Auger Emitter Conjugated PARP Inhibitor for Therapy in Triple Negative Breast Cancers: A Comparative In-Vitro Study.

Ramya Sankaranarayanan,Felix Mottaghy,Matthias Bauwens,Carsten Bolm,Jennifer Peil,Steven Terhorst,Andreas Vogg,Jochen Maurer,Arno Classen,Agnieszka Morgenroth

doi:10.3390/cancers14010230

Abstract

Simple SummaryTriple negative breast cancer (TNBC) is an aggressive subtype of breast cancer, with a high recurrence rate. Since treatment of BRCAmut TNBC patients with PARP inhibitor (PARPi), targeting the nuclear protein PARP1, shows varied responses, its therapeutic efficacy is currently evaluated in combination with chemotherapy. Auger emitters (AEs) are radionuclides that can cause DNA damage when delivered close to the DNA. Due to the nuclear location of PARP1, radiolabelling of PARPi with AEs provide an efficient nuclear delivery mechanism. This study shows the radiosynthesis of an AE radiolabelled PARPi ([125I]-PARPi-01) and its therapeutic effect as monotherapy or in combination with chemotherapeutics in a panel of TNBC cell lines. We found that [125I]-PARPi-01 efficiently induces DNA damage with therapeutic effect irrespective of BRCA mutation. All responsive cell lines have homologous recombination deficiency. Short pretreatment with doxorubicin significantly reduces clonogenic survival of both responsive and resistant cell lines.PARP1 inhibitors (PARPi) are currently approved for BRCAmut metastatic breast cancer, but they have shown limited response in triple negative breast cancer (TNBC) patients. Combination of an Auger emitter with PARPis enables PARP inhibition and DNA strand break induction simultaneously. This will enhance cytotoxicity and additionally allow a theranostic approach. This study presents the radiosynthesis of the Auger emitter [125I] coupled olaparib derivative: [125I]-PARPi-01, and its therapeutic evaluation in a panel of TNBC cell lines. Specificity was tested by a blocking assay. DNA strand break induction was analysed by γH2AX immunofluorescence staining. Cell cycle analysis and apoptosis assays were studied using flow cytometry in TNBC cell lines (BRCAwt/mut). Anchorage independent growth potential was evaluated using soft agar assay. [125I]-PARPi-01 showed PARP1-specificity and higher cytotoxicity than olaparib in TNBC cell lines irrespective of BRCA their status. Cell lines harbouring DNA repair deficiency showed response to [125I]-PARPi-01 monotherapy. Combined treatment with Dox-NP further enhanced therapeutic efficiency in metastatic resistant BRCAwt cell lines. The clonogenic survival was significantly reduced after treatment with [125I]-PARPi-01 in all TNBC lines investigated. Therapeutic efficacy was further enhanced after combined treatment with chemotherapeutics. [125I]-PARPi-01 is a promising radiotherapeutic agent for low radiation dosages, and mono/combined therapies of TNBC.

Highlights

Triple negative breast cancer (TNBC) is the most aggressive and heterogenous subtype of breast cancer, accounting for 15–20% of all breast cancers [1]
We reported the synthesis of a PARP1 specific, olaparib derived Auger emitting PARP1 inhibitors (PARPis): [125 I]-PARPi-01
Among the current HRD screening tests in clinic only two (Myriad’s myChoice® CDx and HRD Focus Panel) screen for telomeric allelic imbalance (TAI) in patient samples to account for HRD [59]. Taken together these results show that [125 I]-PARPi-01 mono-therapy significantly induces γH2AX formation, mitotic phase arrest higher than olaparib and blocks the tumorigenicity in triple negative breast cancer (TNBC) cells regardless of BRCA status, PI3K mutations and RAS family mutations

Summary

Introduction

Triple negative breast cancer (TNBC) is the most aggressive and heterogenous subtype of breast cancer (the most common female cancer), accounting for 15–20% of all breast cancers [1]. It is characterised by the lack of hormone receptors (progesterone and oestrogen receptors) and reduced expression of human epidermal growth factor receptor (HER2). Current FDA approved targeted therapies for TNBC include immunotherapy using the PDL1 inhibitor (PDL1i) atezolizumab (Tecentriq) in combination with nab-paclitaxel [2] or PD1 inhibitor pembrolizumab (Keytruda) combined with chemotherapy in PDL1/PD1 positive patients [3].

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