Gastric Cancer Metastatic Lymph Nodes Research Articles

Single-cell RNA-seq dissecting heterogeneity of tumor cells and comprehensive dynamics in tumor microenvironment during lymph nodes metastasis in gastric cancer.

Lymph node metastasis is the common metastasis route of gastric cancer. However, until now, heterogeneities of tumor cells and tumor microenvironment in primary tumors (PT) and metastatic lymph nodes (MLN) of gastric cancer (GC) remains uncharacterized. In our study, single cell RNA sequencing was performed on tissues from PT and MLN of gastric cancer. Trajectory analysis and function enrichment analyses were conducted to decode the underlying mechanisms contributing to LN metastasis of gastric cancer. Heterogeneous composition of immune cells and distinct intercellular interactions in PT and MLN were analyzed. Based on the generated single cell transcriptome profiles, dynamics of gene expressions in cancer cells between PT and MLN were characterized. Moreover, we reconstructed the developmental trajectory of GC cells' metastasis to LN and identified two subtypes of GC cells with distinct potentials of having malignant biological behaviors. We characterized the repression of neutrophil polarization associated genes, like LCN2, which would contribute to LN metastasis, and histochemistry experiments validated our findings. Additionally, heterogeneity in neutrophils, rather than macrophages, was characterized. Immune checkpoint associated interaction of SPP1 was found active in MLN. In conclusion, we decode the dynamics of tumor cells during LN metastasis in GC and to identify a subtype of GC cells with potentials of LN metastasis. Our data indicated that the disordering the neutrophils polarization and maturation and the activation of immune checkpoint SPP1 might contribute to LN metastasis in GC, providing a novel insight on the mechanism and potential therapeutic targets of LN metastasis in GC.

Deep neural network trained on gigapixel images improves lymph node metastasis detection in clinical settings

The pathological identification of lymph node (LN) metastasis is demanding and tedious. Although convolutional neural networks (CNNs) possess considerable potential in improving the process, the ultrahigh-resolution of whole slide images hinders the development of a clinically applicable solution. We design an artificial-intelligence-assisted LN assessment workflow to facilitate the routine counting of metastatic LNs. Unlike previous patch-based approaches, our proposed method trains CNNs by using 5-gigapixel images, obviating the need for lesion-level annotations. Trained on 5907 LN images, our algorithm identifies metastatic LNs in gastric cancer with a slide-level area under the receiver operating characteristic curve (AUC) of 0.9936. Clinical experiments reveal that the workflow significantly improves the sensitivity of micrometastasis identification (81.94% to 95.83%, P < .001) and isolated tumor cells (67.95% to 96.15%, P < .001) in a significantly shorter review time (−31.5%, P < .001). Cross-site evaluation indicates that the algorithm is highly robust (AUC = 0.9829).

Tumor-associated macrophages induce capillary morphogenesis of lymphatic endothelial cells derived from human gastric cancer.

Tumor lymphangiogenesis is a major prognostic indicator of gastric cancer. Tumor‐induced inflammation has been shown to attract tumor‐associated macrophages that affect lymphangiogenesis. However, detailed mechanisms of macrophage‐induced lymphangiogenesis have not been elucidated. Here, we evaluated the interaction between tumor‐associated macrophages and lymphatic endothelial cells (LECs) derived from lymph nodes (LNs) of human gastric cancer. Lymphatic endothelial cells were directly or indirectly cocultured with macrophages from healthy human blood, with or without the supernatant of the gastric cancer cell line, OCUM‐12. We analyzed the effect of cancer pretreated macrophages and of macrophages from metastatic LNs of gastric cancer on LECs. We observed morphological changes of LECs in coculture and assessed the gene expression of possible lymphangiogenic molecules of macrophages and LECs after contact coculture, and of cancer pretreated macrophages, by quantitative RT‐PCR. Specimens of metastatic LN of gastric cancer were immunofluorescently stained. We found that tubulogenesis of LECs was observed only in the contact coculture model. OCUM‐12 cells promoted macrophage‐induced tubulogenesis of LECs. Relative gene expression of MMP and adhesion molecules was significantly upregulated in both capillary‐forming LECs and cocultured macrophages. Cancer pretreated macrophages upregulated lymphangiogenic factors including inflammatory cytokines, MMPs, adhesion molecules, and vascular endothelial growth factor‐C. Blocking of intercellular adhesion molecule‐1 and macrophage activation suppressed tubulogenesis of LECs. Immunohistochemistry showed macrophages localized around lymphatic vessels. Our results suggested that interaction between LECs and macrophages may be an important initial step of tumor lymphangiogenesis developing LN metastasis. Understanding of its mechanisms could be useful for future therapeutics of gastric cancer.

Usefulness of Two-Dimensional Values Measured Using Preoperative Multidetector Computed Tomography in Predicting Lymph Node Metastasis of Gastric Cancer.

Multidetector computed tomography (MDCT) is essential for the prediction of lymph node (LN) metastasis in gastric cancer. However, the measurement method and size criteria for metastatic LNs using MDCT are unclear. MDCTs of gastric cancer patients who underwent surgery and had pathological staging were reviewed by radiologists. The two-dimensional cutoff values for LNs with suspected metastasis were calculated, and clinicopathological data were analyzed using those cutoff values. The total number of enrolled patients was 327. The cutoff values of the maximal area with metastatic LNs were obtained significantly at stations 3, 4, and 6, and those values were 112.09, 33.79, and 85.88 mm(2), respectively. The common cutoff value was 112.09 mm(2), and the area under the curve was 0.617 (P = 0.002). The overall survival rate of the patients with LNs less than 112.09 mm(2) was significantly better than those with LNs greater than 112.09 mm(2) (P < 0.001). In multivariate analysis, the maximal LN area was an independent prognostic factor (adjusted hazard ratio, 1.697 [95% confidence interval 1.116-2.582]). Using two-dimensional values for LNs measured by MDCT is a practical method of predicting metastatic LNs in gastric cancer. The maximal LN area value would be useful in both the preoperative staging and prognosis prediction of gastric cancer.

Colonic and gastric cancer metastatic lymph nodes: applications of autofluorescence-based techniques

Colonic and gastric cancer metastatic lymph nodes: applications of autofluorescence-based techniques

Colonic and gastric cancer metastatic lymph nodes: applications of autofluorescence-based techniques

Lymphadenectomy and extended lymphadenectomy are still controversial issues in surgical oncology. The detection methods for metastatic lymph nodes include lymphoscintigraphy and radiolabelled antibody, but immunohistochemical sentinel lymph node (SLN) identification is commonly used. The potential diagnostic use of cell and tissue autofluorescence (AF) is well known. Here we review our studies on the application of AF-based techniques for diagnosing metastatic lymph nodes. We had previously demonstrated that AF imaging allows discrimination between hyperplastic and primary neoplastic lymph nodes. Our more recent studies show that the combination of autofluorescence microspectroscopy and multispectral imaging autofluorescence microscopy can be applied to the diagnosis of secondary neoplastic lymph nodes in gastric and colorectal cancer. These techniques have been validated by histochemical and immunohistochemical methods. Studies are in progress to select the best dye tracer for SLN identification, to improve image quality and to develop a software for automatic analysis of tissue AF to help the decision-making process during surgery.

Colonic and gastric cancer metastatic lymph nodes: applications of autofluorescence-based techniques

Lymphadenectomy and extended lymphadenectomy are still controversial issues in surgical oncology. The detection methods for metastatic lymph nodes include lymphoscintigraphy and radiolabelled antibody, but immunohistochemical sentinel lymph node (SLN) identification is commonly used. The potential diagnostic use of cell and tissue autofluorescence (AF) is well known. Here we review our studies on the application of AF-based techniques for diagnosing metastatic lymph nodes. We had previously demonstrated that AF imaging allows discrimination between hyperplastic and primary neoplastic lymph nodes. Our more recent studies show that the combination of autofluorescence microspectroscopy and multispectral imaging autofluorescence microscopy can be applied to the diagnosis of secondary neoplastic lymph nodes in gastric and colorectal cancer. These techniques have been validated by histochemical and immunohistochemical methods. Studies are in progress to select the best dye tracer for SLN identification, to improve image quality and to develop a software for automatic analysis of tissue AF to help the decision-making process during surgery.

Colonic and gastric cancer metastatic lymph nodes: applications of autofluorescence-based techniques

Lymphadenectomy and extended lymphadenectomy are still controversial issues in surgical oncology. The detection methods for metastatic lymph nodes include lymphoscintigraphy and radiolabelled antibody, but immunohistochemical sentinel lymph node (SLN) identification is commonly used. The potential diagnostic use of cell and tissue autofluorescence (AF) is well known. Here we review our studies on the application of AF-based techniques for diagnosing metastatic lymph nodes. We had previously demonstrated that AF imaging allows discrimination between hyperplastic and primary neoplastic lymph nodes. Our more recent studies show that the combination of autofluorescence microspectroscopy and multispectral imaging autofluorescence microscopy can be applied to the diagnosis of secondary neoplastic lymph nodes in gastric and colorectal cancer. These techniques have been validated by histochemical and immunohistochemical methods. Studies are in progress to select the best dye tracer for SLN identification, to improve image quality and to develop a software for automatic analysis of tissue AF to help the decision-making process during surgery.

Multispectral Imaging Autofluorescence Microscopy in Colonic and Gastric Cancer Metastatic Lymph Nodes

The lymphadenectomy and extended lymphadenectomy procedures have been points of controversy in surgical oncology. The methods available for the detection of metastatic lymph nodes are numerous. These include lymphoscintigraphy and radiolabeled antibody detection, but in most cancers the currently used technique is sentinel lymph node identification, performed primarily through the use of immunohistochemistry. We propose the application of autofluorescence (AF)-based techniques for lymph node evaluation in colorectal and gastric tumors. We studied 30 clinical cases: 15 colorectal cancers and 15 gastric cancers. All of the patients were in the advanced stages of the disease and were candidates for adjuvant therapy. Autofluorescence microspectroscopy and multispectral imaging autofluorescence microscopy have been used to analyze the AF emission of metastatic lymph node sections, excited with 365-nm wavelength radiation. The AF spectra were recorded in the range of 400-700 nm. Monochrome AF images were acquired sequentially through interference filters peaked at 450, 550, and 650 nm, and then combined together in a single red-green-blue image. The AF pattern and the emission spectrum of metastatic lymph nodes have unique characteristics that can be used to distinguish them from the normal ones. The results, compared with standard histopathologic procedures and with specific staining methods, supplied a satisfactory validation of the proposed technique, revealing the possibility of improving the actual diagnostic procedures for malignant lymph node alterations. With the development of appropriate instrumentation, the proposed technique could be particularly suitable in intrasurgical diagnosis of metastatic lymph nodes.