GSTM1 and GSTT1 double null genotypes determining cell fate and proliferation as potential risk factors of relapse in children with hematological malignancies after hematopoietic stem cell transplantation

• Published in: Journal of cancer research and clinical oncology 2022-01, Vol.148 (1), p.71-86
• Main Authors: Jurkovic Mlakar, Simona, Uppugunduri, Satyanarayana Chakradhara Rao, Nava, Tiago, Mlakar, Vid, Golay, Hadrien, Robin, Shannon, Waespe, Nicolas, Rezgui, Mohamed Aziz, Chalandon, Yves, Boelens, Jaap Jan, Bredius, Robert G. M., Dalle, Jean-Hugues, Peters, Christina, Corbacioglu, Selim, Bittencourt, Henrique, Krajinovic, Maja, Ansari, Marc
• Format: Article
• Language: English
• Subjects: Adolescent; Apoptosis; Biomarkers, Tumor - genetics; Busulfan; Busulfan - therapeutic use; Cancer Research; Cell culture; Cell death; Cell fate; Cell Line, Tumor; Cell proliferation; Cell Proliferation - genetics; Child; Child, Preschool; Children; CRISPR; Cytotoxicity; Drug Resistance, Neoplasm - genetics; Female; Gene Deletion; Genetic Predisposition to Disease - genetics; Genotype; Genotypes; Glutathione; Glutathione - analysis; Glutathione - metabolism; Glutathione Transferase - genetics; GSTM1 protein; GSTT1 protein; Hematologic Neoplasms - genetics; Hematologic Neoplasms - pathology; Hematologic Neoplasms - therapy; Hematology; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Humans; Infant; Internal Medicine; Leukemia - genetics; Leukemia - pathology; Leukemia - therapy; Lymphoblastoid cell lines; Male; Medicine; Medicine & Public Health; NCT; NCT01257854; Neoplasm Recurrence, Local - genetics; Null cells; Oncology; Original Article – Cancer Research; Original – Cancer Research; Retrospective Studies; Risk Factors; Stem cell transplantation; Tumor cell lines; Tumors
• ISSN: 0171-5216; 1432-1335
• DOI: 10.1007/s00432-021-03769-2

Purpose This study aimed to retrospectively evaluate the genetic association of null variants of glutathione S-transferases GSTM1 and GSTT1 with relapse incidence in children with hematological malignancies (HMs) undergoing busulfan (BU)- containing allogeneic hematopoietic stem cell transplantation (HSCT) and to assess the impact of these variants on BU-induced cytotoxicity on the immortalized lymphoblastoid cell lines (LCLs) and tumor THP1 GST gene-edited cell models. Methods GSTM1- and GSTT1-null alleles were genotyped using germline DNA from whole blood prior to a conditioning BU-based regimen. Association of GSTM1- and GSTT1-null variants with relapse incidence was analyzed using multivariable competing risk analysis. BU-induced cell death studies were conducted in GSTs - null and non-null LCLs and CRISPR–Cas9 gene-edited THP1 leukemia cell lines. Results Carrying GSTM1/GSTT1 double null genotype was found to be an independent risk factor for post-HSCT relapse in 86 children (adjusted HR: 6.52 [95% Cl, 2.76–15.42; p  = 1.9 × 10 –5 ]). BU-induced cell death preferentially in THP1 GSTM1(non−null) and LCLs GSTM1(non−null) as shown by decreased viability, increased necrosis and levels of the oxidized form of glutathione compared to null cells, while GSTT1 non-null cells showed increased baseline proliferation. Conclusion The clinical association suggests that GSTM1 / GSTT1 double null genotype could serve as genetic stratification biomarker for the high risk of post-HSCT relapse. Functional studies have indicated that GSTM1 status modulates BU-induced cell death. On the other hand, GSTT1 is proposed to be involved in baseline cell proliferation.