American Association for Cancer Research
10780432ccr130595-sup-supp_fig_5.pdf (48.68 kB)

Figure S5 from Targeting Treatment-Resistant Breast Cancer Stem Cells with FKBPL and Its Peptide Derivative, AD-01, via the CD44 Pathway

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journal contribution
posted on 2023-03-31, 17:24 authored by Lana McClements, Anita Yakkundi, Angelos Papaspyropoulos, Hannah Harrison, Matthew P. Ablett, Puthen V. Jithesh, Hayley D. McKeen, Rachel Bennett, Christopher Donley, Adrien Kissenpfennig, Stuart McIntosh, Helen O. McCarthy, Eric O'Neill, Robert B. Clarke, Tracy Robson

Figure S5 - PDF file 48K, Figure S5. High FKBPL and low Nanog expression correlates with better DMFS and shows a trend towards better RFS. Samples with opposing FKBPL and Nanog expression (high FKBPL/low Nanog and low FKBPL/high Nanog) were analysed using microarray data from publically available data set (GSE7390). Samples were stratified based on median expression values of NANOG and FKBPL. Kaplan-Meier survival curves of breast cancer patients (n=94) presented



Purpose: FK506-binding protein like (FKBPL) and its peptide derivative, AD-01, have already shown tumor growth inhibition and CD44-dependent antiangiogenic activity. Here, we explore the ability of AD-01 to target CD44-positive breast cancer stem cells (BCSC).Experimental Design: Mammosphere assays and flow cytometry were used to analyze the effect of FKBPL overexpression/knockdown and AD-01 treatment ± other anticancer agents on BCSCs using breast cancer cell lines (MCF-7/MDA-231/ZR-75), primary patient samples, and xenografts. Delays in tumor initiation were evaluated in vivo. The anti–stem cell mechanisms were determined using clonogenic assays, quantitative PCR (qPCR), and immunofluorescence.Results: AD-01 treatment was highly effective at inhibiting the BCSC population by reducing mammosphere-forming efficiency and ESA+/CD44+/CD24− or aldehyde dehydrogenase (ALDH)+ cell subpopulations in vitro and tumor initiation in vivo. The ability of AD-01 to inhibit the self-renewal capacity of BCSCs was confirmed; mammospheres were completely eradicated by the third generation. The mechanism seems to be due to AD-01–mediated BCSC differentiation shown by a significant decrease in the number of holoclones and an associated increase in meroclones/paraclones; the stem cell markers, Nanog, Oct4, and Sox2, were also significantly reduced. Furthermore, we showed additive inhibitory effects when AD-01 was combined with the Notch inhibitor, DAPT. AD-01 was also able to abrogate a chemo- and radiotherapy-induced enrichment in BCSCs. Finally, FKBPL knockdown led to an increase in Nanog/Oct4/Sox2 and an increase in BCSCs, highlighting a role for endogenous FKBPL in stem cell signaling.Conclusions: AD-01 has dual antiangiogenic and anti-BCSC activity, which will be advantageous as this agent enters clinical trial. Clin Cancer Res; 19(14); 3881–93. ©2013 AACR.