A residual population of quiescent Phgdhhigh DTCs that survived initial innate immune clearance and became transiently enriched in micrometastases are identified, providing insights into the development of micrometastasis-targeting regimens.
Abstract
The mechanisms underlying the interactions between disseminated tumor cells (DTCs) and their tissue microenvironment during metastatic colonization are currently poorly understood. We integrated multimodal single-cell and spatial profiling from liver cancer mouse models and human metastases to track the spatiotemporal dynamics of DTCs and their microenvironments from single-cell seeding to overt lung metastasis. We identified a residual population of quiescent Phgdhhigh DTCs that survived initial innate immune clearance and became transiently enriched in micrometastases. These cells shaped an immune-scarce microenvironment through PHGDH-dependent, H3K27me3-mediated epigenetic silencing of chemokine transcription, thereby promoting metastatic expansion. Cx3cr1high interstitial macrophages were also transiently enriched before DTC expansion, creating an immune-privileged niche for metastatic outgrowth by recruiting immunosuppressive cells. Inactivating the PHGDH-H3K27me3 axis in DTCs or depleting interstitial macrophages restored immune surveillance and inhibited metastatic colonization. These findings provide insights into the development of micrometastasis-targeting regimens.
Intrahepatic metastasis in multifocal hepatocellular carcinoma is associated with poor prognosis and therapeutic resistance, yet the immune mechanisms driving disease progression remain unclear. Here, we analyzed genetic and immune differences between primary tumors and intrahepatic metastatic lesions using sequencing approaches and spatial validation methods. We found that metastatic lesions shared key genomic features with primary tumors but exhibited a distinct immunosuppressive environment enriched in myeloid and T cell populations. In particular, a subset of macrophages expressing glycoprotein nonmetastatic melanoma protein B (GPNMB) was consistently enriched in metastatic niches across multiple independent cohorts. These macrophages were spatially colocalized with CD8+ T cells exhibiting features of terminal exhaustion. Mechanistically, integrated multiomics and functional analyses revealed that GPNMB overexpression triggers lipid metabolic rewiring via the phosphatidylinositol 3-kinase/AKT-cyclooxygenase-2 cascade, leading to elevated prostaglandin E2 secretion, which directly suppresses CD8+ T cell cytotoxicity. Specific silencing of this subset using a dual-targeted, lipid-polymer nanoparticle (APLsiGpnmb) effectively reversed T cell exhaustion, inhibited metastasis, and synergized with anti-programmed death 1 immunotherapy in mouse models without inducing systemic toxicity. These findings identify GPNMB-positive macrophages as key metabolic and immune regulatory hubs, suggesting that targeting the GPNMB–prostaglandin E2 axis provides a promising precision therapeutic strategy for intrahepatic metastasis in multifocal hepatocellular carcinoma.
Yuyan Xu, Cheng Zhang, Zhuo-Cheng Ji et al.· Research· 0 citations
Advanced renal cell carcinoma frequently acquires resistance to immune checkpoint blockade (ICB), underscoring the pivotal influence of the tumor immune microenvironment (TME) on therapeutic efficacy. While recent studies have implicated multicellular crosstalk within the TME as a central driver of ICB resistance, the precise multicellular programs (MCPs) that orchestrate this process remain poorly defined. Here, through integrative single-cell and spatial transcriptomic profiling of clear cell RCC (ccRCC) cohorts, we delineated a previously unrecognized MCP associated with ICB resistance, distinguished by heightened lysosomal activity, adipogenic signaling, and rewired fatty acid metabolism. Within this program, we uncover a coordinated interplay among TAM_APOE, ccRCC_CXCL14, and endothelial cells, whereby ccRCC_CXCL14 recruits TAM_APOE, which subsequently promotes tumor lipid metabolic reprogramming and angiogenesis, forming a pro-tumorigenic feedforward loop. Spatial mapping revealed a malignant gene topic colocalizing with this MCP in tumor cores, which robustly predicted both unfavorable survival and resistance in ICB-treated patients. Functional assays confirmed that the CXCL14-TAM axis promotes metabolic reprogramming, while dual CXCR4 and PD-1 blockade synergistically reverses the resistant phenotype by restoring CD8⁺ T-cell cytotoxicity. Multiplex immunofluorescence further validated the enrichment of this MCP specifically in non-responders. Collectively, our study defines a spatially organized, functionally coordinated multicellular niche that drives ICB resistance in ccRCC, establishing both a predictive biomarker for patient stratification and a mechanistic framework for therapeutic intervention.
Immunotherapy has transformed the therapeutic landscape of advanced cervical cancer, yet clinical benefit remains limited by a highly heterogeneous and immunosuppressive tumor microenvironment. Traditional paradigms, including binary M1/M2 macrophage polarization and models that interpret T-cell dysfunction solely through checkpoint expression, are insufficient to capture the localized intercellular dynamics that drive immune evasion. Recent advances in single-cell and spatial multi-omics have fundamentally reshaped our understanding of this landscape. In this review, we synthesize emerging high-dimensional atlases to reframe macrophage–T-cell crosstalk from simple ligand–receptor interactions into a spatially organized ecological model. We highlight the paradigm shift toward highly resolved myeloid programs, particularly SPP1+ and C1QC+ macrophage states, and discuss how these programs interact with stromal barriers, regulatory T cells, and metabolic checkpoints to restrict, exclude, or functionally constrain effector T cells within suppressive niches. Crucially, we position persistent high-risk human papillomavirus infection not merely as an initiating carcinogenic trigger, but as an upstream and continuous programmer that rewires innate immune sensing, including context-dependent cGAS–STING-related circuits, to stabilize local immune tolerance throughout disease progression. Finally, we propose translational strategies for distilling complex multi-omic atlases into pathology-compatible prognostic and predictive biomarker signatures. Ultimately, by deciphering these spatially organized networks, this review aims to provide actionable translational insights for targeting macrophage vulnerabilities, guiding biomarker-driven combinatorial immunotherapies, and overcoming immune resistance in cervical cancer.
Li Huang, Nan Lin, Hao Li et al.· Frontiers in Immunology· 0 citations
Cholangiocarcinoma is an aggressive biliary tract malignancy in which metastatic relapse and primary or acquired resistance to immunotherapy remain major causes of mortality. Although immune checkpoint inhibitors have improved first-line treatment for advanced biliary tract cancer, most patients do not achieve durable benefit, indicating that immune failure is not explained by a single checkpoint pathway. In this Review, we propose a spatial immune-regulatory continuum for cholangiocarcinoma dissemination. Most direct single-cell and spatial evidence currently derives from intrahepatic cholangiocarcinoma, and its applicability to perihilar and distal disease remains to be established. This continuum begins in the tumor core and invasive front, where malignant cells, cancer-associated fibroblasts, tumor-associated macrophages, endothelial and lymphatic cells, regulatory T cells, immature neutrophils and excluded or dysfunctional cytotoxic T cells form a pro-invasive ecosystem. It then extends through extracellular vesicles, soluble mediators and lymphovascular routes that may educate organotropic pre-metastatic niches. Finally, lymph node, lung, liver, peritoneal and bone microenvironments provide organ-specific extracellular matrix, myeloid and stromal programs that enable immune evasion and metastatic colonization. By integrating clinical evidence, multi-omics studies, single-cell and spatial transcriptomics, extracellular vesicle biology, pre-metastatic niche concepts and emerging therapeutic strategies, we argue that cholangiocarcinoma metastasis should be targeted before overt dissemination whenever possible. In this Review, “metastasis-intercepting immunotherapy” is used as an author-defined conceptual framework for strategies intended to prevent or disrupt the immune–stromal conditions that enable dissemination and colonization, rather than merely shrink established metastatic lesions. Metastasis-intercepting immunotherapy will likely require rational combinations that reprogram the invasive front, restore dendritic-cell-mediated antigen presentation, block tumor-stroma-myeloid circuits, disrupt EV-mediated communication that may contribute to niche formation and select patients using spatial biomarkers rather than bulk immune markers alone.
Longhao Zhang, Kai Zhang, Zhihong Chen et al.· Frontiers in Immunology· 0 citations
Colitis-associated colorectal cancer (C-CRC) develops in chronic inflammatory settings, where tumor—microenvironment interactions critically influence progression. Spatial transcriptomics enables analysis of these interactions while preserving tissue architecture. We aimed to define tumor cell states and associated microenvironmental features in C-CRC.
Seven tissue samples from three C-CRC patients were analyzed using consecutive sections processed by 10x Xenium and 10x Visium. Single-cell spatial coordinates from Xenium were integrated with Visium spots for combined spatial transcriptomic analysis.
Xenium profiling identified 3,506,705 cells comprising epithelial, fibroblast, and immune populations. Tumor subclustering revealed two states: proliferating tumor cells enriched for cell-cycle genes (MKI67, TOP2A) and invasive tumor cells with elevated epithelial mesenchymal transition (EMT)-associated genes (CD44, CXCL1). Spatial neighborhood analysis identified a niche unique to invasive tumor cells characterized by proximity to cancer-associated fibroblasts (CAF) and SPP1+ macrophages. Invasive regions showed enrichment of angiogenesis, hypoxia, and inflammatory pathways. Receptor—ligand analysis identified SPP1—CD44 signaling between SPP1+ macrophages and invasive tumor cells. A gene signature distinguishing invasive and proliferating tumor cells stratified TCGA sporadic colorectal cancer samples into invasive-like and proliferating-like groups, with invasive-like tumors enriched for consensus molecular subtype 4.
This integrated spatial transcriptomics study identifies two tumor populations in C-CRC with distinct microenvironmental contexts. The invasive tumor phenotype is associated with CAF and SPP1+ macrophage proximity and SPP1—CD44-mediated signaling, highlighting the tumor microenvironment as a potential therapeutic target in C-CRC.
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Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
Yoonho Lee, Yongjae Kim, J. Baek et al.· Journal of Immunology· 0 citations
Metastasis is the principal cause of death from colorectal cancer (CRC), yet the cellular states that enable tumor dissemination remain poorly defined. Disseminated tumor cells (DTCs) are rare, transient, and clinically inaccessible, limiting mechanistic insight into their biology. Here we show that CRC cells transiently adopt a wound-healing program normally used by epidermal keratinocytes during tissue repair to enable metastatic dissemination. Using serial orthotopic transplantation of patient-derived organoids to model metastasis, we find that DTCs lose cancer stem cell features and instead express wound-inducible keratins, including KRT17, before metastatic outgrowth. This state is reversible, as cells reacquire primary tumor–like characteristics upon colonization of distant organs. Mechanistically, this transition is associated with reduced EZH2 activity and activation of YAP signaling. Clinically, KRT17⁺ cells localize to the invasive front of primary CRCs and are absent from adjacent normal tissue. These findings uncover unexpected lineage plasticity across distinct developmental origins and identify a transient, targetable state critical for metastatic progression. The plasticity of disseminated tumor cells (DTCs) remains to be understood. Here the authors use serial orthotopic transplantation of colorectal patient-derived organoids to mimic cancer metastasis, and observe that DTCs have transiently adopted a wound-healing program of epidermal keratinocytes to express keratins before the metastatic outgrowth in distant organs.
M. Sakahara, Takuya Okamoto, Kohei Kumegawa et al.· Nature Communications· 0 citations
