Single Cell RNA-sequencing of BCG naïve and recurrent non-muscle invasive bladder cancer reveals a CD6/ALCAM-mediated immune-suppressive pathway
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PROVIDER: PRJNA1148227 | ENA |
REPOSITORIES: ENA
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Project description:Bacillus Calmette-Guérin (BCG) is the mainstay of treatment for intermediate- and high-risk non-muscle invasive bladder cancer (NMIBC), yet recurrence rates remain high. To improve the efficacy of BCG, a better understanding of the immune landscape underlying BCG resistance is critical. Here, we performed single-cell RNA-sequencing (scRNA-seq) and whole-exome sequencing on tumors from NMIBC patients before and after BCG treatment. Our analysis revealed a marked increase in CD6/ALCAM interactions between T cells and urothelial cells in BCG recurrent tumors. CD6-high T cells were enriched in recurrent tumors and exhibited downregulation of activation-related genes, indicative of functional impairment. These observations were supported by analysis of an independent BCG-treated NMIBC cohort, in which CD6/ALCAM signaling was correlated with shorter recurrence-free survival (p = 0.00059). Our findings reveal a previously unrecognized association between CD6/ALCAM signaling and BCG resistance in NMIBC patients and highlight this pathway as a potential therapeutic target to enhance response to BCG.
2025-08-12 | GSE269877 | GEO
Project description:Single Cell RNA-sequencing of BCG naïve and recurrent non-muscle invasive bladder cancer reveals a CD6/ALCAM-mediated immune-suppressive pathway
| PRJNA1148226 | ENA
Project description: Not available
| S-EPMC12475466 | biostudies-literature
Project description:Throughout the development of the GC-B cell subset, CD9+ B cells are present, and their differentiation may be controlled by FOXP1. Additionally, CD9+ B cells had a significant impact on the activation of death signals in adenoid tissue during the immune development stage. Adenoid hypertrophy was closely related to an increase in Tfh cells, along with a decrease in CD9+ B cells. The absence of CD9 can result in a significant decrease in serum IgG levels in mice, as indicated by Cd9 knockout mice. Our findings indicate that the CD9+ B cell subset causes Tfh cells to apoptosis via the ALCAM-CD6 signaling pathway, leading to the natural atrophy of adenoids. These findings provide novel clues in the mechanisms that regulate GC regression and termination.
2026-09-09 | GSE262822 | GEO
Project description:Endometrial cancer is the most common gynaecological cancer in western countries, being the endometrioid tumours the most common subtype. Most patients are diagnosed at an early stage and present an excellent prognosis. However, a consistent number of those continue to suffer recurrence, without means to be identified by risk classification systems. Thus, finding a reliable marker to predict recurrence becomes an important unmet clinical issue. ALCAM is a cell–cell adhesion molecule, member of the Immunoglobulin superfamily, which has been associated to the carcinogenesis of many cancers. In here, we first determined the value of ALCAM as marker of recurrence in endometrioid endometrial cancer by conducting a retrospective multicenter study in 174 primary tumours. In early stage patients (N=134), recurrence-free survival was poorer in patients with ALCAM-positive compared to ALCAM-negative tumours (HR 4.237; 95%CI 1.01-17.76). This difference was more significant in patients with early stage moderately-poorly differentiated tumours (HR 9,259; 95%CI 2.12-53.47). In multivariate analysis, ALCAM-positivity was an independent prognostic factor in early stage disease (HR 6.027, 95% CI 1.41-25.74). Then, we demonstrated in vitro a role for ALCAM in cell migration and invasion by using a loss-of-function model in two endometrial cancer cell lines. Moreover, ALCAM depletion resulted in a reduced primary tumour size and reduced metastatic local spread in an orthotopic murine model. Gene expression analysis of ALCAM-depleted cell lines supported that motility, invasiveness, cellular assembly and organization were the most deregulated functions associated to ALCAM. Finally, we evidenced some of the downstream effector genes that are involved in ALCAM mediated cell migration; specifically FLNB, TXNRD1 and LAMC2 were validated. In conclusion, our results highlight the potential of ALCAM as a recurrent biomarker in early stage endometrioid endometrial cancer and point to ALCAM as an important molecule in endometrial cancer dissemination by regulating cell migration, invasion and metastasis.
2016-09-08 | GSE86543 | GEO
Project description:Signal differences between TMRM-hi and ALCAM-low vs. TMRM-hi and ALCAM-hi in representative cardiac and hepatic gene expression in mRNA microarray analysis. INTRODUCTION: Non-genetic purification methods for pluripotent stem cell-derived hepatocyte-like cells are useful for liver regenerative therapy and pharmaceutical applications. METHODS: Fluorescent activated cell sorting (FACS) was used to separate cells by combining two parameters: cellular mitochondrial content evaluated by the mitochondrial membrane potential-dependent fluorescent probe (TMRM) and immunocytochemical detection of activated leukocyte cell adhesion molecule (ALCAM). This method was applied to murine fetal, human embryonic stem cell (ESC)-derived, and human induced pluripotent stem cell (iPSC)-derived cell-mixtures. Separately sorted cell fractions were evaluated by quantitative PCR, immunohistochemistry, and cytochemistry for HNF4a, AFP, and albumin mRNA and/or protein expression. RESULTS: Hepatocyte-like cells were highly segregated into the high TMRM signal and ALCAM-positive population. The purity of hepatocyte-like cells derived from human iPSCs was 97 ± 0.38% (n = 5). CONCLUSIONS: This hepatocyte-like cell purification method may be applicable to quality control of cells for liver regenerative cell therapy and pharmaceutical development.
2020-03-01 | GSE126812 | GEO
Project description:BCG vaccination has well‑documented off‑target effects in early life, but the underlying molecular mechanisms remain incompletely understood. This study investigated whether BCG reprograms neonatal myeloid‑derived suppressor cells (MDSCs) through oxidative phosphorylation (OXPHOS). Neonatal mice (3-4 days old) received a single subcutaneous injection of BCG (5×10⁵ CFU) or PBS. Splenic monocytic (M‑) and polymorphonuclear (PMN‑) MDSCs were analyzed using single‑cell RNA‑seq, bulk RNA‑seq, qRT‑PCR, flow cytometry, and Seahorse metabolic analysis. Immunosuppressive function was assessed with T‑cell proliferation assays in the presence or absence of oligomycin, an OXPHOS inhibitor. BCG vaccination significantly increased the expression of OXPHOS‑related genes (Ndufab1, Sdhd, Uqcrfs1, Cox4i) and mitochondrial mass in both MDSC subsets. Seahorse analysis revealed a higher oxygen consumption rate in BCG‑exposed PMN‑MDSCs. Functional assays showed that BCG impaired MDSC‑mediated T‑cell suppression; oligomycin treatment partially restored this suppressive capacity. Together, these findings indicate that BCG‑induced OXPHOS potentiation contributes to the loss of MDSC immunosuppressive function in neonatal mice, providing a molecular mechanism for the non‑specific immunomodulatory effects of BCG vaccination early in life.
2026-09-02 | GSE330508 | GEO
Project description:Although the intravesical instillation of Bacillus Calmette-Guerin (BCG) is widely used as adjuvant treatment for nonmuscle-invasive bladder cancers, the clinical benefit is variable across patients, and the molecular mechanisms underlying the sensitivity to BCG administration and disease progression are poorly understood. To establish the molecular signatures that predict the responsiveness and disease progression of bladder cancers treated with BCG, we performed transcriptome sequencing (RNA-seq) for 13 treatment-naïve and 22 post-treatment specimens obtained from 14 bladder cancer patients. To overcome disease heterogeneity, we used non-negative matrix factorization to identify the latent molecular features associated with drug responsiveness and disease progression. At least 12 molecular features were present, among which the immune-related feature was associated with drug responsiveness, indicating that pre-treatment anti-cancer immunity might dictate BCG responsiveness. We also identified disease progression-associated molecular features indicative of elevated cellular proliferation in post-treatment specimens. The progression-associated molecular features were validated in an extended cohort of BCG-treated bladder cancers. Our study advances understanding of the molecular mechanisms of BCG activity in bladder cancers and provides clinically relevant gene markers for evaluating and monitoring patients.
2023-12-25 | GSE244895 | GEO
Project description:The primary therapy for high-risk bladder cancer (BCa) is repeated instillations of the tuberculosis vaccine, Bacillus Calmette-Guerin (BCG). While BCG decreases the risk of recurrence by more than half, the concerted mechanisms of immune activation from BCG are unknown. Our objective was to investigate how the immune response differs between responders and non-responders to BCG therapy. We performed single-cell RNA-sequencing of isolated immune cells adjacent to high-risk bladders before and after BCG in BCG responders and non-responders. We identify an increase in Th17-like Th1 cells in BCG responders, characterized by greater expression of pro-inflammatory cytokines. Alternatively, non-responders had increased CD8+ T-cell exhaustion and T-regulatory cells. We identify that the primary mechanism of divergent T cell activity is driven by altered polarization and immunosuppressive signaling with myeloid cells. Through a machine-learning-based approach, we identified a Th17-like Th1 cytokines, such as IL17, IL21, and IL26, were predictive of a response, which were then validated in a separate BCG-treated BCa cohort. Together, this suggests that dynamic regulation of myeloid-T cell interactions can be targeted to improve BCG activity.
2026-03-30 | GSE322495 | GEO
Project description:Patients with high-risk non-muscle-invasive bladder cancer (NMIBC) frequently relapse after standard intravesical BCG therapy and may have a dismal outcome. Resistance mechanisms to such immunotherapy remain misunderstood. Here, using cancer cell lines, freshly resected human bladder tumors and cohorts of bladder cancer patients pre- and post-BCG therapy, we demonstrate two distinct patterns of immune subversion upon BCG relapse. In the first pattern, intracellular BCG infection of cancer cells induced a post-transcriptional downregulation of HLA-I membrane expression via an inhibition of the autophagy flux. Patients with HLA-I deficient cancer cells post-BCG therapy displayed a myeloid immunosuppressive tumor microenvironment with epithelial-to-mesenchymal transition (EMT) characteristics and dismal outcomes. Conversely, patients with HLA-I proficient cancer cells post-BCG therapy presented with CD8+ T cell tumor infiltrates, upregulation of inflammatory cytokines and inhibitory immune checkpoint molecules. Those patients had a very favorable outcome. We surmise that HLA-I expression in bladder cancers at relapse post-BCG does not result from immunoediting but rather from an immune subversion process directly induced by BCG on cancer cells, which predicts dismal prognosis. Cancer cells HLA-I scoring by immunohistochemistry (IHC) staining can be easily implemented by pathologists in routine practice to stratify future urothelial cancer patient treatment strategies.
2022-05-05 | GSE199616 | GEO