Abstract 4075: Proportion of breast cancer stem cells in fine needle aspirates co-relates with the marker of metastatic outcome TMEM
Authors: Oktay Maja H. , Grunblatt Eli , Roy Sweta , Agi Nathan , Adler Esther , Jones Joan G. , Condeelis John S. , Goswami Sumanta 
Published: August 2015

Abstract
Cell surface biomarkers CD44, CD24 and CD133 and intracellular marker ALDH1 have recently been used to identify breast cancer stem cells. A very small number of these cancer stem cells can initiate a tumor growth and cause it to metastasize. The micro-anatomical sites of hematogenous dissemination of breast cancer cells called Tumor MicroEnvironment of Metastasis (TMEM) have been described in formalin-fixed paraffin-embedded invasive ductal carcinomas from patients. These sites are defined as the direct apposition of tumor cells and perivascular macrophages and are visualized using triple immunohistochemistry for CD68, CD31 and Mena to identify the macrophages, endothelial cells and invasive tumor cells respectively. The identification of TMEM sites is clinically important because TMEM density is a predictor of metastatic outcome. Migratory subpopulation of tumor cells has been shown to be enriched for cancer stem cells.
We have performed flow cytometry to identify CD44high/CD24low cells, mRNA fluorescent in situ hybridization (FISH) and qRT-PCR to identify CD133 and ALDH1 expression in breast cancer cells in 50 samples of invasive ductal carcinoma obtained from patients’ cancer excisions by fine needle aspiration (FNA) before formalin fixation. FNA is a minimally invasive method that uses 26 gauge needles to collect discohesive cancer cells primarily by capillary action. The stem cell marker expression was then compared to TMEM scores obtained from the very same cancer excisions after formalin fixation and paraffin embedding.
Our results show very strong correlation between the percentage of CD44high/CD24low cells with TMEM scores (r = 0.91), as well as the percentage of CD133 and ALDH1 expressing cells with TMEM scores (r = 0.88 and 0.86 respectively). We validated the results obtained by FISH with the direct qRT-PCR for CD133 and ALDH1 in the FNA samples. The correlation of CD133 and ALDH1 expression obtained by qRT-PCR with TMEM score was again very strong (r = 0.76 and 0.73 respectively).
Our findings here and those previously reported for migratory tumor cells demonstrate that TMEM rich microenvironments are enriched for cancer stem cells and partially explain positive correlation of TMEM scores with metastasis in patients. They also suggest that the assessment of stem cell proportion in breast cancers can be assessed in FNA samples. The clinical value of stem cell assessment using this approach for prognostic and predictive is under investigation.
Citation Format: Maja H. Oktay, Eli Grunblatt, Sweta Roy, Nathan Agi, Esther Adler, Joan G. Jones, John S. Condeelis, Sumanta Goswami. Proportion of breast cancer stem cells in fine needle aspirates co-relates with the marker of metastatic outcome TMEM. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4075. doi:10.1158/1538-7445.AM2015-4075