Journal Article

FKBPL and its peptide derivatives inhibit endocrine therapy resistant cancer stem cells and breast cancer metastasis by downregulating DLL4 and Notch4

Clarke, Robert B., Robson, Tracy, Simões, Bruno M., Moustafa, Nermeen, Annett, Stephanie, McIntosh, Stuart A., Alqudah, Abdelrahim, Short, Amy, McCarthy, Helen O., Valentine, Andrea, O’Rourke, Martin, Yakkundi, Anita, Furlong, Fiona, McClements, Lana

2019

Overview

Background Optimising breast cancer treatment remains a challenge. Resistance to therapy is a major problem in both ER- and ER+ breast cancer. Tumour recurrence after chemotherapy and/or targeted therapy leads to more aggressive tumours with enhanced metastatic ability. Self-renewing cancer stem cells (CSCs) have been implicated in treatment resistance, recurrence and the development of metastatic disease. Methods In this study, we utilised in vitro, in vivo and ex vivo breast cancer models using ER+ MCF-7 and ER- MDA-MB-231 cells, as well as solid and metastatic breast cancer patient samples, to interrogate the effects of FKBPL and its peptide therapeutics on metastasis, endocrine therapy resistant CSCs and DLL4 and Notch4 expression. The effects of FKBPL overexpression or peptide treatment were assessed using a t-test or one-way ANOVA with Dunnett’s multiple comparison test. Results We demonstrated that FKBPL overexpression or treatment with FKBPL-based therapeutics (AD-01, pre-clinical peptide /ALM201, clinical peptide) inhibit i) CSCs in both ER+ and ER- breast cancer, ii) cancer metastasis in a triple negative breast cancer metastasis model and iii) endocrine therapy resistant CSCs in ER+ breast cancer, via modulation of the DLL4 and Notch4 protein and/or mRNA expression. AD-01 was effective at reducing triple negative MDA-MB-231 breast cancer cell migration ( n  ≥ 3, p  < 0.05) and invasion ( n  ≥ 3, p  < 0.001) and this was translated in vivo where AD-01 inhibited breast cancer metastasis in MDA-MB-231-lucD3H1 in vivo model ( p  < 0.05). In ER+ MCF-7 cells and primary breast tumour samples, we demonstrated that ALM201 inhibits endocrine therapy resistant mammospheres, representative of CSC content ( n  ≥ 3, p  < 0.05). Whilst an in vivo limiting dilution assay, using SCID mice, demonstrated that ALM201 alone or in combination with tamoxifen was very effective at delaying tumour recurrence by 12 ( p  < 0.05) or 21 days ( p  < 0.001), respectively, by reducing the number of CSCs. The potential mechanism of action, in addition to CD44, involves downregulation of DLL4 and Notch4. Conclusion This study demonstrates, for the first time, the pre-clinical activity of novel systemic anti-cancer therapeutic peptides, ALM201 and AD-01, in the metastatic setting, and highlights their impact on endocrine therapy resistant CSCs; both areas of unmet clinical need.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Adaptor Proteins, Signal Transducing / Analysis / Angiogenesis / Animal models / Animals / Antineoplastic Agents, Hormonal - pharmacology / Antineoplastic Agents, Hormonal - therapeutic use / Antineoplastic Combined Chemotherapy Protocols - pharmacology / Antineoplastic Combined Chemotherapy Protocols - therapeutic use / Aromatase inhibitors / Biomedical and Life Sciences / Biomedicine / Breast - pathology / Breast cancer / Breast cancer stem cells / Breast Neoplasms - drug therapy / Breast Neoplasms - pathology / Calcium-Binding Proteins / Cancer metastasis / Cancer Research / Cancer therapies / Cancer treatment / Care and treatment / CD44 antigen / Cell adhesion & migration / Cell cycle / Cell Line, Tumor / Cell migration / Chemotherapy / Development and progression / Down-Regulation - drug effects / Drug Resistance, Neoplasm / Endocrine therapy / Estrogen receptor / Estrogens / Experimental therapeutics and drug development / Female / FKBPL / Gene expression / Gene Expression Regulation, Neoplastic - drug effects / Genetic aspects / Health aspects / Health Promotion and Disease Prevention / Humans / Hypoxia / Immunophilins - pharmacology / Immunophilins - therapeutic use / Intercellular Signaling Peptides and Proteins - metabolism / Internet / Letrozole / Medicine/Public Health / Messenger RNA / Metastases / Metastasis / Mice / Mice, SCID / Neoplasm Recurrence, Local - prevention & control / Neoplastic Stem Cells - drug effects / Neoplastic Stem Cells - pathology / Notch4 protein / Novels / Oncology / Peptides / Peptides - pharmacology / Peptides - therapeutic use / Phenols (Class of compounds) / Physiological aspects / Proteins / Receptor, Notch4 - metabolism / Recurrence (Disease) / Research Article / RNA / Signal Transduction - drug effects / Stem cell transplantation / Stem cells / Surgical Oncology / Tacrolimus Binding Proteins / Tamoxifen / Tamoxifen - pharmacology / Tamoxifen - therapeutic use / Therapeutics / Treatment Outcome / Treatment resistance / Triple negative breast cancer / Tumors / Xenograft Model Antitumor Assays