This study investigated the role of long-chain acyl-CoA dehydrogenase (ACADL) in lung adenocarcinoma (LUAD). ACADL was significantly downregulated in human LUAD tissues compared to normal lung tissues. In vitro, ectopic expression of ACADL in murine LLC cells decreased cell viability, migration, and invasion, while ACADL knockdown exhibited the opposite effect. In vivo, ACADL overexpression impeded tumor growth and metastasis. Mechanistically, ACADL hindered tumor progression by inducing cell cycle arrest, promoting apoptosis, and suppressing the epithelial-mesenchymal transition (EMT) process. These findings suggest ACADL acts as a tumor suppressor in LUAD progression.

Cellular pliancy refers to the unique disposition of different stages of cellular differentiation to transform when exposed to specific oncogenic insults. This concept highlights a strong interconnection between cellular identity and tumorigenesis, and implies overcoming of epigenetic barriers defining cellular states. Emerging evidence suggests that the cell-type-specific response to intrinsic and extrinsic stresses is modulated by accessibility to certain areas of the genome. Understanding the interplay between epigenetic mechanisms, cellular differentiation, and oncogenic insults is crucial for deciphering the complex nature of tumorigenesis and developing targeted therapies. Hence, cellular pliancy relies on a dynamic cooperation between the cellular identity and the cellular context through epigenetic control, including the reactivation of cellular mechanisms, such as epithelial-to-mesenchymal transition (EMT). Such mechanisms and pathways confer plasticity to the cell allowing it to adapt to a hostile environment in a context of tumor initiation, thus changing its cellular identity. Indeed, growing evidence suggests that cancer is a disease of cell identity crisis, whereby differentiated cells lose their defined identity and gain progenitor characteristics. The loss of cell fate commitment is a central feature of tumorigenesis and appears to be a prerequisite for neoplastic transformation. In this context, EMT-inducing transcription factors (EMT-TFs) cooperate with mitogenic oncoproteins to foster malignant transformation. The aberrant activation of EMT-TFs plays an active role in tumor initiation by alleviating key oncosuppressive mechanisms and by endowing cancer cells with stem cell-like properties, including the ability to self-renew, thus changing the course of tumorigenesis. This highly dynamic phenotypic change occurs concomitantly to major epigenome reorganization, a key component of cell differentiation and cancer cell plasticity regulation. The concept of pliancy was initially proposed to address a fundamental question in cancer biology: why are some cells more likely to become cancerous in response to specific oncogenic events at particular developmental stages? We propose the concept of epipliancy, whereby a difference in epigenetic configuration leads to malignant transformation following an oncogenic insult. Here, we present recent studies furthering our understanding of how the epigenetic landscape may impact the modulation of cellular pliancy during early stages of cancer initiation.

Necrotizing sialometaplasia (NSM) is a nonneoplastic lesion listed in the World Health Organization Classification of Tumours-Head and Neck Tumours. In early NSM lesion, there is infarction and necrosis of the acinar cells, and squamous metaplasia of the salivary ducts occurs as the lesion matures. Differentiation from squamous cell carcinoma and other malignancies is sometimes required clinically and histopathologically. Local hypoxia caused by trauma and vascular compromise is a proposed etiology of NSM. However, the mechanisms underlying the pathogenesis are unclear. This study focused on the early stages of NSM. Histopathologic observations revealed that the region showing acinar necrosis contained myoepithelial cells with reticular arrangement. Hypoxic in vitro experiments using mouse salivary gland organoids revealed that myoepithelial and basal cells were more tolerant to hypoxia than acinar cells. Moreover, the residual myoepithelial cells in NSM and hypoxia-tolerant cells in organoids expressed transforming growth factor-β3 (TGFB3), which plays a critical role in cell proliferation and squamous metaplasia. Organoid experiments have also replicated the process of squamous metaplasia in NSM during hypoxia and the resolution of hypoxia. Thus, this study demonstrated that hypoxia is a possible etiology of NSM based on the results of histopathologic and in vitro experimental observations.

Breast cancer (BC) and gynecological malignancies, including cervical, ovarian, and uterine cancers, are significant global health challenges due to their high prevalence, complex nature, and elevated mortality rates. Dysregulation of the Wnt/β-catenin signaling pathway is a common feature in gynecological malignancies, contributing to cancer cell growth, progression, migration, and metastasis. Recent studies have highlighted the pivotal role of non-coding RNAs (ncRNAs), particularly circular RNAs (circRNAs), in modulating the Wnt/β-catenin signaling pathway. Acting as sponges for microRNAs (miRNAs), circRNAs regulate key oncogenic and tumor-suppressive processes by influencing Wnt-related components. This research explores the role of circRNAs in breast and gynecological malignancies, focusing on their regulatory effects on the Wnt/β-catenin pathway. The findings reveal that circRNAs modulate critical cellular processes such as proliferation, apoptosis, autophagy, and metastasis, with potential implications for therapeutic interventions. Targeting circRNA-mediated dysregulation of Wnt signaling could offer novel strategies for improving diagnostic precision, treatment efficacy, and survival outcomes in breast and gynecological cancers.

We sought to evaluate prognostic factors in human epidermal growth factor receptor 2 (HER2)-positive metastatic breast cancer (MBC) and their relationship with short- and long-term overall survival (OS).

Using the Surveillance, Epidemiology, and End Results (SEER) database, we evaluated patients with de novo HER2-positive MBC diagnosed from 2010 to 2018. Univariate analyses were performed to determine effect of each variable on OS. Significant variables were included in a multivariate Cox model for OS. Univariate and multivariate logistic regression were used to evaluate the association of each variable with short- (<2 years) and long- (≥5 years) term OS.

Overall, 5576 patients were included. Median follow up was 48 months (interquartile range 25-73 months), and median OS was 41 months. The proportion alive at 2, 5, and 8 years was 63.3% (95% confidence interval [CI] 62.0%-64.7%), 37.8% (95% CI, 36.2%-39.4%), and 26.8% (95% CI, 24.8%-28.9%), respectively. Factors associated with short-term OS were older age; Black race; nonductal nonlobular; brain, liver, or lung metastases; estrogen/progesterone receptor (ER/PR)-negative disease, and lower income (all P < .04). Number of metastatic organ sites was not significant. Factors associated with long-term OS were younger age, White race, fewer metastatic organ sites, ER/PR-positive disease, and higher income (all P < .02). Specific organ sites were not significant.

In this cohort with de novo HER2-positive MBC, OS improved significantly over the study period. We identified patient-specific and tumor-specific factors that were associated with short- and long-term survival in HER2-positive MBC.

Paediatric bone sarcomas (e.g. Ewing sarcoma, osteosarcoma) comprise significant biological and clinical heterogeneity. This extreme heterogeneity affects response to systemic therapy, facilitates inherent and acquired drug resistance and possibly underpins the origins of metastatic disease, a key component implicit in cancer related death. Across all cancers, metastatic models have offered competing accounts on when dissemination occurs, either early or late during tumorigenesis, whether metastases at different foci arise independently and directly from the primary tumour or give rise to each other, i.e. metastases-to-metastases dissemination, and whether cell exchange occurs between synchronously growing lesions. Although it is probable that all the above mechanisms can lead to metastatic disease, clinical observations indicate that distinct modes of metastasis might predominate in different cancers. Around 70% of patients with bone sarcoma experience metastasis during their disease course but the fundamental molecular and cell mechanisms underlying spread are equivocal. Newer therapies such as tyrosine kinase inhibitors have shown promise in reducing metastatic relapse in trials, nonetheless, not all patients respond and 5-year overall survival remains at ~ 50%. Better understanding of potential bone sarcoma biological subgroups, the role of the tumour immune microenvironment, factors that promote metastasis and clinical biomarkers of prognosis and drug response are required to make progress. In this review, we provide a comprehensive overview of the approaches to manage paediatric patients with metastatic Ewing sarcoma and osteosarcoma. We describe the molecular basis of the tumour immune microenvironment, cell plasticity, circulating tumour cells and the development of the pre-metastatic niche, all required for successful distant colonisation. Finally, we discuss ongoing and upcoming patient clinical trials, biomarkers and gene regulatory networks amenable to the development of anti-metastasis medicines.

To investigate the effect of low-intensity pulsed ultrasound (LIPUS) on epithelial-mesenchymal transition (EMT) in lens epithelial cells. EMT was induced using high glucose (HG) in SRA01/04 cells. Optimal parameters for LIPUS irradiation were determined by cell counting kit-8 assays and flow cytometry. Cell morphology was assessed by light microscopy, while cell migration ability was analyzed by a wound healing assay. Levels of specific proteins and the relationship between autophagy and the cytoskeleton were examined by immunofluorescence (IF) staining and Western blot (WB). Cytoskeletal structures were visualized by phalloidin staining and autophagosomes were quantified by transmission electron microscopy. EMT was successfully induced by HG treatment. Compared to the model group, LIPUS irradiation resulted in a change in cell morphology from spindle to oval, a significant decrease in cell migration area, and an increase in E-cadherin and LC3B/LC3A levels. In contrast, α-SMA and SQSTM1/P62 levels decreased, the number of autophagosomes increased and F-actin levels decreased in the LIPUS group. SRA01/04 cells treated with LIPUS irradiation after autophagy inhibitors 3-MA and CQ showed increased cell migration area compared to the 3-MA/CQ group; LC3B/LC3A levels decreased; SQSTM1/P62 and F-actin levels increased in the LIPUS + 3-MA/CQ group compared to 3-MA/CQ treatment alone. Colocalization of the cytoskeletal marker Arpc2 with the autophagy marker SQSTM1/P62 was also observed. After treatment with the cytoskeletal inhibitor CK666+LIPUS combination therapy, SQSTM1/P62 levels increased while LC3B/LC3A levels decreased. LIPUS inhibited HG-induced EMT by restoring autophagy, which appears to be associated with cytoskeletal remodeling.

The leading factor contributing to patient mortality is the local invasion and metastasis of tumors, which are influenced by the malignant progression of tumor cells. The epithelial-mesenchymal transition (EMT) is key to understanding malignancy development. EMT is a critical regulatory mechanism for differentiating cell populations initially observed during the neural crest and embryonic gastrulation formation. This process is closely associated with tumor metastasis in cancer and is also related to the maintenance of cancer stem cells. Flavonoids, known for their antioxidant properties, have been widely studied for their anticancer potential to protect plants from harmful environmental conditions. They have attracted considerable attention and have been the focus of numerous experimental and epidemiological studies to evaluate their potential in cancer treatment. In vitro and in vivo research has demonstrated that flavonoids can significantly impact cancer-related EMT. They may inhibit the EMT process by reducing the levels of Twist1, N-cadherin, ZEB1, integrins, SNAI1/2, CD44, MMPs, and vimentin while increasing E-cadherin levels and targeting the PI3K/AKT, NF-κB p65, and JAK2/STAT3 signaling pathways. In order to suppress the transcription of the E-cadherin promoter, several Zn-finger transcription factors, such as SNAI2, ZEB1, and ZEB2, and basic helix-loop-helix (bHLH) factors, such as Twist, may directly bind to its E-boxes. Overall, clinical cancer research should integrate the anticancer properties of flavonoids, which address all phases of carcinogenesis, including EMT, to improve the prospects for targeted cancer therapies in patients suffering from aggressive forms of tumors.

Glycosylation occurs mainly in the Golgi apparatus, whereas the synthesis of nucleotide sugars occurs in the cytoplasm or nucleus. GDP-fucose in mammalian cells could be produced via de novo and salvage pathways in the cytoplasm; the first one is responsible for about 90% of GDP-fucose in the total pool of this nucleotide sugar in the cell. SLC35C1 (C1) is the primary transporter of GDP-fucose to the Golgi apparatus. In the absence of this transporter, it was proposed that nucleotide sugar could still reach the Golgi apparatus via a SLC35C2, the homologue of SLC35C1. However, simultaneous inactivation of the two transporters did not influence GDP-fucose transport across the Golgi apparatus membranes after external fucose supplementation. In this study, we combined the inactivation of SLC35C1 and enzymes of the GDP-fucose biosynthesis pathways (FCSK, GMDS and TSTA3) to study the impact of double inactivation on the production of nucleotide sugar and fucosylated glycans. We found that a lack of SLC35C1 changed the level of enzymes of both de novo and salvage pathways. Upon fucose supplementation, stimulation of the salvage pathway was remarkably high in the absence of the TSTA3 protein, and the concentration of GDP-fucose increased to millimolar values. In this work, we discovered that simultaneous deficiency of the SLC35C1 protein and TSTA3 enzyme increased GDP-fucose production via the salvage pathway to an even higher level. Finally, we found that nucleotide sugar still accessed the Golgi apparatus and had differential effects on N- and O-glycans.

Breast cancer has a high global incidence, and benzo[a]pyrene (B[a]P) is considered a contributing factor that increases carcinogenic risk. This study examined B[a]P's oncogenic mechanisms in mammary epithelial cells. Chronic B[a]P exposure induced morphological changes and enhanced proliferative/clonogenic capacity in MCF-10A cells. Chronic B[a]P exposure altered gene expression in MCF-10A cells, revealing differential levels of circRNAs, lncRNAs, miRNAs, and mRNAs. qRT-PCR validation demonstrated strong alignment with RNA-seq results, ensuring sequencing reliability. Additionally, chronic B[a]P exposure upregulated the protein expression of AhR and ARNT, as well as TGF-β, pSmad2/3, and KRT14, while increasing Vimentin expression and decreasing E-cadherin expression. Notably, treatment with the TGF-β inhibitor SB431542 reversed these protein expression changes in transformed cells. These results show that exposure to Chronic B[a]P induces MCF-10A cell transformation. The underlying mechanisms involve significant transcriptional alterations, AhR/ARNT expression regulation, TGF-β signaling pathway activation, KRT14 protein modulation, and EMT. Furthermore, Chronic B[a]P exposure may drive transformation through TGF-β modulation. Chronic B[a]P exposure promotes breast carcinogenesis, revealing mechanistic insights and potential preventive biomarkers.

The proper distribution of nutrients and metabolites between the mother and fetus is one important factor for successful pregnancy. As a bridge, the placenta plays a key role in sensing the nutritional needs of the fetus, coordinating the maternal nutrition supply, and enhancing its nutritional transport capabilities. Imperfect placental development can lead to pregnancy-related disorders such as preeclampsia, recurrent miscarriage, and/or fetal growth restriction, posing risks to both mother and child in the short and long term. However, current understanding of the human placenta remains as a "black box", and its developmental control mechanisms for adaptive pregnant regulation still needs to be elucidated. As one form of post-translational modification (PTM), ubiquitination plays an important role in regulating cellular functions and is regarded as a valuable drug target. Particularly, ubiquitination related to placenta development has been discovered in recent years. Placental development processes closely associated with pregnant complications, such as blastocyst implantation, syncytiotrophoblast cell differentiation, and immune barrier maintenance, have been reported to be affected by ubiquitination. However, the diagnosis and intervention of pregnancy diseases also urgently need to be improved. Thus, aiming to comprehensive summarize and further exploring the molecular mechanism, target and regulatory mechanism of pregnancy complications, we have herein reviewed genes and pathways regulating pregnancy progress and diseases and focusing on ubiquitin-related physiological process in placenta.

Tubulointerstitial fibrosis (TIF), a critical pathological hallmark in progressive chronic kidney disease (CKD), may be potentiated by renal lipid metabolism dysregulation and ectopic lipid deposition, though these processes likely exhibit bidirectional interactions with fibrotic progression Lipophagy is a type of selective autophagy that specifically recognizes lipid droplets and is accountable for lipid stability and metabolism. It serves as a link between lipid metabolism and autophagy. It was found that a positive correlation between elevated LARS1 expression and the severity of renal interstitial fibrosis in CKD patients. In Lars1+/- mice, we observed that the absence of LARS1 significantly reduced lipid deposition and TIF. Mechanistically, stimulation of HK-2 cells with TGF-β1 resulted in LARS1-mediated activation of mTORC1 and suppression of lipophagy, consequently leading to increased lipid accumulation and epithelial mesenchymal transition (EMT) through a defined mechanistic pathway. Collectively, our studies demonstrate that LARS1 plays a pivotal role in renal fibrosis at least in part by inhibiting lipophagy, suggesting that targeting LARS1 may represent a novel therapeutic strategy for patients with CKD.

microRNAs have received wide attention as potential therapeutic targets. This study explored the action of miR-128-3p in acute myeloid leukemia (AML). miR-128-3p expression in AML was determined by quantitative PCR method. MTT proliferation assay and immunoblot assay were employed to detect proteins related to proliferation and apoptosis in THP-1 cells overexpressing miR-128-3p. RNA immunoprecipitation and dual luciferase reporting system were utilized to verify downstream targets of miR-128-3p. Flow cytometry was conducted to analyze the apoptosis rate and immune escape of THP-1 cells in the T-cell co-culture system. miR-128-3p was lowly expressed in AML patients (reduced by 41.6%). Overexpression of miR-128-3p inhibited THP-1 cell proliferation and immune escape, and stimulated apoptosis. ZEB1 was a downstream target of miR-128-3p, and up-regulation of miR-128-3p inhibited ZEB1 mRNA and protein expression (respectively reduced by 65.8% and 42.0%). Upregulating ZEB1 reversed the inhibitory effect of upregulating miR-128-3p on THP-1 cell proliferation and immune escape. Upregulating ZEB1 promoted PD-L1 protein expression (increased by 0.75-fold). Blocking PD-L1 reversed the promotion of THP-1 cell proliferation and immune escape by upregulating ZEB1. The miR-128-3p/ZEB1/PD-L1 axis is involved in regulating the proliferation and immune escape of AML cells, providing new insights into the molecular mechanism of miR-128-3p in AML and, more importantly, a new target for immunotherapy of AML.

Breast cancer (BRCA) is the most common malignant tumor in women, and distant metastasis is an important cause of death. Epithelial mesenchymal transition (EMT) is an important factor in tumor cell metastasis, in which TGF-β signaling pathway plays an important role. SMAD7 can inhibit TGF-β pathway. Previously, we found that ovarian tumor domain-containing protein 3(OTUD3) could maintain the stability of multiple molecules through deubiquitination. In this study, multiple experiments were conducted to verify whether OTUD3 can inhibit TGF-β pathway by deubiquitinating SMAD7.

Firstly, bioinformatics was used to search the expression of OTUD3 in breast cancer and its correlation with SMAD7 in the TCGA database. The correlation between the protein and mRNA expression levels of OTUD3 and SMAD7 in multiple BRCA cell lines was verified. Also, the OTUD3 and SMAD7 expression in human BRCA samples and its influence on prognosis were verified by immunohistochemical experiments. Then, the CO-IP experiment was performed by transfecting OTUD3 and SMAD7 in HEK293T cells to confirm whether OTUD3 could maintain SMAD7 protein stability through deubiquitination. Furthermore, luciferase reporting assay, in vitro protein interaction, and transwell assay were used to verify whether OTUD3 could inhibit TGF-β pathway by deubiquitinating SMAD7 and affect cell invasion. Western blot and RT-qPCR were used to detect the correlation between OTUD3 and molecules regulated by the TGF-β pathway. Finally, the effect of OTUD3 on tumor cells was determined by 3D matrigel cell culture.

The expression of OTUD3 was low in BRCA and positively correlated with SMAD7. Cytological experiments and immunohistochemistry confirmed that OTUD3 was positively correlated with the expression of SMAD7, and the patients with a low expression of OTUD3 had a short recurrence-free survival (RFS). Cell experiments confirmed that OTUD3 could regulate the TGF-β pathway by deubiquitinating SMAD7, which affected EMT and inhibited cell invasion. OTUD3 was found to inhibit the stemness of tumor cells by 3D matrigel cell culture.

Our findings indicated OTUD3 inhibited BRCA metastasis associated with TGF-β signaling by deubiquitination to stabilize SMAD7 protein levels.

We investigated the efficacy of quantitative evaluation of tumor signal heterogeneity on gadoxetic acid-enhanced magnetic resonance imaging (EOB-MRI) to predict prognosis and antitumor immunity in patients with hepatocellular carcinoma (HCC) undergoing liver resection.

A total of 297 patients who underwent curative resection for primary HCC were included. Tumor signal heterogeneity in the hepatobiliary phase (HBP) of EOB-MRI was quantified as the coefficient of variation (CV), calculated as the standard deviation divided by the mean signal intensity. Patients were classified into homogeneous (low CV) and heterogeneous (high CV) groups based on a cutoff value of 0.16 from receiver operating characteristic curve analysis. Tumor-infiltrating CD4+ and CD8+ T cells and PD-L1 expression were assayed by immunohistochemistry, and their associations with tumor signal heterogeneity were evaluated.

Among the 297 patients, 116 (39.1%) were classified into the heterogeneous group. The overall survival (OS) and recurrence-free survival (RFS) rates were significantly lower in the heterogeneous group (p < 0.001 for both). Multivariate analysis identified heterogeneous group as an independent prognostic factor for OS and RFS (p < 0.001 and p = 0.012, respectively). Extrahepatic recurrence was significantly more frequent in the heterogeneous group (18.1% vs. 7.7%, p = 0.024). CD4+ and CD8+ T cells were significantly decreased, and the PD-L1 positivity rate was significantly lower in the heterogeneous group (p < 0.001 for all).

The quantitative evaluation of tumor signal heterogeneity in the HBP of EOB-MRI using CV is useful for predicting postoperative prognosis in patients with HCC. Tumor signal heterogeneity may also reflect impaired local immunity and an immunologically "cold" tumor.

Cancer initiation and early tumour growth are complex processes influenced by multiple cellular and microenvironmental factors. A critical aspect of tumour progression is the dynamic interplay between cancer cells and the extracellular matrix (ECM), which undergoes significant alterations to support malignancy. The loss of cell polarity is an early hallmark of tumour progression, disrupting normal tissue architecture and fostering cancerous transformation. Circumstantially, cancer-associated microRNAs (miRNAs) regulate key oncogenic processes, including ECM remodelling, epithelial-to-mesenchymal transition (EMT), and tumorigenic vascular development, further driving tumour growth. ECM alterations, particularly changes in stiffness and mechanotransduction signals, create a supportive niche for cancer cells, enhancing their survival, proliferation, and invasion. EMT and its subtype, epithelial-to-endothelial transition (EET), contribute to tumour plasticity, promote the generation of cancer stem cells (CSCs), and support tumour vascularisation. Furthermore, processes of vascular development like vasculogenesis and angiogenesis are critical for sustaining early tumour growth, supplying oxygen and nutrients to hypoxic malignant cells within the evolving cancerous microenvironments. This review explores key mechanisms underlying these changes in tumorigenic microenvironments, with an emphasis on their collective role for tumour initiation and early tumour growth. It will further delve into present in vitro modelling strategies developed to closely mimic early cancer pathophysiology. Understanding these processes is crucial for developing targeted therapies aimed at disrupting key cancer-promoting pathways and improving clinical outcomes.

Epithelial to Mesenchymal transitions (EMT) drive cell plasticity and are associated with cell features such as invasiveness, migration and stemness. They are orchestrated by select families of EMT-associated transcription factors, which exhibit pleiotropic roles in the malignant progression of various cancer types, such as breast and colorectal cancer (CRC). This has spurred interest in EMT as a promising target for the development of novel therapeutic strategies. In this study, we developed a phenotypic dual EMT Sensor screening assay, amendable to efficient high-throughput identification of small molecules interfering with EMT. In a proof-of-concept screening we identified anti-EMT repurposing drugs. From these, we validated RepSox, a selective inhibitor of the TGF-β type I receptor ALK5, and demonstrated that it is potently blocking EMT in both breast and colorectal cancer cell lines in vitro. In addition, utilizing a Drosophila melanogaster metastatic CRC model we confirmed the ability of the identified anti-EMT hits to suppress metastatic behavior in vivo.

Epithelial-to-mesenchymal transition (EMT), a key process in cancer metastasis and fibrosis, disrupts cellular adhesion by replacing epithelial E-cadherin with mesenchymal N-cadherin. While, how the shift from E-cadherin to N-cadherin impacts molecular-scale adhesion mechanics and cluster dynamics-and how these changes weaken adhesion under varying mechanical and environmental conditions-remains poorly understood, limiting our ability to target EMT-driven pathological adhesion dynamics. Here, we developed a unified lattice-clutch model to investigate cadherin clustering, cortical tension, and adhesion strength during EMT. Using atomic force microscopy experiments, we measured the mechanical properties of single cadherin trans-bonds and cadherin-mediated cell-cell and cell-matrix adhesions across varying conditions. Our results demonstrate that N-cadherin trans-bonds are mechanically weaker than E-cadherin trans-bonds, leading to reduced adhesion strength during EMT. Computational modeling and experimental validation further revealed that EMT impairs cadherin clustering and cortical tension regulation, which collectively weaken both cell-cell and cell-matrix adhesions, particularly on stiff substrates. These findings highlight how EMT disrupts adhesion strength at multiple scales-from individual cadherin bonds to collective cluster dynamics. Our study elucidates how EMT-driven changes in cadherin type weaken adhesion strength and mechanotransduction, providing insights into cellular adhesion mechanics and potential therapeutic strategies for targeting EMT-associated diseases such as cancer metastasis and tissue remodeling.

Human pluripotent stem cell-derived tissue engineering offers great promise for designer cell-based personalized therapeutics, but harnessing such potential requires a deeper understanding of tissue-level interactions. We previously developed a cell replacement manufacturing method for ectoderm-derived skin epithelium. However, it remains challenging to manufacture the endoderm-derived esophageal epithelium despite possessing a similar stratified epithelial structure. Here, we employ single-cell and spatial technologies to generate a spatiotemporal multi-omics cell census for human esophageal development. We identify the cellular diversity, dynamics, and signal communications for the developing esophageal epithelium and stroma. Using Manatee, a machine-learning algorithm, we prioritize the combinations of candidate human developmental signals for in vitro derivation of esophageal basal cells. Functional validation of Manatee predictions leads to a clinically compatible system for manufacturing human esophageal mucosa.

Lysyl oxidase-like-1 (LOXL1) is a copper-dependent amine oxidase that maintains the structural integrity of the extracellular matrix (ECM) by catalyzing the cross-linking of collagen and elastin. However, aberrations in LOXL1 expression can contribute to diseases like glaucoma, tissue fibrosis, and cancer. LOXL1 has been found to be overexpressed in various malignancies, playing a pivotal role in tumor growth and metastasis. Although some studies suggest tumor-suppressive attributes of LOXL1, its role in tumorigenesis remains controversial. Research on LOXL1 has been primarily focused on pseudoexfoliation syndrome/glaucoma, with limited reviews on its impact on cancer. This review aims to explore LOXL1 comprehensively, including its structure, biological effects, and regulatory processes. Emphasis is placed on understanding the relationship between LOXL1 and tumorigenesis, specifically how LOXL1 influences tumor microenvironment remodeling, tumorigenesis, and metastasis. The review also discusses potential therapeutic strategies targeting LOXL1 for anti-fibrosis and anti-tumor interventions.

The process of epithelial-mesenchymal transition (EMT) is crucial in various physiological/pathological circumstances such as development, wound healing, stem cell behavior, and cancer progression. It involves the conversion of epithelial cells into a mesenchymal phenotype, which causes the cells to become highly motile. This reprogramming is initiated and controlled by various signaling pathways and governed by several key transcription factors, including Snail 1, Snail 2 (Slug), TWIST 1, TWIST2, ZEB1, ZEB2, PRRX1, GOOSECOID, E47, FOXC2, SOX4, SOX9, HAND1, and HAND2. The intracellular signaling pathways are activated/inactivated by signals received from the extracellular environment and the transcription factors are carefully regulated at the transcriptional, translational, and post-translational levels to maintain tight regulatory control of EMT. One of the most important pathways involved in this process is the transforming growth factor-β (TGFβ) family signaling pathway. This review will discuss the role of EMT in promoting epithelial cancer progression and the convergence/interplay of multiple signaling pathways and transcription factors that regulate this phenomenon.

X-box binding protein 1 (XBP1), as a transcription factor, plays pivotal role in unfolded protein response (UPR), which is activated in response to endoplasmic reticulum (ER) stress to restore ER homeostasis. IRE1α/XBP1 pathway is a key component of UPR, and the expression levels of XBP1 can dictate the fate of cells under ER stress, either promoting survival or driving apoptosis. High expression of XBP1 in breast tumors is closely associated with poor prognosis. The paper elucidates the biological functions of XBP1 and its involvement in UPR, while also surveying the latest research on how XBP1 influences immunity, metabolism, apoptosis, angiogenesis, and the invasive and migratory behaviors of breast cancer cells. Moreover, it contemplates the potential of XBP1 as a therapeutic target for breast cancer treatment.

Pathological epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs) plays a crucial role in the formation of lens fibrosis, particularly in fibrotic posterior capsular opacification and anterior subcapsular cataract (ASC). Here we investigated the potential roles of endoplasmic reticulum (ER) stress in the development of lens fibrosis.

RNA sequencing was performed to examine global gene expression changes in patients with ASC, as well as in TGFβ2-induced human lens explants and rabbit primary LECs. Rabbit LECs were treated with TGFβ2 in the presence or absence of the ER stress modulator, PERK inhibitor ISRIB, and autophagy inducer for in vitro studies. In vivo investigations were carried out using a mouse model of injury-induced capsular fibrosis, with ISRIB administration. To uncover the underlying mechanisms, we conducted lipidomics analysis, transmission electron microscopy, immunostaining, quantitative PCR, Western blot, and capillary Western immunoassay.

ER stress genes were upregulated in patients with ASC, TGFβ2-stimulated human explants and primary LECs. Pharmacologic ER stress induction promoted EMT, while its inhibition reduced TGFβ2-induced mesenchymal gene levels. Blocking the PERK axis of ER stress with ISRIB or targeting downstream factor ATF4 suppressed EMT, whereas the IRE1 axis showed no effect. Consistent with these in vitro observations, anterior chamber injection of ISRIB also reduced subcapsular plaque formation in a mouse model of lens fibrosis by suppressing SMAD2/3 activation. Mechanistically, ISRIB suppressed LC3-II conversion and P62 degradation, indicating autophagy suppression. Lipidomics revealed phosphatidylethanolamine (PE), essential for autophagosome formation, was downregulated in TGFβ2-treated LECs and upregulated with ISRIB cotreatment. Inducing autophagy with rapamycin significantly rescued the mesenchymal gene suppression by ISRIB, whereas autophagy inhibitor CQ produced opposite effects.

ER stress, particularly the PERK axis, promotes LECs' EMT through autophagy and PE metabolism, offering potential therapeutic targets for the treatment of lens fibrosis.

Locally advanced esophageal adenocarcinoma remains difficult to treat and the ecological and evolutionary dynamics responsible for resistance and recurrence are incompletely understood. Here, we performed longitudinal multiomic analysis of patients with esophageal adenocarcinoma in the MEMORI trial. Multi-region multi-timepoint whole-exome and paired transcriptome sequencing was performed on 27 patients before, during and after neoadjuvant treatment. We found major transcriptomic changes during treatment with upregulation of immune, stromal and oncogenic pathways. Genetic data revealed that clonal sweeps through treatment were rare. Imaging mass cytometry and T cell receptor sequencing revealed remodeling of the tumor microenvironment during treatment. The presence of genetic immune escape, a less-cytotoxic T cell phenotype and a lack of clonal T cell expansions were linked to poor treatment response. In summary, there were widespread transcriptional and environmental changes through treatment, with limited clonal replacement, suggestive of phenotypic plasticity.

Airway remodelling significantly contributes to the progressive loss of lung function and heightened symptom severity in chronic asthma. Additionally, it often persists and demonstrates reduced responsiveness to the mainstay treatments. The excessive deposition of collagen and extracellular matrix proteins leads to subepithelial fibrosis and airway remodelling, resulting in increased stiffness and decreased elasticity in the airway. Studies have emphasized the crucial role of subepithelial fibrosis in the pathogenesis of asthma. Fibrotic processes eventually cause airway narrowing, reduced lung function, and exacerbation of asthma symptoms. Macrophages play a crucial role in this process by producing pro-fibrotic cytokines, growth factors, and enzymes such as matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Additionally, identification of novel genetic markers has provided evidence for a strong genetic component in fibrosis within macrophage regulated fibrosis. Although macrophages contribute to the progression of airway remodelling and subepithelial fibrosis, interventions targeting macrophage-driven fibrotic changes have not yet been developed. This review synthesizes research on the intricate pathways through which macrophages contribute to subepithelial fibrosis in chronic asthma and its' pathological features. Understanding the interplay between macrophages, fibrosis, and asthma pathogenesis is essential for developing effective therapeutic strategies to manage severe asthma and improve patient outcomes.

The appearance of hybrid epithelial-mesenchymal (E/M) cells expressing E-cadherin is favourable for the establishment of pro-invasive function. Although the potential role of E-cadherin in cancer invasion is now accepted, the molecular mechanisms involved in this process are not completely elucidated. To gain further insight, we focused our analysis on invadopodia formation, an early event in the invasion process. We used models of E/M hybrid cell lines, tissue sections and patient-derived xenografts from a multi-centre clinical trial. E-cadherin involvement in invadopodia formation was assessed using a gelatin-FITC degradation assay. Mechanistic studies were performed by using proteomic analysis, siRNA strategy and proximity ligation assay. We showed that E-cadherin is a critical component of invadopodia. This unexpected localization results from a synergistic trafficking of E-cadherin and MT1-MMP through a Rab vesicle-dependent pathway. Modulation of E-cadherin expression or activation impacted invadopodia formation. Moreover, colocalization of E-cadherin and Actin in "ring structures" as precursors of invadopodia reveals that E-cadherin is required for invadopodia structuration. E-cadherin, initially localised in the adherens junctions, could be recycled to nascent invadopodia where it will interact with several components enriched in invadopodia, such as Arp2/3, Cortactin or MT1-MMP. The trans-adhesive properties of E-cadherin are therefore essential for structuring invadopodia. This new localisation of E-cadherin and its unexpected role in cell invasion shine a new light on hybrid E/M transition features in tumoral invasion.

Despite significant advancements in identifying novel therapeutic targets and compounds, cancer stem cells (CSCs) remain pivotal in driving therapeutic resistance and tumor progression in gastric cancer (GC). High-resolution knowledge of the transcriptional programs underlying the role of CSC niche in driving tumor stemness and progression is still lacking. Herein, spatial and single-cell RNA sequencing of 32 human gastric mucosa tissues at various stages of malignancy, illuminating the phenotypic plasticity of tumor epithelium and transcriptional trajectory from mature gastric chief cells to the CSC state, which is associated with activation of EGFR and WNT signaling pathways, is conducted. Moreover, the CSCs interact with not only the immunosuppressive CXCL13+ T cells and CCL18+ M2 macrophages to evade immune surveillance, but also the inflammatory cancer-associated fibroblasts (iCAFs) to promote tumorigenesis and maintain stemness, which construct the CSC niche leading to inferior prognosis. Notably, it is uncovered that amphiregulin (AREG) derived from iCAFs promotes tumor stemness by upregulating the expression of SOX9 in tumor cells, and contributes to drug resistance via the AREG-ERBB2 axis. This study provides valuable insight into the characteristics of CSC niche in driving tumor stemness and progression, offering novel perspective for designing effective strategies to overcome GC therapy resistance.

Lymphatic metastasis of gastric cancer is a challenging issue in clinical practice. Recently, copper single-atom nanozymes (SAZ) have gained tremendous attention due to its superior peroxidase (POD) activity that has good nonocatalytic tumor therapy (NCT) capabilities, and photothermal properties. Therefore, using a high-expressing P-selectin platelet membrane (PM) to encapsulate SAZ and cisplatin is proposed, forming PSC nanoparticles. Due to their exquisite nanoscale size and the unique structure of lymphatic vessels, PSC can highly target cancer cells in invasive primary tumors and metastatic lymph nodes that both highly express CD44. It is noteworthy that cisplatin can simultaneously perform chemotherapy and generate H₂O₂ under the action of NADPH oxidases (NOXs) that further enhance the catalytic activity of SAZ and increase intracellular reactive oxygen species (ROS) production. Both in vitro and vivo experiments have demonstrated the superior targeting and elimination capability of the PCS system in primary and metastatic tumor cells. In addition, transcriptomic analysis reveals that PSC + NIR induced apoptosis in MFC cells. This marks the first proposal of combining single-atom nanozymes and chemotherapy drugs for dual-targeting in gastric cancer and lymphatic metastasis, providing new insights into a challenging clinical issue in the treatment of gastric cancer lymphatic metastasis.

The formation of gastric precancerous lesions (GPLs) has been identified as a critical step in tumorigenesis, and patients with GPLs have an increased risk of gastric cancer (GC). Magnolol is the primary biphenolic compound in Magnolia officinalis. It possesses various pharmacological properties, such as cardio- and neuroprotective properties, and inhibits tumor growth. However, its therapeutic effects on GPL treatment and the related mechanisms have not yet been studied. To address this, the mechanisms by which magnolol affects GPLs were elucidated via protein-chemical interaction prediction analysis, molecular docking, molecular dynamics (MD) simulation, and experimental verification. GPL-related targets were obtained from the GeneCards database, whereas magnolol targets were obtained from the STITCH database. The two groups of targets were compared by constructing a Venn diagram, and potential key GPL-related targets of magnolol were identified. Next, the interactions between the active compounds of magnolol and various epithelial-mesenchymal transition (EMT)-related proteins were evaluated via molecular docking. The protein-compound complexes with the optimal binding affinity were analyzed via MD simulation. The efficacy of magnolol in the treatment of GPLs and the related mechanisms was further assessed using in vitro models. In this study, five core GPL-related targets of magnolol were identified. Molecular docking revealed that magnolol and ERBB2 had the strongest binding affinity, suggesting that ERBB2 is a potentially important target for the treatment of GPLs. Similarly, MD simulations revealed a strong affinity between magnolol and ERBB2. Overall, this study showed that magnolol can inhibit the malignant behavior of precancerous lesions in GC cells. Magnolol exerts its pharmacological effects by acting on multiple targets. ERBB2 might be a potential target of magnolol in GPL treatment.

Prostate cancer (PCa) is the most prevalent cancer among men globally. Although, there are various therapeutic methods for the localized or advanced cancers, there is still a high rate of mortality among PCa patients that is mainly associated with bone metastasis in advanced tumors. There are few options available for treating bone metastasis in PCa, which only provide symptom relief without curing the disease. Therefore, it is crucial to evaluate the molecular mechanisms associated with bone metastasis of PCa cells to suggest the novel diagnostic and therapeutic approaches that could lower the morbidity and mortality rates in PCa patients. MicroRNAs (miRNAs) are involved in regulation of various pathophysiological processes such as tumor growth and osteoblasts/osteoclasts formation. Since, miRNA deregulation has been also frequently observed in PCa patients with bone metastasis, we discussed the role of miRNAs in bone metastasis during PCa progression. It has been reported that miRNAs mainly reduced the ability of PCa tumor cells for the bone metastasis through the regulation of WNT, NF-kB, PI3K/AKT, and TGF-β signaling pathways. They also affected the EMT process, transcription factors, and structural proteins to regulate the bone metastasis during PCa progression. This review paves the way to suggest the miRNAs as the reliable markers not only for the non-invasive early diagnosis, but also for the targeted therapy of PCa tumors with bone metastasis.

The development of early human tooth primordia is not well understood. Here, we linked single-cell RNA sequencing, spatial transcriptomics, and secretome analysis to characterize human fetal tooth development over time. A spatiotemporal atlas of human tooth development at multiple levels was mapped, identifying previously uncharacterized epithelial subpopulations with distinct gene expression profiles and spatial localization. Dynamic changes in epithelial-mesenchymal interactions across developmental stages were characterized. Secretome analysis confirmed the extensive paracrine signaling from the epithelial to mesenchymal compartments and uncovered signaling factors produced by dental epithelium (DE) that regulate mesenchymal cell fate and differentiation. Integration of these datasets highlighted the crucial role of the DE in orchestrating tooth morphogenesis. Our multi-omics approach not only provides unprecedented insights into the cellular and molecular mechanisms of ectoderm-derived tissue development but also serves as a valuable resource, which is publicly available online, for future studies on human tooth regeneration and related diseases.

Aberrant methylation of the EphA7 promoter has been observed in cervical cancer (CC); however, its precise function and role in CC remain largely unknown. In this study, we investigated the role and molecular mechanisms of EphA7 promoter methylation in cervical carcinogenesis. First, our results indicated that the reactivation of EphA7 expression via a CRISPR demethylation tool (dCas9-Tet1) had antitumor effects. It restrained tumor proliferation and invasion while promoting apoptosis via the PI3K/AKT signaling pathway in both CaSki and SiHa cells. The upstream interacting factors were subsequently captured by CRISPR-mediated pull-down in situ, and the result revealed that SP1 and MAZ interacted with the promoter of EphA7. However, the perturbation results revealed that EphA7 expression was associated with SP1/DNMT1 but not MAZ. Furthermore, 17-β-estradiol (E2) can upregulate EphA7 expression through demethylation via the SP1/DNMT1 axis. A rescue experiment revealed that interference with SP1 expression could restore the effect of E2 on increasing the expression of EphA7 by upregulating estrogen receptor expression. In addition, EphA7 demethylation reduced the half-maximal inhibitory concentration (IC50) of cisplatin and paclitaxel. Pooled analysis revealed that EphA7 promoter hypermethylation was positively correlated with tumor purity but negatively correlated with immune cell infiltration, cytotoxic T lymphocyte (CTL) and immune checkpoint (IC) activity, and the expression of EphA7 was significantly positively correlated with tumor mutational burden (TMB), microsatellite instability (MSI) and the presence of single nucleotide variant (SNV) neoantigens, suggesting a better prognosis for patients with EphA7 promoter hypomethylation and high expression. Collectively, these findings indicate that targeted demethylation of the EphA7 promoter and restoration of endogenous EphA7 expression by dCas9-Tet1 are promising therapeutic approaches and are favorable for the prognosis of CC patients.

The relationship between circulating tumor cells (CTCs) and patients with advanced gallbladder adenocarcinoma (aGA) has been rarely studied. This article was to demonstrate the enumeration, classification, and clinical application of CTCs in patients with aGA.

Peripheral blood samples were collected and CTCs were detected using the CanPatrol® technique. T test, χ2 test, Wilcoxon rank sum test or Kruskal-Wallis test, log-rank test and Cox regression analysis were performed to conduct statistical analysis.

CTCs were detected at pre-treatment in 75.00% (27/36) of the patients. Both CTCs positive rate and CTCs enumeration at pre-treatment were significantly associated with clinicopathological parameters including Ca199 level (P = 0.014, P < 0.001 respectively), tumor differentiation (P = 0.007, P = 0.002 respectively), lymph infiltration (P = 0.010, P = 0.025 respectively), vascular infiltration (P = 0.007, P < 0.001 respectively), and distant metastasis (P = 0.015, P = 0.002 respectively). CTCs-positive patients had a significantly shorter OS (HR 0.335, 95% CI 0.165-0.678, P = 0.0023) and PFS (HR 0.364, 95% CI 0.179-0.739, P = 0.0024) than CTCs-negative patients. Mesenchymal CTCs enumeration was closely related to the chemotherapy response, and CTCs programmed cell death ligand-1 (PD-L1) was highly correlated with the immunotherapy response. Positive CTCs at pre-treatment was closely related to the poor OS (HR 0.089, 95% CI 0.020-0.399, P = 0.002) as well as distant metastasis (HR 0.159, 95% CI 0.041-0.610, P = 0.007), untreated with chemotherapy (HR 4.510, 95% CI 1.403-14.499, P = 0.011) and untreated with immunotherapy (HR 6.845, 95% CI 1.894-24.738, P = 0.003).

Pretreatment-positive CTCs was closely related to the poor prognosis in patients with aGA. Monitoring the subtype and phenotype of CTCs may be one of the means to assess tumor treatment response.

Sodium-glucose cotransporter 2 (SGLT2) inhibitors have been shown to prevent the progression of diabetic kidney disease (DKD). However, their impact on renal fibrosis remains largely uninvestigated. This study aimed to explore the effect of SGLT2 inhibitor empagliflozin on renal fibrosis in DKD patients and DKD models, and the molecular mechanisms involved.

Kidney samples of DKD patients and DKD models were used in this study. DKD mouse models included STZ-treated CD-1 mice and HFD-fed C57BL/6 mice were all treated with empagliflozin for 6 to 12 weeks. Kidney pathological changes were analysed and fibrotic factors were detected. HK-2 cells were treated with normal glucose (NG), high glucose (HG), or HG with empagliflozin. RNA sequencing was employed to identify the differentially expressed genes. Epithelial-mesenchymal transition (EMT) markers were detected. Binding of transcription factor and target gene was determined using a dual-luciferase reporter assay.

Empagliflozin significantly ameliorated kidney fibrosis in DKD patients and DKD models. This was evidenced by tubulointerstitial fibrosis reduction observed through PAS and Masson staining, along with fibrotic factors downregulation. RNA sequencing and the subsequent in vitro and in vivo validation identified PKM2 as the most significantly upregulated glycolytic enzyme in DKD patients and models. Empagliflozin downregulated PKM2 and alleviated EMT and renal fibrosis. Importantly, empagliflozin improves fibrosis by downregulating PKM2. The downregulation of PKM2 by empagliflozin was achieved by inhibiting the binding of estrogen-related receptor α at the promoter.

Empagliflozin ameliorates kidney fibrosis via downregulating PKM2 in DKD.