
==== Front
Cancer Diagn Progn
Cancer Diagn Progn
Cancer Diagnosis & Prognosis
2732-7787
International Institute of Anticancer Research

10.21873/cdp.10368
Research Article
Association of PARP1 Expression Levels and Clinical Parameters in Different Leukemic Subtypes With BCR::ABL1 p190+ Translocation
DE MORAIS GUILHERME PASSOS 1
MACHADO CAIO BEZERRA 2
DIAS NOGUEIRA BEATRIZ MARIA 2
DE PINHO PESSOA FLÁVIA MELO CUNHA 2
DE SOUSA OLIVEIRA DEIVIDE 3
RIBEIRO RODRIGO MONTEIRO 3
DA SILVA JEAN BRENO SILVEIRA 3
SEABRA ALINE DAMASCENO 4
MELLO JÚNIOR FERNANDO AUGUSTO RODRIGUES 4
BURBANO ROMMEL RODRIGUEZ 45
KHAYAT ANDRÉ SALIM 5
DE MORAES FILHO MANOEL ODORICO 2
DE MORAES MARIA ELISABETE AMARAL 1
MOREIRA-NUNES CAROLINE AQUINO 1256
1 Unichristus University Center, Faculty of Biomedicine, Fortaleza, Brazil
2 Pharmacogenetics Laboratory, Drug Research and Development Center (NPDM), Department of Physiology and Pharmacology, Federal University of Ceará, Fortaleza, Brazil
3 Department of Hematology, Hospital Geral de Fortaleza (HGF), Fortaleza, Brazil
4 Molecular Biology Laboratory, Ophir Loyola Hospital, Belem, Brazil
5 Department of Biological Sciences, Oncology Research Center, Federal University of Pará, Belem, Brazil
6 Clementino Fraga Group, Central Unity, Molecular Biology Laboratory, Fortaleza, Brazil
Caroline Aquino Moreira-Nunes, Federal University of Ceará, Coronel Nunes de Melo st, n 1000, Rodolfo Teófilo, CEP: 60416-000 Fortaleza, CE, Brazil. Tel: +55 8533668033, email: carolfam@gmail.com
1 9 2024
Sep-Oct 2024
4 5 592598
22 5 2024
21 6 2024
12 7 2024
©2024 The Author(s). Published by the International Institute of Anticancer Research.
2024
https://creativecommons.org/licenses/by-nc-nd/4.0/ This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license
Background/Aim

Although the reciprocal translocation t(9;22)(q34;q11) is a hallmark of chronic myeloid leukemia (CML), it is also present in acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). Depending on the gene's breakpoint, it is possible to obtain three isoforms, among which p190 stands out for the poor prognosis it induces whenever it appears. Due to the genomic instability induced by BCR::ABL1, it is proposed to expand the applicability of poly-ADP-ribose polymerase-1 (PARP1) and its inhibitors in hematological neoplasms.

Materials and Methods

We measured the expression levels of PARP1 by quantitative real-time PCR (qPCR) using TaqMan®, correlating its expression with BCR::ABL1 p190+, to evaluate its influence in the clinic of adult patients.

Results

We found that PARP1 is expressed differently in ALL, AML and CML and that p190 transcripts do not follow a linear pattern in these populations. We also found that PARP1 expression is not correlated with age, white blood cell and the amount of p190 transcripts.

Conclusion

Despite the lack of statistical correlation between the variables analyzed, the role of PARP1 in BCR::ABL1 leukemia cannot be ruled out, given the instability profile promoted by this translocation. Finally, further studies involving a larger sample of patients are needed, as well as investigations into other molecular pathways that may impact on the pathogenesis of different BCR::ABL1 leukemic subtypes.

Biomarker
PARP polymerase
hematological malignancies
BCR::ABL
==== Body
pmcLeukemia is a group of hematological malignancies, presenting a high clonal expansion of malignant cells, originating from myeloid and lymphoid lineages (1). Despite the higher frequency of the t(9;22)(q34;q11) chromosomal translocation in patients with chronic myeloid leukemia (CML) (2), the worst prognosis is observed in patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL), when they harbor the Philadelphia chromosome (Ph+), being classified as high risk, which tends to worsen with age (3,4).

This cytogenetic alteration leads to creation of the chimeric gene BCR activator of RhoGEF and GTPase::ABL proto-oncogene 1 (BCR::ABL1). The gene has three primary isoforms, namely p190BCR::ABL1, p210BCR::ABL1, and p230BCR::ABL1, depending on the different gene breakpoints (5,6). The p210BCR::ABL1 is the main isoform responsible for pathogenesis in CML patients (7). Although p190 is detected in only 1-2% of CML patients (8,9), the p190 transcript confers a higher risk of treatment failure in chronic-phase patients, as well as being associated with distinct clinical characteristics (9). Among the acute leukemias, ALL exhibits the highest frequency of p190BCR::ABL1 (10). Although treatment with tyrosine kinase inhibitors (TKIs) has shown a significant benefit for Ph+ patients, adult patients still suffer from low response of the p190 isoform to TKIs (10-12).

Regardless of leukemia type, the isoform p190BCR::ABL1 produces a protein with 190 kDa, the lowest molecular weight compared to the others. This is associated with a more aggressive leukemia phenotype, due to shorter transcripts causing the loss of important BCR regulatory sequences (13,14).

The presence of BCR::ABL1 induces genomic instability in neoplastic clones through deregulation of mitochondrial membrane potential by the accumulation of reactive oxygen species. In addition, BCR::ABL1 induces greater dependence on non-homologous repair pathways, contributing to increased genomic instability and decreasing the ability of these neoplastic clones to reliably repair damage to the DNA molecule (15-17).

The enzyme poly-ADP-ribose polymerase-1 (PARP1) has emerged as a potential therapeutic target in certain types of solid tumors (18–21) with excessive dependence on non-conservative repair pathways, with the aim of exploiting a distinctive feature of cancer proposed by Hanahan, genomic instability (22). Among a family of 17 enzymes, PARP1 is the main enzyme responsible for molecular signaling for DNA repair by adding ADP-ribose chains in a process known as PARylation (23).

PARP inhibitors (PARPi) act by the principle of synthetic lethality, increasing damage to the genetic material through the accumulation of DNA damage and genomic instability. In addition to the cytotoxicity exerted by direct inhibition of the PARylation process, PARPi acts on the PARP trapping process, forming insoluble complexes between DNA molecules and PARP. This trapping prevents DNA repair and induces greater genetic damage to neoplastic clones (23,24).

PARP1 expression may play an important role in the pathogenesis of adult patients with BCR::ABL1 p190+, as the Ph+ chromosome confers a profile of greater genetic instability in tumor cells by inducing DNA damage (15,16). In this study, we measured PARP1 expression levels in patients and correlated its expression with BCR::ABL1 p190+, to assess its influence on the clinical outcome of adult patients and propose PARP1 as a potential molecular biomarker for future therapeutic applications.

Materials and Methods

Patients and ethical issues. The study group consisted of a total of 59 adult patients diagnosed with CML, AML and ALL who had the BCR::ABL1 p190+ from the Ophir Loyola Hospital, Para, Brazil and Fortaleza General Hospital, Ceara, Brazil. Peripheral blood and bone marrow samples from the patients were used. The analysis also included a control group with 10 peripheral blood samples from healthy individuals, for statistical comparison of gene expression. The patients were brought into the study only after understanding and accepting the terms and informed written consent was obtained from each of them. All methods were carried out in accordance with the Helsinki guidelines and regulations. The research was submitted to the Brazil Platform and was approved by the Research Ethics Committee under registration number 5.207.521.

Expression analysis. Total RNA was extracted from the leucocyte layer using TRIzol Reagent® (Invitrogen™, Carlsbad, CA, USA). Extracted RNA was quantified utilizing the equipment NanoDrop (Thermo Scientific, Carlsbad, CA, USA) and 20 ng were used for cDNA confection using the High-Capacity cDNA Reverse Transcriptase kit (Life Technologies, Carlsbad, CA, USA). The samples were quantified by quantitative real-time PCR (qPCR) using TaqMan® Gene expression assays (Applied Biosystems®, Foster City, CA, USA) for the PARP1 (Hs00242302_m1), BCR::ABL1 p190+ (e1a2) (Hs03024844_ft) and ABL (Hs_01104728_m1) and ACTB (Hs01060665_g1) genes, which were used as endogenous controls.

The commercial kit TaqMan® Expression Master Mix (Applied Biosystems®, Foster City, CA, USA) was used to detect expression analysis by qPCR begin performed using QuantStudio 5 Real-Time PCR system (Applied Biosystems®). The experiments were performed in triplicate and the standard requirements for performing the technique were followed (25). For calculating relative expression levels of PARP1 the 2–∆∆CQ (delta delta cycle quantification) method was used (26), using the health donor samples as the calibrator/control, considering that the amplification of the target gene and the amplification efficiency of the internal control should be approximately equal. A standard curve was applied to determine the p190/ng copy number values.

Statistical analysis. The Chi-square test was conducted to assess the correlation between clinical variables. The Shapiro–Wilk test was used to evaluate the sample’s distribution. Comparison tests were performed by Analysis of Variance (ANOVA) Kruskal–Wallis test and Dunn’s Multiple Comparison post-test were compared in non-parametric tests. The data is based on the mean±standard deviation of the mean of the experimental groups in biological triplicates. The Spearman rank test was applied for correlation between PARP expression and p190/ng copy number values. All statistics were performed using GraphPad Prism (version 8.0.1) and significant differences were considered with an interval of confidence of 95% (p<0.05).

Results

A total of 59 samples from adults diagnosed with BCR::ABL1 p190+ leukemia were used, distributed as ALL (n=28), AML (n=7) and CML (n=24). Among the patients with ALL, 21 were males (75.0%) and 7 females (25.0%), with an average age of 46 years. The average leukometry in peripheral blood was 75.36 (109/l) with an average of 50.1% blasts in the bone marrow.

The patients with AML were 4 males (57.14%) and 3 females (42.85%), with a mean age of 47.5 years. The average leukometry in peripheral blood was 10.59 (109/l) with an average of 72% blasts in the bone marrow. Of the patients with CML, 12 were males (50.0%) and 12 females (50.0%), with a mean age of 56 years. The average leukometry in peripheral blood was 7.15 (109/l) with an average of 3.6% blasts in the bone marrow.

Using the Chi-Square test, we evaluated the influence of clinical variables on the type of leukemia. A significant correlation was identified between age (p=0.0111) on the types of ALL, AML, and CML. However, no significant correlations were found within the other comparisons (Table I).

Table I Clinical data of patients included in the study.

WBC: White blood cell; BM: bone marrow; CK: complex karyotype; ALL: acute lymphoblastic leukemia; AML: acute myeloid leukemia; CML: chronic myeloid leukemia.

Due to the high prevalence of the p190 transcript in the ALL population (10), we decided to evaluate the clinical variables of this isolated group. We stratified these patients into two groups according to PARP1 expression (Table II), to assess the influence of the patient clinical status on PARP1 expression. Similarly, no significant correlations were found between the variables analyzed.

Table II Chi-square analysis of clinical data of patients with acute lymphoblastic leukemia, separated by fold change.

WBC: White blood cell; BM: bone marrow; ALL: acute lymphoblastic leukemia.

The differential expression of PARP1 (F [2,56] = 5,949; p < 0.005; ŋp2, = 0,175 "large effect") concerning the types of leukemia was analyzed and statistically significant values were found between the expression in the group with ALL and CML (Δ: 19,29 [37,91; 18,63] p=0.0002) and between AML and CML (Δ: 18,73 [37,36; 18,63] p=0.0334) (Figure 1).

Figure 1 Expression of poly-ADP-ribose polymerase 1 (PARP1) in patients by type of leukemia BCR::ABL p190+. Expression in patients with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) was compared using analysis of Kruskal's variance and multiple comparisons of Dunn's.. PARP1 expression was normalized using the endogenous gene actin beta (ACTB). Comparisons between CML, AML and ALL were statistically significant with *p=0.0334 and ***p=0.0002. ALL: Acute lymphoblastic leukemia; AML: acute myeloid leukemia; CML: chronic myeloid leukemia.

Subsequently, to analyze the correlation between PARP1 and p190 transcript, we stratified the patients who had a PARP1 expression higher than 1.5-Fold Change. All these patients were diagnosed with ALL and AML, representing 22.03% of the total patients used in the study. We found that in this group with a Fold-Change greater than 1.5, the number of copies of p190/ng of RNA transcripts did not show direct proportionality with PARP1 expression.

In CML, all the patients had a Fold Change of less than 1.5, showing reduced PARP1 expression. In addition, most of these patients had a low concentration of p190 copies/ng RNA.

Using the Spearman’s rank test, we compared all the PARP1, p190/ng copies, age and white blood cell (WBC) variables with each other. No statistically significant correlations were found between PARP1 relative expression and the number of p190/ng copies (r=–0.093), age (r=–0.099) and WBC (r=0.247) (Table III).

Table III Spearman correlation analyses of PARP1, BCR::ABL1 p190+ transcripts, age and WBC of patients with increased expression in more than 1.5-fold of PARP1.

BCR::ABL1 p190+: copy number of p190/nanogram of RNA, WBC: white blood cell (109/l).

Discussion

BCR::ABL1 p190+ leukemia continues to be a complex disease with the worst prognosis in ALL, AML and CML, posing a greater risk and aggressiveness to patients due to p190 (3,4,9). ALL patients exhibit the highest frequency of p190 (10). Due to various advances in the diagnosis and treatment of pediatric ALL, mortality rates have fallen (27,28). However, adult patients continue to suffer from a poor prognosis that tends to worsen with age and the lack of targeted therapies (29).

Given that BCR::ABL1 is an important inducer of genetic instability (15,16), it is hypothesized that patients afflicted with malignant hematological tumors harboring this translocation might benefit from the applicability of PARP as a therapeutic target (30,31). Considering this, we investigated PARP1 expression as a potential biomarker for prognosis in this population.

Although the Chi-square test showed no significant associations between the clinical variables and types of leukemia, the clinical heterogeneity observed suggests that the relationship between these factors may be more complex. The literature describes that patients with p190+ CML have distinct clinical characteristics, increased leukemic progression and, especially high rates of resistance/failure to treatment using first-generation TKIs (32–34).

BCR::ABL1 in AML is still a rare entity (35) and for this reason data on its main clinical characteristics and responses to TKIs are still scarce. Some case reports with the presence of p190+ in adults describe that the presence of mutations and co-expression of other rare transcripts can affect the response to treatment and induce a more aggressive leukemia (36,37).

Epidemiologically corroborating our results in ALL, one study showed a higher frequency of the p190 isoform in adult male patients than in female patients (38). Due to the heterogeneity of clinical characteristics and responses to treatment, we have tried to evaluate alterations in the genome of these patients. The most recurrent alterations are deletions in the IKZF1, PAX5 and CDKN2A/B genes, which are negatively associated with patient outcomes (39,40).

When we evaluated the expression of PARP1 between leukemia types (Figure 1), we found significant differences, showing that the expression in patients with ALL was higher than in patients with AML (p=0.0334) and substantially higher than in patients with CML (p=0.0002). Indeed, the differential expression of PARP1 found a 2-fold increase in ALL compared to healthy patients, and reduced expression in CML (31).

As for AML, studies have shown that high PARP1 expression is associated with complex cytogenetic abnormalities (41). Another study revealed that AML patients with high PARP1 expression had a shorter overall survival and had high levels of blasts, WBC in peripheral blood and were associated with a higher frequency of mutations in FLT3 (42). In addition, a cell culture study showed that PARP2 is also overexpressed in AML cell lines. This suggests that in addition to PARP1, PARP2 may contribute to the pathogenesis of AML (43).

Although PARP1 is overexpressed in CML cell lines (30), our results show that there were no statistically significant correlations between the relative expression of PARP1 and the amount of p190 transcripts in patients with CML, demonstrating that the two events are apparently independent. Although this lack of statistical correlation is evident in this study, the role of PARP1 in BCR::ABL1 leukemia cannot be ruled out, given that cells expressing BCR::ABL1 have homologous recombination (HR) defects and that this expression increases DNA damage and reactive oxygen species (44). In addition, it has been described that this oncogene is capable of inducing leukemia on its own in Ph+ tumors that tend to be extremely dependent on BCR::ABL1 expression (45,46), giving rise to an increased anti-leukemic effect in BCR::ABL1 positive cells in combination with TKI and PARPi therapies (47).

Finally, the characterization of the genetic background of mutations, cytogenetic alterations and DNA repair capacity must be considered so that PARPi therapy can benefit leukemia patients in the future (48).

Conclusion

BCR::ABL1 p190+ leukemia remains a complex disease with a poor prognosis and distinct clinical features. We found that PARP1 is differentially expressed in ALL, AML and CML and that p190 transcripts do not follow a linear pattern in these populations. We also found that PARP1 expression is not correlated with age, white blood cell and the amount of p190 transcripts. Despite the lack of statistical correlation between the variables analyzed, the role of PARP1 in BCR::ABL1 leukemia cannot be ruled out, given the instability profile promoted by this translocation. Finally, further studies involving a larger sample of patients are needed, as well as investigations into other molecular pathways that may impact on the pathogenesis of different BCR::ABL1 leukemic subtypes.

Conflicts of Interest

The Authors declare no conflicts of interest regarding this study.

Authors’ Contributions

Morais GP, Machado CB and Moreira-Nunes CA, performed the study design; Morais GP, Machado CB and JBSS performed the performed gene expression analysis; Morais GP, Machado CB, FMCPP and Nogueira BMD performed the molecular and statistical analysis; Morais GP, Machado CB, and Nogueira BMD, Moraes-Filho MO, Moraes MEA, and Moreira-Nunes CA wrote the article. All Authors read and approved the final article.

Acknowledgements

This study was supported by Brazilian funding agencies National Counsel of Technological and Scientific Development (CNPq; to ASK; MEAM, MOMF and CAMN).
==== Refs
1 Juliusson G Hough R Leukemia Prog Tumor Res 2016 43 87 100 10.1159/000447076 27595359
2 Rashid K Hassan A Tanvir I Ehsan K Significance of Philadelphia chromosome in chronic myeloid leukemia patients of Anmol Hospital, Lahore, Pakistan Pakistan Biomed J 2018 1(1) 10.52229/pbmj.v1i1.47
3 Döhner H Wei AH Appelbaum FR Craddock C Dinardo CD Dombret H Ebert BL Fenaux P Godley LA Hasserjian RP Larson RA Levine RL Miyazaki Y Niederwieser D Ossenkoppele G Röllig C Sierra J Stein EM Tallman MS Tien H Wang J Wierzbowska A Löwenberg B Diagnosis and management of AML in adults: 2022 recommendations from an international expert panel on behalf of the ELN Blood 2022 140(12) 1345 1377 10.1182/blood.2022016867 35797463
4 DeAngelo DJ Jabbour E Advani A Recent advances in managing acute lymphoblastic leukemia Am Soc Clin Oncol Educ Book 2020 (40) 330 342 10.1200/edbk_280175 32421447
5 Ren R Mechanisms of BCR–ABL in the pathogenesis of chronic myelogenous leukaemia Nat Rev Cancer 2005 5(3) 172 183 10.1038/nrc1567 15719031
6 Hantschel O Structure, regulation, signaling, and targeting of abl kinases in cancer Genes Cancer 2012 3(5-6) 436 446 10.1177/1947601912458584 23226581
7 Molica M Zacheo I Diverio D Alimena G Breccia M Long-term outcome of chronic myeloid leukaemia patients with p210 and p190 co-expression at baseline Br J Haematol 2015 169(1) 148 150 10.1111/bjh.13184 25296902
8 Verma D Kantarjian HM Jones D Luthra R Borthakur G Verstovsek S Rios MB Cortes J Chronic myeloid leukemia (CML) with P190 BCR-ABL: analysis of characteristics, outcomes, and prognostic significance Blood 2009 114(11) 2232 2235 10.1182/blood-2009-02-204693 19531657
9 Awad SA Hohtari H Javarappa KK Brandstoetter T Kim D Potdar S Heckman CA Kytölä S Porkka K Doma E Sexl V Kankainen M Mustjoki S BCR-ABL1 p190 in CML: a minor breakpoint with a major impact Blood 2019 134(Suppl_1) 190 10.1182/blood-2019-126584
10 El Fakih R Jabbour E Ravandi F Hassanein M Anjum F Ahmed S Kantarjian H Current paradigms in the management of Philadelphia chromosome positive acute lymphoblastic leukemia in adults Am J Hematol 2018 93(2) 286 295 10.1002/ajh.24926 28971501
11 Jiao Q Bi L Ren Y Song S Wang Q Wang YS Advances in studies of tyrosine kinase inhibitors and their acquired resistance Mol Cancer 2018 17(1) 36 10.1186/s12943-018-0801-5 29455664
12 Mishra S Zhang B Cunnick JM Heisterkamp N Groffen J Resistance to imatinib of Bcr/Abl P190 lymphoblastic leukemia cells Cancer Res 2006 66(10) 5387 5393 10.1158/0008-5472.CAN-05-3058 16707466
13 Komorowski L Fidyt K Patkowska E Firczuk M Philadelphia chromosome-positive leukemia in the lymphoid lineage-similarities and differences with the myeloid lineage and specific vulnerabilities Int J Mol Sci 2020 21(16) 5776 10.3390/ijms21165776 32806528
14 Qin R Wang T He W Wei W Liu S Gao M Huang Z Jak2/STAT6/c-Myc pathway is vital to the pathogenicity of Philadelphia-positive acute lymphoblastic leukemia caused by P190(BCR-ABL) Cell Commun Signal 2023 21(1) 27 10.1186/s12964-023-01039-x 36721266
15 Pawlowska E Blasiak J DNA repair—a double-edged sword in the genomic stability of cancer cells—the case of chronic myeloid leukemia Int J Mol Sci 2015 16(11) 27535 27549 10.3390/ijms161126049 26593906
16 Popp HD Kohl V Naumann N Flach J Brendel S Kleiner H Weiss C Seifarth W Saussele S Hofmann WK Fabarius A DNA damage and DNA damage response in chronic myeloid leukemia Int J Mol Sci 2020 21(4) 1177 10.3390/ijms21041177 32053969
17 Abdulmawjood B Costa B Roma-Rodrigues C Baptista PV Fernandes AR Genetic biomarkers in chronic myeloid leukemia: what have we learned so far Int J Mol Sci 2021 22(22) 12516 10.3390/ijms222212516 34830398
18 O’Malley DM Krivak TC Kabil N Munley J Moore KN PARP inhibitors in ovarian cancer: a review Target Oncol 2023 18(4) 471 503 10.1007/s11523-023-00970-w 37268756
19 Mateo J Lord CJ Serra V Tutt A Balmaña J Castroviejo-Bermejo M Cruz C Oaknin A Kaye SB de Bono JS A decade of clinical development of PARP inhibitors in perspective Ann Oncol 2019 30(9) 1437 1447 10.1093/annonc/mdz192 31218365
20 Taylor AK Kosoff D Emamekhoo H Lang JM Kyriakopoulos CE PARP inhibitors in metastatic prostate cancer Front Oncol 2023 13 1159557 10.3389/fonc.2023.1159557 37168382
21 Gout J Perkhofer L Morawe M Arnold F Ihle M Biber S Lange S Roger E Kraus JM Stifter K Hahn SA Zamperone A Engleitner T Müller M Walter K Rodriguez-Aznar E Sainz B Jr Hermann PC Hessmann E Müller S Azoitei N Lechel A Liebau S Wagner M Simeone DM Kestler HA Seufferlein T Wiesmüller L Rad R Frappart PO Kleger A Synergistic targeting and resistance to PARP inhibition in DNA damage repair-deficient pancreatic cancer Gut 2021 70(4) 743 760 10.1136/gutjnl-2019-319970 32873698
22 Hanahan D Hallmarks of Cancer: New dimensions Cancer Discov 2022 12(1) 31 46 10.1158/2159-8290.CD-21-1059 35022204
23 Lord CJ Ashworth A PARP inhibitors: Synthetic lethality in the clinic Science 2017 355(6330) 1152 1158 10.1126/science.aam7344 28302823
24 Murai J Huang SY Renaud A Zhang Y Ji J Takeda S Morris J Teicher B Doroshow JH Pommier Y Stereospecific PARP trapping by BMN 673 and comparison with olaparib and rucaparib Mol Cancer Ther 2014 13(2) 433 443 10.1158/1535-7163.MCT-13-0803 24356813
25 Bustin SA Benes V Garson JA Hellemans J Huggett J Kubista M Mueller R Nolan T Pfaffl MW Shipley GL Vandesompele J Wittwer CT The MIQE Guidelines: Minimum information for publication of quantitative real-time PCR experiments Clin Chem 2009 55(4) 611 622 10.1373/clinchem.2008.112797 19246619
26 Schmittgen TD Livak KJ Analyzing real-time PCR data by the comparative CT method Nat Protoc 2008 3(6) 1101 1108 10.1038/nprot.2008.73 18546601
27 Hunger SP Lu X Devidas M Camitta BM Gaynon PS Winick NJ Reaman GH Carroll WL Improved survival for children and adolescents with acute lymphoblastic leukemia between 1990 and 2005: a report from the children’s oncology group J Clin Oncol 2012 30(14) 1663 1669 10.1200/JCO.2011.37.8018 22412151
28 Inaba H Mullighan CG Pediatric acute lymphoblastic leukemia Haematologica 2020 105(11) 2524 2539 10.3324/haematol.2020.247031 33054110
29 Aldoss I Stein AS Advances in adult acute lymphoblastic leukemia therapy Leuk Lymphoma 2018 59(5) 1033 1050 10.1080/10428194.2017.1354372 28745565
30 Machado CB DA Silva EL Dias Nogueira BM DE Moraes Filho MO Montenegro RC DE Moraes MEA Moreira-Nunes CA PARP1 is overexpressed in hematological malignant cell lines: a framework for experimental oncology Anticancer Res 2021 41(5) 2397 2402 10.21873/anticanres.15014 33952464
31 Machado CB da Silva EL Ferreira WAS Pessoa FMCP de Quadros AU Fantacini DMC Furtado IP Rossetti R Silveira RM de Lima SCG Mello Júnior FAR Seabra AD Moreira ECO de Moraes Filho MO de Moraes MEA Montenegro RC Ribeiro RM Khayat AS Burbano RMR de Oliveira EHC Covas DT de Souza LEB Moreira-Nunes CFA PARP1 characterization as a potential biomarker for BCR::ABL1 p190+ acute lymphoblastic leukemia Cancers (Basel) 2023 15(23) 5510 10.3390/cancers15235510 38067214
32 Adnan-Awad S Kim D Hohtari H Javarappa KK Brandstoetter T Mayer I Potdar S Heckman CA Kytölä S Porkka K Doma E Sexl V Kankainen M Mustjoki S Characterization of p190-Bcr-Abl chronic myeloid leukemia reveals specific signaling pathways and therapeutic targets Leukemia 2021 35(7) 1964 1975 10.1038/s41375-020-01082-4 33168949
33 Arun AK Senthamizhselvi A Mani S Vinodhini K Janet NB Lakshmi KM Abraham A George B Srivastava A Srivastava VM Mathews V Balasubramanian P Frequency of rare BCR-ABL1 fusion transcripts in chronic myeloid leukemia patients Int J Lab Hematol 2017 39(3) 235 242 10.1111/ijlh.12616 28035733
34 Xue M Wang Q Huo L Wen L Yang X Wu Q Pan J Cen J Ruan C Wu D Chen S Clinical characteristics and prognostic significance of chronic myeloid leukemia with rare BCR-ABL1 transcripts Leuk Lymphoma 2019 60(12) 3051 3057 10.1080/10428194.2019.1607329 31258010
35 Arber DA Orazi A Hasserjian R Thiele J Borowitz MJ Le Beau MM Bloomfield CD Cazzola M Vardiman JW The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia Blood 2016 127(20) 2391 2405 10.1182/blood-2016-03-643544 27069254
36 Piedimonte M Ottone T Alfonso V Ferrari A Conte E Divona M Bianchi MP Ricciardi MR Mirabilii S Licchetta R Campagna A Cicconi L Galassi G Pelliccia S Leporace A Lo Coco F Tafuri A A rare BCR-ABL1 transcript in Philadelphia-positive acute myeloid leukemia: case report and literature review BMC Cancer 2019 19(1) 50 10.1186/s12885-019-5265-5 30630459
37 Kim H Kim IS Kim H Emergence of BCR-ABL1 (p190) in acute myeloid leukemia post-gilteritinib therapy Ann Lab Med 2023 43(4) 386 388 10.3343/alm.2023.43.4.386 36843408
38 Haider S Mahmood A Akhtar F Mahmood R Muzafar S Batool M BCR-ABL1 gene mutation in acute lymphoblastic leukemia Ann Pak Inst Med Sci 2022 18(3) 181 185 10.48036/apims.v18i3.629
39 Fedullo AL Messina M Elia L Piciocchi A Gianfelici V Lauretti A Soddu S Puzzolo MC Minotti C Ferrara F Martino B Chiusolo P Calafiore V Paolini S Vignetti M Vitale A Guarini A Foà R Chiaretti S Prognostic implications of additional genomic lesions in adult Philadelphia chromosome-positive acute lymphoblastic leukemia Haematologica 2019 104(2) 312 318 10.3324/haematol.2018.196055 30190342
40 Mullighan CG Miller CB Radtke I Phillips LA Dalton J Ma J White D Hughes TP Le Beau MM Pui C Relling MV Shurtleff SA Downing JR BCR–ABL1 lymphoblastic leukaemia is characterized by the deletion of Ikaros Nature 2008 453(7191) 110 114 10.1038/nature06866 18408710
41 Pashaiefar H Yaghmaie M Tavakkoly-Bazzaz J Ghaffari SH Alimoghaddam K Momeny M Izadifard M Kasaeian A Ghavamzadeh A Significance of PARP1 expression levels in patients with acute myeloid leukemia Ann Oncol 2018 29 ix87 10.1093/annonc/mdy437
42 Li X Li C Jin J Wang J Huang J Ma Z Huang X He X Zhou Y Xu Y Yu M Huang S Yan X Li F Pan J Wang Y Yu Y Jin J High PARP-1 expression predicts poor survival in acute myeloid leukemia and PARP-1 inhibitor and SAHA-bendamustine hybrid inhibitor combination treatment synergistically enhances anti-tumor effects EBioMedicine 2018 38 47 56 10.1016/j.ebiom.2018.11.025 30472087
43 Gil-Kulik P Dudzińska E Radzikowska-Büchner E Wawer J Jojczuk M Nogalski A Wawer GA Feldo M Kocki W Cioch M Bogucka-Kocka A Rahnama M Kocki J Different regulation of PARP1, PARP2, PARP3 and TRPM2 genes expression in acute myeloid leukemia cells BMC Cancer 2020 20(1) 435 10.1186/s12885-020-06903-4 32423430
44 Hiroki H Ishii Y Piao J Namikawa Y Masutani M Honda H Akahane K Inukai T Morio T Takagi M Targeting Poly(ADP)ribose polymerase in BCR/ABL1-positive cells Sci Rep 2023 13(1) 7588 10.1038/s41598-023-33852-2 37165001
45 Bavaro L Martelli M Cavo M Soverini S Mechanisms of disease progression and resistance to tyrosine kinase inhibitor therapy in chronic myeloid leukemia: an update Int J Mol Sci 2019 20(24) 6141 10.3390/ijms20246141 31817512
46 Stepanenko AA Dmitrenko VV Pitfalls of the MTT assay: Direct and off-target effects of inhibitors can result in over/underestimation of cell viability Gene 2015 574(2) 193 203 10.1016/j.gene.2015.08.009 26260013
47 Nieborowska-Skorska M Sullivan K Dasgupta Y Podszywalow-Bartnicka P Hoser G Maifrede S Martinez E Di Marcantonio D Bolton-Gillespie E Cramer-Morales K Lee J Li M Slupianek A Gritsyuk D Cerny-Reiterer S Seferynska I Stoklosa T Bullinger L Zhao H Gorbunova V Piwocka K Valent P Civin CI Muschen M Dick JE Wang JC Bhatia S Bhatia R Eppert K Minden MD Sykes SM Skorski T Gene expression and mutation-guided synthetic lethality eradicates proliferating and quiescent leukemia cells J Clin Invest 2017 127(6) 2392 2406 10.1172/JCI90825 28481221
48 Padella A Ghelli Luserna Di Rorà A Marconi G Ghetti M Martinelli G Simonetti G Targeting PARP proteins in acute leukemia: DNA damage response inhibition and therapeutic strategies J Hematol Oncol 2022 15(1) 10 10.1186/s13045-022-01228-0 35065680
