Targeting treatment-resistant breast cancer stem cells with FKBPL and Its peptide derivative, AD-01, via the CD44 pathway

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

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

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 radiotherapyinduced 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.

Original languageEnglish
Pages (from-to)3881-3893
Number of pages13
JournalClinical Cancer Research
Volume19
Issue number14
DOIs
Publication statusPublished - 15 Jul 2013
Externally publishedYes

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Tacrolimus Binding Proteins
Neoplastic Stem Cells
Breast Neoplasms
Stem Cells
Therapeutics
Neoplasms
Aldehyde Dehydrogenase
Heterografts
Antineoplastic Agents
Fluorescent Antibody Technique
Cell Differentiation
Flow Cytometry
Research Design
AD-01 peptide
Clinical Trials
Cell Line
Polymerase Chain Reaction
Growth
Population

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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Targeting treatment-resistant breast cancer stem cells with FKBPL and Its peptide derivative, AD-01, via the CD44 pathway. / McClements, Lana; Yakkundi, Anita; Papaspyropoulos, Angelos; Harrison, Hannah; Ablett, Matthew P.; Jithesh, Puthen V.; McKeen, Hayley D.; Bennett, Rachel; Donley, Christopher; Kissenpfennig, Adrien; McIntosh, Stuart; McCarthy, Helen O.; O'Neill, Eric; Clarke, Robert B.; Robson, Tracy.

In: Clinical Cancer Research, Vol. 19, No. 14, 15.07.2013, p. 3881-3893.

Research output: Contribution to journalArticle

McClements, L, Yakkundi, A, Papaspyropoulos, A, Harrison, H, Ablett, MP, Jithesh, PV, McKeen, HD, Bennett, R, Donley, C, Kissenpfennig, A, McIntosh, S, McCarthy, HO, O'Neill, E, Clarke, RB & Robson, T 2013, 'Targeting treatment-resistant breast cancer stem cells with FKBPL and Its peptide derivative, AD-01, via the CD44 pathway', Clinical Cancer Research, vol. 19, no. 14, pp. 3881-3893. https://doi.org/10.1158/1078-0432.CCR-13-0595
McClements, Lana ; Yakkundi, Anita ; Papaspyropoulos, Angelos ; Harrison, Hannah ; Ablett, Matthew P. ; Jithesh, Puthen V. ; McKeen, Hayley D. ; Bennett, Rachel ; Donley, Christopher ; Kissenpfennig, Adrien ; McIntosh, Stuart ; McCarthy, Helen O. ; O'Neill, Eric ; Clarke, Robert B. ; Robson, Tracy. / Targeting treatment-resistant breast cancer stem cells with FKBPL and Its peptide derivative, AD-01, via the CD44 pathway. In: Clinical Cancer Research. 2013 ; Vol. 19, No. 14. pp. 3881-3893.
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abstract = "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 radiotherapyinduced 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.",
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T1 - Targeting treatment-resistant breast cancer stem cells with FKBPL and Its peptide derivative, AD-01, via the CD44 pathway

AU - McClements, Lana

AU - Yakkundi, Anita

AU - Papaspyropoulos, Angelos

AU - Harrison, Hannah

AU - Ablett, Matthew P.

AU - Jithesh, Puthen V.

AU - McKeen, Hayley D.

AU - Bennett, Rachel

AU - Donley, Christopher

AU - Kissenpfennig, Adrien

AU - McIntosh, Stuart

AU - McCarthy, Helen O.

AU - O'Neill, Eric

AU - Clarke, Robert B.

AU - Robson, Tracy

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