Irreversible inhibition of TRF2TRFH recruiting functions by a covalent cyclic peptide induces telomeric replication stress in cancer cells

Alexander P Sobinoff; Salvatore Di Maro; Ronnie R J Low; Rosaria Benedetti; Stefano Tomassi; Antonia D'Aniello; Rosita Russo; Ilaria Baglivo; Ugo Chianese; Paolo V Pedone; Angela Chambery; Anthony J Cesare; Lucia Altucci; Hilda A Pickett; Sandro Cosconati

doi:10.1016/j.chembiol.2023.11.008

Irreversible inhibition of TRF2_TRFH recruiting functions by a covalent cyclic peptide induces telomeric replication stress in cancer cells

Cell Chem Biol. 2023 Dec 21;30(12):1652-1665.e6. doi: 10.1016/j.chembiol.2023.11.008. Epub 2023 Dec 7.

Authors

Affiliations

¹ Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, University of Sydney, Westmead, NSW 2145, Australia.
² DiSTABiF, University of Campania "Luigi Vanvitelli", Via Vivaldi 43, 81100 Caserta, Italy.
³ Genome Integrity Unit, Children's Medical Research Institute, Faculty of Medicine and Health, University of Sydney, Westmead, NSW 2145, Australia.
⁴ Department of Precision Medicine, University of Campania "Luigi Vanvitelli", Vico L. De Crecchio 7, 80138 Naples, Italy; Azienda Ospedaliera Universitaria "Luigi Vanvitelli", Medical Epigenetics Program.
⁵ Department of Pharmacy, University of Naples "Federico II", Via D. Montesano 49, 80131 Naples, Italy.
⁶ Department of Precision Medicine, University of Campania "Luigi Vanvitelli", Vico L. De Crecchio 7, 80138 Naples, Italy.
⁷ Department of Precision Medicine, University of Campania "Luigi Vanvitelli", Vico L. De Crecchio 7, 80138 Naples, Italy; BIOGEM, 83031 Ariano Irpino, Italy; Azienda Ospedaliera Universitaria "Luigi Vanvitelli", Medical Epigenetics Program; IEOS CNR, Napoli, Italy.
⁸ Telomere Length Regulation Unit, Children's Medical Research Institute, Faculty of Medicine and Health, University of Sydney, Westmead, NSW 2145, Australia. Electronic address: [email protected].
⁹ DiSTABiF, University of Campania "Luigi Vanvitelli", Via Vivaldi 43, 81100 Caserta, Italy. Electronic address: [email protected].

PMID: 38065101
DOI: 10.1016/j.chembiol.2023.11.008

Abstract

The TRF2 shelterin component is an essential regulator of telomere homeostasis and genomic stability. Mutations in the TRF2_TRFH domain physically impair t-loop formation and prevent the recruitment of several factors that promote efficient telomere replication, causing telomeric DNA damage. Here, we design, synthesize, and biologically test covalent cyclic peptides that irreversibly target the TRF2_TRFH domain. We identify APOD53 as our most promising compound, as it consistently induces a telomeric DNA damage response in cancer cell lines. APOD53 forms a covalent adduct with a reactive cysteine residue present in the TRF2_TRFH domain and induces phenotypes consistent with TRF2_TRFH domain mutants. These include induction of a telomeric DNA damage response, increased telomeric replication stress, and impaired recruitment of RTEL1 and SLX4 to telomeres. We demonstrate that APOD53 impairs cancer cell growth and find that co-treatment with APOD53 can exacerbate telomere replication stress caused by the G4 stabilizer RHPS4 and low dose aphidicolin (APH).

Keywords: RTEL1; SLX4; TRF2; covalent binder; cyclic peptides; replication stress; shelterin; telomere.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line, Tumor
DNA Damage
Peptides, Cyclic* / pharmacology
Protein Domains
Telomere
Telomeric Repeat Binding Protein 2* / antagonists & inhibitors
Telomeric Repeat Binding Protein 2* / chemistry
Telomeric Repeat Binding Protein 2* / genetics

Substances

Peptides, Cyclic
Telomeric Repeat Binding Protein 2