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  2. Exploring the ATR-CHK1 pathway in the response of doxorubicin-induced DNA damages in acute lymphoblastic leukemia cells

Exploring the ATR-CHK1 pathway in the response of doxorubicin-induced DNA damages in acute lymphoblastic leukemia cells

  • Cell Biol Toxicol. 2021 Sep 14. doi: 10.1007/s10565-021-09640-x.
Andrea Ghelli Luserna Di Rorà 1 Martina Ghetti 2 Lorenzo Ledda 2 Anna Ferrari 2 Matteo Bocconcelli 3 Antonella Padella 2 Roberta Napolitano 2 Maria Chiara Fontana 2 Chiara Liverani 2 Enrica Imbrogno 2 Maria Teresa Bochicchio 2 Matteo Paganelli 2 Valentina Robustelli 3 Seydou Sanogo 2 Claudio Cerchione 4 Monica Fumagalli 5 Michela Rondoni 6 Annalisa Imovilli 7 Gerardo Musuraca 4 Giovanni Martinelli 8 Giorgia Simonetti 2
Affiliations

Affiliations

  • 1 Biosciences Laboratory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) "Dino Amadori", Via Piero Maroncelli, 40, 47014, Meldola, FC, Italy. andrea.ghellilusernadirora@irst.emr.it.
  • 2 Biosciences Laboratory, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) "Dino Amadori", Via Piero Maroncelli, 40, 47014, Meldola, FC, Italy.
  • 3 Department of Experimental, Diagnostic and Specialty Medicine, Institute of Hematology "L. e A. Seràgnoli", University of Bologna, Bologna, Italy.
  • 4 Hematology Unit, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) "Dino Amadori", Meldola, FC, Italy.
  • 5 Hematology Division and Bone Marrow Transplantation Unit, San Gerardo Hospital, Monza, Italy.
  • 6 Hematology Unit, Ospedale Santa Maria delle Croci, Ravenna, Italy.
  • 7 Hematology, AUSL-IRCCS di Reggio Emilia, Reggio Emilia, Italy.
  • 8 Scientific Directorate, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) "Dino Amadori", Meldola, FC, Italy.
Abstract

Doxorubicin (Dox) is one of the most commonly used anthracyclines for the treatment of solid and hematological tumors such as B-/T cell acute lymphoblastic leukemia (ALL). Dox compromises Topoisomerase II Enzyme functionality, thus inducing structural damages during DNA replication and causes direct damages intercalating into DNA double helix. Eukaryotic cells respond to DNA damages by activating the ATM-CHK2 and/or ATR-CHK1 pathway, whose function is to regulate cell cycle progression, to promote damage repair, and to control Apoptosis. We evaluated the efficacy of a new drug schedule combining Dox and specific ATR (VE-821) or Chk1 (prexasertib, PX) inhibitors in the treatment of human B-/T cell precursor ALL cell lines and primary ALL leukemic cells. We found that ALL cell lines respond to Dox activating the G2/M cell cycle checkpoint. Exposure of Dox-pretreated ALL cell lines to VE-821 or PX enhanced Dox cytotoxic effect. This phenomenon was associated with the abrogation of the G2/M cell cycle checkpoint with changes in the expression pCDK1 and cyclin B1, and cell entry in mitosis, followed by the induction of Apoptosis. Indeed, the inhibition of the G2/M checkpoint led to a significant increment of normal and aberrant mitotic cells, including those showing tripolar spindles, metaphases with lagging chromosomes, and massive chromosomes fragmentation. In conclusion, we found that the ATR-CHK1 pathway is involved in the response to Dox-induced DNA damages and we demonstrated that our new in vitro drug schedule that combines Dox followed by ATR/Chk1 inhibitors can increase Dox cytotoxicity against ALL cells, while using lower drug doses. • Doxorubicin activates the G2/M cell cycle checkpoint in acute lymphoblastic leukemia (ALL) cells. • ALL cells respond to doxorubicin-induced DNA damages by activating the ATR-CHK1 pathway. • The inhibition of the ATR-CHK1 pathway synergizes with doxorubicin in the induction of cytotoxicity in ALL cells. • The inhibition of ATR-CHK1 pathway induces aberrant chromosome segregation and mitotic spindle defects in doxorubicin-pretreated ALL cells.

Keywords

ATR; Acute lymphoblastic leukemia; CHK1; Cell cycle; Doxorubicin.

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