Biochimica et biophysica acta. Molecular basis of disease最新文献

{"title":"Targeting Ca2+-activated K+ channels in glioma","authors":"Yuequ Zhang ,&nbsp;Hennrique Taborda Ribas ,&nbsp;Sheila M.B. Winnischofer ,&nbsp;Martina Schmidt ,&nbsp;Marina Trombetta-Lima ,&nbsp;Amalia M. Dolga","doi":"10.1016/j.bbadis.2025.167950","DOIUrl":"10.1016/j.bbadis.2025.167950","url":null,"abstract":"<div><div>Glioma affects millions of people worldwide and there is a lack of effective therapies. Glioblastoma multiforme (GBM) is the most common and deadliest form of primary brain tumor in adults. Emerging evidence indicated that targeting ion channels may be a promising therapeutic approach for GBM. Altered expression and activity of the Ca²⁺- activated K⁺ (K_Ca) channels have been reported in GBM patients. Generally, large-conductance K_Ca (BK_Ca) channels and intermediate-conductance K_Ca (IK_Ca or SK4) channels are highly expressed in glioma samples compared to healthy control brain tissue. Analyzing TCGA database, the expression of <em>KCNMB1</em> (encoding the BK_Ca channel protein) and <em>KCNN4</em> (encoding the K_Ca3.1/IK_Ca protein) genes was upregulated, while <em>KCNN1</em> (encoding the K_Ca2.1 protein) gene expression was downregulated in GBM patients grade IV compared to GBM patients grade I or II. The gene expression and activity of K_Ca channels may contribute to survival outcomes, by regulating cellular processes like cell proliferation and migration. Importantly, modulation of the activity of K_Ca channels reduced the proliferation and migration of GBM cells and suppressed glioma progression both in vivo and in vitro cell models for GBM. Herein, we aim to review how modulation of the activity of K_Ca channels impacts tumor development in terms of proliferation, cell death, invasion, metabolism and immune system in GBM.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167950"},"PeriodicalIF":4.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144242231","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of the CEACAM7-JAK2/STAT3-BST2 Axis in the migration of nasopharyngeal carcinoma cells","authors":"Ju-Pi Li ,&nbsp;Chiao-Wen Lin ,&nbsp;Yen-Ting Lu ,&nbsp;Cheng-Chen Huang ,&nbsp;Yu-Ting Ho ,&nbsp;Chung-Han Hsin ,&nbsp;Shun-Fa Yang","doi":"10.1016/j.bbadis.2025.167948","DOIUrl":"10.1016/j.bbadis.2025.167948","url":null,"abstract":"<div><h3>Objective</h3><div>Nasopharyngeal carcinoma (NPC) is a rare malignancy of the head and neck, with recurrence and distant metastasis being the primary causes of mortality among affected patients. Carcinoembryonic antigen cellular adhesion molecules (CEACAMs) are implicated in various physiological and pathophysiological processes. However, the role of CEACAM7 in NPC progression remains unclear. This study investigated the influence of CEACAM7 on NPC progression.</div></div><div><h3>Methods</h3><div>Cell migration and invasion ability were conducted using a Boyden chamber assay. Regulation of signaling pathways was evaluated by immunoblotting.</div></div><div><h3>Results</h3><div>The findings revealed that CEACAM7 is highly expressed in human NPC tissues, with particularly elevated levels observed in malignant tissues and high-grade squamous cell carcinoma tissues. Furthermore, CEACAM7 was demonstrated to regulate cell migration and invasion in human NPC cells but did not influence cell proliferation. Moreover, CEACAM7 enhanced cell motility through the activation of the JAK2/STAT3/BST2 signaling pathway. Notably, cotreatment with WP1066, a JAK2/STAT3 inhibitor, suppressed CEACAM7-induced cell migratory ability and downregulated BST2 expression in NPC cells.</div></div><div><h3>Conclusions</h3><div>These findings indicate that the CEACAM7/JAK2/STAT3/BST2 axis plays a crucial role in promoting NPC cell migration and underscore its potential as a therapeutic target for mitigating NPC metastasis.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167948"},"PeriodicalIF":4.2,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144242230","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Calreticulin (crt-1) silencing reduces Aß1–42-induced toxicity and restores muscle function in C. elegans.","authors":"Elena Caldero-Escudero,&nbsp;Silvia Romero-Sanz,&nbsp;Pilar Álvarez-Illera,&nbsp;Sergio De la Fuente,&nbsp;Paloma García-Casas,&nbsp;Rosalba I. Fonteriz,&nbsp;Mayte Montero,&nbsp;Javier Álvarez,&nbsp;Jaime Santo-Domingo","doi":"10.1016/j.bbadis.2025.167946","DOIUrl":"10.1016/j.bbadis.2025.167946","url":null,"abstract":"<div><div>Accumulation of aggregated β-amyloid peptide is a key histopathological feature of Alzheimer's Disease (AD). Experimental models of AD based on β-amyloid peptide display calcium (Ca²⁺) signaling alterations, and targeting key components of the cellular Ca²⁺ signaling system has been postulated to modulate AD onset and progression. Here we have taken advantage of a <em>C. elegans</em> strain that over-expresses the most toxic human ß-amyloid peptide (Aß₁_–₄₂) in body-wall muscle cells, to study the impact of calreticulin (<em>crt-1</em>) silencing on body-wall muscle performance. <em>Crt-1</em> knockdown reduced the percentage of paralyzed worms in a dose-dependent manner and improved locomotion parameters in free-mobility assays in Aß₁_–₄₂-overexpressing worms. At the cellular level, <em>crt-1</em> silencing prevented Aß₁_–₄₂-induced exacerbated mitochondrial respiration and mitochondrial ROS production without impacting mitochondrial sarcomere organization. <em>Crt-1</em> knockdown reduced the number and size of Aß₁_–₄₂ aggregates in body-wall muscle cells and prevented the formation of Aß₁_–₄₂ oligomers. We propose that <em>crt-1</em> depletion reduces the number of Aß₁_–₄₂ aggregates, precluding Aß₁_–₄₂-induced mitochondrial toxicity and improving muscle function. We identify <em>C. elegans crt-1</em> as a gene involved in the toxicity associated with the expression of human Aß₁_–₄₂, and thus a potential new target for treatment.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167946"},"PeriodicalIF":4.2,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144222247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microglia-drive IRF8 upregulates complement pathway in Parkinson's disease","authors":"Hongkai Yao ,&nbsp;Chenming Liu ,&nbsp;Lingjing Jin ,&nbsp;Yunping Song","doi":"10.1016/j.bbadis.2025.167928","DOIUrl":"10.1016/j.bbadis.2025.167928","url":null,"abstract":"<div><div>Parkinson's disease (PD) is a widespread degenerative disorder of the central nervous system. The gradual degeneration of dopaminergic neurons in the substantia nigra region is one of the primary pathological features of PD. Glial cells in SN are also linked to the pathological PD alterations. To discern the role of neurons and glial cells as well as their corresponding genetic modifications in PD, we utilized diverse bioinformatics techniques and performed biological experiments on cell and animal models. Several transcriptome datasets of the substantia nigra region were collected from the Gene Expression Omnibus dataset. Cibersort was used to deconvolute the data into proportions of brain cell types. WGCNA was used to analyze the association between modules and traits. Machine learning was used to select the hub genes from WGCNA results. Based on the results of transcriptome analysis, microglia were the most related cell type. Through machine learning, IRF8 was identified as the hub gene associated with PD and microglia. Furthermore, an increased ratio of IRF8+ microglia was observed in PD mice, along with an elevated expression of IRF8 in primary microglia cultures treated with α-synuclein preformed fibril (PFFs). To explore the function of IRF8 in microglia under disease condition, we conducted siRNA of IRF8 and found it was highly associated with complement pathway, which may cause the activation of microglia. In conclusion, our research indicated IRF8 may be involved in the functional regulation of microglia in PD.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167928"},"PeriodicalIF":4.2,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144190486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-cell RNA-seq reveals the immune profile changes in patients with diarrhoeal-irritable bowel syndrome","authors":"Xiaoxuan Xue ,&nbsp;Juncong Hu ,&nbsp;Wenjing Pei ,&nbsp;Tangyou Mao ,&nbsp;Yali Yuan ,&nbsp;Yang Zhang ,&nbsp;Yupu Yao ,&nbsp;Lei Wang ,&nbsp;Chune Xie ,&nbsp;Junxiang Li ,&nbsp;Xingjie Zhao","doi":"10.1016/j.bbadis.2025.167945","DOIUrl":"10.1016/j.bbadis.2025.167945","url":null,"abstract":"<div><h3>Background</h3><div>Diarrhea-predominant irritable bowel syndrome (IBS-D) is characterized by recurrent abdominal pain and chronic diarrhea. T lymphocytes, which play a crucial role in gut inflammation and immune responses, may significantly contribute to the pathophysiology of IBS-D. However, the exact mechanisms by which T lymphocytes affect IBS-D remain unclear. The precise pathways and interactions involved in IBS-D are still to be determined.</div></div><div><h3>Methods</h3><div>We conducted single-cell RNA sequencing on blood samples from 4 IBS-D patients and 4 healthy controls. Following data preprocessing, we conducted subsequence bioinformatics analysis. Additionally, serum and colon tissue samples from IBS-D rat models were analyzed using ELISA and three-parameter fluorescence to further elucidate the T lymphocytes landscape associated with IBS-D.</div></div><div><h3>Results</h3><div>A total of 45,649 cells were classified into four distinct cell types. Among them, T lymphocytes were further subdivided into 20 unique clusters. Novel markers that were highly expressed in T lymphocytes were identified. Dysregulation of HIF-1α pathway, NF-kappa B pathway, and IL-17 signaling pathway, were observed through trajectory analysis. Additionally, single-cell regulatory network inference and clustering analysis revealed the FOS signaling pathway as a potential therapeutic target for IBS-D. Furthermore, we detected abnormally elevated levels of PLK3 and NFKBIZ in the serum and colon tissues of the IBS-D rat model. Our study mapped the communication atlas of T lymphocytes that may influence the pathophysiology of IBS-D.</div></div><div><h3>Conclusions</h3><div>This study uncovers novel molecular features and identifies potential therapeutic targets of T lymphocytes in IBS-D, thereby advancing our understanding of the disease and expanding treatment options.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167945"},"PeriodicalIF":4.2,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144210472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular vesicle-dependent crosstalk between hepatic stellate cells and Kupffer cells promotes their mutual activation","authors":"Junyu Wang ,&nbsp;Jia Li ,&nbsp;Manon Buist-Homan ,&nbsp;Martin C. Harmsen ,&nbsp;Han Moshage","doi":"10.1016/j.bbadis.2025.167914","DOIUrl":"10.1016/j.bbadis.2025.167914","url":null,"abstract":"<div><h3>Background &amp; aims</h3><div>Hepatic fibrosis results from hepatic stellate cell (HSC) activation and excessive extracellular matrix (ECM) deposition, driven by chronic inflammation. Kupffer cells (KCs) play a central role in HSC activation. We previously showed that HSC-secreted factors, particularly extracellular vesicles (EVs), activate KCs. However, the reciprocal effects of activated KCs on HSCs remain poorly understood. This study investigates the bidirectional crosstalk between HSCs and KCs, focusing on the role of KC-derived EVs in regulating HSC activation and fibrosis progression.</div></div><div><h3>Methods</h3><div>Primary HSCs and KCs were isolated from male Wistar rats. HSCs were co-cultured with KCs for 24 h to assess inflammatory and activation markers. LPS-stimulated KC-derived EVs and controls were administered to HSCs on day 1. LPS and the Toll-like receptor 4 (TLR4) inhibitor TAK-242 were used to investigate the intercellular communication in detail.</div></div><div><h3>Results</h3><div>Co-cultured HSCs and KCs showed mutual activation, demonstrated by elevated inflammatory markers in both cell types and enhanced HSC pro-fibrotic activation. Pro-inflammatory (LPS)-activated KCs amplified HSC activation in a TLR4-dependent fashion. Part of this augmented HSC activation was attributed to EVs.</div></div><div><h3>Conclusions</h3><div>In co-culture, KCs and HSCs show mutual activation in a TLR4-dependent fashion. This bidirectional activation is augmented by pro-inflammatory mediators. KC-derived EVs (partially) activate HSCs, which might contribute to progression of liver fibrosis <em>in vivo</em>. Modulating KC activation, such as by blocking TLR4 signaling, may alter EV secretion or cargo composition, reducing HSC activation and fibrosis progression. Targeting this EV-mediated crosstalk could provide novel therapeutic strategies for liver fibrosis.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167914"},"PeriodicalIF":4.2,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144168478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combined inhibition of EGFR and FGFRs with Cetuximab and Infigratinib showed effectiveness and relevance in proliferation and migration of HNSCC cell lines","authors":"Monika Orsolic,&nbsp;Marc Diensthuber,&nbsp;Timo Stöver,&nbsp;Christin Geißler","doi":"10.1016/j.bbadis.2025.167940","DOIUrl":"10.1016/j.bbadis.2025.167940","url":null,"abstract":"<div><div>Head and neck squamous cell carcinoma (HNSCC) represents a significant oncological challenge, necessitating novel treatments due to high global mortality. The current study focuses on the potential of combining Cetuximab (CTX), targeting EGFR, and Infigratinib (BGJ), targeting FGFRs, to improve treatment outcomes.</div><div>The effects of CTX and BGJ on HNSCC cell lines were investigated by analyzing cell count and gap closure to assess proliferation and migration. Immunohistochemistry was used to assess EGFR and FGFRs expressions, and responses to related growth factors were studied.</div><div>CTX primarily reduced gap closure in the migration assay, while BGJ reduced cell counts more clearly. Combined application enhanced performance in three out of four cell lines. All cell lines exhibited high EGFR expression, while KGFR expression was observed in a subpopulation. EGF stimulation led to cell elongation and an increase in size in three cell lines, accompanied by notable changes in migration. KGF affected cell morphology and migration in one cell line. This study shows that the combination of CTX and BGJ was most effective in the cell lines, highlighting the crucial roles of EGFR and KGFR, with KGFR potentially mediating the effects of BGJ. The findings suggest that adding targeted therapies for receptors on relevant cell subpopulations may enhance outcomes in therapy.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167940"},"PeriodicalIF":4.2,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144192602","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deficiency in the conserved ECHS1 gene causes Leigh syndrome by impairing mitochondrial respiration efficiency and suppressing ADRB2-PKA signaling","authors":"Baojiang Wang ,&nbsp;Yueyuan Qin ,&nbsp;Yantao Bao ,&nbsp;Shiguo Chen ,&nbsp;Junge Zheng ,&nbsp;Sheng Lin ,&nbsp;Kaifeng Zheng ,&nbsp;Shan Duan","doi":"10.1016/j.bbadis.2025.167930","DOIUrl":"10.1016/j.bbadis.2025.167930","url":null,"abstract":"<div><div>Deficiency in the short-chain enoyl-CoA hydratase 1 (ECHS1) gene causes Leigh syndrome (LS), a rare inherited metabolic disorder. Despite LS that arises as a result of inborn errors of energy metabolism, the specific contributions of ECHS1 deficiency to energy metabolism processes, developmental delay, and its mediated signaling mechanism remain unclear. Here, we identify a novel compound heterozygous variant [c.724G &gt; A (p.Glu242Lys) and c.865G &gt; A (Asp289Asn)] in the <em>ECHS1</em> gene from a family of Han Chinese descent, with the affected individual displaying typical LS symptoms. The ECHS1 variants exhibit reduced 2-enoyl-CoA hydratase activity, resulting in a restricted ATP production rate, but the cellular ATP levels remains unchanged. ECHS1 deficiency also decreases cell viability and proliferation. Mechanistically, ECHS1 interacts with ADRB2, and its variants suppress the ADRB2/protein kinase A (PKA) signaling. Treatment with PKA signaling agonists or overexpression of PKA subunits in ECHS1-deficient cells can rescue the ATP production rate and restore cell viability. Additionally, the mitochondrial E3 ligase MUL1 mediates the ubiquitylation and degradation of ECHS1 protein variants. In conclusion, our study suggests that ECHS1 deficiency impairs mitochondrial respiratory efficiency, thereby lowering the ATP production rate, and reveals a promising therapeutic approach by targeting ADRB2/PKA signaling to combat ECHS1 deficiency-induced LS.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167930"},"PeriodicalIF":4.2,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144188655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tumor-secreted TNF induces NCoA3 upregulation in breast cancer-associated adipocytes via TNFR1/NF-κB signaling","authors":"María Cecilia Lira ,&nbsp;Francisco D. Rosa ,&nbsp;Juliana Lourdes Bernacchia ,&nbsp;Alejandra G. Palma ,&nbsp;Natalia Paladino ,&nbsp;Mónica A. Costas ,&nbsp;María Fernanda Rubio","doi":"10.1016/j.bbadis.2025.167931","DOIUrl":"10.1016/j.bbadis.2025.167931","url":null,"abstract":"<div><div>In breast cancer, adipocytes are the predominant cell type in the microenvironment, and the continuous communication between these tissues alters the adipose phenotype. However, molecular mechanisms promoting these changes are still poorly understood. Previously, we demonstrated that NCoA3 expression is increased in adipose tissue adjacent to breast cancer and that this increase is associated with an inflammatory profile. This study aimed to investigate the mechanisms underlying NCoA3 expression in adipocytes within the breast tumor microenvironment.</div><div>We demonstrated that breast cancer-secreted TNF increases NCoA3 expression in adipocytes, and this upregulation is dependent on NF-κB transcriptional activity. Furthermore, the use of a TNF blocker prevented both coactivator overexpression and macrophages recruitment, mimicking the effects observed when NCoA3 expression was downregulated using a short hairpin RNA.</div><div>These findings shed light on the molecular mechanisms by which breast cancer cells modulate adipocyte behavior, identifying NCoA3 as a key mediator in the tumor-adipose tissue crosstalk. Targeting this pathway through TNF inhibition offers promising therapeutic strategy to attenuate tumor-associated inflammation and potentially improve outcomes in breast cancer patients.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167931"},"PeriodicalIF":4.2,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144185072","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improving neuronal recovery in spinal cord injury with NEP1-40-modified neural stem cells through RhoA/ROCK signaling pathway modulation","authors":"Wenbo Gu ,&nbsp;Xiao Zhang ,&nbsp;Xiaoya Yuan ,&nbsp;Donghui Cao ,&nbsp;Hongyang Zhao ,&nbsp;Jinlong Hu ,&nbsp;Haifeng Yuan","doi":"10.1016/j.bbadis.2025.167929","DOIUrl":"10.1016/j.bbadis.2025.167929","url":null,"abstract":"<div><div>To evaluate the therapeutic potential of NEP1-40-expressing neural stem cells (NSCs) in improving outcomes in spinal cord injury (SCI) by modulating the RhoA/ROCK signaling pathway. NEP1-40 was overexpressed by transfecting isolated NSCs with NEP1-40 lentivirus, and their differentiation following Nogo-A treatment was assessed <em>via</em> Tuj-1 immunofluorescence. The effects of NEP1-40 overexpression on axonal damage of differentiated NSCs and DRG neurons were examined using Tuj-1 and F-actin immunofluorescence, along with Western blotting for GAP-43, MAP-2, and APP. The activation of RhoA/ROCK and its downstream pathways, namely, PTEN/PI3K/AKT and MLC/LIMK, were analyzed by Western blotting. In a rat SCI model, following NSC transplantation, was assessed by examining neuronal regeneration markers. Furthermore, the Basso, Beattie, and Bresnahan (BBB) scale behavioral tests were used to assess motor function recovery. NEP1-40 overexpression inhibited the RhoA/ROCK signaling, followed by the stimulating the PI3K/AKT pathway and promoting neuronal differentiation under Nogo-A treatment. The inhibition of RhoA/ROCK signaling suppressed the MLC/LIMK pathway to enhance neurite outgrowth in NSC-differentiated and DRG neurons. In SCI rats, NEP1-40 overexpression increased the number of NSCs and neurons, whereas NEP1-40 overexpression in NSCs significantly increased the number of 5-HT-, ChAT-, and CGRP-positive neurons, as well as the levels of p-GAP-43, GAP-43 and MAP-2, and reduced the level of APP. These changes were accompanied by improved BBB scores. NEP1-40-modified NSCs enhance SCI recovery by activating the PI3K/AKT pathway and inhibiting the MLC/LIMK cascade, thereby promoting neuronal differentiation and reducing axonal damage. Altogether, this approach demonstrates promising therapeutic potential to improve NSC efficacy in SCI.</div></div>","PeriodicalId":8821,"journal":{"name":"Biochimica et biophysica acta. Molecular basis of disease","volume":"1871 7","pages":"Article 167929"},"PeriodicalIF":4.2,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144188656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}