Treatment Of Chronic Myeloid Leukemia Research Articles

Association between plasma imatinib levels and response to treatment of chronic myeloid leukemia in patients from Manaus, Brazil.

Imatinib mesylate (IM) is the drug of choice for the treatment of chronic myeloid leukemia (CML). However, despite most of the results obtained with this therapy being positive, some patients still present a suboptimal therapeutic response or still develop some type of resistance. Therefore, the aim of this study was to evaluate IM plasma levels in CML patients treated at a referral unit in Manaus and correlate them with variables that might interfere with these levels. Data from 52 patients were obtained through a standardized questionnaire containing clinical, sociodemographic, lifestyle, and use of other medication information, as well as an estimate of therapeutic adherence. Additionally, blood collection was performed to measure the plasma concentration of the drug using the HPLC-UV technique. Molecular studies were done to identify the presence of polymorphism in the ABCG2 C421A membrane transporter. Most patients were male with a mean age of 52 ± 12.3 years (95% CI 49.0-55.9). There was a high variation in drug concentrations in the range from 0 to 4694 ng/mL, with a mean of 1558.59 ± 989.79 ng/mL (95% CI 1283.0-1834.1). Approximately two-thirds of patients were classified in the drug-level range considered therapeutic, and there was a correlation between plasma concentration and higher molecular response. Additionally, most individuals had the normal genotype for the ABCG2 C421A polymorphism but further studies should be performed to reveal the role of this variable in the outcome of the disease in this population.

Myocardial Infarction in Chronic Myeloid Leukemia: Results from Nationwide Readmission Database.

Background and Objective Chronic myeloid leukemia (CML) is a hematological malignancy with an excellent prognostic outcome. After advancements in CML treatment and the introduction of different tyrosine kinase inhibitors (TKI), the life expectancy of CML patients has become equivalent to that of the general population. As a result, coronary artery disease is anticipated to be the leading cause of death among CML patients. Moreover, TKI use is associated with a risk of endothelial dysfunction, thrombosis, and cardiovascular events, including myocardial infarction. In this study, we compare the outcomes of percutaneous coronary intervention (PCI) in patients with CML to their matched non-CML counterparts. and compare the events reported with different TKI. Method The Nationwide Readmission Database (NRD) was searched from January 2016 to December 2020. for the study population. Adults of age ≥ 18 years with or without CML hospitalized for with a primary diagnosis of acute myocardial infarction, ST-segment elevation myocardial infarction (STEMI), and non-ST-segment elevation myocardial infarction (NSTEMI) and underwent PCI were included. The patients were identified using ICD-10 codes. The baseline characteristics and outcomes of the CML and non-CML patients who underwent PCI were compared. The primary outcomes were in-hospital mortality and 30-day readmission rates. The secondary outcomes were the PCI complications rates. Data regarding TKI reports of STEMI and acute coronary syndrome were obtained from The FDA Adverse Event Reporting System (FARES). Results Out of 2,727,619 patients with myocardial infarction, 2,124 CML patients were identified. A total of 888 CML patients underwent PCI. The mean age was 68.34 ± 11.14, with 62.46 % above 65 years. The analysis showed no significant difference between CML and non-CML patients after PCI for death (OR? 0.93 (95% CI 0.49-1.80), P=0.527) and 30-day readmission (OR? 1.41 (95% CI 0.99-2.01), P=0.056) rates. CML patients were significantly older (mean age 68.34 ± 11.14 versus 64.40 ± 12.61, p < 0.001) than non-CML patients without a difference in sex distribution. Hypertension (85.45% versus 78.64%), diabetes (45.48% versus 37.29), stroke (11.84% versus 7.78) at baseline were significantly higher in CML group. Prior myocardial infarction events (20.51% versus 15.17%) and prior PCI procedure (24.47% versus 16.89%) were significantly higher in CML group. CML had a significantly longer hospital stay (4.66 ± 4.40 versus 3.75 ± 4.62 days, p = 0.001). The primary outcomes did not differ between the comparison groups. Conclusion This analysis showed no statistically significant difference in mortality, 30-day readmission, and complication rates post PCI between CML and non-CML patients. However, interestingly, CML patients may experience lower coronary artery dissection and ischemic stroke rates than those without CML diagnosis.

Treatment of chronic-phase chronic myeloid leukemia in patients randomized to dasatinib or imatinib after suboptimal responses to three months of imatinib therapy: final 5-year results from DASCERN.

Early molecular response (EMR) at 3 months is predictive of improved overall survival (OS) and progression-free survival (PFS) in patients with chronic myeloid leukemia in the chronic phase (CML-CP). Although about one-third of patients treated with first-line imatinib do not achieve EMR, long-term OS and PFS are still observed in most patients. DASCERN (NCT01593254) is a prospective, phase IIb, randomized trial evaluating a switch to dasatinib in patients who have not achieved EMR after 3 months of treatment with first-line imatinib. Early analysis demonstrated an improved major molecular response (MMR) rate at 12 months with dasatinib versus imatinib (29% vs. 13%, P=0.005). Here, we report results from the final 5-year follow-up. In total, 174 patients were randomized to dasatinib and 86 to remain on imatinib. Forty-six (53%) patients who remained on imatinib but subsequently experienced failure were allowed to cross over to dasatinib per protocol. At a minimum follow-up of 60 months, the cumulative MMR rate was significantly higher in patients randomized to dasatinib versus imatinib (77% vs. 44%, P.

Global DNA Methylation analysis of imatinib resistant and sensitive K562 cells

OBJECTIVE: Chronic myeloid leukemia (CML) is a hematological disease which is known by the presence of Philadelphia chromosome (Ph+). BCR-ABL protein is expressed by Ph+ chromosome, represents constant increased tyrosine kinase activity. Imatinib (IMA) is a tyrosine kinase inhibitor (TKI) which is utilized as a first line treatment in CML. Emergence of IMA resistance at some point of therapy leads to treatment failure. DNA methylation is considered to be the most stable epigenetic change and several studies have shown that epigenetic changes may play a role in drug resistance. We investigated the global methylation profile of IMA-sensitive K562S, IMA-resistant K562R and IMA-resistant and adherent K562R (K562R-adh) cells to determine whether epigenetic reprogramming is involved in the resistance to IMA and the change in phenotype due to this resistance. MATERIAL AND METHODS: In this study, morphologically distinct, IMA-sensitive K562S and 5µM IMA-resistant K562R and K562R-adh in-vitro CML cell models were used to analyze the global DNA methylation profile. After DNA was isolated from the cells, global 5mC DNA methylation profiles were investigated by ELISA using equal amounts of DNA. RESULTS: Compared to K562S, the global methylation of K562R showed an increase in DNA methylation profile, but this increase in methylation was not statistically significant. Whereas, a slight hypermethylation was observed in the DNA of the K562R-adh vs K562S and K562R-adh vs K562R which is statistically significant. We observed slight hypermethylation in IMA-resistant cells lines versus to the IMA-sensitive cell line. CONCLUSION: Our observed differences in 5methyl-Cytosine on CpG islands (5mC) in K562S versus K562R and K562R-adh cell lines suggest that the DNA methylation alteration in resistant cells may partly contributed in phenotype switching.

The Depth of the Molecular Response in Patients with Chronic Myeloid Leukemia Correlates with Changes in Humoral Immunity.

Background and Objectives: The effective treatment of chronic myeloid leukemia leads to the restoration of proper immune system function. We aimed to investigate fluctuations in circulating cytokines, angiogenic factors and complement components in patients with CML during the first year of treatment with TKI and correlate them with the degree of achieved molecular response. Material and Methods: We recruited 31 patients with newly diagnosed CML. Peripheral blood and bone marrow samples were obtained, and concentrations of serum proteins were measured using an immunology multiplex assay. Results: The study cohort was divided into two groups of optimal or non-optimal in accordance with the European Leukemia Net (ELN) guidelines. We found significantly higher concentrations of C1q, C4 and C5a in serum after 3 months of TKI treatment in patients who achieved optimal responses in the 6 months after diagnosis. The most alterations were observed during 12 months of therapy. Patients in the optimal response group were characterized by higher serum concentrations of TGF-β, EGF, VEGF, Angiopoietin 1, IFN-γ and IL-8. Conclusions: The later plasma concentrations of complement components were significantly increased in patients with optimal responses. The changes after 12 months of treatment were particularly significant. Similar changes in bone marrow samples were observed.

Discovery of a Meisoindigo-Derived PROTAC as the ATM Degrader: Revolutionizing Colorectal Cancer Therapy via Synthetic Lethality with ATR Inhibitors.

Meisoindigo (Mei) has long been recognized in chronic myeloid leukemia (CML) treatment. To elucidate its molecular target and mechanisms, we embarked on designing and synthesizing a series of Mei-derived PROTACs. Through this endeavor, VHL-type PROTAC 9b was identified to be highly cytotoxic against SW620, SW480, and K562 cells. Employing DiaPASEF-based quantitative proteomic analysis, in combination with extensive validation assays, we unveiled that 9b potently and selectively degraded ATM across SW620 and SW480 cells in a ubiquitin-proteasome-dependent manner. 9b-induced selective ATM degradation prompted DNA damage response cascades, thereby leading to the cell cycle arrest and cell apoptosis. This pioneering discovery renders the advent of ATM degradation for anti-cancer therapy. Notably, 9b-induced ATM degradation synergistically enhanced the efficacy of ATR inhibitor AZD6738 both in vitro and in vivo. This work establishes the synthetic lethality-inducing properties of ATR inhibitors in the ATM-deficient context, thereby providing new avenues to innovative therapies for colorectal cancer.

Oxidative Stress and Chronic Myeloid Leukemia: A Balance between ROS-Mediated Pro- and Anti-Apoptotic Effects of Tyrosine Kinase Inhibitors.

The balanced reciprocal translocation t (9; 22) (q34; q11) and the BCR-ABL fusion gene, which produce p210 bcr-abl protein production with high tyrosine kinase activity, are characteristics of chronic myeloid leukemia, a myeloproliferative neoplasm. This aberrant protein affects several signaling pathways connected to both apoptosis and cell proliferation. It has been demonstrated that tyrosine kinase inhibitor treatment in chronic myeloid leukemia acts by inducing oxidative stress and, depending on its level, can activate signaling pathways responsible for either apoptosis or survival in leukemic cells. Additionally, oxidative stress and reactive oxygen species generation also mediate apoptosis through genomic activation. Furthermore, it was shown that oxidative stress has a role in both BCR-ABL-independent and BCR-ABL-dependent resistance pathways to tyrosine kinases, while patients with chronic myeloid leukemia were found to have a significantly reduced antioxidant level. The ideal environment for tyrosine kinase inhibitor therapy is produced by a favorable oxidative status. We discuss the latest studies that aim to manipulate the redox system to alter the apoptosis of cancerous cells.

Changes in chronic myeloid leukemia treatment modalities and outcomes after introduction of second-generation tyrosine kinase inhibitors as first-line therapy: a multi-institutional retrospective study by the CML Cooperative Study Group.

This study investigated changes in treatment modalities and outcomes of chronic myeloid leukemia in the chronic phase (CP-CML) after the approval of second-generation tyrosine kinase inhibitors (2G-TKIs) for first-line therapy. Patients were grouped into those who underwent TKI therapy up to December 2010 (imatinib era group, n=185) and after January 2011 (2G-TKI era group, n=425). All patients in the imatinib era group were initially treated with imatinib, whereas patients in the 2G-TKI era group were mostly treated with dasatinib (55%) or nilotinib (36%). However, outcomes including progression-free survival, overall survival, and CML-related death (CRD) did not differ significantly between groups. When stratified by risk scores, the prognostic performance of the ELTS score was superior to that of the Sokal score. Even though both scoring systems predicted CRD in the imatinib era, only the ELTS score predicted CRD in the 2G-TKI era. Notably, the outcome of patients classified as high-risk by ELTS score was more favorable in the 2G-TKI era group than in the imatinib era group. Thus, expanding treatment options may have improved patient outcomes in CP-CML, particularly in patients classified as high-risk by ELTS score.

Abstract LB415: Association between waist circumference, body mass index, high-density lipoprotein cholesterol level, and risk of chronic myeloid leukemia

Abstract Advances in the treatment of chronic myeloid leukemia (CML) have resulted in increased prevalence. However, a more comprehensive understanding of its pathogenesis and epidemiology is necessary. Recent studies suggested the association between altered lipid metabolism and carcinogenesis, prompting an investigation of the relationship between metabolic disorders and incidence of CML. Thus, we conducted a study to examine the association between waist circumference, body mass index (BMI), high-density lipoprotein cholesterol (HDL-C), and the incidence of CML.Using data from the Korean National Health Insurance Service from 2009 to 2017, a competing risks regression was conducted to examine the hazard ratios of CML in 3,879,560 individuals aged 20 years and above. During a mean of 10.13±1.24 years of follow-up, there were a total of 848 cases of CML, including 539 male and 309 female patients.1) Individuals in the group with a waist circumference equal to or greater than 100cm had the greatest risk of CML (adjusted hazard ratio [aHR], 95% CI = 1.593, 1.058-2.398), compared to those with a waist circumference from 80 to less than 85cm.2) Those with a BMI less than 18.5 showed the lowest incidence of CML ([aHR], 95% CI = 0.509, 0.278–0.931) in comparison to the reference population with a BMI between 18.5 and 22.9.3) Individuals in the highest quartile of HDL-C had the lowest risk of CML ([aHR], 95% CI = 0.795, 0.657–0.961) compared to those in the lowest HDL-C quartile.A large waist circumference was significantly associated with increased risk of CML, whereas a low BMI and high HDL-C were associated with a reduced risk of CML. This suggests that a large waist circumference may be an independent risk factor of CML, whereas a low BMI and high HDL-C may be association with a protective effect against CML. Table 1. Risk of chronic myeloid leukemia by subgroups of waist circumference, bodey mass index, and high-density lipoprotein cholesterol level. Waist circumference (cm) N Case Duration IR (1000,000 PY) HR (95% CI) Model 1 Model 2 Model 3 Model 4 Model 5 <80.0 1,424,747 236 14493864.7 16.2830 0.644 (0.533, 0.778) 0.858 (0.707, 1.041) 0.859 (0.707, 1.045) 0.836 (0.662, 1.055) 0.853 (0.675, 1.078) 80.0 - 84.9 921,040 193 9344290.4 20.6540 0.816 (0.669, 0.995) 0.876 (0.718, 1.069) 0.876 (0.717, 1.069) 0.866 (0.705, 1.063) 0.843 (0.685, 1.037) 85.0 - 89.9 769,528 197 7785119.0 25.3050 1 (Ref.) 1 (Ref.) 1 (Ref.) 1 (Ref.) 1 (Ref.) 90.0 - 94.9 451,608 116 4551043.2 25.4890 1.008 (0.801, 1.267) 0.963 (0.766, 1.212) 0.965 (0.767, 1.215) 0.976 (0.771, 1.234) 1.016 (0.802, 1.286) 95.0 - 99.9 201,541 68 2022854.6 33.6160 1.329 (1.009, 1.751) 1.267 (0.962, 1.670) 1.274 (0.966, 1.681) 1.302 (0.970, 1.747) 1.401 (1.044, 1.880) ≥100 111,096 38 1108876.8 34.2690 1.356 (0.958, 1.919) 1.352 (0.955, 1.915) 1.365 (0.962, 1.938) 1.421 (0.959, 2.105) 1.593 (1.058, 2.398) Model 1 was not adjusted.Model 2 was adjusted for age and sex.Model 3 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, and chronic kidney disease.Model 4 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, chronic kidney disease, and BMI (Continuous).Model 5 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, chronic kidney disease, and BMI (5 level). Body mass index N Case Duration IR (1000,000 PY) HR (95% CI) Model 1 Model 2 Model 3 Model 4 Model 5 <18.5 143,144 11 1421140.8 0.0077 0.410 (0.225, 0.749) 0.496 (0.272, 0.906) 0.505 (0.276, 0.921) 0.586 (0.319, 1.077) 0.509 (0.278, 0.931) 18.5 - 22.9 1,512,449 290 15308105.8 0.0189 1 (Ref.) 1 (Ref.) 1 (Ref.) 1 (Ref.) 1 (Ref.) 23.0 - 24.9 955,515 239 9697732.1 0.0246 1.300 (1.096, 1.543) 1.131 (0.952, 1.343) 1.120 (0.942, 1.331) 0.998 (0.828, 1.203) 1.060 (0.873, 1.287) 25.0 - 29.9 1,130,207 274 11476143.9 0.0239 1.260 (1.068, 1.486) 1.075 (0.910, 1.269) 1.060 (0.895, 1.256) 0.842 (0.675, 1.051) 0.878 (0.699, 1.104) ≥30.0 138,245 34 1402926.2 0.0242 1.280 (0.897, 1.826) 1.234 (0.865, 1.761) 1.218 (0.849, 1.747) 0.786 (0.500, 1.236) 0.765 (0.483, 1.211) Model 1 was not adjusted.Model 2 was adjusted for age and sex.Model 3 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, and chronic kidney disease.Model 4 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, chronic kidney disease, and waist circumference (Continuous).Model 5 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, chronic kidney disease, and waist circumference (5 level). HDL-C level N Case Duration IR (1000,000 PY) HR (95% CI) Model 1 Model 2 Model 3 Model 4 Q1 (Lowest) 1,007,163 277 10156736.52 0.0273 1 (Ref.) 1 (Ref.) 1 (Ref.) 1 (Ref.) Q2 910,627 202 9242549.22 0.0219 0.802 (0.669, 0.961) 0.851 (0.710, 1.021) 0.853 (0.711, 1.023) 0.851 (0.709, 1.021) Q3 979,148 177 9948872.05 0.0178 0.653 (0.540, 0.788) 0.720 (0.596, 0.870) 0.724 (0.598, 0.876) 0.721 (0.596, 0.873) Q4 (Highest) 982,622 192 9957890.96 0.0193 0.708 (0.589, 0.851) 0.789 (0.656, 0.949) 0.798 (0.660, 0.965) 0.795 (0.657, 0.961) Model 1 was not adjusted.Model 2 was adjusted for age and sex.Model 3 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, chronic kidney disease, and BMI (5 level).Model 4 was adjusted for age, sex, household income, smoking, alcohol consumption, regular exercise, diabetes mellitus, hypertension, dyslipidemia, chronic kidney disease, and dyslipidemia medication. HDL-C high-density lipoprotein cholesterl, IR incidence rate, PY person-years, HR hazard ratio, CI confidence interval. Citation Format: Ka Young Kim, Kyung Do Han, Sung-Eun Lee. Association between waist circumference, body mass index, high-density lipoprotein cholesterol level, and risk of chronic myeloid leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB415.

Endothelial function in patients with chronic myeloid leukemia treated with tyrosine kinase inhibitors is not related to cardiovascular risk assessed by SCORE2 algorithm.

Tyrosine kinase inhibitors (TKIs) revolutionized the treatment of chronic myeloid leukemia (CML) but are endowed with negative effects on endothelial function. To characterize endothelial function in patients with CML treated with various TKIs. Forty-eight patients diagnosed with chronic phase CML treated with TKIs, such as imatinib, bosutinib, nilotinib, ponatinib, and asciminib were included. Endothelial function was assessed in the brachial artery and microcirculation based on flow-mediated dilation (FMD), reactive hyperemia peripheral arterial tonometry (RH-PAT) and Flow Mediated Skin Fluorescence (FMSF). Reactive Hyperemia Index (RHI), FMD [%], Reactive Hyperemia Response (RHR [%]), Normoxia Oscillatory index (NOI [%]) and Hyperemic Response index (HR index [%]) did not differentiate between the group of patients with low / moderate risk in the Systemic Coronary Risk Estimation 2 (SCORE2), SCORE2-Older Persons (SCORE2-OP) and those with high / very high-risk scores. Among patients with low/intermediate risk based on the SCORE2 algorithms, some had lower (<Q1) values of endothelial parameters, reflecting impaired endothelial function, when compared with the whole population. Lower values of endothelial function parameters were associated with overall long-term treatment with TKIs or ponatinib treatment. Importantly, endothelial function assessed by FMSF (RHR [%]) negatively correlated with the total duration of TKI treatment, also after adjustment for age. Endothelial function in CML patients treated with TKIs was not related to cardiovascular risk based on SCORE2/SCORE2-OP algorithms but CML-specific factors, including duration of TKI treatment. FMSF-based assessment of skin microcirculation was a sensitive method for detecting the vascular effects of TKIs.

The Progress of Small Molecule Targeting BCR-ABL in the Treatment of Chronic Myeloid Leukemia.

Chronic myelogenous leukemia (CML) is a malignant myeloproliferative disease. According to the American Cancer Society's 2021 cancer data report, new cases of CML account for about 15% of all leukemias. CML is generally divided into three stages: chronic phase, accelerated phase, and blast phase. Nearly 90% of patients are diagnosed as a chronic phase. Allogeneic stem cell transplantation and chemotherapeutic drugs, such as interferon IFN-α were used as the earliest treatments for CML. However, they could generate obvious side effects, and scientists had to seek new treatments for CML. A new era of targeted therapy for CML began with the introduction of imatinib, the first-generation BCR-ABL kinase inhibitor. However, the ensuing drug resistance and mutant strains led by T315I limited the further use of imatinib. With the continuous advancement of research, tyrosine kinase inhibitors (TKI) and BCR-ABL protein degraders with novel structures and therapeutic mechanisms have been discovered. From biological macromolecules to classical target protein inhibitors, a growing number of compounds are being developed to treat chronic myelogenous leukemia. In this review, we focus on summarizing the current situation of a series of candidate small-molecule drugs in CML therapy, including TKIs and BCR-ABL protein degrader. The examples provided herein describe the pharmacology activity of small-molecule drugs. These drugs will provide new enlightenment for future treatment directions.

Integrated drug profiling and CRISPR screening identify BCR::ABL1-independent vulnerabilities in chronic myeloid leukemia.

BCR::ABL1-independent pathways contribute to primary resistance to tyrosine kinase inhibitor (TKI) treatment in chronic myeloid leukemia (CML) and play a role in leukemic stem cell persistence. Here, we perform exvivodrug screening of CML CD34+ leukemic stem/progenitor cells using 100 single drugs and TKI-drugcombinations and identify sensitivities to Wee1, MDM2, and BCL2 inhibitors. These agents effectively inhibit primitive CD34+CD38- CML cells and demonstrate potent synergies when combined with TKIs. Flow-cytometry-baseddrug screening identifies mepacrine to induce differentiation of CD34+CD38- cells. We employ genome-wide CRISPR-Cas9 screening for six drugs, and mediator complex, apoptosis, and erythroid-lineage-related genes are identified as key resistance hits for TKIs, whereas the Wee1 inhibitor AZD1775 and mepacrine exhibit distinct resistance profiles. KCTD5, a consistent TKI-resistance-conferring gene, is found to mediate TKI-induced BCR::ABL1 ubiquitination. In summary, we delineate potential mechanismsfor primary TKI resistance and non-BCR::ABL1-targeting drugs, offering insights for optimizing CML treatment.

Proteomic Analysis Reveals Trilaciclib-Induced Senescence

Trilaciclib, a CDK4/6 inhibitor, was approved as a myeloprotective agent for protecting bone marrow from chemotherapy-induced damage in extensive-stage small cell lung cancer (ES-SCLC). This is achieved through the induction of a temporary halt in the cell cycle of bone marrow cells. While it has been studied in various cancer types, its potential in haematological cancers remains unexplored. This research aimed to investigate the efficacy of trilaciclib in haematological cancers. Utilizing mass spectrometry-based proteomics, we examined the alterations induced by trilaciclib in the chronic myeloid leukaemia (CML) cell line, K562. Interestingly, trilaciclib promoted senescence in these cells rather than cell death, as observed in acute myeloid leukaemia (AML), acute lymphoblastic leukaemia (ALL), and myeloma cells. In K562 cells, trilaciclib hindered cell cycle progression and proliferation by stabilising CDK4/6 and downregulating cell cycle-related proteins, along with the concomitant activation of autophagy pathways. Additionally, trilaciclib-induced senescence was also observed in the non-small cell lung carcinoma cell line (NSCLC), A549. These findings highlight trilaciclib's potential as a therapeutic option for haematological cancers and underscore the need to carefully balance senescence induction and autophagy modulation in CML treatment, as well as in NSCLC.

Ponatinib and STAT5 Inhibitor Pimozide Combined Synergistic Treatment Applications Potentially Overcome Drug Resistance via Regulating the Cytokine Expressional Network in Chronic Myeloid Leukemia Cells.

Chronic myeloid leukemia (CML) is a clonal myeloproliferative hematological disease characterized by the chimeric breakpoint-cluster region/Abelson kinase1 (BCR::ABL1) oncoprotein; playing a pivotal role in CML molecular pathology, diagnosis, treatment, and possible resistance arising from the success and tolerance of tyrosine kinase inhibitor (TKI)-based therapy. The transcription factor STAT5 constitutive signaling, which is influenced by the cytokine signaling network, triggers BCR::ABL1-based CML pathogenesis and is also relevant to acquired TKI resistance. The unsuccessful therapeutic approaches targeting BCR::ABL1, in particular third-line therapy with ponatinib, still need to be further developed with alternative combination strategies to overcome drug resistance. As treatment with the STAT5 inhibitor pimozide in combination with ponatinib resulted in an efficient and synergistic therapeutic approach in TKI-resistant CML cells, this study focused on identifying the underlying amplification of ponatinib response mechanisms by determining different cytokine expression profiles in parental and ponatinib-resistant CML cells, in vitro. The results showed that expression of interleukin (IL) 1B, IL9, and IL12A-B was increased by 2-fold, while IL18 was downregulated by 2-fold in the ponatinib-resistant cells compared to sensitive ones. Importantly, ponatinib treatment upregulated the expression of 21 of the 23 interferon and IL genes in the ponatinib-resistant cells, while treatment with pimozide or a combination dose resulted in a reduction in the expression of 19 different cytokine genes, such as for example, inflammatory cytokines, IL1A-B and IL6 or cytokine genes associated with supporting tumor progression, leukemia stem cell growth or poor survival, such as IL3, IL8, IL9, IL10, IL12, or IL15. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis results showed that the genes were mainly enriched in the regulation of receptor signaling through the Janus kinase/signal transducer and activator of transcription pathway, cytokine-cytokine receptor interaction, and hematopoietic cell lineage. Protein-protein interaction analysis showed that IL2, IL6, IL15, IFNG, and others appeared in the top lists of pathways, indicating their high centrality and importance in the network. Therefore, pimozide could be a promising agent to support TKI therapies in ponatinib resistance. This research would help to clarify the role of cytokines in ponatinib resistance and advance the development of new therapeutics to utilize the STAT5 inhibitor pimozide in combination with TKIs.

Nilotinib-induced alterations in endothelial cell function recapitulate clinical vascular phenotypes independent of ABL1

Nilotinib is a highly effective treatment for chronic myeloid leukemia but has been consistently associated with the development of nilotinib-induced arterial disease (NAD) in a subset of patients. To date, which cell types mediate this effect and whether NAD results from on-target mechanisms is unknown. We utilized human induced pluripotent stem cells (hiPSCs) to generate endothelial cells and vascular smooth muscle cells for in vitro study of NAD. We found that nilotinib adversely affects endothelial proliferation and migration, in addition to increasing intracellular nitric oxide. Nilotinib did not alter endothelial barrier function or lipid uptake. No effect of nilotinib was observed in vascular smooth muscle cells, suggesting that NAD is primarily mediated through endothelial cells. To evaluate whether NAD results from enhanced inhibition of ABL1, we generated multiple ABL1 knockout lines. The effects of nilotinib remained unchanged in the absence of ABL1, suggesting that NAD results from off- rather than on-target signaling. The model established in the present study can be applied to future mechanistic and patient-specific pharmacogenomic studies.

FL118 Is a Potent Therapeutic Agent against Chronic Myeloid Leukemia Resistant to BCR-ABL Inhibitors through Targeting RNA Helicase DDX5.

Chronic myeloid leukemia (CML) is induced by the expression of the fused tyrosine kinase BCR-ABL, which is caused by a chromosomal translocation. BCR-ABL inhibitors have been used to treat CML; however, the acquisition of resistance by CML cells during treatment is a serious issue. We herein demonstrated that BCR-ABL induced the expression of the RNA helicase DDX5 in K562 cells derived from CML patients in a manner that was dependent on its kinase activity, which resulted in cell proliferation and survival. The knockout of DDX5 decreased the expression of BIRC5 (survivin) and activated caspase 3, leading to apoptosis in K562 cells. Similar results were obtained in cells treated with FL118, an inhibitor of DDX5 and a derivative compound of camptothecin (CPT). Furthermore, FL118 potently induced apoptosis not only in Ba/F3 cells expressing BCR-ABL, but also in those expressing the BCR-ABL T315I mutant, which is resistant to BCR-ABL inhibitors. Collectively, these results revealed that DDX5 is a critical therapeutic target in CML and that FL118 is an effective candidate compound for the treatment of BCR-ABL inhibitor-resistant CML.

Computer-aided drug discovery of c-Abl kinase inhibitors from plant compounds against chronic myeloid leukemia

Chronic myeloid leukemia (CML) is a hematological malignancy characterized by the neoplastic transformation of hematopoietic stem cells, driven by the Philadelphia (Ph) chromosome resulting from a translocation between chromosomes 9 and 22. This Ph chromosome harbors the breakpoint cluster region (BCR) and the Abelson (ABL) oncogene (BCR-ABL1) which have a constitutive tyrosine kinase activity. However, the tyrosine kinase activity of BCR-ABL1 have been identified as a key player in CML initiation and maintenance through c-Abl kinase. Despite advancements in tyrosine kinase inhibitors, challenges such as efficacy, safety concerns, and recurring drug resistance persist. This study aims to discover potential c-Abl kinase inhibitors from plant compounds with anti-leukemic properties, employing drug-likeness assessment, molecular docking, in silico pharmacokinetics (ADMET) screening, density function theory (DFT), and molecular dynamics simulations (MDS). Out of 58 screened compounds for drug-likeness, 44 were docked against c-Abl kinase. The top hit compound (isovitexin) and nilotinib (control drug) were subjected to rigorous analyses, including ADMET profiling, DFT evaluation, and MDS for 100 ns. Isovitexin demonstrated a notable binding affinity (-15.492 kcal/mol), closely comparable to nilotinib (-16.826 kcal/mol), showcasing a similar binding pose and superior structural stability and reactivity. While these findings suggest isovitexin as a potential c-Abl kinase inhibitor, further validation through urgent in vitro and in vivo experiments is imperative. This research holds promise for providing an alternative avenue to address existing CML treatment and management challenges. Communicated by Ramaswamy H. Sarma

Regulation of Cell Cycle Progression through RB Phosphorylation by Nilotinib and AT-9283 in Human Melanoma A375P Cells.

BCR-ABL tyrosine kinase inhibitors are commonly employed for the treatment of chronic myeloid leukemia, yet their impact on human malignant melanoma remains uncertain. In this study, we delved into the underlying mechanisms of specific BCR-ABL tyrosine kinase inhibitors (imatinib, nilotinib, ZM-306416, and AT-9283) in human melanoma A375P cells. We first evaluated the influence of these inhibitors on cell growth using cell proliferation and wound-healing assays. Subsequently, we scrutinized cell cycle regulation in drug-treated A375P cells using flow cytometry and Western blot assays. Notably, imatinib, nilotinib, ZM-306416, and AT-9283 significantly reduced cell proliferation and migration in A375P cells. In particular, nilotinib and AT-9283 impeded the G1/S transition of the cell cycle by down-regulating cell cycle-associated proteins, including cyclin E, cyclin A, and CDK2. Moreover, these inhibitors reduced RB phosphorylation, subsequently inhibiting E2F transcriptional activity. Consequently, the expression of the E2F target genes (CCNA2, CCNE1, POLA1, and TK-1) was markedly suppressed in nilotinib and AT9283-treated A375P cells. In summary, our findings suggest that BCR-ABL tyrosine kinase inhibitors may regulate the G1-to-S transition in human melanoma A375P cells by modulating the RB-E2F complex.

Utility of T315I mutation in Egyptian patients with chronic myeloid leukemia

ABSTRACT Chronic myeloid leukemia(CML) is a myeloproliferative neoplasm (MPN) characterized by the presence of the Philadelphia chromosome (BCR:ABL), and there is a higher risk in untreated CML to be transformed to blast crisis phase. The development of tyrosine kinase inhibitor (TKI) imatinib was a remarkable step in CML treatment. However, patients showed resistance due to some mutations including T315I which still be the most challenging mutation in CML. Therefore, the aim of our study is to screen T315I in our Egyptian CML patients to predict and determine the frequency of T315I in patients with TKIs resistance. AS-PCR performed on 50 CML patients resistant to the first-line TKIs and 50 CML patients respond to the first-line TKIs. Positive-T315I was found in 15 (30%) of TKIs resistant cases. T315I was significantly associated with higher WBC, platelet count, cellularity of BM aspiration and biopsy when compared to those with negative-T315I. Resistant positive-T315I was significantly associated with lower cumulative OS 55.9% and shorter mean OS 93.4 months when compared to resistant negative-T315I. We conclude that T315I can exist at the diagnosis before starting treatment. Therefore, screening test for T315I can be performed at the time of diagnosis to help in choosing the suitable treatment protocol.

Comorbidities and outcomes of patients with chronic myeloid leukemia treated with tyrosine kinase inhibitors: a real-world, nationwide, retrospective study from Hungary.

Purpose: This study aimed to provide real-world evidence on the characteristics, treatment patterns, and outcomes of patients with chronic myeloid leukemia (CML) receiving tyrosine kinase inhibitor (TKI) treatment in Hungary between 2011 and 2019. Patients and methods: This nationwide, retrospective study included patients who were newly diagnosed with CML in Hungarian clinical practice between January 2011 and December 2019. The analysis was based on the reimbursed prescription claims for imatinib, bosutinib, dasatinib, nilotinib, or ponatinib with the ICD-10 code C9210 in a public pharmacy between January 2009 and December 2019 using data from the National Health Insurance Fund (NHIF) of Hungary. CML incidence and prevalence, TKI treatment patterns, comorbidities, and overall survival (OS) were examined. Results: Between 2011 and 2019, altogether 1,407 patients were diagnosed with CML, with an annual average of 156 patients. The number of patients newly initiating first-line TKI therapy for CML significantly increased between 2011 and 2019 (2011: n = 136 vs. 2019: n = 191; p = 0.0043). Nilotinib was typically prescribed for younger patients (≤64years), while older patients (≥65years) mostly received imatinib. The most common comorbidity of CML patients was hypertension, and the proportion of patients with other malignancies was relatively high in all treatment groups. 5-year OS was 77.1% during the whole study period. Patients initiating first-line TKI treatment for CML in 2015 had significantly better 4-year OS compared to those starting treatment in 2011 (82.4% vs. 73.5%, respectively, (HR 0.53 (95%CI 0.32-0.87) p = 0.0118). Conclusion: This study is the first to provide insights into the characteristics, treatment patterns, and outcomes of CML patients treated with TKIs in Hungarian clinical practice between 2011 and 2019. We found slightly lower OS rates compared to other European countries, however, there was a statistically significant improvement in 4-year OS during the study period. The management of CML was in line with international guidelines and recommendations.