Three-Dimensional Human Liver-Chip Emulating Premetastatic Niche Formation by Breast Cancer-Derived Extracellular Vesicles

Abstract

The liver is one of the most common sites of breast cancer metastasis and is associated with high lethality. Although the interaction between tumor cells and their microenvironment at metastatic sites has been recognized as a key regulator of tumor progression, the underlying mechanism is not fully elucidated. Here, we describe a three-dimensional (3D) microfluidic human liver-on-a-chip (liver-chip) that emulates the formation of a premetastatic niche to investigate the roles of breast cancerderived extracellular vesicles (EVs) in liver metastasis. We demonstrate that breast cancer-derived EVs activate liver sinusoidal endothelial cells (LSECs) in the liver-chip, inducing endothelial to mesenchymal transition and destruction of vessel barriers. In addition, we show that transforming growth factor β1 (TGFβ1) in breast cancer-derived EVs upregulates fibronectin, an adhesive extracellular matrix protein, on LSECs, which facilitates the adhesion of breast cancer cells to the liver microenvironment. Furthermore, we observed that EVs isolated from triple-negative breast cancer (TNBC) patients with liver metastasis contain higher TGFβ1 levels and induce adhesion of more breast cancer cells to the 3D human liver-chip than do EVs isolated from healthy donors or nonmetastatic TNBC patients. These findings provide a better understanding of the mechanisms through which breast cancer-derived EVs guide secondary metastasis to the liver. Furthermore, the 3D human liver-chip described in this study provides a platform to investigate the mechanisms underlying secondary metastasis to the liver and possible therapeutic strategies.

Endothelial Extracellular Vesicles: From Keepers of Health to Messengers of Disease

“…By using a modified microfluidic-based approach, one can combine the flow conditions with a system designed to capture cells and visualize, in real time, the secretion of EVs [ 147 ]. One group, using a liver on a chip and breast cancer derived EVs, was able to induce and confirm EndMT in liver sinusoidal endothelial cells [ 148 ]. Another group recently used a heart on a chip model to demonstrate the role of EVs derived from human microvascular endothelial cells in protection and rescue from ischemia-reperfusion injury [ 149 ].…”

Advances in Micro/Nanoporous Membranes for Biomedical Engineering

“…The transmembrane transportation of neutrophils was observed in the chip when an external infection induced liver inflammation. In addition, PDMS TH membranes played a similar role in another liver‐on‐a‐chip constructed by Cho's group [ 130 ] and a human gut‐on‐a‐chip established by Ingber's group (Figure 5D). [ 114 ]…”

Biomimetic Microfluidic Platforms for the Assessment of Breast Cancer Metastasis

“…As illustrated by these representative studies, the microfluidics-based metastasis-on-chip approach could expand our fundamental knowledge of breast TME and enable a more accurate in vitro representation of breast cancer metastasis processes. In recent years, a myriad of studies has used similar microengineering principles to investigate the molecular mechanisms of breast cancer cell invasion (Gioiella et al, 2016;Blaha et al, 2017;Truong et al, 2019;Yankaskas et al, 2019), intravasation (Cui et al, 2017;Nagaraju et al, 2018;Shirure et al, 2018), extravasation (Chen M. B. et al, 2016;Chen et al, 2017;Song et al, 2018;Boussommier-Calleja et al, 2019), breast cancer metastasis organotropism and metastasis niche formation (Bersini et al, 2014;Wheeler et al, 2014;Jeon et al, 2015;Clark et al, 2016b;Xu et al, 2016;Narkhede et al, 2017;Shumakovich et al, 2017;Marturano-Kruik et al, 2018;Mei et al, 2019;Oliver et al, 2019;Kim et al, 2020;Ribeiro et al, 2020;Tian et al, 2020).…”

“…In addition to the soluble factors, EVs have been recognized as intercellular messengers between breast cancer cells and the PMN microenvironment at the distant organ. Recently, several research groups utilized microfluidics approaches to explore the role of breast cancer-derived EVs in the formation of liver PMN (Kim et al, 2020) and breast cancer liver metastasis organotropism (Tian et al, 2020). In the liver-on-chip microfluidic model developed by Kim, Cho and colleagues immortalized human liver sinusoidal endothelial cells were cocultured with immortalized human hepatocytes.…”

“…In addition, the authors show that an upregulation of fibronectin, caused by cytokine TGFβ1 released from breast cancer-derived EVs, facilitated the adhesion of breast cancer cells to the liver microenvironment. As a result, TNBC patients with liver metastasis can attract more breast cancer cells to the liver niche as they produce EVs with higher TGFβ1 levels than do healthy donors or TNBC patients without liver metastasis (Kim et al, 2020). Compared to Kim's system, Tian et al (2020) took a slightly different approach.…”