Cytomorphological Characterization of Individual Metastatic Tumor Cells from Gastrointestinal Cancer Patient Lymph Nodes with Imaging Flow Cytometry

Winter Winter,Thierry Thierry,Ruszkiewicz Ruszkiewicz,Gibson Gibson

doi:10.3390/gidisord1040030

Abstract

The presence or absence of tumor cells within patient lymph nodes is an important prognostic indicator in a number of cancer types and an essential element of the staging process. However, patients with the same pathological stage will not necessarily have the same outcome. Therefore, additional factors may aid in identifying patients at a greater risk of developing metastasis. In this proof of principle study, initially, spiked tumor cells in rat lymph nodes were used to mimic a node with a small cancer deposit. Next, human lymph nodes were obtained from cancer patients for morphological characterization. Nodes were dissociated with a manual tissue homogenizer and stained with fluorescent antibodies against CD45 and Pan-Cytokeratin and then imaging flow cytometry (AMNIS ImageStreamX Mark II) was performed. We show here that imaging flow cytometry can be used for the detection and characterization of small numbers of cancer cells in lymph nodes and we also demonstrate the phenotypical and morphological characterization of cancer cells in gastrointestinal cancer patient lymph nodes. When used in addition to conventional histological techniques, this high throughput detection of tumor cells in lymph nodes may offer additional information assisting in the staging process with therapeutic and prognostic applications.

Highlights

The presence of metastatic cancer cells within regional lymph nodes, including in sentinel lymph nodes (SLNs), is a significant prognostic indicator in a number of cancer types including gastrointestinal (GI) cancers [1]
Determining which patients with positive nodes require Axillary Lymph Node Dissection (ALND) and that would benefit from adjuvant radiotherapy and systemic treatment is a problem [8] as some lymph node metastasis may never develop into clinically detectable disease [9]
We aimed to demonstrate the feasibility of using IFC to detect small numbers of cancer cells in lymph node tissue to represent the clinical setting where patients may present with Isolated tumor cells (ITCs) or micrometastatic lesions (Figure S2)

Summary

Introduction

The presence of metastatic cancer cells within regional lymph nodes, including in sentinel lymph nodes (SLNs), is a significant prognostic indicator in a number of cancer types including gastrointestinal (GI) cancers [1]. Positive identification of metastatic tumor cells in lymph nodes may indicate the need for adjuvant therapy [2]. The size of cancer deposits in lymph nodes is clinically relevant. These can be defined as either (1): Macrometastasis (>2 mm), (2): Micrometastasis (0.2–2 mm), or (3): Isolated tumor cells (ITCs;

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Conclusion