Reduced SLIT2 is Associated with Increased Cell Proliferation and Arsenic Trioxide Resistance in Acute Promyelocytic Leukemia

Isabel Weinhäuser; Diego A Pereira-Martins; Cesar Ortiz; Douglas R Silveira; Luíse A A Simões; Thiago M Bianco; Cleide L Araujo; Luisa C Koury; Raul A M Melo; Rosane I Bittencourt; Katia Pagnano; Ricardo Pasquini; Elenaide C Nunes; Evandro M Fagundes; Ana B Gloria; Fábio Kerbauy; Maria de Lourdes Chauffaille; Armand Keating; Martin S Tallman; Raul C Ribeiro; Richard Dillon; Arnold Ganser; Bob Löwenberg; Peter Valk; Francesco Lo-Coco; Miguel A Sanz; Nancy Berliner; Emanuele Ammatuna; Antonio R Lucena-Araujo; Jan Jacob Schuringa; Eduardo M Rego

doi:10.3390/cancers12113134

Reduced SLIT2 is Associated with Increased Cell Proliferation and Arsenic Trioxide Resistance in Acute Promyelocytic Leukemia

Cancers (Basel). 2020 Oct 27;12(11):3134. doi: 10.3390/cancers12113134.

Authors

Isabel Weinhäuser^{1

2

3}, Diego A Pereira-Martins^{1

2

3}, Cesar Ortiz^{1

2}, Douglas R Silveira^{4

5}, Luíse A A Simões^{2

4}, Thiago M Bianco¹, Cleide L Araujo², Luisa C Koury¹, Raul A M Melo⁶, Rosane I Bittencourt⁷, Katia Pagnano⁸, Ricardo Pasquini⁹, Elenaide C Nunes⁹, Evandro M Fagundes¹⁰, Ana B Gloria¹⁰, Fábio Kerbauy¹¹, Maria de Lourdes Chauffaille¹¹, Armand Keating¹², Martin S Tallman¹³, Raul C Ribeiro¹⁴, Richard Dillon¹⁵, Arnold Ganser¹⁶, Bob Löwenberg¹⁷, Peter Valk¹⁷, Francesco Lo-Coco^{18

19}, Miguel A Sanz^{20

21}, Nancy Berliner²², Emanuele Ammatuna³, Antonio R Lucena-Araujo²³, Jan Jacob Schuringa³, Eduardo M Rego^{2

4}

Affiliations

¹ Department of Internal Medicine, Department of Medical Images, Hematology, and Clinical Oncology, Medical School of Ribeirao Preto, University of Sao Paulo, Ribeirao Preto 14048-900, Brazil.
² Center for Cell Based Therapy, University of São Paulo, Ribeirao Preto 14051-060, Brazil.
³ Department of Experimental Hematology, Cancer Research Centre Groningen, University Medical Centre Groningen, University of Groningen, 9700 RB Groningen, The Netherlands.
⁴ Hematology Division, LIM31, Faculdade de Medicina, University of Sao Paulo, Sao Paulo 05403-000, Brazil.
⁵ Department of Hematology, AC Camargo Cancer Center, Sao Paulo 01525-001, Brazil.
⁶ Department of Internal Medicine, University of Pernambuco, Recife 50100-130, Brazil.
⁷ Hematology Division, Federal University of Rio Grande do Sul, Porto Alegre 96.200-190, Brazil.
⁸ Hematology and Hemotherapy Center, University of Campinas, Campinas 13083-878, Brazil.
⁹ Hematology Division, Federal University of Parana, Curitiba 13083-878, Brazil.
¹⁰ Hematology Division, Federal University of Minas Gerais, Belo Horizonte 30130-100, Brazil.
¹¹ Hematology Division, Federal University of Sao Paulo, Sao Paulo 04023-062, Brazil.
¹² Princess Margaret Cancer Centre, Toronto, ON M5G 2C1, Canada.
¹³ Leukemia Service, Memorial Sloan-Kettering Cancer Center/Weill Cornell Medical College, New York, NY 10065, USA.
¹⁴ Department of Oncology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
¹⁵ Department of Medical and Molecular Genetics, King's College London School of Medicine, London WC2R 2LS, UK.
¹⁶ Department of Hematology, Hemostasis, Oncology, and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, Germany.
¹⁷ Department of Hematology, Erasmus University Medical Center, 3015 GD Rotterdam, The Netherlands.
¹⁸ Department of Biopathology, University Tor Vergata, 00133 Rome, Italy.
¹⁹ Santa Lucia Foundation, 00179 Rome, Italy.
²⁰ Department of Hematology, Hospital Universitari I Politècnic La Fe, 46-009 Valencia, Spain.
²¹ CIBERONC, Instituto Carlos III, 28029 Madrid, Spain.
²² Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
²³ Department of Genetics, Federal University of Pernambuco, Recife 50670-901, Brazil.

Abstract

The SLIT-ROBO axis plays an important role in normal stem-cell biology, with possible repercussions on cancer stem cell emergence. Although the Promyelocytic Leukemia (PML) protein can regulate SLIT2 expression in the central nervous system, little is known about SLIT2 in acute promyelocytic leukemia. Hence, we aimed to investigate the levels of SLIT2 in acute promyelocytic leukemia (APL) and assess its biological activity in vitro and in vivo. Our analysis indicated that blasts with SLIT2^high transcript levels were associated with cell cycle arrest, while SLIT2^low APL blasts displayed a more stem-cell like phenotype. In a retrospective analysis using a cohort of patients treated with all-trans retinoic acid (ATRA) and anthracyclines, high SLIT2 expression was correlated with reduced leukocyte count (p = 0.024), and independently associated with improved overall survival (hazard ratio: 0.94; 95% confidence interval: 0.92-0.97; p < 0.001). Functionally, SLIT2-knockdown in primary APL blasts and cell lines led to increased cell proliferation and resistance to arsenic trioxide induced apoptosis. Finally, in vivo transplant of Slit2-silenced primary APL blasts promoted increased leukocyte count (p = 0.001) and decreased overall survival (p = 0.002) compared with the control. In summary, our data highlight the tumor suppressive function of SLIT2 in APL and its deteriorating effects on disease progression when downregulated.

Keywords: ATRA; SLIT2; acute promyelocytic leukemia; treatment outcomes.

Abstract

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