Aug 2026· Biochemical Pharmacology· pp.
118329
· 0 citations· 40 references
Medicine
TL;DR
Hypoxia-activated endothelial cells drive NSCLC aggressiveness and VM through a paracrine axis involving ITPR3-mediated ER calcium activation, and this HIF-1α/ITPR3 axis represents a potential therapeutic target for disrupting the tumor-vascular niche in NSCLC.
Abstract
Vasculogenic mimicry (VM) predicts poor prognosis in non-small cell lung cancer (NSCLC). Hypoxia is a critical tumor microenvironmental regulator, yet its detailed mechanism linking to VM formation remains poorly defined. This study aimed to elucidate the role of hypoxic endothelial cells in promoting NSCLC aggressiveness and VM formation. We integrated clinical data analysis, single-cell RNA sequencing (scRNA-seq), transcriptomic profiling, and multiple in vitro and in vivo functional validations to consolidate our findings. Clinical NSCLC data revealed a positive correlation between endothelial expression of the hypoxia marker carbonic anhydrase IX (CA9) and VM activity. Conditioned medium from hypoxic HUVECs promoted NSCLC cell proliferation, migration, invasion, stemness, and VM network formation in vitro. Endothelial HIF-1α overexpression accelerated tumor growth and VM in vivo. Transcriptomics screening pinpointed the calcium channel gene Inositol 1,4,5-trisphosphate receptor type 3 (ITPR3) as the core downstream effector in this hypoxic endothelial paracrine signaling. Follow-up mechanistic assays revealed that hypoxic endothelial supernatant and the ITPR3 inhibitor 2-aminoethoxydiphenyl borate (2-APB) activated calcium/endoplasmic reticulum (ER) stress signaling to promote malignant phenotypes in NSCLC, and these effects were further validated by ITPR3 knockdown. In conclusion, hypoxia-activated endothelial cells drive NSCLC aggressiveness and VM through a paracrine axis involving ITPR3-mediated ER calcium activation. This HIF-1α/ITPR3 axis represents a potential therapeutic target for disrupting the tumor-vascular niche in NSCLC.
Vascular invasion (VascI) is a critical step in colorectal cancer (CRC) metastasis, but how specific cells in the tumor microenvironment (TME) regulate this process remains unclear, particularly the role of endothelial cells (ECs). Here, we found that vascular invasion is strongly associated with distant metastasis and poor prognosis in CRC patients. Analysis of single-cell RNA sequencing (scRNA-seq) data from 8 colorectal cancer liver metastases (CRLMs), included 4 VascI
+
CRLMs and 4 VascI
-
CRLMs, and identified 10 distinct EC subpopulations. Notably, an IGFBP5_tipECs significantly enriched in VascI
+
CRLMs. High IGFBP5 expression correlated with VascI
+
status and poor patient prognosis. Functionally, IGFBP5 overexpression (OE) in human umbilical vein endothelial cells (HUVECs) enhanced EC migration, tube formation, barrier disruption, and endothelial-to-mesenchymal transition (EndMT). In co-culture system, IGFBP5-OE ECs promoted malignant epithelial cells (MCs) proliferation, migration, and adhesion, and accelerated the growth of patient-derived CRC organoids. Mechanistically, RNA sequencing nominated ITGB3 as a key downstream effector, with activation of the PI3K/AKT signaling pathway. We confirmed a direct ligand-receptor interaction between EC-derived IGFBP5 and ITGB3 on MCs using multiplex immunofluorescence (mIF), co-immunoprecipitation (Co-IP), and proximity ligation assays (PLA). This interaction activated PI3K/AKT signaling in MCs, driving aggressive behavior both in vitro and in vivo, an effect that was significantly attenuated by ITGB3 silencing in MCs. Our findings identify IGFBP5-OE ECs as a key driver of VascI and liver metastasis in CRC via the ITGB3/PI3K/AKT axis, highlighting a potential therapeutic target to suppress metastatic progression.
Nuersimanguli Maimaitiming, Yiqiao Deng, Huayuan Liu et al.· Cell Death & Disease· 0 citations
Hypoxia is a defining feature of the tumor microenvironment and a key driver of malignant progression through transcriptional reprogramming of metabolic, angiogenic, and survival pathways. In this study, hypoxia-induced molecular responses were investigated in human hepatocellular carcinoma (HepG2) and breast adenocarcinoma (MCF-7) cells using cobalt(II) chloride (CoCl2) as a hypoxia mimetic. Cell viability profiling was performed to determine the optimal experimental concentration, followed by quantitative PCR analysis of hypoxia-related genes and miRNA-gene interaction network analysis. Cell viability profiling identified 100 µM CoCl2 as the optimal condition for downstream analyses. Quantitative PCR revealed robust induction of HIF-1α in both cell lines, indicating activation of hypoxic signaling. However, downstream responses differed markedly between cell types. MCF-7 cells exhibited significant upregulation of SLC2A1 and increased VEGFA and NFKB1 expression, consistent with activation of glycolytic and pro-angiogenic pathways. In contrast, HepG2 cells showed limited downstream transcriptional engagement despite elevated HIF1A expression. Furthermore, miRNA-gene interaction analysis revealed a dense regulatory network in MCF-7 cells, whereas HepG2 cells displayed a more restricted, VEGFA-centered miRNA profile. These findings demonstrate that chemical hypoxia induces cell-type-specific transcriptional and post-transcriptional responses in breast and liver cancer cells. The results suggest that miRNA-mediated regulation may contribute to differential tumor adaptation to hypoxic stress and highlight the importance of cellular context in hypoxia-associated signaling pathways.
Okan Sancer, Pınar Aslan Koşar, Uğur Şahin et al.· Drug and chemical toxicology...· 0 citations
Triple-negative breast cancer (TNBC) is an aggressive subtype with limited therapeutic options. Although anti-angiogenic agents like Axitinib are used in cancers, resistance emerges through poorly understood mechanisms involving stemness and vasculogenic mimicry (VM). Here, we investigated the regulatory role of thrombospondin-1 (THBS1), an anti-angiogenic protein, in Axitinib-mediated signaling and its impact on TNBC. The effects of Axitinib on VM, angiogenesis, and stemness in TNBC were investigated through in vivo models and in vitro endothelial-supernatant cultures. Bioinformatics analyses (GSE230643 and CellChat), immunohistochemistry of clinical samples, exosome transfer assays, and xenograft models were employed to explore how Axitinib regulates THBS1-mediated endothelial-TNBC intercellular communication. Further bioinformatics analyses, combined with in vitro and in vivo experimental validations, were performed to elucidate the mechanism of the THBS1-CD47 axis in TNBC. Axitinib treatment increased stemness and promoted VM in TNBC models, despite tumor growth inhibition. Bioinformatics analyses revealed the Axitinib-dependent THBS pathway between endothelial and endothelial-like tumor cells. Axitinib-induced downregulation of THBS1 expression in HUVEC cells promoted stemness and angiogenesis of recipient TNBC cells via exosomes. Mechanistically, endothelial-derived THBS1 promoted CD47 expression in recipient TNBC cells, thereby attenuating the activity of PI3K/Akt and MAPK/ERK signaling pathways, and inhibiting stemness, angiogenesis, and VM. Importantly, Axitinib disrupted this THBS1-CD47 axis by downregulating endothelial-derived THBS1 expression. Axitinib induces the suppression of THBS1 expression in endothelial cells, thereby enhancing stemness and VM in recipient TNBC via disruption of the THBS1-CD47 axis. Restoring THBS1 signaling may represent a promising strategy to overcome Axitinib resistance.
L. Lei, Huanhuan Zhou, Xiaojing Lai et al.· Apoptosis· 0 citations
It is demonstrated that PDCD10 loss promotes GBM neo-angiogenesis involving complementary paracrine and GSC-like plasticity mechanisms, highlighting PDCD10 as a potential therapeutic target to suppress neo-angiogenesis in GBM.
Zhong-Rong Chen, Zhen Chen, X. Wan et al.· Cells· 0 citations
Tumor-associated macrophage (TAM) infiltration is a critical characteristic of triple-negative breast cancer (TNBC) related to drug resistance and poor prognosis. Integrating macrophages into TNBC spheroids is crucial to improve the accuracy of 3D in vitro models that mimic the complexity of the tumor microenvironment (TME) and assess treatment response. However, this remains challenging since the reciprocal effects of these two cell types on each other are not fully understood. In this study, we used the TNBC cell line, MDA-MB-231, and polarized M1-like or M2-like macrophages derived from THP-1 monocytes to establish 3D co-culture spheroids to examine bidirectional interactions between these cells and responses to chemotherapy. Drug efficacy, epithelial-mesenchymal transition (EMT) in cancer cells, macrophage phenotypes, and RNA sequencing, including pathway enrichment analysis, were performed in 3D spheroids. CIBERSORTx deconvolution of RNA sequencing results facilitated the separation of cell types within mixtures to estimate their corresponding cell fractions. We observed that M2 macrophages increased the viability of MDA-MB-231 cells in 3D spheroids, while both M1 and M2 macrophages increased the chemosensitivity of 3D spheroids to doxorubicin and paclitaxel. Interestingly, instead of maintaining their phenotypes, both M1 and M2 macrophages lost some polarization and formed a mixed M1-M2 phenotype when co-cultured with MDA-MB-231 cells in 3D spheroids, a phenomenon further supported by RNA-seq deconvolution analysis. However, the fraction of M1-like macrophages shifting to M2-like was much lower than the fraction of M2-like macrophages shifting to M1-like in the 3D co-cultures. Compared with 2D cultures, an expected mesenchymal transition, numerous differentially expressed genes (DEGs) and various pathways, including both tumor-promoting and tumor-suppressing genes, were observed in 3D spheroid MDA-MB-231 cells. However, both M1- and M2-like macrophages induced only partial EMT phenotype changes of cancer cells in co-cultures. Furthermore, a coexistence of pro-inflammatory and anti-inflammatory DEGs was observed in both M1 and M2-like co-cultured cancer spheroids. In conclusion, our findings present an effective 3D co-culture system of breast cancer cells and integrated macrophages for studying dynamic cellular phenotype changes and reciprocal interactions in a heterogeneous environment to mimic aspects of the TME and enhance the accuracy of preclinical in vitro treatment response studies.
Chen Cheng, Brett A. McGregor, Junguk Hur et al.· PLoS ONE· 0 citations
These results highlight novel prognostic markers and therapeutic vulnerabilities with translational potential for treating TNBC lung metastasis and establish Exoc4, the Hif-2α-Cd36 vasculogenic mimicry axis, Cxcrl1/2-Cxcr2 signaling, and Sftpc as critical mediators of metastatic adaptation.
Hsiao-Chen Lee, C. Kuo, Fang-Ming Chen et al.· Cell Death and Disease· 0 citations