biomarkers for risk stratification are urgently needed for PaCa. Noncoding RNAs, including circRNAs, function as key ceRNAs (sponges) to regulate the expression of mRNAs, and their discovery greatly expanded the functional genetic information in carcinogenesis.&lt;span&gt;&lt;sup&gt;6-8&lt;/sup&gt;&lt;/span&gt; CircRNAs are highly stable, are not easily digested by RNase, and can be detected in the saliva, blood and other body fluids of patients with cancer.&lt;span&gt;&lt;sup&gt;9, 10&lt;/sup&gt;&lt;/span&gt;&lt;/p&gt;&lt;p&gt;In this study, we revealed that a novel circular RNA (circLIPH/circ_0068398) was upregulated in pancreatic tumour tissue (Figure 1A). A high expression level of circLIPH was significantly correlated with tumour size, tumour stage and the percentage of Ki67-positive tumours (&lt;i&gt;p&lt;/i&gt; &lt; .05; Table S1). circLIPH and lipase H (LIPH) expression levels were also significantly greater in most PaCa cells than in the hTERT-immortalized epithelial (HPNE) control cells (Figure 1C,D). circLIPH is derived from exons 2 to 5 of the LIPH gene and has a length of 669 bp (Figure 1E); we amplified it from cDNA (not from gDNA) via divergent primers (Figure 1F). RNA fluorescence in situ hybridisation revealed circLIPH localisation primarily in the cytoplasm of PaCa cells (Figure 1G). We also demonstrated that circLIPH overexpression promoted the growth of BXPC-3 and PANC-1 cells via CCK-8 assays (Figure S1E,F), whereas si-circLIPH treatment significantly inhibited the proliferation of PaCa cells (Figure S1G,H). Colony formation assays (Figure S1I) revealed that circLIPH overexpression could effectively promote the growth of PaCa cells (Figure S1J) and that si-circLIPH treatment inhibited cancer cell proliferation (Figure S1K). Wound healing (Figure S1L) and Transwell invasion assays (Figure S1O) demonstrated that circLIPH overexpression markedly enhanced the migration (Figure S1M) and invasion abilities (Figure S1P) of PaCa cells, whereas si-circLIPH treatment impaired the migration (Figure S1N) and invasion (Figure S1Q) capabilities of PaCa cells. The protein levels of vimentin and Snail increased after circLIPH overexpression in PaCa cells, whereas the protein level of E-cadherin decreased significantly (Figure S1R), which suggests that circLIPH may serve as an oncogene that facilitates cancer cell progression and promotes the epithelial–mesenchymal transition (EMT) of PaCa cells.&lt;/p&gt;&lt;p&gt;To elucidate the molecular mechanism of circLIPH, seven candidate miRNAs were identified from the starBase, circBank and circInteractome databases (Figure 2A). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses revealed that the level of miR-7","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.70003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142013499","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":"Targeting the chromatin remodelling protein Brahma-related gene 1 for intervention of pulmonary fibrosis","authors":"Teng Wu,&nbsp;Bingshu Wang,&nbsp;Xianhua Gui,&nbsp;Ruiqi Liu,&nbsp;Dong Wei,&nbsp;Yong Xu,&nbsp;Shaojiang Zheng,&nbsp;Nan Li,&nbsp;Ming Kong","doi":"10.1002/ctm2.1775","DOIUrl":"https://doi.org/10.1002/ctm2.1775","url":null,"abstract":"&lt;p&gt;Dear Editor,&lt;/p&gt;&lt;p&gt;We describe in this letter a novel mechanism whereby the chromatin remodelling protein Brahma-related gene 1 (BRG1) contributes to pulmonary fibrosis.&lt;/p&gt;&lt;p&gt;Pulmonary fibrosis is a common manifestation of interstitial lung disease (ILD) that affects over 40 million people worldwide.&lt;span&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/span&gt; Although for a majority of patients with pulmonary fibrosis, one or another underlying cause including radiation, hypersensitivity pneumonitis, and pneumoconiosis have been identified, pulmonary fibrosis can occur in certain individuals with no ascribable aetiology; the latter patient group is categorized as idiopathic pulmonary fibrosis (IPF).&lt;span&gt;&lt;sup&gt;2&lt;/sup&gt;&lt;/span&gt; Regardless of aetiology, extracellular matrix (ECM)-producing myofibroblasts is the principal mediator of pulmonary fibrosis.&lt;span&gt;&lt;sup&gt;3&lt;/sup&gt;&lt;/span&gt; Compared to quiescent fibroblasts from which they are derived, myofibroblasts are highly proliferative and migratory, able to perform muscle-like contraction, and markedly more potent in producing ECM proteins.&lt;span&gt;&lt;sup&gt;4&lt;/sup&gt;&lt;/span&gt; BRG1 is part of the epigenetic machinery that shapes the transcriptomic landscape in mammalian cells.&lt;span&gt;&lt;sup&gt;5&lt;/sup&gt;&lt;/span&gt; In the present study, we sought to determine the role of BRG1 in pulmonary fibrosis.&lt;/p&gt;&lt;p&gt;In the first set of experiments, C57/BL6 mice were given bleomycin to induce pulmonary fibrosis followed by isolation of primary pulmonary fibroblasts. Rapid induction of both BRG1 and periostin, a marker for mature myofibroblast, was observed in the fibroblasts isolated from the lungs 1 week after bleomycin instillation (Figure S1A,B). When primary murine pulmonary fibroblasts or human pulmonary fibroblasts (MRC5) were exposed to transforming growth factor-β (TGF-β), BRG1 expression was up-regulated with a similar kinetics as periostin (Figure S1C,D). BRG1 levels were substantially elevated in the lung tissues of IPF patients compared to the healthy individuals (Figure S1E). In addition, a significant correlation was identified between BRG1 expression and periostin expression (Figure S1F).&lt;/p&gt;&lt;p&gt;Next, primary pulmonary fibroblasts were isolated from BRG1&lt;sup&gt;f/f&lt;/sup&gt; mice and induced to differentiate into myofibroblasts by TGF-β treatment; BRG1 deletion by transduction with Cre-delivering adenovirus significantly attenuated myofibroblast marker genes (Figure 1A), cell proliferation (Figure 1B), cell migration (Figure 1C), and cell contraction (Figure 1D). Similarly, BRG1 knockdown by small interfering RNAs in pulmonary fibroblasts from IPF patients markedly decreased myofibroblast marker gene expression and attenuated cell proliferation/migration/contraction (Figure S2). To verify whether BRG1 deletion in myofibroblasts would alter pulmonary fibrosis in vivo, BRG1&lt;sup&gt;f/f&lt;/sup&gt; mice were crossbred with &lt;i&gt;Postn&lt;/i&gt;-Cre&lt;sup&gt;ERT2&lt;/sup&gt; mice to generate myofibroblast conditional BRG1 knockout mice (BRG1&lt;sup&gt;ΔMF&lt;/sup&gt;, Figure 1E). Pulmonary fibrosis, as measured by ","PeriodicalId":10189,"journal":{"name":"Clinical and Translational Medicine","volume":"14 8","pages":""},"PeriodicalIF":7.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/ctm2.1775","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142013662","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}