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RNA sequencing of sorted immune cells from WT and CD4Cre-Yapfloxed mice from tumors and tumor-draining lymph nodes

ABSTRACT: We report the effect of T cell lineage-specific Yap deletion on tumor immunity in the B16F10 melanoma mouse model.

ORGANISM(S): Mus musculus

PROVIDER: GSE139883 | GEO | 2019/12/11

REPOSITORIES: GEO

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Single-Cell Gene Expression Profiling of CD45+ Cells from Tumors and Draining Lymph Nodes in Trp1 CD4+ T Cell-Treated B16F10 Melanoma-Bearing Mice

Project description:CD4+ T cells play a crucial role in tumor eradication. Employing single-cell RNA sequencing (scRNA-seq), we analyzed the cell states of CD45+ immune cells from tumors and draining lymph nodes in Trp1 CD4+ T cell-treated B16F10 melanoma-bearing mice. This investigation aims to enhance our understanding of how CD4 T cells mediate tumor regression.

2023-12-18 | GSE250511 | GEO

Melanoma-derived extracellular vesicles mediate lymphatic remodeling and impair tumor immunity in draining lymph nodes

Project description:Transcriptional comparison of B16F10 cells with B16F10 cell-derived extracellular vesicles (EVs) to identify transcripts enriched or de-enriched in EVs compared to their donor cells.

2022-03-01 | E-MTAB-11025 | biostudies-arrayexpress

Pharmacokinetics and pharmacodynamics of therapeutic antibodies in tumors and tumor-draining lymph nodes.

Project description:The signaling axis from the primary tumor to the tumor-draining lymph node (TDLN) has emerged as a crucial mediator for the efficacy of immunotherapies in neoadjuvant settings, challenging the primary use of immunotherapy in adjuvant settings. TDLNs are regarded as highly opportunistic sites for cancer cell dissemination and promote further spread via several primary tumor-dependent mechanisms. Lesion-level mixed responses to antibody immunotherapy have been traced to local immune signatures present in the TDLN and the organ-specific primary tumors that they drain. However, the pharmacokinetics (PK) and biodistribution gradients of antibodies in primary tumors and TDLNs have not been systemically evaluated. These concentration gradients are critical in ensuring adequate antibody pharmacodynamic (PD) T-cell activation and/or anti-tumor response. The current work reviews the knowledge for developing physiologically-based PK and pharmacodynamic (PBPK/PD) models to quantify antibody biodistribution gradients in anatomically distinct primary tumors and TDLNs as a means to characterize the clinically observed heterogeneous responses to antibody therapies. Several clinical and pathophysiological considerations in modeling the primary tumor-TDLN axis, as well as a summary of both preclinical and clinical PK/PD lymphatic antibody disposition studies, will be provided.

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Chemotherapy but Not the Tumor Draining Lymph Nodes Determine the Immunotherapy Response in Secondary Tumors.

Project description:Immunotherapies are used as adjuvant therapies for cancers. However, knowledge of how traditional cancer treatments affect immunotherapies is limited. Using mouse models, we demonstrate that tumor-draining lymph nodes (TdLNs) are critical for tumor antigen-specific T cell response. However, removing TdLNs concurrently with established primary tumors did not affect the immune checkpoint blockade (ICB) response on localized secondary tumor due to immunotolerance in TdLNs and distribution of antigen-specific T cells in peripheral lymphatic organs. Notably, treatment response improved with sequential administration of 5-fluorouracil (5-FU) and ICB compared with concurrent administration of ICB with 5-FU. Immune profiling revealed that using 5-FU as induction treatment increased tumor visibility to immune cells, decreased immunosuppressive cells in the tumor microenvironment, and limited chemotherapy-induced T cell depletion. We show that the effect of traditional cytotoxic treatment, not TdLNs, influences immunotherapy response in localized secondary tumors. We postulate essential considerations for successful immunotherapy strategies in clinical conditions.

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Transcriptional remodeling of lymphatic endothelial cells from tumor-draining lymph nodes in melanoma

Project description:We investigated transcriptional changes in lymphatic endothelial cells lining tumor-draining lymph nodes in melanoma using single-cell RNA sequencing.

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Melanoma growth effects on molecular clearance from tumors and biodistribution into systemic tissues versus draining lymph nodes.

Project description:Factors produced within or administered directly into the tumor interstitium, such as cytokines, chemokines, proteases, exosomes, microvesicles, or therapeutic agents, play important and multifaceted roles in the regulation of malignant disease progression. Their bioavailability to mediate signaling in distributed tissues outside of the tumor microenvironment, however, has not been well described. We therefore sought to elucidate the relative extent to which factors from within the primary tumor disseminate to systemic tissues as well as how these distribution profiles are influenced by both hydrodynamic size and the remodeling tumor vasculature. To accomplish this goal, we intratumorally co-infused into the dermal lesions of B16F10 melanoma-bearing mice at prescribed times post tumor implantation a near infrared fluorescent tracer panel ranging from 5 to 500nm in hydrodynamic diameter and compared the in vivo clearance and biodistribution profiles to that of naïve animals. Our results indicate that tumor growth reduces tumor-draining lymph node accumulation and alters the distribution of tumor-derived factors amongst systemic tissues. Despite these changes, previously developed principles of size-dependent lymph node drug targeting are conserved in melanomas, suggesting their applicability to sentinel lymph node-targeted drug delivery. Tumor progression was also found to result in a significant increase in the hydrodynamic size of factors originating from the tumor that accumulated within systemic tissues. This suggests that tumor vascular remodeling may redirect the organism-wide signaling activity of tumor-derived factors and may negatively contribute to disease progression by altering the bioavailability of molecules important to the regulation of pre-metastatic niche formation and the induction of anti-tumor immunity.

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RNA sequencing of sorted immune cells from WT and CD4Cre-Yapfloxed mice from tumors and tumor-draining lymph nodes

Project description:RNA sequencing of sorted immune cells from WT and CD4Cre-Yapfloxed mice from tumors and tumor-draining lymph nodes

| PRJNA587462 | ENA

Mechanisms of Tumor-Induced Lymphovascular Niche Formation in Draining Lymph Nodes.

Project description:Enlargement of the lymphatic vascular network in tumor-draining lymph nodes (LNs) often precedes LN metastasis, likely providing a lymphovascular niche for tumor cells. We investigated morphological and molecular changes associated with the lymphatic remodeling process, using the 4T1 breast cancer and B16F10 melanoma models. Lymphatic expansion in tumor-draining LNs is mediated by sprouting and proliferation of lymphatic endothelial cells (LECs) as early as 4 days after tumor implantation. RNA sequencing revealed an altered transcriptional profile of LECs from tumor-draining compared to naive LNs with similar changes in both tumor models. Integrin ?IIb is upregulated in LECs of tumor-draining LNs and mediates LEC adhesion to fibrinogen in vitro. LEC-associated fibrinogen was also detected in LNs in vivo, suggesting a role of integrin ?IIb in lymphatic remodeling. Together, our results identify specific responses of LN LECs to tumor stimuli and provide insights into the mechanisms of lymphovascular niche formation in tumor-draining LNs.

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Melanoma-derived extracellular vesicles mediate lymphatic remodeling and impair tumor immunity in draining lymph nodes

Project description:We investigated transcriptional changes in lymph node resident lymphatic endothelial cells and medullary sinus macrophages upon interactions with melanoma cell-derived extracellular vesicles using single-cell RNA sequencing.

2022-03-10 | E-MTAB-10434 | biostudies-arrayexpress

Quantitative, architectural analysis of immune cell subsets in tumor-draining lymph nodes from breast cancer patients and healthy lymph nodes.

Project description:BackgroundTo date, pathological examination of specimens remains largely qualitative. Quantitative measures of tissue spatial features are generally not captured. To gain additional mechanistic and prognostic insights, a need for quantitative architectural analysis arises in studying immune cell-cancer interactions within the tumor microenvironment and tumor-draining lymph nodes (TDLNs).Methodology/principal findingsWe present a novel, quantitative image analysis approach incorporating 1) multi-color tissue staining, 2) high-resolution, automated whole-section imaging, 3) custom image analysis software that identifies cell types and locations, and 4) spatial statistical analysis. As a proof of concept, we applied this approach to study the architectural patterns of T and B cells within tumor-draining lymph nodes from breast cancer patients versus healthy lymph nodes. We found that the spatial grouping patterns of T and B cells differed between healthy and breast cancer lymph nodes, and this could be attributed to the lack of B cell localization in the extrafollicular region of the TDLNs.Conclusions/significanceOur integrative approach has made quantitative analysis of complex visual data possible. Our results highlight spatial alterations of immune cells within lymph nodes from breast cancer patients as an independent variable from numerical changes. This opens up new areas of investigations in research and medicine. Future application of this approach will lead to a better understanding of immune changes in the tumor microenvironment and TDLNs, and how they affect clinical outcomes.

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