Circulating and disseminated tumor cells from breast cancer patient-derived xenograft-bearing mice as a novel model to study metastasis

Introduction

Real-time monitoring of biologic changes in tumors may be possible by investigating the transitional cells such as circulating tumor cells (CTCs) and disseminated tumor cells in bone marrow (BM-DTCs). However, the small numbers of CTCs and the limited access to bone marrow aspirates in cancer patients pose major hurdles. The goal of this study was to determine whether breast cancer (BC) patient-derived xenograft (PDX) mice could provide a constant and renewable source of CTCs and BM-DTCs, thereby representing a unique system for the study of metastatic processes.

Methods

CTCs and BM-DTCs, isolated from BC PDX-bearing mice, were identified by immunostaining for human pan-cytokeratin and nuclear counterstaining of red blood cell-lysed blood and bone marrow fractions, respectively. The rate of lung metastases (LM) was previously reported in these lines. Associations between the presence of CTCs, BM-DTCs, and LM were assessed by the Fisher’s Exact and Cochran-Mantel-Haenszel tests. Two separate genetic signatures associated with the presence of CTC clusters and with lung metastatic potential were computed by using the expression arrays of primary tumors from different PDX lines and subsequently overlapped to identify common genes.

Results

In total, 18 BC PDX lines were evaluated. CTCs and BM-DTCs, present as either single cells or clusters, were detected in 83% (15 of 18) and 62.5% (10 to16) of the lines, respectively. A positive association was noted between the presence of CTCs and BM-DTCs within the same mice. LM was previously found in 9 of 18 (50%) lines, of which all nine had detectable CTCs. The presence of LM was strongly associated with the detection of CTC clusters but not with individual cells or detection of BM-DTCs. Overlapping of the two genetic signatures of the primary PDX tumors associated with the presence of CTC clusters and with lung metastatic potential identified four genes (HLA-DP1A, GJA1, PEG3, and XIST). This four-gene profile predicted distant metastases-free survival in publicly available datasets of early BC patients.

Conclusion

This study suggests that CTCs and BM-DTCs detected in BC PDX-bearing mice may represent a valuable and unique preclinical model for investigating the role of these rare cells in tumor metastases.

Electronic supplementary material

The online version of this article (doi:10.1186/s13058-014-0508-5) contains supplementary material, which is available to authorized users.     

The Evolving Role of the Estrogen Receptor Mutations in Endocrine Therapy-Resistant Breast Cancer

Recurrent ligand-binding domain ESR1 mutations have recently been detected in a substantial number of patients with metastatic ER+ breast cancer and evolve under the selective pressure of endocrine treatments. In this review, we evaluate the current understanding of the biological and clinical significance of these mutations. The preclinical studies revealed that these mutations lead to constitutive ligand-independent activity, indicating resistance to aromatase inhibitors and decreased sensitivity to tamoxifen and fulvestrant. Retrospective analyses of ESR1 mutations in baseline plasma circulating tumor DNA from completed clinical trials suggest that these mutations are prognostic and predictive of resistance to aromatase inhibitors in metastatic disease. Currently, we are lacking prospective studies to confirm these results and to determine the optimal treatment combinations for patients with the ESR1 mutations. In addition, the clinical development of novel agents to overcome resistance engendered by these mutations is also needed.     

Allele-Specific Chromatin Recruitment and Therapeutic Vulnerabilities of ESR1 Activating Mutations

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Quantitative experimental assessment of the factors contributing to hearing loss in serous otitis media.

OBJECTIVE: This study was designed to quantitatively assess the contribution of various factors to the conductive hearing loss in otitis media. BACKGROUND: In the conductive hearing loss seen in cases of serous otitis media, various volumes of fluid of different viscosities along with subatmospheric (negative) pressure are found in the middle ear. METHODS: To evaluate the contribution of each of these factors to hearing loss, various volumes of saline, whole blood, or glycerol were applied to the open middle ear cavity of guinea pigs for short periods of time and auditory function was evaluated by recording the threshold of auditory nerve-brainstem evoked responses. In some of the saline experiments, the bulla cavity was also sealed, allowing a subatmospheric (negative) pressure to develop in the cavity as water was osmotically absorbed because of the gradient in colloid osmotic pressure between saline and blood in the vessels lining the middle ear cavity. In other experiments, a thoracic drainage system was connected to the middle ear cavity to induce desired negative middle ear pressures. RESULTS: The degree of hearing loss increased as larger volumes of fluid were introduced into the middle ear, reaching a maximum of 15 to 16 dB. There was no difference in the degree of hearing loss induced by saline or by fluids with viscosities up to 1,000 times greater than that of water (glycerol). A subatmospheric pressure in the middle ear contributed only a small additional (1-2 dB) threshold elevation. CONCLUSION: The major factor contributing to hearing loss in serous otitis media is the volume of fluid in the middle ear, irrespective of its viscosity. The contribution of negative middle ear pressure is much smaller.