Disseminated Tumor Cells In Lymph Nodes Research Articles

Sweeping lymph node micrometastases off their feet: an engineered model to evaluate natural killer cell mediated therapeutic intervention of circulating tumor cells that disseminate to the lymph nodes

Approximately 90% of cancer related deaths are due to metastasis. Cells from the primary tumor can metastasize through either the vascular or lymphatic circulation. Cancer cells in circulation are called circulating tumor cells (CTCs) and it has been shown that bone marrow is a niche for homing of blood borne CTCs from several epithelial tumors. Cancer cells found in bone marrow are termed disseminated tumor cells (DTCs). Likewise, CTCs in the lymphatic circulation are more often seeded in the sentinel lymph nodes (SLN) that drain the tumor. Micrometastases (<2 mm) occur after the arrest and implantation of DTCs in lymph nodes over time. This paper presents a cell culture platform termed microbubbles formed in polydimethylsiloxane (PDMS) from a microfabricated silicon wafer for mimicking lymph node micrometastases. We cultured lymph node seeking cancer cells in microbubbles to evaluate the efficacy of natural killer (NK) mediated therapy for targeting lymph node micrometastasis. The microbubble platform consists of an array of microcavities that provides a unique microenvironment for mimicking the deep cortical unit of the lymph nodes. We show that cancer cells cultured in microbubbles with therapeutic NK cells undergo apoptosis after 24 h in culture. Since lymph node metastases are prevalent across several types of cancer, this platform may be useful for developing improved cancer therapies for targeting lymph node micrometastases.

Prognostic Significance of EpCAM-Positive Disseminated Tumor Cells in Rectal Cancer Patients With Stage I Disease

Here we evaluated the prevalence and prognostic impact of epithelial cell adhesion molecule (EpCAM)-positive disseminated tumor cells (DTCs) in stage I rectal cancer. Further we tested the association of these single tumor cells or small tumor cell groups with the extent of peritumoral lymphangiogenesis. A total of 845 regional lymph nodes (LN) of 44 patients classified as negative on conventional histopathology were retrospectively reanalyzed with immunohistochemistry (IHC) using the monoclonal antibody Ber-Ep4 directed against EpCAM for the detection of DTCs. The degree of lymphangiogenesis in the primary tumors was assessed by IHC of the primary tumor tissue using the monoclonal antibody D2-40, which reacts with the lymphatic endothelium. The IHC results were correlated with clinico-pathologic parameters and clinical follow-up data. EpCAM-positive DTCs in LNs were detected in 8 (18.2%) of the 44 patients. During a median follow-up of 59 months, 3 (37.5%) of the 8 patients with EpCAM-positive DTCs relapsed, whereas none of the DTC-negative patients developed tumor recurrence (P=0.004). Survival analysis revealed a significant effect of the prevalence of DTCs on overall survival (P=0.0009) and on recurrence-free survival (P=0.0001). Finally, the prevalence of EpCAM-positive DTCs in perirectal LNs was significantly correlated with a high density of peritumoral lymphatic vessels (P=0.015). Our results show that DTCs may occur in stage I of rectal cancer and are associated with poor prognosis. Their occurrence seems to be linked to a high density of newly formed lymphatic vessel at the primary tumor site. According to our data, patients with DTCs in their LN might benefit from adjuvant therapy.

Tu2056 Sevofluorane Reduces Liver Damage Secondary to Ischemic/Reperfusion Injury by a Mechanism Not Related to a Preconditioning Effect

Introduction: Recent analyses uncovered genetic variations between paired samples from primary gastrointestinal tumors, lymph node metastases and disseminated tumor cells (DTCs). These findings might help to explain individually variable responses to standard (neo-)adjuvant therapies and further suggest thatmultimodality treatment options in gastrointestinal cancer should be guided by these individual genetic tumor characteristics. Therefore, we assessed the genetic variations in the primary tumor, lymph node metastases and DTCs of patients with esophageal cancer. Patients and Methods: In this translational analysis 86 patients with esophageal cancer undergoing multimodality therapy were included. Initially, we established a protocol for double immunofluorescence labeling for simultaneous visualization of epithelial cell adhesion molecule (EpCAM) expression on cytokeratin positive cells for the detection of DTCs in bone marrow and lymph nodes. After isolation of positively stained cells, their genomic DNA was globally amplified using the MSE-adapter PCR method. Finally, we applied comparative genomic hybridization (CGH) for the genome-wide screening of DNA-gains/-losses on paired samples from primary tumors, lymph node metastases and DTCs of the study patients. Results: DTCs were detected in 25% of the bone marrow and 38% of the lymph node samples. Interestingly, CGH analysis revealed differences between the numbers of chromosal aberrations in DTCs of the bone marrow compared to the lymph node samples with a higher frequency of aberrations in DTCs in the lymph node samples. In addition, genomic analysis revealed differences in the nature of chromosomal aberrations between primary tumors and corresponding lymph node metastases. Moreover, cluster analysis demonstrated similarities of the aberration spectrum between the DTCs and lymph node metastases while primary tumors showed distinct profiles. Conclusion: Chromosal aberration patterns in lymph node metastases and disseminated tumor cells of patients with esophageal cancer undergoing multimodality therapy are very similar while primary tumors show a different genomic aberration pattern. These individual genetic tumor characteristics might guide future multimodality treatment options in esophageal cancer.

Tu2057 Anti-Inflammatory Effects of Hypertonic Saline Solution in Pancreatic Ischemia/Reperfusion Injuries

Introduction: Recent analyses uncovered genetic variations between paired samples from primary gastrointestinal tumors, lymph node metastases and disseminated tumor cells (DTCs). These findings might help to explain individually variable responses to standard (neo-)adjuvant therapies and further suggest thatmultimodality treatment options in gastrointestinal cancer should be guided by these individual genetic tumor characteristics. Therefore, we assessed the genetic variations in the primary tumor, lymph node metastases and DTCs of patients with esophageal cancer. Patients and Methods: In this translational analysis 86 patients with esophageal cancer undergoing multimodality therapy were included. Initially, we established a protocol for double immunofluorescence labeling for simultaneous visualization of epithelial cell adhesion molecule (EpCAM) expression on cytokeratin positive cells for the detection of DTCs in bone marrow and lymph nodes. After isolation of positively stained cells, their genomic DNA was globally amplified using the MSE-adapter PCR method. Finally, we applied comparative genomic hybridization (CGH) for the genome-wide screening of DNA-gains/-losses on paired samples from primary tumors, lymph node metastases and DTCs of the study patients. Results: DTCs were detected in 25% of the bone marrow and 38% of the lymph node samples. Interestingly, CGH analysis revealed differences between the numbers of chromosal aberrations in DTCs of the bone marrow compared to the lymph node samples with a higher frequency of aberrations in DTCs in the lymph node samples. In addition, genomic analysis revealed differences in the nature of chromosomal aberrations between primary tumors and corresponding lymph node metastases. Moreover, cluster analysis demonstrated similarities of the aberration spectrum between the DTCs and lymph node metastases while primary tumors showed distinct profiles. Conclusion: Chromosal aberration patterns in lymph node metastases and disseminated tumor cells of patients with esophageal cancer undergoing multimodality therapy are very similar while primary tumors show a different genomic aberration pattern. These individual genetic tumor characteristics might guide future multimodality treatment options in esophageal cancer.

Methylation-specific PCR for DNA-based detection of occult tumor cells in lymph nodes of prostate cancer patients.

To assess the feasibility of methylation-specific PCR (MSP) for the detection of promoter hypermethylation of the detoxifying glutathione-S-transferase P1 gene (GSTP1) to detect occult prostate cancer cells in lymph nodes (LNs). Paraffin-embedded pelvic LNs from 20 patients with pT2pN0M0R0 prostate cancer who developed PSA relapse were assessed by MSP. In 18 of the patients, samples of the primary tumor were obtained for MSP. In 19 patients, bone marrow (BM) aspirates were analyzed preoperatively for disseminated tumor cells by immunocytochemistry (mAb A45-B/B3). In 16 patients, biopsies of the anastomotic region were performed following PSA relapse. As a negative control GSTP1 methylation status was also assessed in LNs from 9 patients for whom an autopsy was performed for non-cancer-related causes. All primary tumors displayed GSTP1 hypermethylation (HM). Preoperative BM assessment showed disseminated tumor cells in 8/20 cases (40%). In 4 patients, biopsies of the vesico-urethral anastomosis showed local tumor recurrence. The LNs in the cancer patients showed GSTP1 HM in 18/20 cases (90%) versus 1/9 patients (11.1%) in the non-cancer cohort (p<0.0001). GSTP1 HM appears to be well suited for molecular staging of prostate cancer and accurately detects disseminated tumor cells in LNs, which was seen in 90% of the patients with PSA relapse. However, the limited number of patients and the finding of benign and malignant prostatic tissue at the vesico-urethral anastomosis as a putative local source of PSA recurrence does not allow us to draw conclusions on the prognostic significance of our findings, yet.