Vol. 28 (2023)

No. 01  DOI: 10.1186/s11658-022-00409-6 Volume 28 (2023) - 28:01
Authors Linlin Zhang1†, Miaomiao Xu1†, Wanchun Zhang2†, Chuanying Zhu3, Zhilei Cui4, Hongliang Fu1, Yufei Ma1, Shuo Huang1, Jian Cui5*, Sheng Liang1*, Lei Huang6,7* and Hui Wang1*
Abstract Background: Spatial chromatin structure is intricately linked with somatic aberrations, and somatic mutations of various cancer-related genes, termed co-mutations (CoMuts), occur in certain patterns during cancer initiation and progression. The functional mechanisms underlying these genetic events remain largely unclear in thyroid cancer (TC). With discrepant diferentiation, papillary thyroid cancer (PTC) and anaplastic thyroid cancer (ATC) difer greatly in characteristics and prognosis. We aimed to reveal the spatial gene alterations and regulations between the two TC subtypes.
Methods: We systematically investigated and compared the spatial co-mutations between ATC (8305C), PTC (BCPAP and TPC-1), and normal thyroid cells (Nthy-ori-3–1). We constructed a framework integrating whole-genome sequencing (WGS), high-throughput chromosome conformation capture (Hi-C), and transcriptome sequencing, to systematically detect the associations between the somatic co-mutations of cancer-related genes, structural variations (SVs), copy number variations (CNVs), and high-order chromatin conformation.
Results: Spatial co-mutation hotspots were enriched around topologically associating domains (TADs) in TC. A common set of 227 boundaries were identifed in both ATC and PTC, with signifcant overlaps between them. The spatial proximities of the co-mutated gene pairs in the two TC types were signifcantly greater than in the gene-level and overall backgrounds, and ATC cells had higher TAD contact frequency with CoMuts>10 compared with PTC cells. Compared with normal thyroid cells, in ATC the number of the created novel three-dimensional chromatin structural domains increased by 10%, and the number of shifted TADs decreased by 7%. We found fve TAD blocks with CoMut genes/events specifc to ATC with certain mutations in genes including MAST-NSUN4, AM129B/TRUB2, COL5A1/PPP1R26, PPP1R26/GPSM1/CCDC183, and PRAC2/DLX4. For the majority of ATC and PTC cells, the HOXA10 and HIF2α signals close to the transcription start sites of CoMut genes within TADs were signifcantly stronger than those at the background. CNV breakpoints signifcantly overlapped with TAD boundaries in both TC subtypes. ATCs had more CNV losses overlapping with TAD boundaries, and noncoding SVs involved in intrachromosomal SVs, amplifed inversions, and tandem duplication difered between ATC and PTC. TADs with short range were more abundant in ATC than PTC. More switches of A/B compartment types existed in ATC cells compared with PTC. Gene expression was signifcantly synchronized, and orchestrated by complex epigenetics and regulatory elements.
Conclusion: Chromatin interactions and gene alterations and regulations are largely heterogeneous in TC. CNVs and complex SVs may function in the TC genome by interplaying with TADs, and are largely diferent between ATC and PTC. Complexity of TC genomes, which are highly organized by 3D genome-wide interactions mediating mutational and structural variations and gene activation, may have been largely underappreciated. Our comprehensive analysis may provide key evidence and targets for more customized diagnosis and treatment of TC.
Keywords Papillary thyroid cancer, Anaplastic thyroid cancer, Genomic heterogeneity, Topologically associating domains (TADs), High-throughput chromosome conformation capture (Hi-C), A/B compartment switches, Somatic hotspot mutations, structural variations, Copy number variations, RNA sequencing, Whole-genome sequencing
Address and Contact Information 1 Department of Nuclear Medicine, Xinhua Hospital Afliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China.
2 Department of Nuclear Medicine, Shanxi Bethune Hospital (Shanxi Academy of Medical Sciences), Taiyuan 03003, China.
3 Department of Oncology, Xin Hua Hospital Afliated to Shanghai Jiaotong University School of Medicine, Shanghai 200092, China.
4 Department of Respiratory Medicine, XinHua Hospital Afliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China.
5 BioGenius Bioinformatics Institute, Shanghai 200050, People’s Republic of China.
6 Department of Oncology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
7 Medical Center on Aging of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
* Corresponding author: michael@biogenius.cn; liangsheng364214@163.com; lei.huang@alumni.dkfz.de; huiwangshanghai@163.com
Linlin Zhang, Miaomiao Xu, and Wanchun Zhang contributed equally to this work.
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No.02  DOI: 10.1186/s11658-022-00413-w Volume 28 (2023) - 28:02
Authors Seyram Yao Adzraku1,2,3,4†, Guozhang Wang1,2,3,4†, Can Cao1,2,3,4†, Yurong Bao1,2,3, Yizhou Wang1,2,3, Alhaji Osman Smith1,2,3, Yuwei Du1,2,3, Haiyang Wang1,2,3, Yue Li1,2,3, Kailin Xu1,2,3, Jianlin Qiao1,2,3*, Wen Ju1,2,3,4* and Lingyu Zeng1,2,3*
Abstract Background: Hematopoietic stem cell transplantation involves irradiation preconditioning which causes bone marrow endothelial cell dysfunction. While much emphasis is on the reconstitution of hematopoietic stem cells in the bone marrow microenvironment, endothelial cell preservation is indispensable to overcome the preconditioning damages. This study aims to ascertain the role of Roundabout 4 (Robo4) in regulating irradiation-induced damage to the endothelium.
Methods: Microvascular endothelial cells were treated with γ-radiation to establish an endothelial cell injury model. Robo4 expression in the endothelial cells was manipulated employing lentiviral-mediated RNAi and gene overexpression technology before irradiation treatment. The permeability of endothelial cells was measured using qPCR, immunocytochemistry, and immunoblotting to analyze the efect on the expression and distribution of junctional molecules, adherens junctions, tight junctions, and gap junctions. Using Transwell endothelial monolayer staining, FITC-Dextran permeability, and gap junction-mediated intercellular communication (GJIC) assays, we determined the changes in endothelial functions after Robo4 gene manipulation and irradiation. Moreover, we measured the proportion of CD31 expression in endothelial cells by fow cytometry. We analyzed variations between two or multiple groups using Student’s t-tests and ANOVA.
Results: Ionizing radiation upregulates Robo4 expression but disrupts endothelial junctional molecules. Robo4 deletion causes further degradation of endothelial junctions hence increasing the permeability of the endothelial cell monolayer. Robo4 knockdown in microvascular endothelial cells increases the degradation and delocalization of ZO-1, PECAM-1, occludin, and claudin-5 molecules after irradiation. Conversely, connexin 43 expression increases after silencing Robo4 in endothelial cells to induce permeability but are readily destroyed when exposed to 10 Gy of gamma radiation. Also, Robo4 knockdown enhances Y731-VE-cadherin phosphorylation leading to the depletion and destabilization of VE-cadherin at the endothelial junctions following irradiation. However, Robo4 overexpression mitigates irradiation-induced degradation of tight junctional proteins and stabilizes claudin-5 and ZO-1 distribution. Finally, the enhanced expression of Robo4 ameliorates the irradiation-induced depletion of VE-cadherin and connexin 43, improves the integrity of microvascular endothelial cell junctions, and decreases permeability.
Conclusion: This study reveals that Robo4 maintains microvascular integrity after radiation preconditioning treatment by regulating endothelial permeability and protecting endothelial functions. Our results also provided a potential mechanism to repair the bone marrow vascular niche after irradiation by modulating Robo4 expression.
Keywords Robo4, Microvascular endothelial cells, Endothelial junctions, Irradiation, Permeability
Address and Contact Information 1 Blood Diseases Institute, Xuzhou Medical University, Xuzhou 221002, China
2 Key Laboratory of Bone Marrow Stem Cell, Xuzhou 221002, Jiangsu, China
3 Department of Hematology, The Afliated Hospital of Xuzhou Medical University, Xuzhou 221002, China
4 Xuzhou Ruihu Health Management Consulting Co., Ltd, Xuzhou 221002, China
*Corresponding author: jlqiao85@126.com; juwen1987-10@163.com; zengly2000@163.com
Seyram Yao Adzraku, Guozhang Wang and Can Cao have contributed equally to this work
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No. 03 DOI: 10.1186/s11658-023-00419-y Volume 28 (2023) - 28:03
Authors Junqiang Li1†, Yang Song, Chao Zhang1†, Ronglin Wang1, Lei Hua1, Yongdong Guo, Dongxue Gan1, Liaoliao Zhu1, Shanshan Li1, Peixiang Ma1, Cheng Yang1, Hong Li1, Jing Yang1, Jingjie Shi1, Xiaonan Liu2* and Haichuan Su1*
Abstract Correction: Cellular & Molecular Biology Letters (2022) 27:24
Following publication of the original article [1], the authors identifed an error in Fig. 3O. Te invasion image in the shTMEM43+RAP2B group of Fig. 3O was inadvertently placed by mistake. We have double checked the original data and found that the inadvertent errors occurred during picture compilation, and this correction does not change the scientifc conclusions of the article. Te incorrect and the correct figure is given in PDF below.
The online version of the original article can be found at https://doi.org/10.1186/s11658-022-00321-z.
Address and Contact Information 1 Department of Oncology, Tangdu Hospital, Air Force Medical University, Xi’an 710038, Shaanxi, China
2 Ambulatory Surgery Center, Xijing Hospital, Air Force Medical University, Xi’an 710032, Shaanxi, China
*Corresponding author: 15353589999@163.com; suhc@fmmu.edu.cn
Junqiang Li, Yang Song and Chao Zhang contributed equally
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No. 04 DOI: 10.1186/s11658-023-00416-1 Volume 28 (2023) - 28:04
Authors Yirui Cao1,2†, Juntao Chen1,2†, Feng Liu4†, Guisheng Qi1, Yufeng Zhao1,2, Shihao Xu1,2, Jiyan Wang1,2, Tongyu Zhu1,2*†, Yi Zhang3*† and Yichen Jia1*†
Abstract Background: Ischemia–reperfusion injury (IRI) is an inevitable process in renal transplantation that signifcantly increases the risk of delayed graft function, acute rejection, and even graft loss. Formyl peptide receptor 2 (FPR2) is an important receptor in multiple septic and aseptic injuries, but its functions in kidney IRI are still unclear. This study was designed to reveal the pathological role of FPR2 in kidney IRI and its functional mechanisms.
Methods: To explore the mechanism of FPR2 in kidney IRI, the model rats were sacrifced after IRI surgery. Immunofuorescence, enzyme-linked immunosorbent assays, and western blotting were used to detect diferences in the expression of FPR2 and its ligands between the IRI and control groups. WRW4 (WRWWWW-NH2), a specifc antagonist of FPR2, was administered to kidney IRI rats. Kidney function and pathological damage were detected to assess kidney injury and recovery. Flow cytometry was used to quantitatively compare neutrophil infltration among the experimental groups. Mitochondrial formyl peptides (mtFPs) were synthesized and administered to primary rat neutrophils together with the specifc FPR family antagonist WRW4 to verify our hypothesis in vitro. Western blotting and cell function assays were used to examine the functions and signaling pathways that FPR2 mediates in neutrophils.
Results: FPR2 was activated mainly by mtFPs during the acute phase of IRI, mediating neutrophil migration and reactive oxygen species production in the rat kidney through the ERK1/2 pathway. FPR2 blockade in the early phase protected rat kidneys from IRI.
Conclusions: mtFPs activated FPR2 during the acute phase of IRI and mediated rat kidney injury by activating the migration and reactive oxygen species generation of neutrophils through the ERK1/2 pathway.
Keywords Formyl peptide receptor 2, Neutrophil, Kidney IRI, Migration, Mitochondrial-derived formyl peptides
Address and Contact Information 1 Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China
2 Shanghai Key Laboratory of Organ Transplantation, Shanghai, China
3 Zhongshan Hospital Institute of Clinical Science, Zhongshan Hospital, Fudan University, Shanghai, China
4 Department of Integrative Medicine, Huashan Hospital Fudan University, Shanghai, People’s Republic of China
* Correspondence: zs_tyzhu@163.com; yzhang_med@fudan.edu.cn; jia.yichen@zs-hospital.sh.cn
Yirui Cao, Juntao Chen, and Feng Liu contributed equally to this work and share first authorship
Tongyu Zhu, Yi Zhang, and Yichen Jia share last authorship
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No. 05 DOI: 10.1186/s11658-023-00417-0 Volume 28 (2023) - 28:05
Authors Cheng Ju, Yangguang Ma, Xiaoshuang Zuo, Xuankang Wang, Zhiwen Song, Zhihao Zhang, Zhijie Zhu, Xin Li, Zhuowen Liang, Tan Ding, Xueyu Hu* and Zhe Wang*
Abstract Background: Secondary spinal cord injury (SCI) often causes the aggravation of infammatory reaction and nerve injury, which afects the recovery of motor function. Bone-marrow-derived macrophages (BMDMs) were recruited to the injured area after SCI, and the M1 polarization is the key process for inducing infammatory response and neuronal apoptosis. We previously showed that photobiomodulation (PBM) can inhibit the polarization of M1 phenotype of BMDMs and reduce infammation, but the underlying mechanisms are unclear. The purpose of this study is to explore the potential target and mechanism of PBM in treating SCI.
Methods: Transcriptome sequencing and bioinformatics analysis showed that long noncoding RNA taurine upregulated gene 1 (lncRNA TUG1) was a potential target of PBM. The expression and specifc mechanism of lncRNA TUG1 were detected by qPCR, immunofuorescence, fow cytometry, western blotting, fuorescence in situ hybridization, and luciferase assay. The Basso mouse scale (BMS) and gait analysis were used to evaluate the recovery of motor function in mice.
Results: Results showed that lncRNA TUG1 may be a potential target of PBM, regulating the polarization of BMDMs, infammatory response, and the axial growth of DRG. Mechanistically, TUG1 competed with TLR3 for binding to miR-1192 and attenuated the inhibitory efect of miR-1192 on TLR3. This efect protected TLR3 from degradation, enabling the high expression of TLR3, which promoted the activation of downstream NF-κB signal and the release of infammatory cytokines. In vivo, PBM treatment could reduce the expression of TUG1, TLR3, and infammatory cytokines and promoted nerve survival and motor function recovery in SCI mice.
Conclusions: Our study clarifed that the lncRNA TUG1/miR-1192/TLR3 axis is an important pathway for PBM to inhibit M1 macrophage polarization and infammation, which provides theoretical support for its clinical application in patients with SCI.
Keywords Spinal cord injury, Bone-marrow-derived macrophages, Photobiomodulation, Transcriptome sequencing, Infammation, Long noncoding RNA TUG1
Address and Contact Information Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Changle West Road No. 127, Xi’an 710032, Shaanxi, China
*Corresponding author: huxueyu@fmmu.edu.cn; wangzhe@fmmu.edu.cn
Cheng Ju, Yangguang Ma, and Xiaoshuang Zuo have contributed equally to this work and share first authorship
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No. 06 DOI: 10.1186/s11658-023-00418-z Volume 28 (2023) - 28:06
Authors Hamed Hosseinalizadeh1, Omid Mohamadzadeh2, Mohammad Saeed Kahrizi3, Zahra Razaghi Bahabadi4,5, Daniel J. Klionsky6 and Hamed Mirzei7*
Abstract Glioblastoma multiforme (GBM) is an aggressive primary brain tumor and one of the most lethal central nervous system tumors in adults. Despite signifcant breakthroughs in standard treatment, only about 5% of patients survive 5 years or longer. Therefore, much efort has been put into the search for identifying new glioma-associated genes. Tripartite motif-containing (TRIM) family proteins are essential regulators of carcino-genesis. TRIM8, a member of the TRIM superfamily, is abnormally expressed in high-grade gliomas and is associated with poor clinical prognosis in patients with glioma. Recent research has shown that TRIM8 is a molecule of duality (MoD) that can function as both an oncogene and a tumor suppressor gene, making it a “double-edged sword” in glioblastoma development. This characteristic is due to its role in selectively regulating three major cellular signaling pathways: the TP53/p53-mediated tumor suppression pathway, NFKB/NF-κB, and the JAK-STAT pathway essential for stem cell property support in glioma stem cells. In this review, TRIM8 is analyzed in detail in the context of GBM and its involvement in essential signaling and stem cell-related pathways. We also discuss the basic biological activities of TRIM8 in macroautophagy/autophagy, regulation of bipolar spindle formation and chromosomal stability, and regulation of chemoresistance, and as a trigger of infammation.
Keywords Autophagy, Glioblastoma, JAK-STAT, NF-κB, p53, Stem-cell, TRIM8
Address and Contact Information 1 Department of Medical Biotechnology, Faculty of Paramedicine, Guilan University of Medical Sciences, Rasht, Iran
2 Department of Neurosurgery, Tehran University of Medical Science, Tehran, Iran
3 Department of Surgery, Alborz University of Medical Sciences, Karaj, Alborz, Iran
4 School of Medicine, Kashan University of Medical Sciences, Kashan, Iran
5 Student Research Committee, Kashan University of Medical Sciences, Kashan, Iran
6 Life Sciences Institute and Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USA
7 Research Center for Biochemistry and Nutrition in Metabolic Diseases, Institute for Basic Sciences, Kashan University of Medical Sciences, Kashan, Iran
*Corresponding author: mirzaei-h@kaums.ac.ir; h.mirzaei2002@gmail.com
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No. 07 DOI: 10.1186/s11658-023-00420-5 Volume 28 (2023) - 28:07
Authors Jixiao Wang, Huiqi Yang, Xiaobei Ma, Jiani Liu, Lan Li, Lei Chen* and Fulan Wei*
Abstract Background: Mechanotransduction mechanisms whereby periodontal ligament stem cells (PDLSCs) translate mechanical stress into biochemical signals and thereby trigger osteogenic programs necessary for alveolar bone remodeling are being deciphered. Low-density lipoprotein receptor-related protein 6 (LRP6), a Wnt transmembrane receptor, has been qualifed as a key monitor for mechanical cues. However, the role of LRP6 in the mechanotransduction of mechanically induced PDLSCs remains obscure.
Methods: The Tension System and tooth movement model were established to determine the expression profle of LRP6. The loss-of-function assay was used to investigate the role of LRP6 on force-regulated osteogenic commitment in PDLSCs. The ability of osteogenic diferentiation and proliferation was estimated by alkaline phosphatase (ALP) staining, ALP activity assay, western blotting, quantitative real-time PCR (qRT-PCR), and immunofuorescence. Crystalline violet staining was used to visualize cell morphological change. Western blotting, qRT-PCR, and phalloidin staining were adopted to afrm flamentous actin (F-actin) alteration. YAP nucleoplasmic localization was assessed by immunofuorescence and western blotting. YAP transcriptional response was evaluated by qRT-PCR. Cytochalasin D was used to determine the efects of F-actin on osteogenic commitment and YAP switch behavior in mechanically induced PDLSCs.
Results: LRP6 was robustly activated in mechanically induced PDLSCs and PDL tissues. LRP6 defciency impeded force-dependent osteogenic diferentiation and proliferation in PDLSCs. Intriguingly, LRP6 loss caused cell morphological aberration, F-actin dynamics disruption, YAP nucleoplasmic relocation, and subsequent YAP inactivation. Moreover, disrupted F-actin dynamics inhibited osteogenic diferentiation, proliferation, YAP nuclear translocation, and YAP activation in mechanically induced PDLSCs.
Conclusions: We identifed that LRP6 in PDLSCs acted as the mechanosensor regulating mechanical stress-inducible osteogenic commitment via the F-actin/YAP cascade. Targeting LRP6 for controlling alveolar bone remodeling may be a prospective therapy to attenuate relapse of orthodontic treatment.
Keywords Mechanical stress, PDLSCs, Mechanotransduction, LRP6, Filamentous actin, Osteogenic commitment
Address and Contact Information Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, No. 44-1 Wenhua Road West, Jinan 250012, Shandong, China
*Corresponding author: sdcl8006@163.com; weif@email.sdu.edu.cn
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No. 08 DOI: 10.1186/s11658-023-00421-4 Volume 28 (2023) - 28:08
Authors Caroline Diener1*, Martin Hart1, Claudia Fecher‐Trost2, Jessica Knittel1, Stefanie Rheinheimer1, Markus R. Meyer3, Jens Mayer1, Veit Flockerzi2, Andreas Keller4,5 and Eckart Meese1
Abstract Among the concepts in biology that are widely taken granted is a potentiated cooperative efect of multiple miRNAs on the same target. This strong hypothesis contrasts insufcient experimental evidence. The quantity as well as the quality of required side constraints of cooperative binding remain largely hidden. For miR-21-5p and miR-155-5p, two commonly investigated regulators across diseases, we selected 15 joint target genes. These were chosen to represent various neighboring 3′UTR binding site constellations, partially exceeding the distance rules that have been established for over a decade. We identifed diferent cooperative scenarios with the binding of one miRNA enhancing the binding efects of the other miRNA and vice versa. Using both, reporter assays and whole proteome analyses, we observed these cooperative miRNA efects for genes that bear 3′UTR binding sites at distances greater than the previously defned limits. Astonishingly, the experiments provide even stronger evidence for cooperative miRNA efects than originally postulated. In the light of these fndings the defnition of targetomes specifed for single miRNAs need to be refned by a concept that acknowledges the cooperative efects of miRNAs.
Keywords microRNA, Cooperativity, Binding sites, Distances, In silico prediction, RISC, Functional interactions, Networks, Proteomics, Target identifcation
Address and Contact Information 1 Institute of Human Genetics, Saarland University, 66421 Homburg, Germany
2 Department of Experimental and Clinical Pharmacology & Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Center for Molecular Signaling (PZMS), Saarland University, 66421 Homburg, Germany
3 Department of Experimental and Clinical Toxicology & Pharmacology, Institute of Experimental and Clinical Pharmacology and Toxicology, Center for Molecular Signaling (PZMS), Saarland University, 66421 Homburg, Germany
4 Chair for Clinical Bioinformatics, Saarland Informatics Campus, Saarland University, 66123 Saarbrücken, Germany
5 Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz-Centre for Infection Research (HZI), 66123 Saarbrücken, Germany
*Corresponding author: caroline.diener@uni-saarland.de
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No. 09 DOI:10.1186/s11658-023-00422-3 Volume 28 (2023) - 28:09
Authors Xueke Li1,2,3†, Yi Zhong1,2,3†, Wuqi Zhou1,2,3, Yishu Song1,2,3, Wenqu Li1,2,3, Qiaofeng Jin1,2,3, Tang Gao1,2,3, Li Zhang1,2,3* and Mingxing Xie1,2,3*
Abstract Background: Bone marrow-derived mesenchymal stem cells (BMSCs)-derived extracellular vesicles (EVs) have shown potent anti-infammatory function in various pathological conditions, such as osteoarthritis and neurodegenerative diseases. Since the number of EVs naturally secreted by cells is fnite and they usually bear specifc repertoires of bioactive molecules to perform manifold cell–cell communication, but not one particular therapeutic function as expected, their practical application is still limited. Strategies are needed to increase the production of EVs and enhance their therapeutic function. Recent studies have suggested that low-intensity pulsed ultrasound (LIPUS) is a promising non-invasive method to increase the secretion of EVs and promote their anti-infammatory efects. However, the efect of LIPUS stimulation of BMSCs on EVs derived from the cells remains unclear. The objective of this study was to investigate whether LIPUS stimulation on BMSCs could increase the secretion of EVs and enhance their anti-infammatory efects.
Methods: BMSCs were exposed to LIPUS (300 mW/cm2) for 15 min and EVs were isolated by ultracentrifugation. Anti-infammatory efects of EVs were investigated on RAW264.7 cells in vitro and in the allogeneic skin transplantation model. Small RNA-seq was utilized to identify components diference in EVs with/without LIPUS irradiation.
Results: In this study, we found that LIPUS stimulation could lead to a 3.66-fold increase in the EVs release from BMSCs. Moreover, both in vitro and in vivo experimental results suggested that EVs secreted from LIPUS-treated BMSCs (LIPUS-EVs) possessed stronger anti-infammatory function than EVs secreted from BMSCs without LIPUS stimulation (C-EVs). RNA-seq analysis revealed that miR-328-5p and miR-487b-3p were signifcantly up-regulated in LIPUS-EVs compare with C-EVs. The suppression of MAPK signaling pathway by these two up-regulated miRNAs could be the potential mechanism of strengthened anti-infammatory efects of LIPUS-EVs.
Conclusion: LIPUS stimulation on BMSCs could signifcantly increase the secretion of EVs. Moreover, EVs generated from LIPUS-treated BMSCs possessed much stronger antiinfammatory function than C-EVs. Therefore, LIPUS could be a promising non-invasive strategy to promote the production of EVs from BMSCs and augment their anti-infammatory efects.
Keywords Low-intensity pulsed ultrasound, Extracellular vesicles, Infammation, Bone marrow mesenchymal stem cells, microRNA
Address and Contact Information 1 Department of Ultrasound Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
2 Clinical Research Center for Medical Imaging in Hubei Province, Wuhan 430022, China
3 Hubei Province Key Laboratory of Molecular Imaging, Wuhan 430022, China
*Correspondence: zli429@hust.edu.cn; xiemx@hust.edu.cn
Xueke Li and Yi Zhong have equally contributed to this work
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No. 10 DOI: 10.1186/s11658-023-00424-1 Volume 28 (2023) - 28:10
Authors Asfa Soomro1, Mohammad Khajehei1, Renzhong Li1, Kian O’Neil1, Dan Zhang1, Bo Gao1, Melissa MacDonald1, Masao Kakoki2 and Joan C. Krepinsky1,3*
Abstract Background: TGFβ1 is a major profbrotic mediator in chronic kidney disease (CKD). Its direct inhibition, however, is limited by adverse efects. Inhibition of activins, also members of the TGFβ superfamily, blocks TGFβ1 profbrotic efects, but the mechanism underlying this and the specifc activin(s) involved are unknown.
Methods: Cells were treated with TGFβ1 or activins A/B. Activins were inhibited generally with follistatin, or specifcally with neutralizing antibodies or type I receptor downregulation. Cytokine levels, signaling and profbrotic responses were assessed with ELISA, immunofuorescence, immunoblotting and promoter luciferase reporters. Wild-type or TGFβ1-overexpressing mice with unilateral ureteral obstruction (UUO) were treated with an activin A neutralizing antibody.
Results: In primary mesangial cells, TGFβ1 induces secretion primarily of activin A, which enables longer-term profbrotic efects by enhancing Smad3 phosphorylation and transcriptional activity. This results from lack of cell refractoriness to activin A, unlike that for TGFβ1, and promotion of TGFβ type II receptor expression. Activin A also supports transcription through regulating non-canonical MRTF-A activation. TGFβ1 additionally induces secretion of activin A, but not B, from tubular cells, and activin A neutralization prevents the TGFβ1 profbrotic response in renal fbroblasts. Fibrosis induced by UUO is inhibited by activin A neutralization in wild-type mice. Worsened fbrosis in TGFβ1-overexpressing mice is associated with increased renal activin A expression and is inhibited to wild-type levels with activin A neutralization.
Conclusions: Activin A facilitates TGFβ1 profbrotic efects through regulation of both canonical (Smad3) and non-canonical (MRTF-A) signaling, suggesting it may be a novel therapeutic target for preventing fbrosis in CKD.
Keywords Activin A, TGFβ1, Kidney fbrosis, Extracellular matrix
Address and Contact Information 1 Division of Nephrology, Department of Medicine, McMaster University, Hamilton, Canada
2 Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC, USA
3 St. Joseph’s Hospital, 50 Charlton Ave East, Rm T3311, Hamilton, ON L8N 4A6, Canada
*Corresponding author: krepinj@mcmaster.ca
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No. 11 DOI: 10.1186/s11658-023-00426-z Volume 28 (2023) - 28:11
Authors Jiangpeng Wu1†, Xiuping Sun1†, Chunyi Wu1†, Xiaoping Hong2†, Lulin Xie1, Zixu Shiv1, Liang Zhao6,7, Qingfeng Du4*, Wei Xiao3,5*, Jichao Sun1* and Jigang Wang1,4,5,8*
Abstract Background: Glyphosate (GLY), as the active ingredient of the most widely used herbicide worldwide, is commonly detected in the environment and living organisms, including humans. Its toxicity and carcinogenicity in mammals remain controversial. Several studies have demonstrated the hepatotoxicity of GLY; however, the underlying cellular and molecular mechanisms are still largely unknown.
Methods: Using single-cell RNA sequencing (scRNA-seq), immunofuorescent staining, and in vivo animal studies, we analyzed the liver tissues from untreated and GLY-treated mice.
Results: We generated the frst scRNA-seq atlas of GLY-exposed mouse liver. GLY induced varied cell composition, shared or cell-type-specifc transcriptional alterations, and dysregulated cell–cell communication and thus exerted hepatotoxicity efects. The oxidative stress and infammatory response were commonly upregulated in several cell types. We also observed activation and upregulated phagocytosis in macrophages, as well as proliferation and extracellular matrix overproduction in hepatic stellate cells.
Conclusions: Our study provides a comprehensive single-cell transcriptional picture of the toxic efect of GLY in the liver, which ofers novel insights into the molecular mechanisms of the GLY-associated hepatotoxicity.
Keywords Glyphosate, Hepatotoxicity, scRNA-seq
Address and Contact Information 1 Department of Nephrology, Shenzhen Key Laboratory of Kidney Diseases, and Shenzhen Clinical Research Centre for Geriatrics, Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University, The First Afliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China.
2 Department of Rheumatology and Immunology, Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University, The First Afliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China.
3 Key Laboratory of Glucolipid Metabolic Disorder, Ministry of Education, Guangdong Pharmaceutical University, Guangzhou 510006, China.
4 School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China.
5 Dongguan Maternal and Child Health Care Hospital, Postdoctoral Innovation Practice Base of Southern Medical University, Dongguan 523125, Guangdong, China.
6 Department of Pathology, Shunde Hospital, Southern Medical University (The First People’s Hospital of Shunde), Foshan 528300, China.
7 Department of Pathology and Guangdong Province Key Laboratory of Molecular Tumor Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China.
8 Artemisinin Research Center, and Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
*Corresponding author: dqfsmu7@126.com; xw7688@smu.edu.cn; sunjichao@mail.sustech.edu.cn; jgwang@icmm.ac.cn
Jiangpeng Wu, Xiuping Sun, Chunyi Wu, and Xiaoping Hong contributed equally to this work.
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No.  12DOI: 10.1186/s11658-023-00425-0 Volume 28 (2023) - 28:12
Authors Yihang Yu1,2,3, Meiling Chen1,2,3, Qitong Guo1,2,3, Lianju Shen1,2,3, Xing Liu1,2,3, Jianbo Pan4, Yuanyuan Zhang5, Tao Xu6, Deying Zhang1,2,3* and Guanghui Wei1,2,3
Abstract Background: Kidney insults due to various pathogenic factors, such as trauma, infection, and infammation, can cause tubular epithelial cell injury and death, leading to acute kidney injury and the transformation of acute kidney injury to chronic kidney disease. There is no defnitive treatment available. In previous studies, human umbilical cord mesenchymal stem cells have been shown to promote kidney injury. In this preclinical study, we investigate the role and mechanism of human umbilical cord mesenchymal stem cell exosomes (HucMSC-Exos) on the repair of renal tubular epithelial cells after injury.
Methods: C57BL/6 mice underwent unilateral ureteral obstruction, and epithelial cell injury was induced in HK-2 cells by cisplatin. HucMSC-Exos were assessed in vivo and in vitro. The extent of renal cell injury, activation of necroptosis pathway, and mitochondrial quality-control-related factors were determined in diferent groups. We also analyzed the possible regulatory efector molecules in HucMSC-Exos by transcriptomics.
Results: HucMSC-Exo inhibited necroptosis after renal tubular epithelial cell injury and promoted the dephosphorylation of the S637 site of the Drp1 gene by reducing the expression of PGAM5. This subsequently inhibited mitochondrial fission and maintained mitochondrial functional homeostasis, mitigating renal injury and promoting repair. In addition, HucMSC-Exo displayed a regulatory role by targeting RIPK1 through miR-874-3p.
Conclusion: The collective findings of the present study demonstrate that HucMSC-Exos can regulate necroptosis through miR-874-3p to attenuate renal tubular epithelial cell injury and enhance repair, providing new therapeutic modalities and ideas for the treatment of AKI and the process of AKI to CKD transformation to mitigate renal damage.
Keywords Human umbilical cord mesenchymal stem cells, Exosome, Necroptosis, Mitochondrial fusion, miR-874-3p, Kidney tubular epithelial cell damage
Address and Contact Information 1 Department of Urology, Children’s Hospital of Chongqing Medical University, Chongqing 400014, China
2 Chongqing Key Laboratory of Children Urogenital Department and Tissue Engineering, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing 400014, China
3 National Clinical Research Center for Child Health and Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China
4 Center for Novel Target and Therapeutic Intervention, Institute of Life Sciences, Chongqing Medical University, Chongqing 400016, China
5 Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA
6 Biomanufacturing Center, Department of Mechanical Engineering, Tsinghua University, Beijing 100084, China
*Corresponding author: zdy@hospital.cqmu.edu.cn
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No.  13DOI: 10.1186/s11658-022-00410-z Volume 28 (2023) - 28:13
Authors Wenjian Yao1†, Xiangbo Jia1†, Li Zhu2, Lei Xu1, Quan Zhang1, Tian Xia1 and Li Wei1*
Abstract Background: Esophageal squamous carcinoma (ESCC) is a common malignancy that originates in the digestive tract. Lymph node metastasis (LNM) is a complicated process, and tumor lymphangiogenesis has been reported to be associated with the spread of tumor cells to lymph nodes (LNs), including in ESCC. However, little is currently known about the mechanisms involved in lymphangiogenesis in ESCC tumors. According to previous literature, we know that hsa_circ_0026611 expresses at a high level in serum exosomes of patients with ESCC and shows a close association with LNM and poor prognosis. However, details on the functions of circ_0026611 in ESCC remain unclear. We aim to explore the efects of circ_0026611 in ESCC cell-derived exosomes on lymphangiogenesis and its potential molecular mechanism.
Methods: We firstly examined how circ_0026611 may express in ESCC cells and exosomes by quantitative reverse transcription real-time polymerase chain reaction (RT-qPCR). The potential efects circ_0026611 may exert on lymphangiogenesis in ESCC cell-derived exosomes were assessed afterward via mechanism experiments.
Results: circ_0026611 high expression pattern was confrmed in ESCC cells and exosomes. ESCC cell-derived exosomes promoted lymphangiogenesis by transferring circ_0026611. Besides, circ_0026611 interacted with N-α-acetyltransferase 10 (NAA10) to inhibit NAA10-mediated prospero homeobox 1 (PROX1) acetylation with subsequent ubiquitination and degradation. Furthermore, circ_0026611 was verifed to promote lymphangiogenesis in a PROX1-mediated manner.
Conclusions: Exosomal circ_0026611 inhibited PROX1 acetylation and ubiquitination to promote lymphangiogenesis in ESCC.
Keywords Esophageal squamous cell carcinoma, Exosomes, circ_0026611, PROX1, Lymphangiogenesis
Address and Contact Information 1 Department of Thoracic Surgery, Henan Provincial People’s Hospital, People’s Hospital of Zhengzhou University, School of Clinical Medicine, Henan University, No.7, Weiwu Road, Jinshui District, Zhengzhou 450003, Henan, China
2 Department of Thoracic Surgery, Zhengzhou University People’s Hospital, Henan Provincial People’s Hospital, Zhengzhou 450003, Henan, China
*Corresponding author: zhuwei627328429@163.com
Wenjian Yao and Xiangbo Jia co-first authors.
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No.  14DOI: 10.1186/s11658-023-00427-y Volume 28 (2023) - 28:14
Authors Maša Mavri1, Sanja Glišić2, Milan Senćanski2, Milka Vrecl1, Mette M. Rosenkilde3, Katja Spiess3,4 and Valentina Kubale1*
Abstract Background: The viral G-protein-coupled receptor (vGPCR) BILF1 encoded by the Epstein–Barr virus (EBV) is an oncogene and immunoevasin and can downregulate MHC-I molecules at the surface of infected cells. MHC-I downregulation, which presumably occurs through co-internalization with EBV-BILF1, is preserved among BILF1 receptors, including the three BILF1 orthologs encoded by porcine lymphotropic herpesviruses (PLHV BILFs). This study aimed to understand the detailed mechanisms of BILF1 receptor constitutive internalization, to explore the translational potential of PLHV BILFs compared with EBV-BILF1.
Methods: A novel real-time fluorescence resonance energy transfer (FRET)-based internalization assay combined with dominant-negative variants of dynamin-1 (Dyn K44A) and the chemical clathrin inhibitor Pitstop2 in HEK-293A cells was used to study the effect of specific endocytic proteins on BILF1 internalization. Bioluminescence resonance energy transfer (BRET)-saturation analysis was used to study BILF1 receptor interaction with β-arrestin2 and Rab7. In addition, a bioinformatics approach informational spectrum method (ISM) was used to investigate the interaction affinity of BILF1 receptors with β-arrestin2, AP-2, and caveolin-1.
Results: We identified dynamin-dependent, clathrin-mediated constitutive endocytosis for all BILF1 receptors. The observed interaction affinity between BILF1 receptors and caveolin-1 and the decreased internalization in the presence of a dominant-negative variant of caveolin-1 (Cav S80E) indicated the involvement of caveolin-1 in BILF1 trafficking. Furthermore, after BILF1 internalization from the plasma membrane, both the recycling and degradation pathways are proposed for BILF1 receptors.
Conclusions: The similarity in the internalization mechanisms observed for EBV-BILF1 and PLHV1-2 BILF1 provide a foundation for further studies exploring a possible translational potential for PLHVs, as proposed previously, and provides new information about receptor trafficking.
Keywords vGPCR, BILF1, Endocytosis, Internalization, Dynamin, Caveolin, β-Arrestin, EBV, PLHV1-2
Address and Contact Information 1 Institute for preclinical sciences, Veterinary Faculty, Ljubljana, Slovenia
2 Center for Multidisciplinary Research, Institute of Nuclear Sciences VINCA, University of Belgrade, Belgrade, Serbia
3 Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
4 Present Address: Department of Virus and Microbiological Special Diagnostics, Statens Serum Institute, Copenhagen, Denmark
*Corresponding author: valentina.kubale@vf.uni-lj.si
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No.  15DOI: 10.1186/s11658-023-00429-w Volume 28 (2023) - 28:15
Authors Jiangwei Xiao1†, Jingwen Huang1†, Xiaoting Jian1, Han Wang1, Haiqiang Lan1, Zhaohong Liao1, Ruicai Gu2, Jijie Hu3* and Hua Liao1*
Abstract Endoplasmic reticulum stress (ERS) and the unfolded protein response (UPR) are involved in various muscle pathological states. The IRE1α arm of UPR can afect immunological properties of myofber through restraining p38 mitogen-activated protein kinases (MAPK) activation under infammatory milieu. However, the relevant pathway molecules regulating the initiation of the IRE1α arm in myofiber remain unclear. In this work, expression of transforming growth factor-beta (TGF-β) and TGF-β receptor II (TGF-βr2), and UPR pathway activation were examined in cardiotoxin (CTX)-damaged mouse muscle, which revealed the activation of TGF-β signaling and UPR in CTX-damaged muscle and in regenerating myofibers. Using control or transgenic mice with TGF-βr2 deleted in skeletal muscle (SM TGF-βr2−/−) and the derived primary diferentiating myogenic precursor cells (MPCs) treated with/without ERS activator or inhibitor, IRE1α pathway inhibitor, or TGF-β signaling activator, this study further revealed an essential role of intrinsic TGF-β signaling in regulating muscle cell to express infammation-related molecules including H-2Kb, H2-Eα, TLR3, and special myokines. TGF-β signaling prompted UPR IRE1α arm and restrained p38 MAPK activation in myofiber under infammatory milieu. This study uncovers a previously unrecognized function of TGF-β signaling acting as an upstream factor controlling myofiber immune capacities in the infamed state through the UPR–IRE1α–p38 MAPK pathway.
Keywords Myofiber, Infammation, TGF-β, UPR, IRE1α, p38 MAPK
Address and Contact Information 1 Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Department of Anatomy, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, China
2 Department of Cell Biology, University of Oklahoma Health Science Center, Oklahoma City, OK, USA
3 Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China
*Corresponding author: hjj0610@163.com; hua-liao@163.com
Jiangwei Xiao and Jingwen Huang contributed equally to this work
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No.  16DOI: 10.1186/s11658-023-00433-0 Volume 28 (2023) - 28:16
Authors Shouxing Duan1,2, Xuewu Jiang3, Jianhong Li4, Maxian Fu1, Zhuo Li1, Yiyi Cheng2, Yangmu Zhuang2, Ming Yang2, Wenfeng Xiao4, Hongyan Ping3, Yao Xie5* , Xiaojun Xie6* and Xuan Zhang3*
Abstract Background: Testicular hypoplasia can affect the sexual and reproductive ability in adulthood, and even increase the risk of cancer. Abnormal development of the gubernaculum is one of the important factors of testicular hypoplasia. Therefore, a study of the structure and function of the gubernaculum is an important but neglected new breakthrough point for investigating the normal/abnormal development of the testis. Previous findings showed that Insulin like factor 3 (INSL3) is a key factor regulating the growth of gubernaculum, however, the mechanism by which INSL3 acts on the gubernaculum remains unknown. Therefore, we probed the mechanism associated with INSL3-induced the proliferation, migration, and apoptosis of gubernacular cells in mice.
Methods: A culture cell model of neonatal mice gubernaculum is established by INSL3 intervention. We blocked PLC/PKC signaling pathway with U73122 pretreat to investigate the role of the PLC/PKC signaling pathway. The changes of cell proliferation, migration, and apoptosis were detected by molecular biological methods. In addition, the levels of PCNA and F-action were detected by immunofuorescence and western blotting.
Results: We found that INSL3 can promote the proliferation and migration of gubernacular cells and inhibit their apoptosis, meanwhile, INSL3 signifcantly up-regulated PLC/PKC protein phosphorylation. However, treatment with the PLC/PKC signaling pathway inhibitor U73122 significantly inhibited these efects of INSL3. Besides, we found that INSL3 could up-regulate the protein expression level of PCNA and F-actin, while the PCNA and F-actin expression was signifcantly weakened after U73122 pretreatment.
Conclusions: This research revealed that INSL3 binding to RXFP2 may up-regulate the expression levels of PCNA and F-actin by activating the PLC/PKC signaling pathway to promote the proliferation and migration of gubernacular cells. It suggests that the RXFP2-PLC/PKC axis may serve as a novel molecular mechanism by which INSL3 regulates growth of the gubernaculum.
Keywords INSL3, RXFP2, PLC/PKC, Gubernaculum, Signaling pathway
Address and Contact Information 1 Department of Pediatric Surgery, Huazhong University of Science and Technology Union Shenzhen Hospital (Nanshan Hospital), No. 89 Taoyuan Road, Shenzhen 518052, Guangdong, China.
2 Department of Pediatric Surgery, The First Afliated Hospital of Shantou University Medical College, No. 57 Changping Road, Shantou 515041, Guangdong, China.
3 Department of Pediatric Surgery, Pingshan District Maternal and Child Healthcare Hospital of Shenzhen, Pingshan General Hospital of Southern Medical University, No. 6 Longxingnan Road, Shenzhen 518118, Guangdong, China.
4 Department of Pediatric Surgery, The Second Afliated Hospital of Shantou University Medical College, No. 69 Dongxiabei Road, Shantou 515041, Guangdong, China.
5 Department of Radiology, Cancer Hospital of Shantou University Medical College, No. 7 Raoping Road, Shantou 515041, Guangdong, China.
6 Department of General Surgery, The First Afliated Hospital of Shantou University Medical College, No. 57 Changping Road, Shantou 515041, Guangdong, China.
*Corresponding author: stxieyao@163.com; xjxie79@163.com; zhxuan2006@126.com
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No.  17DOI: 10.1186/s11658-023-00431-2 Volume 28 (2023) - 28:17
Authors Aleksandra Polishchuk, Víctor Cilleros‐Mañé, Laia Just‐Borràs, Marta Balanyà‐Segura, Genís Vandellòs Pont, Carolina Silvera Simón, Marta Tomàs, Neus Garcia, Josep Tomàs*† and Maria A. Lanuza*†
Abstract Background: Bidirectional communication between presynaptic and postsynaptic components contribute to the homeostasis of the synapse. In the neuromuscular synapse, the arrival of the nerve impulse at the presynaptic terminal triggers the molecular mechanisms associated with ACh release, which can be retrogradely regulated by the resulting muscle contraction. This retrograde regulation, however, has been poorly studied. At the neuromuscular junction (NMJ), protein kinase A (PKA) enhances neurotransmitter release, and the phosphorylation of the molecules of the release machinery including synaptosomal associated protein of 25 kDa (SNAP-25) and Synapsin-1 could be involved.
Methods: Accordingly, to study the effect of synaptic retrograde regulation of the PKA subunits and its activity, we stimulated the rat phrenic nerve (1 Hz, 30 min) resulting or not in contraction (abolished by μ-conotoxin GIIIB). Changes in protein levels and phosphorylation were detected by western blotting and cytosol/membrane translocation by subcellular fractionation. Synapsin-1 was localized in the levator auris longus (LAL) muscle by immunohistochemistry.
Results: Here we show that synaptic PKA Cβ subunit regulated by RIIβ or RIIα subunits controls activity-dependent phosphorylation of SNAP-25 and Synapsin-1, respectively. Muscle contraction retrogradely downregulates presynaptic activity-induced pSynapsin-1 S9 while that enhances pSNAP-25 T138. Both actions could coordinately contribute to decreasing the neurotransmitter release at the NMJ.
Conclusion: This provides a molecular mechanism of the bidirectional communication between nerve terminals and muscle cells to balance the accurate process of ACh release, which could be important to characterize molecules as a therapy for neuromuscular diseases in which neuromuscular crosstalk is impaired.
Keywords Neuromuscular junction, Synapse, Neurotransmission, ACh release, SNAP-25, Synapsin-1, PKA subunits, Synaptic activity, Electrical stimulation
Address and Contact Information Unitat d’Histologia i Neurobiologia (UHNEUROB), Departament de Ciències Mèdiques Bàsiques, Facultat de Medicina i Ciències de la Salut, Universitat Rovira i Virgili, c/ Sant Llorenç 21, 43201 Reus, Spain
*Corresponding author: josepmaria.tomas@urv.cat; mariaangel.lanuza@urv.cat
Aleksandra Polishchuk and Víctor Cilleros-Mañé contributed equally to this work
Neus Garcia, Josep Tomàs and Maria A. Lanuza contributed equally to this work
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No.  18DOI: 10.1186/s11658-023-00432-1 Volume 28 (2023) - 28:18
Authors Mengqi Zhang1,2*, Qian Wu3, Mimi Tang4,5, Zhuohui Chen1,2 and Haiyue Wu1,2
Abstract Background: Brain microvascular endothelial cell (BMEC) injury can afect neuronal survival by modulating immune responses through the microenvironment. Exosomes are important vehicles of transport between cells. However, the regulation of the subtypes of microglia by BMECs through the exosome transport of microRNAs (miRNAs) has not been established.
Methods: In this study, exosomes from normal and oxygen–glucose deprivation (OGD)-cultured BMECs were collected, and diferentially expressed miRNAs were analyzed. BMEC proliferation, migration, and tube formation were analyzed using MTS, transwell, and tube formation assays. M1 and M2 microglia and apoptosis were analyzed using flow cytometry. miRNA expression was analyzed using real-time polymerase chain reaction (RT-qPCR), and IL-1β, iNOS, IL-6, IL-10, and RC3H1 protein concentrations were analyzed using western blotting.
Results: We found that miR-3613-3p was enriched in BMEC exosome by miRNA GeneChip assay and RT-qPCR analysis. miR-3613-3p knockdown enhanced cell survival, migration, and angiogenesis in the OGD-treated BMECs. In addition, BMECs secrete miR-3613-3p to transfer into microglia via exosomes, and miR-3613-3p binds to the RC3H1 3′ untranslated region (UTR) to reduce RC3H1 protein levels in microglia. Exosomal miR-3613-3p promotes microglial M1 polarization by inhibiting RC3H1 protein levels. BMEC exosomal miR-3613-3p reduces neuronal survival by regulating microglial M1 polarization.
Conclusions: miR-3613-3p knockdown enhances BMEC functions under OGD conditions. Interfering with miR-3613-3p expression in BMSCs reduced the enrichment of miR-3613-3p in exosomes and enhanced M2 polarization of microglia, which contributed to reduced neuronal apoptosis.
Keywords Exosome, Ischemic stroke, Brain microvascular endothelial cell, Macrophage, Neuron
Address and Contact Information 1 Department of Neurology, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha 410008, Hunan, China
2 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, China
3 Department of Neurology, First Afliated Hospital, Kunming Medical University, Kunming 650032, China
4 Department of Pharmacy, Xiangya Hospital, Central South University, Changsha 410008, China
5 Institute of Hospital Pharmacy, Xiangya Hospital, Central South University, Changsha 410008, China
*Corresponding author: zhangmengqi8912@163.com
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No.  19DOI: 10.1186/s11658-023-00428-x Volume 28 (2023) - 28:19
Authors Mengjie Chen1, Xingyu Liu1, Qingyou Liu1,2, Deshun Shi1* and Hui Li1*
Abstract Three-dimensional (3D) genomics is an emerging discipline that studies the three-dimensional structure of chromatin and the three-dimensional and functions of genomes. It mainly studies the three-dimensional conformation and functional regulation of intranuclear genomes, such as DNA replication, DNA recombination, genome folding, gene expression regulation, transcription factor regulation mechanism, and the maintenance of three-dimensional conformation of genomes. Self-chromosomal conformation capture (3C) technology has been developed, and 3D genomics and related fields have developed rapidly. In addition, chromatin interaction analysis techniques developed by 3C technologies, such as paired-end tag sequencing (ChIA-PET) and whole-genome chromosome conformation capture (Hi-C), enable scientists to further study the relationship between chromatin conformation and gene regulation in different species. Thus, the spatial conformation of plant, animal, and microbial genomes, transcriptional regulation mechanisms, interaction patterns of chromosomes, and the formation mechanism of spatiotemporal specificity of genomes are revealed. With the help of new experimental technologies, the identifcation of key genes and signal pathways related to life activities and diseases is sustaining the rapid development of life science, agriculture, and medicine. In this paper, the concept and development of 3D genomics and its application in agricultural science, life science, and medicine are introduced, which provides a theoretical basis for the study of biological life processes.
Keywords 3D genomics, Chromatin conformation, Gene expression regulation, Hi-C, Precision biology, Precision medicine
Address and Contact Information 1 State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Animal Science and Technology, Guangxi University, Nanning 530004, Guangxi Province, China
2 Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan 528225, China
*Corresponding author: ardsshi@gxu.edu.cn; huili@gxu.edu.cn
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No.  20DOI: 10.1186/s11658-023-00430-3 Volume 28 (2023) - 28:20
Authors Ming Ma1,2, Hui Li1,2, Saifu Yin1,2, Tao Lin1,2* and Turun Song1,2*
Abstract Background: Kidney ischemia–reperfusion injury is inevitable in kidney transplantation, and is essential for primary graft dysfunction and delayed graft function. Our previous study has proved that miR-92a could ameliorate kidney ischemia–reperfusion injury, but the mechanism has not been studied.
Methods: This study conducted further research on the role of miR-92a in kidney ischemia–reperfusion injury and organ preservation. In vivo, mice models of bilateral kidney ischemia (30 min), cold preservation after ischemia (cold preservation time of 6, 12, and 24 h), and ischemia–reperfusion (reperfusion time of 24, 48, and 72 h) were established. Before or after modeling, the model mice were injected with miR-92a-ago-mir through the caudal vein. In vitro, the hypoxia–reoxygenation of HK-2 cells was used to simulate ischemia–reperfusion injury.
Results: Kidney ischemia and ischemia–reperfusion signifcantly damaged kidney function, decreased the expression of miR-92a, and increased apoptosis and autophagy in kidneys. miR-92a agomir tail vein injection signifcantly increased the expression of miR-92a in kidneys, improved kidney function, and alleviated kidney injury, and the intervention before modeling achieved a better efect than after. Moreover, miR-92a agomir signifcantly reduced the apoptosis and autophagy in HK-2 cells induced by hypoxia, hypoxia–reoxygenation, and rapamycin, while miR-92a antagomir had opposite efects. Furthermore, mitogen-activated protein kinase, c-Jun NH (2) terminal kinase, caspase 3, Beclin 1, and microtubule-associated protein 1 light chain 3B were inhibited by overexpression of miR-92a both in vivo and in vitro, which in turn reduced apoptosis and autophagy.
Conclusions: Our results prove that overexpression of miR-92a attenuated kidney ischemia–reperfusion injury and improved kidney preservation, and intervention before ischemia–reperfusion provides better protection than after.
Keywords miR-92a, Kidney transplant, Ischemia–reperfusion injury, Autophagy, Apoptosis
Address and Contact Information 1 Department of Urology, West China Hospital, Sichuan University, 37# Guoxue Alley, Chengdu 610041, Sichuan, China
2 Organ Transplantation Center, West China Hospital, Sichuan University, 37# Guoxue Alley, Chengdu 610041, Sichuan, China
*Correspondence: kidney5@163.com; songturun1986@scu.edu.cn
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No.  21DOI: 10.1186/s11658-023-00423-2 Volume 28 (2023) - 28:21
Authors Anna Janaszak‐Jasiecka1, Agata Płoska1, Joanna M. Wierońska2, Lawrence W. Dobrucki1,3,4,5 and Leszek Kalinowski1,6*
Abstract Nitric oxide (NO) is one of the most important molecules released by endothelial cells, and its antiatherogenic properties support cardiovascular homeostasis. Diminished NO bioavailability is a common hallmark of endothelial dysfunction underlying the pathogenesis of the cardiovascular disease. Vascular NO is synthesized by endothelial nitric oxide synthase (eNOS) from the substrate L-arginine (L-Arg), with tetrahydrobiopterin (BH4) as an essential cofactor. Cardiovascular risk factors such as diabetes, dyslipidemia, hypertension, aging, or smoking increase vascular oxidative stress that strongly affects eNOS activity and leads to eNOS uncoupling. Uncoupled eNOS produces superoxide anion (O2) instead of NO, thus becoming a source of harmful free radicals exacerbating the oxidative stress further. eNOS uncoupling is thought to be one of the major underlying causes of endothelial dysfunction observed in the pathogenesis of vascular diseases. Here, we discuss the main mechanisms of eNOS uncoupling, including oxidative depletion of the critical eNOS cofactor BH4, defciency of eNOS substrate L-Arg, or accumulation of its analog asymmetrical dimethylarginine (ADMA), and eNOS S-glutathionylation. Moreover, potential therapeutic approaches that prevent eNOS uncoupling by improving cofactor availability, restoration of L-Arg/ADMA ratio, or modulation of eNOS S-glutathionylation are briefy outlined.
Keywords Cardiovascular disease, Endothelial dysfunction, eNOS uncoupling, Oxidative/nitroxidative stress, Peroxynitrite, Nitric oxide, ADMA, Tetrahydrobiopterin, BH4
Address and Contact Information 1 Department of Medical Laboratory Diagnostics ‐ Fahrenheit Biobank BBMRI.Pl, Medical University of Gdansk, 7 Debinki Street, 80‐211, Gdansk, Poland
2 Department of Neurobiology, Polish Academy of Sciences, Maj Institute of Pharmacology, 12 Smętna Street, 31‐343 Kraków, Poland
3 Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA
4 Beckman Institute for Advanced Science and Technology, 405 N Mathews Ave, MC‐251, Urbana, IL 61801, USA
5 Department of Biomedical and Translational Sciences, Carle-Illinois College of Medicine, Urbana, IL, USA
6 BioTechMed Centre, Department of Mechanics of Materials and Structures, Gdansk University of Technology, 11/12 Gabriela Narutowicza Street, 80‐233 Gdansk, Poland
*Corresponding author: leszek.kalinowski@gumed.edu.pl
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No.  22DOI: 10.1186/s11658-023-00436-x Volume 28 (2023) - 28:22
Authors C. L. Karthika1, Vani Venugopal1, B. J. Sreelakshmi1, S. Krithika1, Jaya Mary Thomas1, Mathew Abraham2, C. C. Kartha3, Arumugam Rajavelu4 and S. Sumi1*
ABSTRACT Background: Cerebral arteriovenous malformations (cAVM) are a signifcant cause of intracranial hemorrhagic stroke and brain damage. The arteriovenous junctions in AVM nidus are known to have hemodynamic disturbances such as altered shear stress, which could lead to endothelial dysfunction. The molecular mechanisms coupling shear stress and endothelial dysfunction in cAVMs are poorly understood. We speculated that disturbed blood fow in artery–vein junctions activates Notch receptors and promotes endothelial mesenchymal plasticity during cAVM formation.
Methods: We investigated the expression profle of endothelial mesenchymal transition (EndMT) and cell adhesion markers, as well as activated Notch receptors, in 18 human cAVM samples and 15 control brain tissues, by quantitative real-time PCR (qRT-PCR) and immunohistochemical evaluation. Employing a combination of a microfuidic system, qRT-PCR, immunofuorescence, as well as invasion and inhibitor assays, the efects of various shear stress conditions on Notch-induced EndMT and invasive potential of human cerebral microvascular endothelial cells (hCMEC/d3) were analyzed.
Results: We found evidence for EndMT and enhanced expression of activated Notch intracellular domain (NICD3 and NICD4) in human AVM nidus samples. The expression of transmembrane adhesion receptor integrin α9/β1 is signifcantly reduced in cAVM nidal vessels. Cell–cell adhesion proteins such as VE-cadherin and N-cadherin were diferentially expressed in AVM nidus compared with control brain tissues. Using well-characterized hCMECs, we show that altered fluid shear stress steers Notch3 nuclear translocation and promotes SNAI1/2 expression and nuclear localization. Oscillatory flow downregulates integrin α9/β1 and VE-cadherin expression, while N-cadherin expression and endothelial cell invasiveness are augmented. Gamma-secretase inhibitor RO4929097, and to a lesser level DAPT, prevent the mesenchymal transition and invasiveness of cerebral microvascular endothelial cells exposed to oscillatory fuid fow.
Conclusions: Our study provides, for the first time, evidence for the role of oscillatory shear stress in mediating the EndMT process and dysregulated expression of cell adhesion molecules, especially multifunctional integrin α9/β1 in human cAVM nidus. Concomitantly, our findings indicate the potential use of small-molecular inhibitors such as RO4929097 in the less-invasive therapeutic management of cAVMs.
Keywords Cerebral arteriovenous malformations, Shear stress, Endothelial cells, EndMT, Notch, Small-molecule inhibitors
Address and Contact Information 1 Cardiovascular Diseases and Diabetes Biology, Rajiv Gandhi Centre for Biotechnology (RGCB), Thiruvananthapuram, Kerala 695014, India
2 Department of Neurosurgery, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala 695011, India
3 Department of Neurology, Amrita Institute of Medical Sciences, Amrita Vishwa Vidyapeetham, Kochi, Kerala 682041, India
4 Department of Biotechnology, Bhupat & Jyoti Mehta School of Biosciences, Indian Institute of Technology, Madras, Chennai, Tamil Nadu 600036, India
* Corresponding author: sumis@rgcb.res.in
C. L. Karthika and Vani Venugopal share first authorship
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No.  23DOI: 10.1186/s11658-023-00437-w Volume 28 (2023) - 28:23
Authors Yazhi Xing1,2†, Kun Peng3†, Qian Yi3, Dongzhen Yu1,2, Haibo Shi1,2, Guang Yang1,2* and Shankai Yin1,2
Abstract Background: Phosphatidylserine is translocated to the inner leaflet of the phospholipid bilayer membrane by the flippase function of type IV P-tape ATPase (P4-ATPase), which is critical to maintain cellular stability and homeostasis. Transmembrane protein 30A (TMEM30A) is the β-subunit of P4-ATPase. Loss of P4-ATPase function causes sensorineural hearing loss and visual dysfunction in human. However, the function of TMEM30A in the auditory system is unclear.
Methods: P4-ATPase subtype expression in the cochlea was detected by immuno-fuorescence staining and quantitative real-time polymerase chain reaction (qRT-PCR) at diferent developmental stages. Hair cell specifc TMEM30A knockout mice and wild-type littermates were used for the following functional and morphological analysis. Auditory function was evaluated by auditory brainstem response. We investigated hair cell and stereocilia morphological changes by immunofuorescence staining. Scanning electron microscopy was applied to observe the stereocilia ultrastructure. Diferentially expressed transcriptomes were analyzed based on RNA-sequencing data from knock-out and wild-type mouse cochleae. Diferentially expressed genes were verifed by qRT-PCR.
Results: TMEM30A and subtypes of P4-ATPase are expressed in the mouse cochlea in a temporal-dependent pattern. Deletion of TMEM30A in hair cells impaired hearing onset due to progressive hair cell loss. The disrupted kinocilia placement and irregular distribution of spectrin-α in cuticular plate indicated the hair cell planar polarity disruption in TMEM30A deletion hair cells. Hair cell degeneration begins at P7 and finishes around P14. Transcriptional analysis indicates that the focal adhesion pathway and stereocilium tip-related genes changed dramatically. Without the TMEM30A chaperone, excessive ATP8A2 accumulated in the cytoplasm, leading to overwhelming endoplasmic reticulum stress, which eventually contributed to hair cell death.
Conclusions: Deletion of TMEM30A led to disrupted planar polarity and stereocilia bundles, and finally led to hair cell loss and auditory dysfunction. TMEM30A is essential for hair cell polarity maintenance and membrane homeostasis. Our study highlights a pivotal role of TMEM30A in the postnatal development of hair cells and reveals the possible mechanisms underlying P4-ATPase-related genetic hearing loss.
Keywords P4-ATPase, Flippases, Planar cell polarity, ER stress, Hearing loss
Address and Contact Information 1 Department of Otolaryngology Head & Neck Surgery, Shanghai Sixth People’s Hospital Afliated to Shanghai Jiao Tong University School of Medicine, 1301 Research Bldg, 600 Yishan Rd, Shanghai, China
2 Otolaryngology Institute of Shanghai Jiao Tong University, 600 Yishan Rd, Shanghai 200233, China
3 Health Management Center, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu 610054, Sichuan, China
*Corresponding author: gyang321@sjtu.edu.cn
Yazhi Xing and Kun Peng contributed equally to this work
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No.  24DOI: 10.1186/s11658-023-00435-y Volume 28 (2023) - 28:24
Authors Guo‐wei Tu1†, Yi Zhang2†, Jie‐fei Ma3†, Jun‐yi Hou1, Guang‐wei Hao, Ying Su1, Jing‐chao Luo1, Lulu Sheng4* and Zhe Luo1,3,5,6*
Abstract Background: Sepsis is an abnormal immune response after infection, wherein the lung is the most susceptible organ to fail, leading to acute lung injury. To overcome the limitations of current therapeutic strategies and develop more specifc treatment, the infammatory process, in which T cell-derived extracellular vesicles (EVs) play a central role, should be explored deeply.
Methods: Liquid chromatography–tandem mass spectrometry was performed for serum EV protein profling. The serum diacylglycerol kinase kappa (DGKK) and endotoxin contents of patients with sepsis-induced lung injury were measured. Apoptosis, oxidative stress, and infammation in A549 cells, bronchoalveolar lavage fuid, and lung tissues of mice were measured by flow cytometry, biochemical analysis, enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction, and western blot.
Results: DGKK, the key regulator of the diacylglycerol (DAG)/protein kinase C (PKC) pathway, exhibited elevated expression in serum EVs of patients with sepsis-induced lung injury and showed strong correlation with sepsis severity and disease progression. DGKK was expressed in CD4+ T cells under regulation of the NF-κB pathway and delivered by EVs to target cells, including alveolar epithelial cells. EVs produced by CD4+ T lymphocytes exerted toxic efects on A549 cells to induce apoptotic cell death, oxidative cell damage, and infammation. In mice with sepsis induced by cecal ligation and puncture, EVs derived from CD4+ T cells also promoted tissue damage, oxidative stress, and infammation in the lungs. These toxic efects of T cell-derived EVs were attenuated by the inhibition of PKC and NOX4, the downstream efectors of DGKK and DAG.
Conclusions: This approach established the mechanism that T-cell-derived EVs carrying DGKK triggered alveolar epithelial cell apoptosis, oxidative stress, infammation, and tissue damage in sepsis-induced lung injury through the DAG/PKC/NOX4 pathway. Thus, T-cell-derived EVs and the elevated distribution of DGKK should be further investigated to develop therapeutic strategies for sepsis-induced lung injury.
Keywords Sepsis-induced lung injury, DGKK, Extracellular vesicles, Oxidative stress, Infammation
Address and Contact Information 1 Cardiac Intensive Care Center, Zhongshan Hospital, Fudan University, Shanghai, China
2 Biomedical Research Center, Institute for Clinical Sciences, Zhongshan Hospital, Fudan University, Shanghai, China
3 Department of Critical Care Medicine, Zhongshan Hospital (Xiamen), Fudan University, Xiamen, China
4 Department of Emergency Medicine, Shanghai Jiao Tong University Afliated Sixth People’s Hospital, Shanghai, China
5 Shanghai Key Laboratory of Lung Infammation and Injury, Shanghai, China
6 Department of Critical Care Medicine, Shanghai Xuhui Central Hospital, Zhongshan-Xuhui Hospital, Fudan University, Shanghai, China
*Corresponding author: yl20111211856@sjtu.edu.cn; luo.zhe@zs-hospital.sh.cn
Guo-wei Tu, Yi Zhang, and Jie-fei Ma contributed equally to this article and are co-frst authors.
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No.  25DOI: 10.1186/s11658-023-00434-z Volume 28 (2023) - 28:25
Authors Xiaohan Jiang, Guoxun Li, Benzhi Zhu, Jingnan Zang, Tian Lan, Rui Jiang* and Bing Wang*
Abstract Background: During cell apoptosis, the C-terminus of BAP31 is cleaved by caspase-8 and generates p20BAP31, which has been shown to induce an apoptotic pathway between the endoplasmic reticulum (ER) and mitochondria. However, the underlying mechanisms of p20BAP31 in cell apoptosis remains unclear.
Methods: We compared the efects of p20BAP31 on cell apoptosis in six cell lines and selected the most sensitive cells. Functional experiments were conducted, including Cell Counting Kit 8 (CCK-8), reactive oxygen species (ROS), and mitochondrial membrane potential (MMP) assay. Then, cell cycle and apoptosis were investigated by flow cytometry and verifed by immunoblotting. Next, NOX inhibitors (ML171 and apocynin), ROS scavenger (NAC), JNK inhibitor (SP600125), and caspase inhibitor (Z-VAD-FMK) were used to further investigate the underlying mechanisms of p20BAP31 on cell apoptosis. Finally, apoptosis-inducing factor (AIF) translocation from the mitochondria to the nuclei was verifed by immunoblotting and immunofuorescence assay.
Results: We found that overexpression of p20BAP31 indeed induced apoptosis and had a much greater sensitivity in HCT116 cells. Furthermore, the overexpression of p20BAP31 inhibited cell proliferation by causing S phase arrest. Further study revealed that p20BAP31 reduced MMP, with a signifcant increase in ROS levels, accompanied by the activation of the MAPK signaling pathway. Importantly, the mechanistic investigation indicated that p20BAP31 induces mitochondrial-dependent apoptosis by activating the ROS/JNK signaling pathway and induces caspase-independent apoptosis by promoting the nuclear translocation of AIF.
Conclusions: p20BAP31 induced cell apoptosis via both the ROS/JNK mitochondrial pathway and AIF caspase-independent pathway. Compared with antitumor drugs that are susceptible to drug resistance, p20BAP31 has unique advantages for tumor therapy.
Keywords p20BAP31, Apoptosis, AIF, ROS/JNK pathway, Colorectal cancer
Address and Contact Information College of Life and Health Science, Northeastern University, 195 Chuangxin Road, Hunnan District, Shenyang, Liaoning Province, China
*Corresponding author: jiangrui@mail.neu.edu.cn; wangbing@mail.neu.edu.cn
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No.  26DOI: 10.1186/s11658-023-00440-1 Volume 28 (2023) - 28:26
Authors Yu‐Min Choi1, Dong Hyun Kim1, Junghwa Jang1, Won Hyeok Choe2 and Bum‐Joon Kim1,3,4,5,6*
Abstract Background: In our previous report, the rt269I type versus the rt269L type in genotype C2 infection led to poor clinical outcomes and enhanced mitochondrial stress in infected hepatocytes. Here, we sought to investigate diferences between the rt269L and rt269I types in mitochondrial functionality in hepatitis B virus (HBV) genotype C2 infection, mainly focusing on endoplasmic reticulum (ER) stress-mediated autophagy induction as an upstream signal.
Methods: Mitochondrial functionality, ER stress signaling, autophagy induction, and apoptotic cell death between rt269L-type and rt269I-type groups were investigated via in vitro and in vivo experiments. Serum samples were collected from 187 chronic hepatitis patients who visited Konkuk or Seoul National University Hospital.
Results: Our data revealed that genotype C rt269L versus rt269I infection led to improved mitochondrial dynamics and enhanced autophagic fux, mainly due to the activation of the PERK–eIF2α–ATF4 axis. Furthermore, we demonstrated that the traits found in genotype C rt269L infection were mainly due to increased stability of the HBx protein after deubiquitination. In addition, clinical data using patient sera from two independent Korean cohorts showed that, compared with rt269I, rt269L in infection led to lower 8-OHdG levels, further supporting its improved mitochondrial quality control.
Conclusion: Our data showed that, compared with the rt269I type, the rt269L type, which presented exclusively in HBV genotype C infection, leads to improved mitochondrial dynamics or bioenergetics, mainly due to autophagy induction via activation of the PERK–eIF2α–ATF4 axis in an HBx protein-dependent manner. This suggests that HBx stability and cellular quality control in the rt269L type predominating in genotype C endemic areas could at least partly contribute to some distinctive traits of genotype C infection, such as higher infectivity or longer duration of the hepatitis B e antigen (HBeAg) positive stage.
Keywords Mitochondrial functionality, ER stress, Autophagy, HBx stability, Deubiquitination
Address and Contact Information 1 Department of Microbiology and Immunology, College of Medicine, Seoul National University, Seoul 110‐799, Republic of Korea
2 Department of Internal Medicine, Konkuk University School of Medicine, Seoul 05030, Republic of Korea
3 Department of Biomedical Sciences, College of Medicine, Seoul National University, Seoul 03080, Republic of Korea
4 Liver Research Institute, College of Medicine, Seoul National University, Seoul 03080, Korea
5 Cancer Research Institute, College of Medicine, Seoul National University, Seoul 03080, Korea
6 Seoul National University Medical Research Center (SNUMRC), Seoul 03080, Korea
*Corresponding author: kbumjoon@snu.ac.kr
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No.  27DOI: 10.1186/s11658-023-00439-8 Volume 28 (2023) - 28:27
Authors Shuzhao Zhuang1,2,3, Aaron Russell3, Yifan Guo1,2, Yingying Xu1,2 and Weihua Xiao1,2*
Abstract Background: Innate immune responses play essential roles in skeletal muscle recovery after injury. Programmed cell death protein 1 (PD-1) contributes to skeletal muscle regeneration by promoting macrophage proinfammatory to anti-infammatory phenotype transition. Interferon (IFN)-γ induces proinfammatory macrophages that appear to hinder myogenesis in vitro. Therefore, we tested the hypothesis that blocking IFN-γ in PD-1 knockout mice may dampen infammation and promote skeletal muscle regeneration via regulating the macrophage phenotype and neutrophils.
Methods: Anti-IFN-γ antibody was administered in PD-1 knockout mice, and cardiotoxin (CTX) injection was performed to induce acute skeletal muscle injury. Hematoxylin and eosin (HE) staining was used to view morphological changes of injured and regenerated skeletal muscle. Masson’s trichrome staining was used to assess the degree of fbrosis. Gene expressions of proinfammatory and anti-infammatory factors, fibrosis-related factors, and myogenic regulator factors were determined by real-time polymerase chain reaction (PCR). Changes in macrophage phenotype were examined by western blot and real-time PCR. Immunofuorescence was used to detect the accumulation of proinfammatory macrophages, anti-infammatory macrophages, and neutrophils.
Results: IFN-γ blockade in PD-1 knockout mice did not alleviate skeletal muscle damage or improve regeneration following acute cardiotoxin-induced injury. Instead, it exacerbated skeletal muscle infammation and fibrosis, and impaired regeneration via inhibiting macrophage accumulation, blocking macrophage proinfammatory to anti-infammatory transition, and enhancing infltration of neutrophils.
Conclusion: IFN-γ is crucial for efcient skeletal muscle regeneration in the absence of PD-1.
Keywords Skeletal muscle, Macrophage, Neutrophil, PD-1, IFN-γ
Address and Contact Information 1 Shanghai Frontiers Science Research Base of Exercise and Metabolic Health, Shanghai University of Sport, Shanghai, China
2 Key Laboratory of Exercise and Health Sciences, Shanghai University of Sport, Ministry of Education, Shanghai, China
3 Institute for Physical Activity and Nutrition, School of Exercise and Nutrition Sciences, Deakin University, Geelong, Australia
*Corresponding author: xiao_weihua@163.com
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No.  28DOI: 10.1186/s11658-023-00442-z Volume 28 (2023) - 28:28
Authors Shijie Li1†, Kerong Xin1†, Shen Pan2†, Yang Wang3†, Jianyi Zheng1, Zeyu Li1, Xuefeng Liu1, Bitian Liu1*, Zhenqun Xu1* and Xiaonan Chen1*
Abstract Bladder cancer (BC) is a clinical challenge worldwide with late clinical presentation, poor prognosis, and low survival rates. Traditional cystoscopy and tissue biopsy are routine methods for the diagnosis, prognosis, and monitoring of BC. However, due to the heterogeneity and limitations of tumors, such as aggressiveness, high cost, and limited applicability of longitudinal surveillance, the identifcation of tumor markers has attracted signifcant attention in BC. Over the past decade, liquid biopsies (e.g., blood) have proven to be highly efcient methods for the discovery of BC biomarkers. This noninvasive sampling method is used to analyze unique tumor components released into the peripheral circulation and allows serial sampling and longitudinal monitoring of tumor progression. Several liquid biopsy biomarkers are being extensively studied and have shown promising results in clinical applications of BC, including early detection, detection of microscopic residual disease, prediction of recurrence, and response to therapy. Therefore, in this review, we aim to provide an update on various novel blood-based liquid biopsy markers and review the advantages and current limitations of liquid biopsy in BC therapy. The role of blood-based circulating tumor cells, circulating tumor DNA, cell-free RNA, exosomes, metabolomics, and proteomics in diagnosis, prognosis, and treatment monitoring, and their applicability to the personalized management of BC, are highlighted.
Keywords Bladder cancer, Liquid biopsy, Circulating tumor cells, Circulating tumor DNA, Cell-free RNA, Exosomes, Metabolomics, Proteomics, Clinical application
Address and Contact Information 1 Department of Urology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, People’s Republic of China
2 Department of Radiology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, People’s Republic of China
3 Department of Gynecology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang 110042, Liaoning, People’s Republic of China
*Corresponding author: liu_bitian@163.com; zqxu@cmu.edu.cn; chenxn@cmu.edu.cn Shijie Li, Kerong Xin, Shen Pan, and Yang Wang contributed equally to this work.
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No.  29DOI: 10.1186/s11658-023-00444-x Volume 28 (2023) - 28:29
Title Correction: Inhibition of PAD4 enhances radiosensitivity and inhibits aggressive phenotypes of nasopharyngeal carcinoma cells
Authors Hao Chen1†, Min Luo1†, Xiangping Wang1, Ting Liang1, Chaoyuan Huang1, Changjie Huang1* and Lining Wei2*
Abstract Correction: Cellular & Molecular Biology Letters 2021, 26(1):9

Following publication of the original article [1], the authors informed us that the images of Transwell assays (Figs. 2G, 3F, 4F) were incorrect. Te correct images are given below. Te replacement of images does not afect the original conclusion.
Address and Contact Information 1 Department of Oncology, The Second Nanning People’s Hospital, No. 13 Dancun Road, Jiangnan District, Nanning 530031, Guangxi, China
2 Department of Endoscopy, The Afliated Tumor Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China
*Corresponding author: hcj1215423068@hotmail.Com; weilininggudy@163.com
Hao Chen and Min Luo contributed equally to this work
The original article can be found online at https://doi.org/10.1186/s11658-021-00251-2.
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No.  30DOI: 10.1186/s11658-023-00441-0 Volume 28 (2023) - 28:30
Authors Shan Wang1,2*, Liu Ying3, Shu‐Yi Yu4, Jie Bai5* and Chunbo Hao6*
Abstract Accurate assessment of the carcinogenic potential of oral mucosal diseases can signifcantly reduce the prevalence of oral cancer. We speculate that precancerous stem cells (pCSCs) arise during the evolution of carcinomas based on long-term experimental findings, published literature, and the cancer stem cell (CSC) theory, wherein pCSCs exist in precancerous lesions and have characteristics of both CSCs and normal stem cells. This apparently contradictory feature may be the foundation of the reversible transformation of precancerous lesions. Predicting malignant transformation in potentially malignant oral illnesses would allow for focused treatment, prognosis, and secondary prevention. Currently available clinical assays for chromosomal instability and DNA aneuploidy have several defciencies. We hope that our study will increase attention to pCSC research and lead to the development of novel strategies for the prevention and treatment of oral cancer by identifying pCSC markers.
Keywords Precancerous stem cells, Cancer stem cells, Oral cancer, Dysplasia, Malignant transformation
Address and Contact Information 1 Department of Oral Pathology, School of Stomatology, Hainan Medical University, Haikou 571199, People’s Republic of China
2 Department of Stomatology, The Second Afliated Hospital of Hainan Medical University, Haikou 570216, People’s Republic of China
3 College of Pharmacy, Hainan Medical University, Haikou 571199, People’s Republic of China
4 Pharmacy Department, First Afliated Hospital of Jiamusi University, Jiamusi 154003, People’s Republic of China
5 Department of Ophthalmology, the Fourth Afliated Hospital, Zhejiang University School of Medicine, Yiwu 322000, People’s Republic of China
6 Department of Stomatology, Hainan General Hospital (Hainan Afliated Hospital of Hainan Medical University), Haikou 570100, People’s Republic of China
*Corresponding author: birchtree20032003@126.com; hy0210032@hainanmc.edu.cn; 8020234@zju.edu.cn; haochunbonuli@sina.com
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No.  31DOI: 10.1186/s11658-023-00443-y Volume 28 (2023) - 28:31
Authors Jiaxian Zhu1,3, Ting Long1, Lingfang Gao1, Yan Zhong1, Ping Wang1,3, Xiaoyan Wang2,3,4, Zuguo Li1,4* and Zhiyan Hu2,3,4*
Abstract Background: Metastasis is the leading cause of death among patients with colorectal cancer (CRC). Therefore, it is important to explore the molecular mechanisms of metastasis to develop efective therapeutic targets for CRC. In the present study, ribosomal protein L21 (RPL21) was considered as being involved in promoting CRC metastasis, yet the underlying mechanism requires further investigation.
Methods: Immunohistochemistry, western blotting, and quantitative reverse transcription polymerase chain reaction were performed to measure the expression of RPL21 and lysosome-associated membrane protein 3 (LAMP3) in CRC tissues and cells. Wound healing, transwell migration, and invasion assays were performed to study the migration and invasion of cultured CRC cells. An orthotopic CRC mouse model was developed to investigate the metastatic ability of CRC. Transcriptome sequencing was conducted to identify the genes related to RPL21. The dual-luciferase reporter gene assay was performed to determine the transcriptional activity of transcription factor EB (TFEB). The GST/His pull-down assay was performed to investigate the specifc binding sites of RPL21 and LAMP3. The cell adhesion assay was performed to determine the adhesion ability of CRC cells. Immunofuorescence staining was performed to observe focal adhesions (FAs).
Results: RPL21 was highly expressed in CRC, contributing to tumor invasiveness and poor patient prognosis. Functionally, RPL21 promoted the migration and invasion of CRC cells in vitro and tumor metastasis in vivo. Moreover, LAMP3 was identifed as being highly related to RPL21 and was essential in promoting the migration and invasion of CRC cells. Mechanistically, RPL21 activated the transcriptional function of TFEB to upregulate LAMP3 expression. RPL21 directly bound to the aa 341–416 domain of LAMP3 via its aa 1–40 and aa 111–160 segments. The combination of RPL21 and LAMP3 enhanced the stability of the RPL21 protein by suppressing the degradation of the ubiquitin–proteasome system. Furthermore, RPL21 and LAMP3 promoted the formation of immature FAs by activating the FAK/paxillin/ERK signaling pathway.
Conclusions: RPL21 promoted invasion and metastasis by regulating FA formation in a LAMP3-dependent manner during CRC progression. The interaction between RPL21 and LAMP3 may function as a potential therapeutic target against CRC.
Keywords RPL21, LAMP3, TFEB, Focal adhesion, Colorectal cancer, Migration, Invasion, Metastasis
Address and Contact Information 1 Department of Pathology, Shenzhen Hospital, Southern Medical University, 1333 Xinhu Road, Shenzhen 518101, Guangdong, People’s Republic of China
2 Department of Pathology, Nanfang Hospital, Southern Medical University, 1838 Guangzhou Avenue North, Guangzhou 510515, Guangdong, People’s Republic of China
3 Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong, People’s Republic of China
4 Key Laboratory of Molecular Tumour Pathology of Guangdong Province, Guangzhou 510515, Guangdong, People’s Republic of China
*Corresponding author: lizg@smu.edu.cn; zhenyucai2011@163.com
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No.  32DOI: 10.1186/s11658-023-00445-w Volume 28 (2023) - 28:32
Authors Xiaomin Chen1, Tiange Lu1, Yiqing Cai1, Yang Han2, Mengfei Ding1, Yurou Chu1, Xiangxiang Zhou1,2,3,4,5* and Xin Wang1,2,3,4,5*
Abstract Background: N6-methyladenosine (m6A) has been shown to participate in various essential biological processes by regulating the level of target genes. However, the function of m6A modifcation mediated by KIAA1429 [alias virus-like m6A methyl-transferase-associated protein (VIRMA)] during the progression of difuse large B-cell lymphoma (DLBCL) remains undefned.
Methods: The expression and clinical signifcance of KIAA1429 were verifed by our clinical data. CRISPR/Cas9 mediated KIAA1429 deletion, and CRISPR/dCas9-VP64 for activating endogenous KIAA1429 was used to evaluate its biological function. RNA sequencing (RNA-seq), methylated RNA immunoprecipitation sequencing (MeRIP-seq), RNA immunoprecipitation (RIP) assays, luciferase activity assay, RNA stability experiments, and co-immunoprecipitation were performed to investigate the regulatory mechanism of KIAA1429 in DLBCL. Tumor xenograft models were established for in vivo experiments.
Results: Dysregulated expression of m6A regulators was observed, and a novel predictive model based on m6A score was established in DLBCL. Additionally, elevated KIAA1429 expression was associated with poor prognosis of patients with DLBCL. Knockout of KIAA1429 repressed DLBCL cell proliferation, facilitated cell cycle arrest in the G2/M phase, induced apoptosis in vitro, and inhibited tumor growth in vivo. Furthermore, carbohydrate sulfotransferase 11 (CHST11) was identifed as a downstream target of KIAA1429, which mediated m6A modifcation of CHST11 mRNA and then recruited YTHDF2 for reducing CHST11 stability and expression. Inhibition of CHST11 diminished MOB1B expression, resulting in inactivation of Hippo–YAP signaling, reprogramming the expression of Hippo target genes.
Conclusions: Our results revealed a new mechanism by which the Hippo–YAP pathway in DLBCL is inactivated by KIAA1429/YTHDF2-coupled epitranscriptional repression of CHST11, highlighting the potential of KIAA1429 as a novel predictive biomarker and therapeutic target for DLBCL progression.
Keywords Difuse large B-cell lymphoma, N6-methyladenosine, KIAA1429, YTHDF2, CHST11, Hippo–YAP
Address and Contact Information 1 Department of Hematology, Shandong Provincial Hospital, Shandong University, No.324, Jingwu Road, Jinan 250021, Shandong, China
2 Department of Hematology, Shandong Provincial Hospital Afliated to Shandong First Medical University, Jinan 250021, Shandong, China
3 4 Branch of National Clinical Research Center for Hematologic Diseases, Jinan 250021, Shandong, China
5 National Clinical Research Center for Hematologic Diseases, The First Afliated Hospital of Soochow University, Suzhou 251006, China
*Corresponding author: xiangxiangzhou@sdu.edu.cn; xinw007@126.com
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No.  33DOI: 10.1186/s11658-023-00438-9 Volume 28 (2023) - 28:33
Authors Mehrdad Hashemi1,2, Eisa Sabouni3, Parham Rahmanian3, Maliheh Entezari1,2, Mahsa Mojtabavi4, Behnaz Raei1, Mohammad Arad Zandieh5, Mitra Behroozaghdam1, Sepideh Mirzaei6, Kiavash Hushmandi5, Noushin Nabavi7, Shokooh Salimimoghadam8, Jun Ren9, Mohsen Rashidi10,11*, Rasoul Raesi12,13*, Afshin Taheriazam1,14*, Athanasios Alexiou15,16, Marios Papadakis17* and Shing Cheng Tan18
Abstract Hepatocellular carcinoma (HCC) is considered one of the greatest challenges to human life and is the most common form of liver cancer. Treatment of HCC depends on chemotherapy, radiotherapy, surgery, and immunotherapy, all of which have their own drawbacks, and patients may develop resistance to these therapies due to the aggressive behavior of HCC cells. New and efective therapies for HCC can be developed by targeting molecular signaling pathways. The expression of signal transducer and activator of transcription 3 (STAT3) in human cancer cells changes, and during cancer progression, the expression tends to increase. After induction of STAT3 signaling by growth factors and cytokines, STAT3 is phosphorylated and translocated to the nucleus to regulate cancer progression. The concept of the current review revolves around the expression and phosphorylation status of STAT3 in HCC, and studies show that the expression of STAT3 is high during the progression of HCC. This review addresses the function of STAT3 as an oncogenic factor in HCC, as STAT3 is able to prevent apoptosis and thus promote the progression of HCC. Moreover, STAT3 regulates both survival- and death-inducing autophagy in HCC and promotes cancer metastasis by inducing the epithelial–mesenchymal transition (EMT). In addition, upregulation of STAT3 is associated with the occurrence of chemoresistance and radioresistance in HCC. Specifcally, non-protein-coding transcripts regulate STAT3 signaling in HCC, and their inhibition by antitumor agents may afect tumor progression. In this review, all these topics are discussed in detail to provide further insight into the role of STAT3 in tumorigenesis, treatment resistance, and pharmacological regulation of HCC.
Keywords Hepatocellular carcinoma, Liver cancer, Noncoding transcripts, STAT3, Molecular signaling
Address and Contact Information 1 Farhikhtegan Medical Convergence Sciences Research Center, Farhikhtegan Hospital Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
2 Department of Genetics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
3 Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran.
4 Mashhad Branch, Islamic Azad University, Mashhad, Iran.
5 Division of Epidemiology, Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
6 Department of Biology, Faculty of Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
7 Department of Urologic Sciences and Vancouver Prostate Centre, University of British Columbia, Vancouver, BC V6H3Z6, Canada.
8 Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
9 Department of Cardiology, Zhongshan Hospital, Shanghai Institute of Cardiovascular Diseases, Fudan University, Shanghai 200032, China.
10 Department Pharmacology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.
11 The Health of Plant and Livestock Products Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
12 Department of Health Services Management, Mashhad University of Medical Sciences, Mashhad, Iran.
13 Department of Medical-Surgical Nursing, Mashhad University of Medical Sciences, Mashhad, Iran.
14 Department of Orthopedics, Faculty of Medicine, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
15 Department of Science and Engineering, Novel Global Community Educational Foundation, Hebersham, Australia.
16 AFNP Med Austria, Vienna, Austria.
17 Department of Surgery II, University Hospital Witten-Herdecke, University of Witten-Herdecke, Heusnerstrasse 40, 42283 Wuppertal, Germany.
18 UKM Medical Molecular Biology Institute, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
*Corresponding author: dr.mohsenrashidi@yahoo.com; raesi.br881@gmail.com; a.taheriazam@iautmu.ac.ir; marios_papadakis@yahoo.gr
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No.  34DOI: 10.1186/s11658-023-00448-7 Volume 28 (2023) - 28:34
Authors Wenfang Zheng1, Xuehui Wang1, Yunhe Yu1, Changle Ji1 and Lin Fang1*
Abstract Background: Breast cancer (BC) is a common threat to women. The continuous activation of nuclear factor kappa B (NF-κB) signaling pathway contributes to the development of BC. This study aimed to investigate the role of a circular RNA (circRNF10) in BC progression and regulating NF-κB signaling pathway.
Methods: Bioinformatics analysis, RT-qPCR, subcellular fractionation, FISH, RNase R treatment, and actinomycin D assay were used to explore the expression and characteristics of circRNF10 in BC. The biological functions of circRNF10 in BC were analyzed by MTT assay, colony formation assay, wound healing assay, and Transwell assay. RNA pulldown and RIP assay were used to identify the interaction between circRNF10 and DEAH (Asp-Glu-Ala-His) box helicase 15 (DHX15). The impact of circRNF10-DHX15 interaction on NF-κB signaling pathway was explored by western blot, IF, and co-IP. Furthermore, dual-luciferase reporter assay, ChIP, and EMSA were performed to assess the efect of NF-κB p65 on DHX15 transcription.
Results: CircRNF10 was downregulated in BC, and lower expression of circRNF10 was related to poor prognosis of patients with BC. CircRNF10 inhibited the proliferation and migration of BC. Mechanically, circRNF10-DHX15 interaction sequestered DHX15 from NF-κB p65, thereby inhibiting the activation of NF-κB signaling pathway. On the other hand, NF-κB p65 enhanced DHX15 transcription by binding to the promoter of DHX15. Altogether, circRNF10 impaired the DHX15-NF-κB p65 positive feedback loop and suppressed the progression of BC.
Conclusion: CircRNF10-DHX15 interaction suppressed the DHX15-NF-κB p65 positive feedback loop, thereby inhibiting BC progression. These findings provide new insights in the continuous activation of NF-κB signaling pathway and raised potential therapeutic approach for BC treatment.
Keywords circRNF10, DHX15, p65, Feedback, Breast cancer
Address and Contact Information 1 Department of Breast and Thyroid Surgery, Shanghai Tenth People’s Hospital, School of Medicine, Tongji University, 301 Yanchangzhong Road, Shanghai 200072, People’s Republic of China
*Corresponding author: 1200038@tongji.edu.cn
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No.  35DOI: 10.1186/s11658-022-00415-8 Volume 28 (2023) - 28:35
Authors Wencheng Di1†, Zhenxiao Jin2†, Wangrui Lei3,4†, Qiong Liu3,4, Wenwen Yang3,4, Shaofei Zhang3,4, Chenxi Lu3,4, Xiaoling Xu3,4* and Huadong Zhao5*
Abstract Background: Heart failure is a common complication of sepsis with a high mortality rate. It has been reported that melatonin can attenuate septic injury due to various properties. On the basis of previous reports, this study will further explore the efects and mechanisms of melatonin pretreatment, posttreatment, and combination with antibiotics in the treatment of sepsis and septic myocardial injury.
Methods and results: Our results showed that melatonin pretreatment showed an obvious protective efect on sepsis and septic myocardial injury, which was related to the attenuation of infammation and oxidative stress, the improvement of mitochondrial function, the regulation of endoplasmic reticulum stress (ERS), and the activation of the AMPK signaling pathway. In particular, AMPK serves as a key efector for melatonin-initiated myocardial benefts. In addition, melatonin posttreatment also had a certain degree of protection, while its efect was not as remarkable as that of pretreatment. The combination of melatonin and classical antibiotics had a slight but limited efect. RNA-seq detection clarifed the cardioprotective mechanism of melatonin.
Conclusion: Altogether, this study provides a theoretical basis for the application strategy and combination of melatonin in septic myocardial injury.
Keywords Melatonin, Septic myocardial injury, RNA-seq, Pretreatment, Posttreatment, Antibiotics
Address and Contact Information 1 Department of Cardiovascular Medicine, National Clinical Research Center for Infectious Disease, Shenzhen Third People’s Hospital, 29 Bulan Road, Shenzhen, Guangdong Province, China
2 Department of Cardiovascular Surgery, Xijing Hospital, The Airforce Military Medical University, 127 Changle West Road, Xi’an, China
3 Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, 229 Taibai North Road, Xi’an, China
4 Faculty of Life Sciences and Medicine, Northwest University, 229 Taibai North Road, Xi’an, China
5 Department of General Surgery, Tangdu Hospital, The Airforce Military Medical University, 1 Xinsi Road, Xi’an, China
*Corresponding author: yang200214yy@nwu.edu.cn; huadongzhaotd@163.com
Wencheng Di,Zhenxiao Jin, Wangrui Lei contributed equally as co-frst authors
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No.  36DOI: 10.1186/s11658-023-00449-6 Volume 28 (2023) - 28:36
Authors Leilei Wang1, Chuan Wang2,3, Xuan Li4, Zhuoying Tao1, Wangyong Zhu5, Yuxiong Su1 and Wing Shan Choi1*
Abstract Background: Oral squamous cell carcinomas are one of the most common cancers worldwide with aggressive behavior and poor prognosis. Reactive oxygen species (ROS) are associated with cancer and cause various types of regulated cell death (RCD). Inducing the RCD pathway by modulating ROS levels is imperative to conquer cancers. The aim of this study is to investigate the synergistic anticancer efects of melatonin and erastin on ROS modulation and subsequent RCD induction.
Methods: Human tongue squamous cell carcinoma cell lines (SCC-15 cells) were treated with melatonin, erastin, or their combination. Cell viability, ROS levels, autophagy, apoptosis, and ferroptosis levels were tested according to the results of the PCR array, which were verifed with/without the induction and inhibition of ROS by H2O2 and N-acetyl-L-cysteine, respectively. In addition, a mouse-based subcutaneous oral cancer xenograft model was constructed to identify the efects of melatonin, erastin, and their combination on the autophagy, apoptosis, and ferroptosis levels in isolated tumor tissues.
Results: ROS levels were increased by the administration of melatonin at high concentrations (mM), and the combination of melatonin with erastin enhanced the levels of malonic dialdehyde, ROS, and lipid ROS, and reduced the levels of glutamate and glutathione. SQSTM1/p62, LC3A/B, cleaved caspase-3, and PARP1 protein levels in SCC-15 cells were also increased by melatoninplus erastin treatment, which further increased as ROS accumulated, and decreased as ROS levels were suppressed. Combined treatment of melatonin and erastin markedly reduced the tumor size in vivo, demonstrated no obvious systemic side efects, and signifcantly enhanced the apoptosis and ferroptosis levels in the tumor tissues, in parallel with decreased autophagy levels.
Conclusions: Melatonin combined with erastin exhibits synergistic anticancer efects without adverse reactions. Herein, this combination might become a promising alternative strategy for oral cancer treatment.
Keywords Reactive oxygen species (ROS), Oral cancer, Melatonin, Erastin, Apoptosis, Autophagy, Ferroptosis
Address and Contact Information 1 Division of Oral and Maxillofacial Surgery, Faculty of Dentistry, The University of Hong Kong, 34 Hospital Road, Hong Kong SAR, China
2 The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST KLOS) and Key Laboratory of Oral Biomedicine Ministry of Education (KLOBME), School and Hospital of Stomatology, Wuhan University, Wuhan, China
3 Department of Periodontology, School and Hospital of Stomatology, Wuhan University, Wuhan, China
4 Division of Periodontology and Implant Dentistry, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
5 Department of Dental Surgery, The University of Hong Kong-Shenzhen Hospital, Shenzhen, Guangdong, China
*Corresponding author: drwchoi@gmail.com
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No.  37DOI: 10.1186/s11658-023-00447-8 Volume 28 (2023) - 28:37
Authors Amir Saf1, Mohammadreza Saberiyan2, Mohammad‐Javad Sanaei3, Samaneh Adelian2, Fateme Davarani Asl2, Mahsa Zeinaly4, Mahdi Shamsi5 and Reza Ahmadi6*
Abstract Metabolic reprogramming is a well-known feature of cancer that allows malignant cells to alter metabolic reactions and nutrient uptake, thereby promoting tumor growth and spread. It has been discovered that noncoding RNAs (ncRNAs), including microRNA (miRNA), long noncoding RNA (lncRNA), and circular RNA (circRNA), have a role in a variety of biological functions, control physiologic and developmental processes, and even infuence disease. They have been recognized in numerous cancer types as tumor suppressors and oncogenic agents. The role of ncRNAs in the metabolic reprogramming of cancer cells has recently been noticed. We examine this subject, with an emphasis on the metabolism of glucose, lipids, and amino acids, and highlight the therapeutic use of targeting ncRNAs in cancer treatment.
Keywords Cancer, Metabolic reprogramming, Noncoding RNAs, MicroRNA, Long noncoding RNA, Circular RNA
Address and Contact Information 1 Clinical Biochemistry Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
2 Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
3 Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
4 Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran
5 Department of Cell and Molecular Biology, Najafabad Branch, Islamic Azad University, Najafabad, Iran
6 Medical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Rahmatiyeh Region, Shahrekord, Iran
*Corresponding author: ahmadi.r@skums.ac.ir
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No.  38DOI: 10.1186/s11658-023-00450-z Volume 28 (2023) - 28:38
Authors Xudong Yu1,2†, Ruijia Liu1†, Wenfeng Gao1, Xuyun Wang3* and Yaosheng Zhang1,2*
Abstract Prostate cancer is one of the more heterogeneous tumour types. In recent years, with the rapid development of single-cell sequencing and spatial transcriptome technologies, researchers have gained a more intuitive and comprehensive understanding of the heterogeneity of prostate cancer. Tumour-associated epithelial cells; cancer-associated fibroblasts; the complexity of the immune microenvironment, and the heterogeneity of the spatial distribution of tumour cells and other cancer-promoting molecules play a crucial role in the growth, invasion, and metastasis of prostate cancer. Single-cell multi-omics biotechnology, especially single-cell transcriptome sequencing, reveals the expression level of single cells with higher resolution and finely dissects the molecular characteristics of diferent tumour cells. We reviewed the recent literature on prostate cancer cells, focusing on single-cell RNA sequencing. And we analysed the heterogeneity and spatial distribution diferences of diferent tumour cell types. We discussed the impact of novel single-cell omics technologies, such as rich omics exploration strategies, multi-omics joint analysis modes, and deep learning models, on future prostate cancer research. In this review, we have constructed a comprehensive catalogue of single-cell omics studies in prostate cancer. This article aimed to provide a more thorough understanding of the diagnosis and treatment of prostate cancer. We summarised and proposed several key issues and directions on applying single-cell multi-omics and spatial transcriptomics to understand the heterogeneity of prostate cancer. Finally, we discussed single-cell omics trends and future directions in prostate cancer.
Keywords Prostate cancer, Single-cell omics, Spatial transcriptomics, Heterogeneity
Address and Contact Information 1 Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
2 Beijing Tumour Minimally Invasive Medical Center of Integrated Traditional Chinese and Western Medicine, Beijing 101121, China
3 Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing 100010, China
*Corresponding author: xuyun1102@sina.com; zhangyaosheng@bucm.edu.cn
Xudong Yu and Ruijia Liu contributed equally to this work and share the frst authorship
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No.  39DOI: 10.1186/s11658-023-00446-9 Volume 28 (2023) - 28:39
Authors Pearlyn Jia Ying Toh1,2, Marius Sudol1,3 and Timothy Edward Saunders1,4,5*
Abstract Background: Tissues need to regenerate to restore function after injury. Yet, this regenerative capacity varies signifcantly between organs and between species. For example, in the heart, some species retain full regenerative capacity throughout their lifespan but human cardiac cells display a limited ability to repair the injury. After a myocardial infarction, the function of cardiomyocytes is impaired and reduces the ability of the heart to pump, causing heart failure. Therefore, there is a need to restore the function of an injured heart post myocardial infarction. We investigate in cell culture the role of the Yes-associated protein (YAP), a transcriptional co-regulator with a pivotal role in growth, in driving repair after injury.
Methods: We express optogenetic YAP (optoYAP) in three diferent cell lines. We characterised the behaviour and function of optoYAP using fluorescence imaging and quantitative real-time PCR of downstream YAP target genes. Mutant constructs were generated using site-directed mutagenesis. Nuclear localised optoYAP was functionally tested using wound healing assay.
Results: Utilising optoYAP, which enables precise control of pathway activation, we show that YAP induces the expression of downstream genes involved in proliferation and migration. optoYAP can increase the speed of wound healing in H9c2 cardiomyoblasts. Interestingly, this is not driven by an increase in proliferation, but by collective cell migration. We subsequently dissect specifc phosphorylation sites in YAP to identify the molecular driver of accelerated healing.
Conclusions: This study shows that optogenetic YAP is functional in H9c2 cardiomyoblasts and its controlled activation can potentially enhance wound healing in a range of conditions.
Keywords Hippo-YAP, Optogenetics, Wound healing
Address and Contact Information 1 Mechanobiology Institute, National University of Singapore, Singapore, Singapore
2 Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark
3 Icahn School of Medicine at Mount Sinai, New York, NY, USA
4 Institute of Molecular and Cell Biology, A*STAR, Singapore, Singapore
5 Warwick Medical School, University of Warwick, Coventry, UK
*Corresponding author: timothy.saunders@warwick.ac.uk
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No.  40DOI: 10.1186/s11658-023-00456-7 Volume 28 (2023) - 28:40
Title Galectin-1 promotes angiogenesis and chondrogenesis during antler regeneration
Authors Xunsheng Li1, Wanwan Shi1, Guanning Wei1, Jinpeng Lv1, Datao Wang1, Baorui Xing1, Jue Zhou1, Jianwei Zhao1 and Hongmei Sun1*
Abstract Background: Deer antlers are the only known mammalian structure that undergoes full regeneration. In addition, it is peculiar because when growing, it contains vascularized cartilage. The diferentiation of antler stem cells (ASCs) into chondrocytes while inducing endochondral extension of blood vessels is necessary to form antler vascularized cartilage. Therefore, antlers provide an unparalleled opportunity to investigate chondrogenesis, angiogenesis, and regenerative medicine. A study found that Galectin-1 (GAL-1), which can be used as a marker in some tumors, is highly expressed in ASCs. This intrigued us to investigate what role GAL-1 could play in antler regeneration.
Methods: We measured the expression level of GAL-1 in antler tissues and cells by immunohistochemistry, WB and QPCR. We constructed antlerogenic periosteal cells (APCs, one cell type of ASCs) with the GAL-1 gene knocked out (APCGAL-1−/−) using CRISPR-CAS9 gene editing system. The efect of GAL-1 on angiogenesis was determined by stimulating human umbilical vein endothelial cells (HUVECs) using APCGAL-1−/− conditioned medium or adding exogenous deer GAL-1 protein. The efect of APCGAL-1−/− on chondrogenic diferentiation was evaluated compared with the APCs under micro-mass culture. The gene expression pattern of APCGAL-1−/− was analyzed by transcriptome sequencing.
Results: Immunohistochemistry revealed that GAL-1 was widely expressed in the antlerogenic periosteum (AP), pedicle periosteum (PP) and antler growth center. Western blot and qRT-PCR analysis using deer cell lines further supports this result. The proliferation, migration, and tube formation assays of human umbilical vein endothelial cells (HUVECs) showed that the proangiogenic activity of APCGAL-1−/− medium was signifcantly decreased (P<0.05) compared with the APCs medium. The proangiogenic activity of deer GAL-1 protein was further confrmed by adding exogenous deer GAL-1 protein (P<0.05). The chondrogenic diferentiation ability of APCGAL-1−/− was impeded under micro-mass culture. The terms of GO and KEGG enrichment of the diferentially expressed genes (DEGs) of APCGAL-1−/− showed that down-regulated expression of pathways associated with deer antler angiogenesis, osteogenesis and stem cell pluripotency, such as the PI3K-AKT signaling pathway, signaling pathways regulating pluripotency of stem cells and TGF-β signaling pathway.
Conclusions: Deer GAL-1, has strong angiogenic activity, is widely and highly expressed in deer antler. The APCs can induce angiogenesis by secreting GAL-1. The knockout of GAL-1 gene of APCs damaged its ability to induce angiogenesis and diferentiate into chondrocytes. This ability is crucial to the formation of deer antler vascularized cartilage. Moreover, Deer antlers ofer a unique model to explore explore how angiogenesis at high levels of GAL-1 expression can be elegantly regulated without becoming cancerous.
Keywords Galectin-1, Antler stem cells, Regeneration, Crispr/cas9, Angiogenesis, Chondrogenesis
Address and Contact Information 1 Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Jilin, China
*Corresponding author: Sunhongmei@caas.cn
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No.  41DOI: 10.1186/s11658-023-00454-9 Volume 28 (2023) - 28:41
Authors Jinhe Li2†, Qinchun Liang2,3†, HuaPing Zhou1, Ming Zhou1* and Hongxin Huang1*
Abstract The plasmid vector platform is the most commonly used vector for the expression of the versatile CRISPR-Cas technique and the promoter is a crucial element for the expression vector, thus profling the impact of the promoters on CRISPR editors provides the basic information for the gene-editing toolkits and can be a guideline for its design. Herein, we made a parallel comparison among four commonly used promoters (CAG,~1700 bp; EF1a core,~210 bp; CMV, ~500 bp; and PGK,~500 bp) in CRISPR-Cas12a system in mammalian cells to explore the impact of promoters on this powerful tool. We found that without badly damaging targeting specifcity, the CAG promoter-driving Cas12a editor exhibited the most active (eficiency takes as 100%, specificity index= ~75%) in genomic cleavage, multiplex editing, transcriptional activation, and base editing, followed by promoter CMV (eficiency=70~90% (vs CAG), specificity index= ~78%), and then EF1a core and PGK (both efciency=40–60%, vs CAG) but with higher specificity (specificity index= ~84% and~82%, respectively). Therefore, CAG is recommended in the CRISPR-Cas12a system for the applications that need a robust editing activity but without size limitation, CMV mostly can be an alternative for CAG when requiring a smaller space, EF1a is similar to PGK with relatively high specificity, but has a smaller size, thus is more suitable for in vivo therapeutic applications. The data outlined the properties of the widely used promoters in the CRISPR-Cas12a system, which can be a guide for its applications and can be a useful resource for the gene-editing feld.
Keywords CRISPR-Cas system, Promoters, Impact, Comparison, Plasmid design
Address and Contact Information 1 Afliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou 510095, China
2 The Third Afliated Hospital of Southern Medical University, Guangzhou 510630, China
3 Guangzhou Key Laboratory of Neuropathic Pain Mechanism at Spinal Cord Level, Guangzhou 510630, China
*Corresponding author: zhouming@gzhmu.edu.cn; huanghx08@gzhmu.edu.cn
Jinhe Li and Qinchun Liang equally contributed to this work.
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