Project description:Analysis of purified immune and breast tumor cells from three major compartments where cancer and immune cells interact: primary tumor, tumor draining lymph nodes (tumor invaded or tumor free), and peripheral blood. The results suggests that node-positive patients’ immune regulation and functionality is down-regulated compared to node-negative patients. CD45+ Immune and ESA+ tumor cells were purified from breast cancer patients' primary tumor, tumor-draining lymph node, and peripheral blood (ficoll) and placed onto Agilent microarrays using the dye-swap method. A universal human reference was used as a reference for the patient samples.
Project description:Analysis of purified immune and breast tumor cells from three major compartments where cancer and immune cells interact: primary tumor, tumor draining lymph nodes (tumor invaded or tumor free), and peripheral blood. The results suggests that node-positive patients’ immune regulation and functionality is down-regulated compared to node-negative patients.
Project description:If we can more accurately predict the likelihood of regional lymph node metastasis (LNM) for endoscopically resected T1-stage colorectal cancers (CRC), the number of unnecessary additional surgeries can be reduced. We aimed of identify miRNA markers that can predict LNM-positive tumors among T1-stage CRCs and we also developed a miRNA classifier set for facilitating the accuracy and applicability.
Project description:Lymphatic endothelial cells (LEC) residing in lymph nodes (LN) have been shown to express genes normally restricted to one or a few tissues, termed peripheral tissue antigens (PTA). The expression of one of these PTA, tyrosinase, by LN-resident LEC has been shown to mediate peripheral T cell tolerance. We used a microarray approach to determine the gene expression profile of LN-resident LEC and blood endothelial cells as a comparison with the objective of determining the global PTA repertoire in these LN stromal populations. Skin draining and mesenteric lymph nodes were pooled from 6 week old adult C57BL/6 mice, minced, and enzymatically digested yielding single cell suspensions. Lymph node stromal cells were purified via CD45 magnetic bead negative selection and pure populations of lymphatic endothelial cells (LEC) and blood endothelial cells (BEC) were obtained via electronic cell sorting according to their expression of gp38 and CD31 (LEC: gp38+ CD31+, BEC: gp38- CD31+). Total RNA was extracted, amplified, and hybridized to Affymetrix microarrays. 3 paired independent samples of purified lymph node LEC and BEC were analyzed.
Project description:Purpose: Lymph node invasion is a hallmark of breast cancer disease progression, but current treatment strategies lack guidance from lymph node biomarkers. We investigated JAGGED1 (JAG1) as a promoter of lymph node metastasis and a prognostic biomarker in metastatic lymph node specimens. Experimental Design: We used mouse models to assess the role of JAG1 expression in human and mouse breast cancer cells on lymphovascular invasion, lymph node metastatic potential, transcriptional profiles, and tumor interactions with lymphatic endothelium. We examined breast and lymph node samples from 284 breast cancer patients to determine the correlative and prognostic value of tumoral JAG1 expression in the lymph node. Results: In matched human breast tumor and lymph node samples, tumor cells that invaded lymph nodes showed higher JAG1 expression than their associated primary tumors (P value < 0.0001). In multiple models, breast cancer cells with high JAG1 expression showed increased lymphovascular invasion, lymph node metastasis, lymph node metastatic outgrowth, and migration through lymphatic endothelium. Transcriptomic analysis indicated that tumoral JAG1 regulates both juxtacrine and paracrine signaling pathways that induce inflammatory and pro-metastatic genes in lymphatic endothelium. When examining patients with identical surgical treatments, patients with lymph node JAG1 H-scorelow showed increased 5-year recurrence free survival rates than patients with lymph node JAG1 H-scorehi (87% vs 70%, P=0.016). Conclusions: JAG1 expression promotes lymphovascular invasion and lymph node metastasis in murine models. In patients, high expression of JAG1 in tumor cells in lymph node metastases predicts reduced recurrence free survival.
Project description:Purpose: Lymph node invasion is a hallmark of breast cancer disease progression, but current treatment strategies lack guidance from lymph node biomarkers. We investigated JAGGED1 (JAG1) as a promoter of lymph node metastasis and a prognostic biomarker in metastatic lymph node specimens. Experimental Design: We used mouse models to assess the role of JAG1 expression in human and mouse breast cancer cells on lymphovascular invasion, lymph node metastatic potential, transcriptional profiles, and tumor interactions with lymphatic endothelium. We examined breast and lymph node samples from 284 breast cancer patients to determine the correlative and prognostic value of tumoral JAG1 expression in the lymph node. Results: In matched human breast tumor and lymph node samples, tumor cells that invaded lymph nodes showed higher JAG1 expression than their associated primary tumors (P value < 0.0001). In multiple models, breast cancer cells with high JAG1 expression showed increased lymphovascular invasion, lymph node metastasis, lymph node metastatic outgrowth, and migration through lymphatic endothelium. Transcriptomic analysis indicated that tumoral JAG1 regulates both juxtacrine and paracrine signaling pathways that induce inflammatory and pro-metastatic genes in lymphatic endothelium. When examining patients with identical surgical treatments, patients with lymph node JAG1 H-scorelow showed increased 5-year recurrence free survival rates than patients with lymph node JAG1 H-scorehi (87% vs 70%, P=0.016). Conclusions: JAG1 expression promotes lymphovascular invasion and lymph node metastasis in murine models. In patients, high expression of JAG1 in tumor cells in lymph node metastases predicts reduced recurrence free survival.