Chronic Myeloid Leukemia Group Research Articles

S1P Signaling Genes as Prominent Drivers of BCR-ABL1-Independent Imatinib Resistance and Six Herbal Compounds as Potential Drugs for Chronic Myeloid Leukemia.

Imatinib (IM), a breakthrough in chronic myeloid leukemia (CML) treatment, is accompanied by discontinuation challenges owing to drug intolerance. Although BCR-ABL1 mutation is a key cause of CML resistance, understanding mechanisms independent of BCR-ABL1 is also important. This study investigated the sphingosine-1-phosphate (S1P) signaling-associated genes (SphK1 and S1PRs) and their role in BCR-ABL1-independent resistant CML, an area currently lacking investigation. Through comprehensive transcriptomic analysis of IM-sensitive and IM-resistant CML groups, we identified the differentially expressed genes and found a notable upregulation of SphK1, S1PR2, and S1PR5 in IM-resistant CML. Functional annotation revealed their roles in critical cellular processes such as proliferation and GPCR activity. Their network analysis uncovered significant clusters, emphasizing the interconnectedness of the S1P signaling genes. Further, we identified interactors such as BIRC3, TRAF6, and SRC genes, with potential implications for IM resistance. Additionally, receiver operator characteristic curve analysis suggested these genes' potential as biomarkers for predicting IM resistance. Network pharmacology analysis identified six herbal compounds-ampelopsin, ellagic acid, colchicine, epigallocatechin-3-gallate, cucurbitacin B, and evodin-as potential drug candidates targeting the S1P signaling genes. In summary, this study contributes to efforts to better understand the molecular mechanisms underlying BCR-ABL1-independent CML resistance. Moreover, the S1P signaling genes are promising therapeutic targets and plausible new innovation avenues to combat IM resistance in cancer clinical care in the future.

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

Introduction: Chronic myeloid leukemia (CML) is a hematological malignancy with an excellent prognostic outcome. After the advancements in CML treatment and the introduction of different tyrosine kinase inhibitors (TKIs), 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. Method: This is a retrospective cohort study using the Nationwide Readmission Database from January 2016 to December 2020. Adults with or without CML hospitalized for acute myocardial infarction and underwent PCI were included. The patients were identified using ICD-10 codes. The primary outcomes were in-hospital mortality and 30-day readmission rates. The secondary outcomes were PCI complications rates. Results: Out of 2,727,619 patients with myocardial infarction, 2,124 CML patients were identified. A total of 888 CML patients underwent PCI. CML patients were significantly older (mean age: 68.34 ± 11.14 vs. 64.40 ± 12.61 years, p < 0.001) than non-CML patients without a difference in sex distribution. Hypertension (85.45% vs. 78.64%), diabetes (45.48% vs. 37.29), stroke (11.84% vs. 7.78) at baseline were significantly higher in the CML group. Prior myocardial infarction events (20.51% vs. 15.17%) and prior PCI procedure (24.47% vs. 16.89%) were significantly higher in the CML group. CML patients had a significantly longer hospital stay (4.66 ± 4.40 vs. 3.75 ± 4.62 days, p = 0.001). The primary outcomes did not differ between the comparison groups. The risk of post-PCI complications did not differ between the comparison groups in the propensity matched analysis except for coronary artery dissection (odds ratio [OR]: 0.10; 95% confidence interval [CI]: 0.02–0.65, p = 0.016) and ischemic stroke (OR: 0.35; 95% CI: 0.14–0.93, p = 0.034) which were lower in the CML group. Conclusion: This analysis showed no statistically significant difference in mortality, 30-day readmission, and post PCI complications rates between CML and non-CML patients. However, interestingly, CML patients may experience lower coronary artery dissection and ischemic stroke events than those without CML diagnosis.

Characteristics, Outcomes, Tfr Rates in Young Adults with Chronic Myeloid Leukemia in the Tyrosine Kinase Inhibitor Era: A French Observational Study

Introduction. Chronic myeloid leukemia (CML) is a myeloproliferative disorder with a median age of approximately 60 years. Our study emphasizes the fact that CML can present at younger ages. These patients have significantly different life challenges compared with older patients. The 2020 update of the ELN recommendations mentioned that“ TFR may be the main goal for any patient, irrespective of age, but it is clear that the younger the patient the stronger the case for achieving TFR”. We report here the main characteristics and TFR data in CML young adults. Methods. The French CML group (FI-LMC) initiated an observational study (CCTIRS no. 14.745) to describe the characteristics of CML in young adult patients in terms of characteristics at diagnosis, treatment choices, tolerance, adherence, therapeutic response, evolution and survival. Patients ranging from 18 to &amp;lt; 30 years with newly diagnosed CML in chronic phase (CP) or accelerated phase (AP) treated with a tyrosine kinase inhibitor (TKI) in first line were eligible for enrolment. Results. The observatory included 253 unselected young adults from 22 centers with newly diagnosed CML in CP (n=246) or in AP (n=7) among which 51.6% were treated within a first line trial. At the time of CML diagnosis, the median patient age was 24 years (range, 18-29 years); 165 patients were males (63%). Median (range) values were: WBC 154.3 G/L (6.5; 841); platelets 374 G/L (51; 1799), hemoglobin 11.3 g/dL (4.1; 18.3), blast cells 1 % (0; 20), spleen below costal margin 4 cm (0; 30). In CP patients, Sokal score was low in 132 (53.6%) patients, intermediate in 46 (18.7%), high in 37 (15.1%) and unknown in 31 (12.6%) and ELTS score was low in 142 (57.7%) patients, intermediate in 48 (19.5%), high in 22 (9%) and unknown in 34 (13.8%). Initial TKI was imatinib alone (52.4%) or in combination with IFN (2.4%) or Ara-C (0.8%), nilotinib alone (24.6%) or in combination with IFN (4%), dasatinib alone (8.2%) or in combination with IFN (0.4%), bosutinib (1.2%), ponatinib (4.8%) or asciminib (1.2%). Median follow-up is 106.2 months (range: 0.03; 279.2). Ninety-nine (39.3%) patients remained under first-line TKI at latest follow-up and 21 pts (8.3%) underwent bone marrow transplantation (BMT) for resistance or transformation. Median overall survival at 9 years is 96.9% [95% CI: 94.5- 99.4]. Event free survival without death, transformation, or BMT at 9 years is 93.1% [95% CI: 89.7- 96.7]. Six patients died, 1 from sarcoma, 5 from CML including 1 before any therapy (brain hemorrhage). We investigated the TFR strategy. In 169 patients treated for at least 5 years, 56 (33.1%) attempted TKI discontinuation. TKI free survival at 24 months is 57.9% [95% CI: 45.8- 73.2] (Figure 1). Complete analysis comparing characteristics and outcome between the 2 groups of patients (TFR vs no TFR), will be available for ASH presentation. Conclusion. Despite aggressive features including prominent splenomegaly, high WBC counts and high blast cell percentage, TFR seems achievable in young adults with CML.

On the Children Side. Overview of 194 Outcomes from Gimema Registry of Conception/Pregnancy in Chronic Myeloid Leukemia (CML)

Pregnancy in patients with chronic myeloid leukemia (CML) is no longer precluded. The literature published to date has mainly evaluated the management of CML and pregnancy with little mention of follow-up of children. This work aims to focus on the effects of the disease and of the drugs taken before conception and/or during pregnancy on fertility, gestation, pre-natal development of the fetus, and the growth of the child. GIMEMA has promoted an observational retrospective/prospective study to harvest and study detailed information on both pregnancies in female CML patients and pregnancies of partners of male patients followed at participating centers. The information requested addressed information on demographic, CML- related (CML therapy before/during/after pregnancy; transcript kinetics; outcome post delivery), pregnancy-related (type of conception; abortion rate; pregnancy progression, delivery, breastfeeding), and baby-related (birth, physical, intellectual, emotional, behavioral, social and sexual development) Data from 83 male patients (109 pregnancies) and 74 female patients (84 pregnancies) were evaluated for a total of 193 pregnancies at term. Fourteen (7 female and 7 male) conceptions were medically assisted, all the others spontaneous. One hundred ninety-four children were born (a twin pregnancy), 15 abortions were registered, 12 spontaneous and 3 voluntary all within the first trimester, while one pregnancy is still in progress. Eleven patients (1 male, 10 female) were diagnosed as having CML during pregnancy. Three patients interrupted pregnancy (2 electives). All male patients, with the exception of two (one undergoing assisted reproduction and one in TFR), conceived during treatment. TKI used were Imatinib (71%), Nilotinib (22%), Dasatinib (2%), Bosutinib (4%) and Ponatinib (1%). The pregnancies carried out by the partners of the male patients did not show any problems, neither pre-conception, nor during gestation, nor at birth. An initial speech delay in a child was reported, with recovery after speech therapy. Most pregnancies result in spontaneous delivery around the 39th week of gestation. Ninety-two/109 babies were breastfed (84%) The last TKI used in the female CML group was imatinib (43%), nilotinib (33%), dasatinib (22%), and Ponatinib (2%). Most patients stopped TKI at the FPT “first pregnancy test” (3-5 weeks). Cesarean deliveries amounted to 41%, just above the national average of 30%, with a mean of 38 weeks of gestation. Of the 84 babies born to women diagnosed with CML, only 20 were breastfed (23%). Three patients treated with IFN at the time of delivery breastfed for about a month, while one breastfed for 9 months and did not resume TKI as she was well-controlled by IFN and eager to start a new pregnancy. The lack of assimilation of IFN through breast milk orally does not contraindicate breastfeeding. In female patients, out of 84 pregnancies, and 1 in progress, 15 required antileukemic therapy during pregnancy. IFN was administered in 13 gestations, and also during organogenesis. In one case the patient introduced IFN at 7 weeks, then stopped due to poor disease control and started Nilotinib at 21 weeks, after placenta formation. In 2 gestations (including the one mentioned above) the patients took nilotinib (starting at 21 and 23 week of gestation). One patient was treated with imatinib at 20 week. The evaluation of the pediatric population under examination was divided into four age groups: 0-3 years for the evaluation of birth and psycho-physical development; 3-6 years for cognitive development; &amp;gt;6 years for socialization and interaction with the surrounding environment, including school performance; &amp;gt;12 years for sexual and hormonal development. Five infants were small for date. Of those, only one was exposed to imatinib from the 20th week of gestation. (Table 1). All children presented a rapid recovery after birth and their development is comparable to that of children belonging to the same age group. The oldest child is 15 years old, and was born at 39 weeks to a father on TFR after suspension of imatinib therapy; the youngest is a 15 days old girl, born at 38 week to a mother treated with IFN during pregnancy. From the results obtained from the study, children born to parents diagnosed with CML do not appear to show any difference compared to their peers in the general population, even if exposed to therapy in the prenatal period.

Role of Flow Cytometry in the Diagnosis of Chronic Myeloid Leukemia

To analyze the immunological phenotype of chronic myeloid leukemia (CML), and explore its characteristics and significance. The immunophenotypes of 40 CML children and 40 controls were analyzed by multicolor flow cytometry. CD45/SSC, as the basic gate, was used to delineate neutrophils. Then, the distribution of cluster differentiation (CD) molecules on the surface of granulocytes was analyzed in three ranges (≥1%, ≥5%, and ≥20%), and the expression rates of CD molecules (≥1% included in the statistical analysis) and the mean fluorescence intensity (MFI) were compared between the two groups. The proportion of granulocytes in the CML group was (82.1±6.4)%, which was significantly higher than (57.8±11.8)% in the control group (P <0.001). The expression rates of CD15/CD11b/CD33/CD13 in CML and control groups were high, and both distributed in the range of ≥20%. The differentiation trajectory of CD33/CD13 was normal and there were no significant differences in the expression rate and MFI between the two groups. However, both the expression rate of CD11b and CD15 MFI in the CML group were significantly lower than those in the control group (P <0.001). There were no significant differences in the expression rate and MFI of CD10 between the two groups, and the expression levels of CD10 between the two groups were consistent in different distributions. In the CML group, there was a large number of cases with abnormal high expression of CD56, 52.5% of the cases had a CD56 expression rate of ≥5%, and 42.5% had a CD56 expression rate of ≥20%, while the control group did not express CD56 (<1%). The expression distribution of CD117 was different between the two groups. In the range of expression rate ≥5%, there were 35.0% cases in the CML group, while only 2.5% in the control group. The expression rate of CD117 in the CML group was higher than that in the control group (P <0.001), though there was no significant difference in MFI. The immunophenotyping of CML is characterized by increased proportion of mature neutrophils, decreased CD15 MFI, decreased proportion of CD11b and abnormal high expression of CD56 and CD117. Flow cytometric analysis of immunophenotype can effectively distinguish normal granulocytes from chronic granulocytes, and help in the diagnosis of CML.

Treg Cells, FoxP3 and TGF-β Expression and Significance in Chronic Myeloid Leukemia

To investigate the expression and significance of regulatory T cells (Tregs), FoxP3 and transforming growth factor-β (TGF-β) in different phase of chronic myeloid leukemia (CML). Peripheral blood of 73 CML patients in Department of Hematology, Heze Municipal Hospital from March 2018 to March 2021 were collected. According to patient's period in CML, they were divided into ND CML group (newly diagnosed), CP CML group (chronic period), and BP CML group (blast phase). The percentage of Tregs, expression level of FoxP3 mRNA and TGF-β were detected by flow cytometry, RT-qPCR, and ELISA, respecitively. The roles of above indices in clinical pathogenesis of patients with CML were analyzed. The proportion of Treg in the ND CML group was slightly higher than the CP CML group, but the difference was not statistically significant (P =0.695), while the BP CML group was significantly higher than the other two groups (P =0.008, P <0.001). The expression levels of FoxP3 mRNA in ND CML group, CP CML group and BP CML group were 11.61±2.21, 6.46±1.35 and 8.54±2.13, respectively. Significant difference in FoxP3 mRNA levels was observed among patients in different phases of CML (F =55.199, P <0.001). The expression levels of FoxP3 mRNA both in ND CML group and BP CML group were significantly higher than that in CP CML group (P <0.001), and the ND CML group was the highest (P <0.001). However, the expression levels of TGF-β in different phases of CML showed no statistical differences (H =0.634, P =0.728). The abnormal distribution of Treg subset in different phases of CML and the significant increase of the expression level of FoxP3 mRNA in the new onset and blast phase of CML suggest that Tregs may promote the occurrence and progression of CML through immune regulation.

Chronic leukemias and COVID-19: An analysis of mortality and morbidity outcomes in the United States.

e18783 Background: COVID-19 pandemic has resulted in a global health crisis with widespread mortality and morbidity. Patients with chronic leukemias (CL) are at an increased risk for severe outcomes due to their compromised immune system. Despite the potential impact of COVID-19 on this population, there is a scarcity of information on the outcomes for CL patients. Our study aims to evaluate the mortality and morbidity of COVID-19 in CL patients in the United States. Methods: In this retrospective cohort study, we employed the National Inpatient Sample database of 2020, the largest inpatient database in the United States, to include adult patients (age ≥ 18 years) with chronic leukemias (CL) who were hospitalized for COVID-19, as identified by ICD-10-CM codes. The primary outcome was all-cause in-hospital mortality, while the secondary outcomes were acute respiratory failure (ARF) and the requirement for mechanical ventilation (MV). To identify the predictors of mortality, we conducted multivariable logistic regression analyses, with a p-value of less than 0.05 being considered statistically significant. Additionally, we conducted sensitivity analyses based on the type of CL (chronic lymphocytic leukemia (CLL) or chronic myeloid leukemia (CML)) and the remission status (in remission, not achieved remission, and relapse). The statistical analysis was performed using STATA software. Results: Among 91,375 patients with CL, 5.2% were hospitalized for COVID-19, with 87% of these patients having CLL. The majority of the CL patients (88.6%) had not achieved remission. The mean age of the patients was 72.7 years, with 62.4% being male and 78.9% being white. The mortality rate among CL-COVID-19 patients was 17%, with no significant difference in mortality rates between patients with and without CL (odds ratio [OR] 0.96, 95% confidence interval [CI] 0.81-1.14, p-value 0.65). About 61.5% of CL-COVID-19 patients had ARF and only 10.5% required MV. Patients with CL showed a 16% increase in the odds of ARF (OR 1.16, 95% CI 1.01-1.32, p-value 0.03) while no significant difference was found in the odds of MV. Sensitivity analysis showed no significant differences in mortality, ARF, or MV rates between CLL and CML groups. Another analysis by remission status revealed that patients who had not achieved remission had higher odds of ARF (OR 1.64, 95% CI 1.09-2.46, p-value 0.018) compared to those in remission or relapse. Conversely, patients in remission had a significantly lower likelihood of ARF (OR 0.63, 95% CI 0.41-0.97, p-value 0.036). No significant differences in mortality and MV odds were found according to remission status. Conclusions: Our findings emphasize the importance of vigilant monitoring and management of CL patients during the COVID-19 pandemic, particularly for those who have not achieved remission, to reduce their risk of severe respiratory outcomes.

Biochemical and breakpoint cluster region-c-ABL oncogene 1 polymorphism study among Iraqi patients with chronic myeloid leukemia

BACKGROUND: Chronic myeloid leukemia (CML) has been well recognized as an exemplary instance of a malignant disease characterized by a distinctive molecular occurrence, namely the presence of the breakpoint cluster region (BCR)-c-ABL oncogene 1 (ABL1) oncogene. The Philadelphia chromosome gives rise to an anomalous fusion gene characterized by atypical kinase activity, resulting in the accumulation of reactive oxygen species and genetic instability that holds significance in the advancement of diseases. OBJECTIVE: The objective of this study was to investigate the detection rate of BCR-ABL1 polymorphism and BCR protein level in a group of Iraqi patients with CML. MATERIALS AND METHODS: This study has been carried out on 150 specimens, 120 patients subjected to CML included 20 patients diagnosed as newly diagnosis CML and 100 patients treated with CML. In addition to 30 apparently healthy persons as a control group (normal persons) from the National Center of Hematology/Mustansiryiah University/Baghdad, 65 out of 100 patients on imatinib while 35 nonimatinib (nilotinib and bosutinib). Fresh whole blood and serum were obtained from all patients and controls. We used total DNA genomic extraction extracted from ethylenediaminetetraacetic acid blood for genetic detection of Bcr/Abl Genes Polymorphism by sequencing technique in patients with CML and apparently control groups and used serum for biochemical tests include urea, lactate dehydrogenase (LDH), aspartate transaminase (AST), alanine transaminase (ALT), and creatinine using biochemicals methods (colorimetric and kinetic), respectively, as well as detection BCR protein level using sandwich enzyme-linked immunosorbent assays technique. RESULTS: According to age and sex, the patients’ groups were matching with the control group. Regarding the biochemical parameters (urea creatinine, ALT, AST, and LDH) serum level, there are no significant differences among new diagnosis CML, patients respond to treatments and failure group except in serum level of creatinine between new diagnosis CML group and failure group, there are significant differences (P = 0.01). The present results showed that DNA polymorphism distribution was according to C\C; G\C; A\T; and A\A were 32%, 26%, 18%, and 24%, respectively, in patients with CML and 28%; 20%;12%; and 40%, respectively, in the control group. There are significant statistical differences (P &lt; 0.05) between different groups according to the genotyping of BCR\ABL, the results obtained from the sequenced 429 bp fragments, and the detailed positions of the observed variations are described in the NCBI reference sequences (rs766724113). The samples were submitted in NCBI, and the accession number of nucleotide sequences of BCR\ABL as new recording: LC 775148, LC 775149, and LC 775150, while regarding with BCR protein, there are significant differences in level between new diagnosis CML and CML on treatment and control groups, P &lt; 0.001 for each comparison while there are no significant differences between treated group and control group (P = 0.729). CONCLUSION: The present results indicate that BCR-ABL1 polymorphism and BCR protein level in a group of Iraqi patients with CML may play a role in the tumor biology of the examined subset of CML and may contributed to their development.

Demographic Characteristics and Outcomes in Patients with ALL and Previous Diagnosis of CML or MDS

Introduction: Acute Lymphocytic Leukemia (ALL) is a malignancy of lymphocytes characterized by uncontrolled proliferation of immature cells which ultimately leads to the replacement of bone marrow and other lymphoid organs. Typically, ALL develops in a previously healthy person however, it can rarely progress from an existing Myelodysplastic syndrome (MDS) or Chronic Myeloid Leukaemia (CML). In this study, we highlight the baseline characteristics and outcomes in patients with ALL and previous diagnoses of MDS or CML. Methodology: Healthcare Cost and Utilization Project National Inpatient Sample (HCUP-NIS) was queried to identify all ALL admissions between 2016-2019 in adult patients with a secondary diagnosis of MDS or CML using International Classification of Disease 10th Revision (ICD-10) codes. We evaluated baseline characteristics including Age, Sex, Race, Insurance type, Median household income, Region of hospital, Hospital Bed size, and Location/teaching status of hospital, in addition to mortality as the primary outcome and length of the hospital stay (LOS) and total hospital charges (THC) as secondary outcomes. Results: A total of 27385 patients with ALL were hospitalized and among them, 195 (0.7%) had a secondary diagnosis of CML. The mean age of the CML group was non-significantly higher (51.3 vs. 48.5) compared to those without CML. In patients with ALL and previous CML, there was a higher proportion of female sex, black race, Medicaid insurance, hospitalizations in south and west region, small sized hospitals in comparison to those with ALL without previous CML. though not statistically significant as reflected in Table (1). There was no difference in, Charlson comorbidity index, and household income. CML group had non-significantly lower odds of mortality in comparison to patients without CML (5.1% vs 5.8 % P=0.84) with adjusted odds ratio (AOR) of 0.79 (p=0.75 CI 0.18-3.34). Patients with a higher Charlson comorbidity index had significantly higher odds of mortality (1.68 with P=0.022). Among secondary outcomes, the CML group had a higher mean length of stay (20.3 vs 18) and lower THC ($240161 vs $279386) although this did not reach statistical significance. Out of all patients admitted with ALL, 180 (0.6%) had a secondary diagnosis of MDS, and the mean age of the MDS group was higher (62.6 vs. 48.4) compared to the other group. In ALL patients with previous MDS group, there was a significantly higher proportion of people with Medicare insurance, and hospitalization in the south and west region. There was a trend towards an increased proportion of Female sex, white Race, lower median household income, small sized hospitals in comparison to ALL without previous MDS group though it was not statistically significant (Table1). Patients with ALL and a previous Diagnosis of MDS had non-significantly higher odds of mortality (11% vs 6 %) in comparison to patients without MDS (AOR 1.7 p=0.32 CI 0.60-6.65). Among the secondary outcomes, the MDS group had a non-significantly lower mean LOS (12 vs 18) and lower THC ($243389 vs $ 279112). Conclusion: In this analysis, we found that sex, race, insurance type, median household income, and region of the hospitalization impacts the risk of developing ALL after MDS or CML. Prospective studies are required to look into the cause of these differences as it can be related to the socioeconomic disparities or specific gene mutations more prevalent in certain races of regions. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Antibody seroprevalence against SARS-Cov-2 among chronic myeloid leukemia patients.

e19061 Background: The rapid spread of SARS-CoV-2 has elicited an equally rapid development of effective vaccines, leading to a reduction of COVID-19 severity and deaths. There is limited data on COVID-19-related immunity in chronic myeloid leukemia (CML) patients. Methods: SPARTA (SARS2 SeroPrevalence And Respiratory Tract Assessment) is an ongoing observational study for participants age ≥18 years to investigate immunity to SARS-CoV-2 after infection and/or vaccination. We included patients with CML and compared them with a non-cancer group. We collected saliva and peripheral blood to measure antigen levels by RT-PCR and antibodies (secretory IgG antibodies and neutralizing antibodies). Results: From October 1, 2021, to February 4, 2022, we prospectively enrolled 49 participants (23 CML, 26 non-cancer). Most were male (56.5%) in the CML group and female in the control group (61.5%), mean age 56.39 y vs. 51.96 y, respectively, and self-identified as white (87% vs. 76.9%). In the CML group, 11 (47.8%) had ≥1 comorbidities, vs 13 (50%) in the control group. Twenty-one (91.3%) CML patients were receiving tyrosine-kinase inhibitors; 4 (18.2%) non-cancer subjects reported taking any medication. Most participants in both groups had received at least one dose of COVID-19 vaccine (73.9% vs. 73.1%); 100% of CML patients received two doses vs. 84.2% of controls; the CML group had a higher percentage of subjects fully vaccinated (66.7% vs. 25%). The CML group had a lower percentage of patients previously diagnosed with COVID-19 (8.8% vs. 57.7%). However, there was no difference in the detection of SARS-CoV-2 antigen at the time of enrollment (0% vs. 4%). SARS-CoV-2 IgG antibodies were detected in most of the participants regardless of cancer status (78.3% in the CML cohort and 88% in the non-cancer cohort), and neutralizing antibodies were detected in 82.6% and 95.6%, respectively. The two groups had comparable IgG (mean 146.3 Ru/ml vs. 148.9 Ru/ml) and neutralizing (mean 1329.1 ng/ml vs. 1112 ng/ml) antibody levels. Conclusions: Our preliminary data comparing concomitant cohorts with similar socio-demographic characteristics and medical history indicate that a diagnosis of CML did not impact the development of antibodies against SARS-CoV-2. We are conducting continuous analysis of antibodies levels over time to assess the evolution of antibody immunity and functional studies including cellular immunity assessments.[Table: see text]

Immune and inflammatory cytokines profile in Iraqi patients with acute and chronic myeloid leukemia

Introduction and Aim: Evaluation of the profile of immunological and inflammatory cytokines, IL-6, IL-2 and TNF? in acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) patients compared their level with healthy controls, and assessment of the association of these cytokines with clinical and hematological variables. Patients and Methods: Thirty AML, thirty CML patients, and thirty healthy control individuals were recruited for this study from the National Hematology Center for Research and Treatment in Baghdad for the period of October 2020 to March 2021. Leukemic patients were under chemotherapy. Complete blood picture routine test and cytokines profile test were done. IL-6, IL-2 and TNF-? were quantitatively measured using enzymes linked immunosorbent assay (ELISA) kits. Results: Hematological myeloid cancers affect more males than female, in AML male/female ratio was 17/13 while in CML was 16/14. The range of duration of the disease for both AML and CML was 1-3 years. in both AML group the level of IL-6 59.66 ± 31.99 pg/mL and in CML group 57.07± 16.52 pg/mL were both significantly higher P value &lt; 0.001 than that of control group 37.95± 6.10 pg/mL. TNF? in AML group was significantly higher P value &lt; 0.001 than that of control group. A high significant difference in serum level of IL-2 P value &lt;0.0001 in AML and CML compared to controls. Conclusion: IL-6, TNF? and IL-2 levels were higher in myeloid malignancy groups than controls. Serum cytokines shown to be positively associated with each other and could signify to poor outcome of the myeloid malignancy patients.

Comparative Assessment of Echocardiographic Patterns Among Chronic Myeloid Leukemia Patients on Tyrosine Kinase Inhibitor and Healthy Controls.

PurposeChronic myeloid leukemia (CML) is one of the common hematological malignancies in Nigeria. Cardiac abnormalities are associated with CML irrespective of treatment with tyrosine kinase inhibitors such as imatinib, which is available gratis in Nigeria.ObjectiveTo assess the prevalence and patterns of cardiac dysfunction among patients with CML irrespective of treatment with imatinib using transthoracic echocardiography, and 12-lead surface electrocardiography.Patients and MethodsCML patients without Imatinib, CML patients with imatinib, and apparently healthy (age- and sex-matched) controls were 70 each in the study. Various echocardiographic parameters were measured and data obtained were analyzed, and the level of significance was taken as p < 0.05.ResultsOf 70 CML patients with imatinib, 54.3% were men and 45.7% were women, while the CML group without imatinib had 62.9% men and 37.1% women, non-CML control had 54.3% men and 45.7% women. The average hematocrit was significantly lower in the CML group without Imatinib compared with the other groups (p<0.001). And, 12.9% and 17.1% of CML groups with and without imatinib had LVH, respectively, and none of the non-CML controls had LVH (P<0.041). Impaired left ventricular relaxation in 25.71% and 28.57% of CML patients with and without imatinib respectively but only 8.57% of the non-CML control had impaired left ventricular relaxation (p=0.236). Mitral valve regurgitation was the most frequent valvular abnormality across the groups. Pulmonary hypertension in 17.4% and 20% of CML patients with and without imatinib, respectively, but none of the non-CML controls had pulmonary hypertension (p<0.001). Pericardial effusion in 32.86% and 45.71% of CML patients with and without imatinib, respectively, but none of the non-CML controls had pericardial effusion (p<0.001). There was no significant difference in the QTC interval across the three groups.ConclusionCardiac abnormalities are present in CML patients with or without Imatinib treatment, with significant prevalence than what is seen in the non-CML control group.

The Outcomes of Ponatinib Therapy in Patients With Chronic Myeloid Leukemia Resistant or Intolerant to Previous Tyrosine Kinase Inhibitors, Treated in Poland Within the Donation Program

IntroductionTyrosine kinase inhibitors (TKIs) have greatly improved the treatment outcome for most patients with chronic myeloid leukemia (CML). Ponatinib is a new pan-inhibitor of TK active in resistant CML. This study aimed to evaluate the efficacy and safety of ponatinib in patients suffering from CML. Patients and methodsThis multicenter, non-randomized, observational, retrospective study evaluated the efficacy and safety of ponatinib administered in adult CML patients in any disease phase, including those with a detected ABL T315I mutation, which were resistant or intolerant to previous-generation TKIs. The study comprised 43 patients benefiting from the ponatinib donation program who were treated in 16 Polish centers. ResultsFor patients who started treatment with ponatinib in chronic phase (CP) (n = 23) and in accelerated phase (AP) (n = 3) the median time on ponatinib was 19.5 months (range: 1.0-35.4), and 31.7 months (range: 31.0-34.1), respectively. All these patients were in CP after 1 month of treatment and at the end of observation – none of them progressed to AP or blastic phase (BP) during the study, meaning that progression-free survival was 100% at the end of observation (35.4 months). The estimated 2-year survival in this group of patients was 84%. For all 43 patients, median survival was not reached (lower quartile 6.3 months), and estimated 2-year survival was 60%. ConclusionOur analysis confirmed ponatinib efficacy in a significant proportion of patients heavily pre-treated with TKIs achieving durable responses in both CP and AP/BP CML groups.

Treatment Free Survival (TFS) in Patients (pts) with Chronic Myeloid Leukemia (CML) Carrying Atypical BCR-ABL1 Fusion Transcripts: The French CML Group (Fi-LMC) Experience

▪AimsLife expectancy of CML pts optimally responding to tyrosine kinase inhibitors (TKI) is close to that of the general population and recently, TFR has been acknowledged as a new goal of CML management. TKI discontinuation in the view of TFR requires the achievement of deep and long-lasting molecular responses (MR). The gold standard BCR-ABL mRNA quantification technology and MR definitions rely on internationally standardized (IS) RT-qPCR but atypical transcripts located outside the Major-BCR region, harbored by 1-2% of pts, cannot be expressed on the IS scale. Thus, most trials and clinical practice recommendations prevent such pts from attempting TFR. The Fi-LMC group retrospectively collected real-life observations to assess TFR likelihood in this rare population.MethodsData from CML pts with precise characterization of atypical transcripts in whom any line TKI was stopped for any reason but after at least 2 years of undetectable molecular residual disease (UMRD) by individualized non-standardized RT-qPCR were collected. RT-qPCR sensitivity varied depending on transcript type and local molecular biology laboratory. TFS was estimated by the Kaplan-Meier method. Relapse was analyzed using the cumulative incidence function, relapse being as UMRD loss at any time and any level during follow-up (FU).ResultsOur series comprised 16 adult CP CML pts with atypical BCR-ABL fusions including 12 males (75%). Median age at CML diagnosis was 56 years (range: 21-75) and that at TKI discontinuation was 67 years [range: 29-82]. Sokal score was low, intermediate and high in 7, 8 and 1 pts, respectively. ELTS score was low and intermediate in 10 and 4 pts, respectively and unknown in 2. Most pts expressed e19a2 (n=6) followed by e6a2 (n=4), b3a3 (n=3), b2a3 (n=2) and e8a2 (n=1). Seven pts discontinued imatinib, 4 stopped dasatinib, 4 nilotinib and 1 bosutinib. Number of lines of therapy was 2 in 8 pts, 1 in 5 pts and 3 in 3 pts. Median TKI treatment duration before discontinuation was 64 months (range: 31-218) and median duration of UMRD was 41 months (range: 21-168). The median FU after TKI discontinuation was 68 months (range: 3-149). Five pts experienced relapse leading to TKI resumption. Four relapses occurred within 3-6 months and included 2 loss of hematologic response in CP, 1 loss of hematologic response in accelerated phase CML and 1 molecular recurrence with BCR-ABL transcripts up to 1.5%. One relapse occurred at 49 months and consisted in loss of a complete cytogenetic response. These 5 pts resumed TKI and regained UMRD within 6 months, including 1 pt who died in UMRD from non-CML-related cause at the age of 82 years and 1 pt who rapidly failed a 2 nd TKI discontinuation attempt.In 1 additional pt, BCR-ABL transcripts became detectable intermittently with maximum transcript level of 0.15% and TKI was not resumed. The median FU of pts who remained treatment-free was 68 months (range: 8-149).Overall, the 5-year cumulative incidence of relapse regardless of whether TKI was resumed was 41.6% (95% confidence interval: 21.9%-78.7%) (Figure 1). The 5-year TFS rate was 65.2% (95% confidence interval: 40.3%-90.2%) (Figure 2).ConclusionsOur observational study of TKI discontinuation in CML pts with atypical BCR-ABL transcripts is the largest reported so far. While effort must be made for proper assessment of deep MR, preliminary results suggest that TFS pattern might favorably compare with that obtained in pts with Major-type BCR-ABL transcripts. However, relapses may be more aggressive and caution is required in order to avoid loss of hematologic responses and progression. Whether the type of atypical fusion gene influences TKI discontinuation outcome, as well as other potential prognostic factors, need to be determined in a larger series. [Display omitted] DisclosuresCharbonnier: Novartis: Speakers Bureau; Incyte: Speakers Bureau. Rea: Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Incyte: Honoraria, Membership on an entity's Board of Directors or advisory committees; Pfizer: Honoraria, Membership on an entity's Board of Directors or advisory committees. Etienne: Novartis: Consultancy, Speakers Bureau; Incyte: Consultancy, Speakers Bureau. Rousselot: Incyte, Pfizer: Consultancy, Research Funding. Nicolini: Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel, accommodations, expenses, Research Funding; Kartos Therapeutics: Consultancy, Membership on an entity's Board of Directors or advisory committees; Sun Pharma Ltd.: Consultancy, Membership on an entity's Board of Directors or advisory committees; Incyte Biosciences: Honoraria, Other: travel, accommodations, expenses, Research Funding, Speakers Bureau; BMS: Honoraria.

Granulocytic and Monocytic Myeloid-Derived Suppressor Cells are Functionally and Prognostically Different in Patients with Chronic Myeloid Leukemia.

Myeloid-derived suppressor cells (MDSCs) represent phenotypically heterogeneous populations that suppress tumor-specific T-cell responses. MDSCs are produced from myeloid precursors in emergent states and are increased in several hematologic malignancies. We evaluated the differences in the levels and prognostic significance of MDSCs according to the clinical status of chronic myeloid leukemia (CML). The percentages and numbers of granulocytic (g)MDSCs and monocytic (m)MDSCs in peripheral blood (PB) and bone marrow (BM) aspirates were determined by five-color flow cytometry (HLA-DR/CD11b/CD15/CD33/CD14). The median BM-gMDSC% and PB-gMDSC% of the CML group were lower than those of the complete hematologic response (CHR) and control groups (P<0.001). In the CHR group, patients with major molecular response (MMR) showed higher median BM-gMDSC% than those without MMR (P=0.039). Conversely, the PB-mMDSC number of the CML group was higher than those of the CHR and control groups (P<0.001). Patients with high PB-gMDSC number exhibited superior survival to those with low PB-gMDSC number (P=0.021), and patients with high PB-mMDSC% showed inferior survival to those with low PB-mMDSC%, but there was no statistical significance (P=0.182). Increased gMDSCs at CHR may reflect non-leukemic granulopoiesis, and a high number of PB-gMDSCs suggests better prognosis in CML. However, mMDSCs may be associated with malignant conditions and poor prognosis.

Assessment of Changes of Membrane Properties and Platelets Function in Patients with Chronic Myeloid Leukaemia

▪Background. Thrombosis is frequent complication in patients with chronic myeloid leukemia (CML). Qualitative and quantitative modifications of platelet membrane receptors are involved in alteration of platelet function. Moreover the properties of cellular membrane may influence the expression and activity of receptors. The aim of our study was to determine whether changes of platelet membrane properties (lipid bilayer fluidity and resting transmembrane potential) may be correlated with the modified expression of platelet receptors and altered platelet function in CML.Materials and methods. This retrospective study included 30 patients with CML as well as 10 healthy volunteers. Platelet function was analyzed by optical platelet aggregometry using as agonists ADP, collagen, ristocetin and epinephrine. Lipid bilayerfluidity (FA) of platelet membrane was evaluated by fluorescence anisotropy technique using as marker 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene p-toluenesulfonate (TMA DPH). The resting transmembrane potential was measured by fluorescence spectroscopy using 3,3‘-dipropyl-2,2‘-thiadicarbocyanine iodide (DiSC3-5). Depending on platelet response to different reagents used for platelet aggregation, we divided the CML patients into two groups: patients with normal response (amplitude > 60%) and patients with absent/low response. Platelet receptor expression was analyzed by flow cytometry for adhesion markers (CD 42a and CD 42b) and aggregation markers (CD61, CD41).Results and discussion. It has been observed a statistically significant difference in FA for advanced-phase of CML patients when compared to controls (P=0.02). Patients with molecular remission presented a significantly lower FA (more fluid lipid bilayer) than patients without molecular remission (median value 0.1445 (95% CI 0.10-0.16) compared to median value 0.1694 (95% CI 0.13-0.29), P=0.05). The inhibitors of tyrosine kinase (TKI) turned into less fluid the lipid bilayer: the value of FA significantly increased between untreated status FA=0.064 vs. after 2 months of treatment FA=0.119. This analysis was performed only for a small group (4 patients) and has to be confirmed on a larger population of patients under treatment. The amplitude and slope of the platelet aggregation curves are significantly lower for CML patients than for healthy volunteers, for all reagents except ristocetin; the platelet response was more influenced in CML patients-accelerated and blastic phase. The CD61/CD41 expression (GP IIbIIIa) presented statistically significant lower values in CML patients (median value CD 61=76.63 (59.18-84.6); CD 41=55.3 (43.7-71.42)) versus controls (median value: CD61= 96.74 (71.36-98.16); CD 41=89.03 (55.67-92.92)).There was not diference in expression of CD42a/CD42b in the CML group compared with control groupPatients in advanced or blastic phase has a lower value of GP IIbIIIA expression (median value: CD 61=70.02 (41.36-83.35); CD 41= 45.45 (29.21-67.78)) than patients in chronic phase (median value: CD 61=84.63 (66.98-94.92); CD 41= 72.82 (59.78-86.85)). There was no statistical significant correlation between the lipid bilayer fluidity and platelet transmembrane resting potential. The average value of membrane potential of platelets from CML patients was -62±15 mV in accordance with values found in literature.Conclusions. CML patients have a low expreser expression of platelet aggregation receptors and consequent lower parameters of amplitude and slope on aggregation curves, but an increased lipid bilayer fluidity. A normal response to ristocetin was observed, although the platelet receptors expressions were low. TKI decreased the lipid bilayer fluidity and confirmation of this result on larger population is undergoing. The influence of platelet transmembrane resting potential on platelet aggregation response needs to be analysed on a higher number of patients. DisclosuresNo relevant conflicts of interest to declare.

Expression and Functional Assessment of Some Featured Coding and Non-coding RNAs Encoded by 8q24 Chromosomal Region in CML Patients

Background: The association between the human chromosomal 8q24 region and cancer development remains dim. The proto-oncogene MYC is known as the most prominent target of this chromosomal region. However, numerous cancer-associated genetic alterations in the region extend beyond the MYC locus. Accordingly, it is likely that the MYC oncogene is not the only target of these carcinogenesis-related alterations. Objectives: In the present study, the expression of MYC and the correlation between MYC and two non-coding RNAs, namely PVT1 (circular and linear forms) and CASC11, which are residents of the 8q24 region in the MYC neighborhood, were investigated in chronic myeloid leukemia (CML). Methods: Real-time polymerase chain reaction (PCR) was used to assess BCR-ABL transcripts and categorize positive and negative (normal) samples for CML. Afterward, real-time PCR was exploited to evaluate the expression of different genes, including MYC, linear PVT1, circular PVT (CircPVT1), CASC11, and ACTB in CML and normal samples. Results: We found that the expression of linear PVT1 is significantly increased in CML compared with normal samples. However, CircPVT1, CASC11, and MYC did not show significantly altered expression between CML and normal groups. The experimental and in silico analyses of the correlation coefficients of gene expressions suggested changes in the correlations between the gene expressions in CML compared with normal samples. We also assessed the miR-trapping potential of PVT1 and CASC11 and the possible effects of these interactions on signaling pathways. Our findings indicated that these lncRNAs could have a possible regulatory link with critical pathways associated with leukemogenesis. Conclusions: Our results indicate that non-coding genes surrounding MYC within the 8q24 region might have regulatory roles in CML carcinogenesis.

Molecular Epidemiology of BCR-ABL Rearrangement Variants in Chronic Myeloid Leukemia and Acute Lymphocytic Leukemia from Major Institute of Pakista

In Pakistan the disease burden of Chronic Myeloid Leukemia (CML) and Acute Lymphoblastic Leukemia (ALL) is quite high, yet there is a lack of scientific evidence regarding the spectrum of BCR-ABL rearrangement variants in CML and ALL. Knowing about BCR-ABL rearrangement is important in determining the prognosis and treatment strategy of disease at the time of diagnosis. This study included a total of 685 patients, out of which there were 644 CML patients and 41 ALL patients, from October 2016-July 2019. From the CML group, 270 patients were reported to have the BCR-ABL1 transcript, from which 50% were males. Whereas in the ALL group, 35 patients were reported to have the BCR-ABL1 transcript out of which 65.7% were males. Proportions of BCR-ABL transcript type differed between the two groups, with b3a2 (63.3%) and b2a2 (34.8%) transcript types having the highest frequency in CML patients, whereas e1a2 (77.1%) and b3a2 (11.3%) transcript types were found to have the highest frequency in ALL patients. Our data shows transcript genotypes in CML and ALL patients in an Asian population, which may be useful to guide the clinical management and assess prognosis. Since the majority of our CML population had the b3a2 transcript, they have a better prognosis and treatment response.

THE PROGNOSTIC SIGNIFICANCE OF TET2 SINGLE NUCLEOTIDE POLYMORPHISM IN EGYPTIAN CHRONIC MYELOID LEUKEMIA

BackgroundChronic myeloid leukemia (CML) is a myeloproliferative neoplasm where pathogenesis is based on the oncoprotein termed BCR-ABL1.1 TET2 initiates DNA demethylation and is frequently mutated in hematological malignancies, including CML. The relation between TET2 acquisition and CML transformation and/or imatinib resistance is needed to be investigated.2AimTo evaluate Ten Eleven Translocation 2 gene (TET2) single nucleotide polymorphism (SNP) (rs2454206, rs34402524, rs61744960) in chronic myeloid leukemia (CML) in relation to the disease prognostic criteria.Materials & MethodThe study included 84 subjects; 54 CML in chronic phase and 30 healthy subjects as control group matched for age and sex. Routine investigations, including CBC, bone marrow aspiration, biochemical investigations, and molecular study, were performed in CML patients to identify the disease stage. DNA extraction and SNP assay for TET2 gene polymorphism were done using (Thermo-Fisher predesigned SNP, USA) PCR prism 7500.ResultsThe mean age was 45.98±15.7 yrs in CML patients and 39.3±6.587 yrs in the control group (p>0.05). TET2 SNP rs 34402524 was either heterozygous or homozygous in CML (48%, 46.2% respectively) but was mainly homozygous among control (80%) group (p=0.012). TET2 SNP rs 2454206 wild type within CML was detected in 65.4% of patients and in controls was 63.3% (p=0.046). TET2 SNP rs 61744960 showed a homozygous pattern among all groups (CML and control) (p=0.528). TET2 SNP in CML cases did not alter the prognostic criteria as no statistical significance was noted (p>0.05) yet, it was significantly related to spleen size in rs 34402524 where the homozygous group had larger spleen size and higher BCR-ABL1 levels six months after starting TKIs (p<0.05).Conclusions/RecommendationTET2 SNP is common among Egyptian chronic myeloid leukemia. TET2 SNP rs 3442524 was associated with larger spleen size and higher BCR-ABL1 levels after six months of starting TKIs suggesting disease progression.

The TKI-Free Duration after a First Discontinuation Attempt That Failed in CP CML Patients Is a Predictive Factor of TKI-Free Remission after a Second Attempt

Legros: Incyte Biosciences: Honoraria, Research Funding; BMS: Honoraria; Pfizer: Honoraria, Research Funding; Novartis: Honoraria. Nicolini:Sun Pharma Ltd: Consultancy; Incyte Biosciences: Honoraria, Research Funding, Speakers Bureau; Novartis: Research Funding, Speakers Bureau. Etienne:BMS: Honoraria, Speakers Bureau; Incyte Biosciences: Honoraria, Speakers Bureau; Pfizer: Honoraria, Speakers Bureau; Novartis: Consultancy, Honoraria, Speakers Bureau. Rousselot:Pfizer: Research Funding; Incyte: Research Funding. Rea:Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees; Pfizer: Honoraria, Membership on an entity's Board of Directors or advisory committees; BMS: Honoraria; Incyte Biosciences: Honoraria. Guerci:INCYTE: Consultancy, Honoraria. Huguet:Incyte Biosciences: Honoraria; Novartis: Honoraria; Servier: Honoraria; Jazz Pharmaceuticals: Honoraria; Amgen: Honoraria; BMS: Honoraria; Pfizer: Honoraria. Coiteux:Novartis: Honoraria, Speakers Bureau; BMS: Honoraria, Speakers Bureau; Incyte Biosciences: Honoraria, Speakers Bureau; Pfizer: Honoraria, Speakers Bureau. Mahon:Novartis: Consultancy, Honoraria, Speakers Bureau; BMS: Honoraria, Speakers Bureau; Pfizer: Honoraria, Speakers Bureau; Incyte Biosciences: Honoraria, Speakers Bureau.