Development Of Myelodysplastic Syndrome Research Articles

Irisin in myelodysplastic syndromes: connecting inflammation, oncogenesis, and atherogenesis

Objective This study aimed to measure irisin level and carotid intima-media thickness (CIMT), and, to unravel their correlations with other variables in patients with myelodysplastic syndromes (MDS). Background Multiple components of the marrow environment and of the MDS cell hierarchy itself can contribute to an inflammatory state which can influence MDS emergence, propagation, and evolution. Irisin is an adipomyokine that is involved.in.the.regulation.of.metabolic.processes.and.also.influences.processes.related.to.inflammation including cancer. Atherosclerosis is a chronic inflammatory vascular disease driven by traditional and nontraditional risk factors. Patients and methods This case–control study was carried out on 40 MDS patients and 40 age-matched and sex-matched healthy controls. Serum irisin level and CIMT were measured. Results The median irisin level and high-density lipoproteins were significantly lower in MDS patients compared with controls whereas CIMT, fasting blood glucose, 2 h postprandial blood sugar, hemoglobin A1c, homeostatic model assessment of insulin resistance, triglycerides, homocysteine, and high sensitive C-reactive protein were significantly higher in patients than controls. Conclusion Irisin level is significantly lower in patients with MDS where as CIMT is significantly higher in MDS patients compared to controls.

Azacitidine combined with venetoclax alleviates AML-MR with TP53 mutation in SDS: a case report and literature review.

Shwachman-Diamond syndrome (SDS) is an autosomal recessive genetic disease, which is prone to transform into myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). TP53 mutation is a driving factor involved in the transformation of SDS into MDS/AML, and in the evolution of MDS to AML. Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) is the only curable approach, however, challenge remains regarding the balance between efficacy and the high risk from treatment-related toxicity and mortality to achieve temporary disease control before transplantation to gain time and opportunities for transplantation. At present, pre-transplant bridging therapy has emerged as one of the important options with improved efficacy, reduced tumor burden, and less treatment-related toxicity. Here we reported azacitidine combined with venetoclax was used as pre-transplant bridging regimen in a TP53-mutant AML-MR case developed from SDS. He achieved complete remission with incomplete recovery and proceeded to Allo-HSCT. We hope to provide some evidence and insight for in-depth research and clinical treatment by presenting this case.

Study on the Changes of Immune Cells in the Evolution of Myelodysplastic Syndromes

ABSTRACT Objective: To analyze the differences of peripheral blood immune cells in healthy people, and patients with myelodysplastic syndromes (MDS) patients or acute myeloid leukemia (AML), and to investigate their correlations with International Prostate Symptom Score(IPSS), blasts and other factors in MDS patients. Methods: 55 patients with MDS(MDS group), 67 patients with AML(AML group) and 25 heathy people(healthy group) were selected as research objects from Dongzhimen Hospital Affiliated with Beijing University of Chinese Medicine from January, 2012 to April, 2022 for this retrospective analysis. By comparing peripheral blood lymphocyte subsets count and peripheral blood lymphocyte subsets percentage, we studied the expression of T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+, CD3+CD4+CD25+FoxP3+), B cells (CD19+) and NK cells (CD3-CD16+CD56+). The correlations between the different indexes and IPSS and blasts were analyzed. Results: (1) Peripheral blood lymphocyte subsets count: There were differences among the three groups. Through pairwise comparison, MDS group had lower peripheral blood lymphocyte subsets count(P＜0.025), probably owing to the low peripheral blood lymphocyte count. (2)Peripheral blood lymphocyte subsets percentage: The percentages of Treg cells, B cells and T cells varied among the three groups. Compared with the healthy group, the percentages of T cells ( P＜0.025) and Treg cells ( P＞0.025) in MDS group were higher, and the percentage of B cells ( P＜0.025) was lower. Compared with MDS group, the percentage of Treg cells ( P＜0.025) in AML group was higher and the proportion of B cells was lower ( P＜0.025). (3)Correlations with IPSS and blasts: The percentage of Treg cells was positively correlated with IPSS and blasts. Conclusion: Treg cells showed an increasing trend in turn by healthy people group, MDS group and AML group. The high expression of Treg cells may have great significance in prognosis and progression of MDS. Key words: Acute myeloid leukemia; Blasts; Immune cells; Myelodysplastic syndromes.

IL-18 and IL-18 Binding Protein Are Related to Disease Severity of Myelodysplastic Syndromes

Background: Myelodysplastic syndromes (MDS) was defined as a clonal hematopoietic disorder disease, although current stratified therapy for MDS has greatly improved overall survival, there is still a subset of them who suffered from progression to AML with a poor prognosis. Immune factors play essential roles in the pathogenesis and evolution of MDS. It is known that the level of plasma IL-18 which has anti-tumor and immunomodulatory effects increased in MDS patients, recent research found that the presence of IL-18BP depresses its effect. In this study, we intended to discuss the importance of IL-18 and IL-18BP on the severity of MDS and the potential mechanisms affecting the efficacy and explore new targets for MDS further therapy. Methods: 43 MDS patients, 14 AML patients, and 16 healthy volunteers were enrolled in our study, the levels of IL-18, IL-18BP of bone marrow supernatant were tested by ELISA and clinical information was collected; the expression of perforin, granzyme B, and IFN-γ, as well as PD-1 and TIM-3 expression on the surface CD8+ T and NK cells, was tested in MDS patients by flow cytometry, correlations among all these data were analyzed by SPSS. Results: We found that the levels of IL-18, IL-18BP, and fIL-18 in the bone marrow supernatants of both MDS and AML were higher than those of healthy controls. fIL-18 were negatively correlated with the severity of MDS. CD8+ T cells in MDS were hypofunctional, with a lower secretion of perforin, granzyme B, and IFN-γ than in healthy controls. The level of fIL-18 was positively correlated with perforin and IFN-γ; the expression of IL-18Rα on the surface of CD8+ T cells was low in MDS patients. The expression of IL-18Rα was negatively correlated with perforin, granzyme B, and IFN-γ. In addition, elevated levels of PD-1 and TIM-3 on the surface of CD8+ T cells in MDS seemed no significant correlation with fIL-18 and IL-18BP. Conclusion: MDS is in a state of immunosuppression as evidenced by the decreased number and depressed function of CD8+ T cells and NK cells. Increased expression of fIL-18 in MDS patients was shown, compared to healthy controls. Due to the presence of IL-18BP, fIL-18is relatively insufficient. Thus antagonizing IL-18BP is a promising new target for MDS.

Impact of the Updated IPSS-Molecular Prognostic Scoring System for Myelodysplastic Syndrome in 10,283 Real World Samples

Introduction The International Prognostic Scoring System-Molecular (IPSS-M) is a validated prognostic method that incorporates molecular information to improve risk stratification for patients with myelodysplastic syndromes (MDS). It is a weighted sum of prognostic variables consisting of clinical, cytogenetic, and somatic mutation information (up to 31 genes), used to generate a patient-specific risk score and associated risk category assignment. To assess the impact of combining additional molecular data into MDS prognosis, we retrospectively analyzed 10,283 real world samples from patients who had been sequenced with a commercially available targeted next-generation sequencing (NGS) panel to determine the number of patients whose risk stratification could potentially be altered by IPSS-M. Methods Next-generation sequencing was performed using a panel capable of detecting and reporting single nucleotide variants and small indels across 50 genes. IPSS-M uses 19 binary molecular features incorporating somatic mutation information from 31 genes, 27 of which are targeted by the NGS panel used. Whole blood or bone marrow samples from patients with cause-for-testing for MDS or peripheral blood cytopenias were submitted for analysis by a clinician. DNA was extracted and assayed by the targeted, NGS panel and sequenced on Illumina DNA sequencers (Illumina, San Diego, CA). Results were reviewed, orthogonally confirmed unless previously validated, and reported by clinical laboratory directors. Disease status or symptoms were abstracted from test requisitions for each patient. TP53 loss of heterozygosity and KMT2A partial tandem duplications (MLLPTD) are molecular features in the IPSS-M but could not be assessed in this study. Results A total 10283 samples were analyzed including 4437 with an indication for MDS. 46.0% (4728/10283) of samples had at least one of 17 IPSS-M molecular features. The mean number of features observed was 0.83 with a range 0-6. In the IPSS-M, multiple mutations in TP53 have the strongest negative effect and this feature was observed in 2.3% of patients (236/10283). SF3B1-α, which provides the strongest positive effect, was observed in 6.5% (666/10283) of patients. The most common features were ASXL1 (15.3%, 1571/10283), DNMT3A (11.7%, 1204/10283) and SRSF2 (10.0%, 1030/10283). 629 MDS or peripheral blood cytopenia patients had serial testing performed allowing us to observe possible changes in IPSS-M risk scores over time. Patients with serial testing had a mean of 2.16 tests performed with a range of 2-5 tests. The mean interval between tests was 225 days (median = 177 days, range 0-1211 days). 63.4% (399/629) of patients with serial tests had at least one molecular feature in at least one sample. Furthermore, a change in IPSS-M molecular features was observed between tests in 44.1% (176/399) of cases potentially leading to a change of IPSS-M risk score and clinical management in these patients. Gains of features between tests were observed in 30.1% (120/399) while losses were observed in 22.1% (88/399). These changes in mutation status may be result of tumor evolution, response to therapeutic intervention, or a combination thereof. A small number of patients (n=23) showed both losses and gains of molecular features over the course of testing revealing insights into the biological evolution of MDS. Efforts are ongoing to gather clinical and cytogenetic data associated with these samples to fully assess changes in IPSS-M scores over time. Conclusion A large proportion (50.1%, 2257/4437) of MDS patients analyzed have molecular findings that allow for further clinical risk stratification. These findings show the benefit of targeting a broad panel of genes using NGS for MDS prognosis and patient risk stratification.

CCUS and Low-Risk MDS Are Inherently Similar, Sharing Clonal and Clinical Features

Background Idiopathic cytopenia of undetermined significance (ICUS) is a condition of unexplained blood cytopenia not meeting the diagnostic criteria of myelodysplastic syndromes (MDS). Recently, the newer designation of clonal cytopenia of undetermined significance (CCUS) has been proposed to characterize patients with ICUS in the presence of one or more somatic mutations or copy number abnormalities. Clinically, CCUS has been associated with an increased risk of developing MDS; however, the mechanism and factors associated with evolution remain unclear. We sought to determine the molecular similarity of cytopenic cases with equivocal morphological dysplasia to bona fide MDS, as well as whether serial sequencing of ICUS and CCUS cases could identify factors that predict evolution to MDS. Methods We compared the incidence and frequency of somatic variants in 48 genes recurrently mutated in myeloid neoplasms (MNs), using targeted Next-Generation Sequencing (NGS) of bone marrow genomic DNA from 193 individuals with confirmed or suspected MDS or MDS/myeloproliferative neoplasm (MDS/MPN), including sequential investigation for 28 individuals at the time of diagnosis and during follow-up. Cytopenias qualifying patients as ICUS were based on WHO laboratory diagnostic criteria: hemoglobin levels <130g/L for males or <120 g/L for females; platelet count <150x109/L; and neutrophils <1.8x109/L. MDS patients were further subclassified as low risk (MDS-LR), including "Very Low” and "Low” International Prognostic Scoring System-Revised (IPSS-R) categories; intermediate risk (MDS-IR), including "Intermediate” category; or high risk (MDS-HR), including "High” and "Very High” risk categories. Results NGS further facilitated the diagnosis of all suspicious cases, either as MN (21%), CCUS (34%), or ICUS (45%). We found that there was no significant difference in most measured clinical features between CCUS and MDS-LR. While we saw significantly increased hemoglobin and platelet counts in individuals with CCUS compared with MDS-HR (Mann-Whitney U test; p=0.003 and p=0.012, respectively), no significant differences in cell counts were seen between CCUS and the MDS-LR or MDS-IR subgroups (Table 1). There was also no difference in overall survival between CCUS and MDS-LR when adjusted for age and sex in multivariable models (HR=0.575; 95% CI=0.256-1.294; p=0.330), while MDS-HR was associated with worse overall survival when compared with CCUS (HR=2.759; 95% CI=1.274-5.975; p=0.010). We saw no significant difference in clonal phenotypes, including number of variants or variant allele frequencies (VAFs), when comparing CCUS and MDS-LR. We further classified genes mutated in at least one patient in our cohort into functional groups for further analysis. These groups included epigenetic regulators: ASXL1, BCOR, DNMT3A, EZH2, IDH1, IDH2, KDM6A, PHF6, RAD21, STAG2, TET2; splicing factors: DDX41, SF3B1, SRSF2, U2AF1, ZRSR2; genes involved in signal transduction: BRAF, CALR, CBL, CSF3R, FLT3, GNB1, JAK2, KIT, KRAS, MPL, NF1, NPM1, NRAS, PTPN11, SETBP1; transcription factors: CEBPA, ETV6, GATA2, RUNX1; and genes involved in DNA damage repair: PPM1D and TP53. A significantly higher number of transcription factor mutations was seen when comparing CCUS to the MDS-IR and MDS-HR groups (p=0.007 and p=0.008, respectively), but not the MDS-LR group (p=0.754). We did, however, see significant significantly lower number of splicing factor mutations in CCUS cases compared with MDS-LR (p<0.001; Figure 1). Serial sequencing revealed no significant associations between number, type, or VAF of variants initially present at ICUS or CCUS diagnosis compared to those later acquired at the time of MDS evolution. Of note, we saw an increased probability of evolution to MDS of individuals with CCUS compared to ICUS over the first 5 years (HR=3.569; 95% CI=1.029-12.380, p=0.045). Conclusions Our analyses revealed no conclusive pattern associating clonal expansion or number of variants with evolution of CCUS to MDS, perhaps further supporting the similarity of these diseases and the proposal that CCUS should be classified as low-risk MDS in the next revision of the WHO guidelines. If future studies can better associate variants in specific types of genes with progression (such as splicing factors), this may also assist in predicting the probability of evolution and thus inform management strategies. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The Mesenchymal Niche in Myelodysplastic Syndromes.

Myelodysplastic syndromes (MDSs) are clonal disorders characterized by ineffective hematopoiesis, resulting in cytopenias and a risk of developing acute myeloid leukemia. In addition to mutations affecting hematopoietic stem cells (HSCs), numerous studies have highlighted the role of the bone marrow microenvironment (BMME) in the development of MDSs. The mesenchymal niche represents a key component of the BMME. In this review, we discuss the role of the mesenchymal niche in the pathophysiology of MDS and provide an overview of currently available in vitro and in vivo models that can be used to study the effects of the mesenchymal niche on HSCs.

GATA2 deficiency and MDS/AML: Experimental strategies for disease modelling and future therapeutic prospects.

The importance of predisposition to leukaemia in clinical practice is being increasingly recognized. This is emphasized by the establishment of a novel WHO disease category in 2016 called "myeloid neoplasms with germline predisposition". A major syndrome within this group is GATA2 deficiency, a heterogeneous immunodeficiency syndrome with a very high lifetime risk to develop myelodysplastic syndrome (MDS) and acute myeloid leukaemia (AML). GATA2 deficiency has been identified as the most common hereditary cause of MDS in adolescents with monosomy 7. Allogenic haematopoietic stem cell transplantation is the only curative option; however, chances of survival decrease with progression of immunodeficiency and MDS evolution. Penetrance and expressivity within families carrying GATA2 mutations is often variable, suggesting that co-operating extrinsic events are required to trigger the disease. Predictive tools are lacking, and intrafamilial heterogeneity is poorly understood; hence there is a clear unmet medical need. On behalf of the ERAPerMed GATA2 HuMo consortium, in this review we describe the genetic, clinical, and biological aspects of familial GATA2-related MDS, highlighting the importance of developing robust disease preclinical models to improve early detection and clinical decision-making of GATA2 carriers.

P749: PROGNOSTIC SIGNIFICANCE OF SERIAL CIRCULATING TUMOR DNA STATUS POST TREATMENT IN MYELODYSPLASTIC SYNDROMES AND ACUTE MYELOID LEUKEMIA

Background: A major advance in our understanding of Myelodysplastic Syndromes (MDS) and Acute Myeloid Leukemia (AML) biology has been the implementation of next generation sequencing (NGS) which influenced diagnostic, prognostic, and therapeutic decisions in myeloid malignancies. Circulating tumor DNA (ctDNA) sequencing, as as a novel and minimally invasive measure, was reported to exhibit excellent correlations with matched bone marrow NGS in MDS and AML. However, the clinical relevance of dynamic ctDNA monitor during active treatment is unknown. Aims: In this study, we assessed the role of ctDNA as a biomarker to monitor therapeutic response and clonal evolution in MDS and AML. Methods: Thirty-one MDS and AML patients who had both bone marrow NGS and ctDNA at baseline were included for concordance analysis. Twenty-seven patients who had at least two serial ctDNA assessment with at least one month interval were included for dynamic ctDNA analysis. Targeted deep sequencing was performed on bone marrow and ctDNA using a customized panel of 165 genes known to be recurrently mutated in MDS and acute myeloid leukemia. Results: Thirty out thirty-one patients were identified with ctDNA mutation at baseline. Diagnostic ctDNA and matched bone marrow DNA exhibited excellent correlations with variant allele frequencies (VAFs) (R2=0.721, p<0.001). For 27 patients who had ctDNA mutation in baseline and treated with HMA or chemotherapy, the average mutation VAFs at the end of treatment was lower in patients achieving a CR/CRi versus those with mCR, mLFS or SD (2.7% vs 24.1%, respectively, p<0.001). As expected, mutation VAFs from all patients with treatment failure did not decrease during treatment. (Figure 1). Similarly, the average decrease in mutation VAFs from pretreatment to end of treatment was greater in patients achieving a CR versus those who did not (87.7% vs 35.5%, respectively, p= 0.001). ctDNA negative post treatment was associated with longer PFS (median PFS not reached (NR) vs. 5.6 (95% CI 4.08-7.13) months; P=0.009)(Fig 2A) and OS (median OS NR vs. 12.0 (95% CI 9.08-15.0) months; P=0.023) (Fig 2B). Seven of ten MDS patients who transformed to AML were identified with lately acquired subclones harboring FLT3 or NF1, mutations involving in signaling pathway. Moreover, new subclones were detected 0.5 to 4 months prior to AML transformation in 4 cases, indicating that genetic progression can predate morphological progression. Image:Summary/Conclusion: ctDNA status post treatment was relevant to clinical response and was of prognostic significance for disease progress and relapse. Dynamic ctDNA changes revealed complex patterns of clonal structure of MDS and AML in response to treatment. ctDNA sequencing could be an attractive, prospective measure for disease monitoring.

The Differential Expression of CD47 may be Related to the Pathogenesis From Myelodysplastic Syndromes to Acute Myeloid Leukemia

Myelodysplastic syndrome (MDS) can lead to the development of peripheral blood cytopenia and abnormal cell morphology. MDS has the potential to evolve into AML and can lead to reduced survival. CD47, a member of the immunoglobulin family, is one molecule that is overexpressed in a variety of cancer cells and is associated with clinical features and poor prognosis in a variety of malignancies. In this study, we analyzed the expression and function of CD47 in MDS and AML, and further analyzed its role in other tumors. Our analysis revealed significantly low CD47 expression in MDS and significantly high expression in AML. Further analysis of the function or pathway of CD47 from different perspectives identified a relationship to the immune response, cell growth, and other related functions or pathways. The relationship between CD47 and other tumors was analyzed from four aspects: DNA methyltransferase, TMB, MSI, and tumor cell stemness. Changes in gene expression levels have a known association with aberrant DNA methylation, and this methylation is the main mechanism of tumor suppressor gene silencing and clonal variation during the evolution of MDS to AML. Taken together, our findings support the hypothesis that the differential expression of CD47 might be related to the transformation of MDS to AML.

"Whole Exome Sequencing or Pan-Myeloid NGS Gene Panel to Assess Leukemic Evolution of Myelodysplastic Syndromes. Advantages and Disadvantages"

"Whole Exome Sequencing or Pan-Myeloid NGS Gene Panel to Assess Leukemic Evolution of Myelodysplastic Syndromes. Advantages and Disadvantages"

Hematopoietic Stem Cells (HSC) and Granulocyte Macrophage Progenitors (GMP) are the Oxidative Stress Targets in Low/Intermediate-1 Risk Myelodysplastic Syndromes.

Myelodysplastic Syndromes (MDS) are a heterogenous group of clonal hematopoietic stem cell malignancies. Previous studies showed that Reactive Oxygen Species (ROS) play a role in the pathogenesis and clinical evolution of MDS, contributing to Hematopoietic Stem and Progenitor Cells (HSPC) genetic instability. Less is known about ROS levels in the various sub-populations of MDS HSPC and how they correlate with clinical data in MDS patients. Our study aims to analyze ROS levels in MDS Hematopoietic Stem Cells (HSC), common myeloid progenitors (CMP), Granulocyte Macrophages Progenitors (GMP) and megakaryocyte-erythrocyte progenitors (MEP); afterwards, we looked at the relationship between ROS levels and clinical data. thirty-eight MDS and 27 Normal Bone Marrow (NBM) samples were collected; ROS levels were analyzed via multicolor flow cytometry. In both NBM and MDS, HSC showed much higher ROS levels than progenitors (3 to 4 folds, p < 0.0001); HSC ROS were significantly more elevated in MDS-no excess blasts versus MDS with excess blasts and versus NBM. GMP from MDS-no excess blasts showed higher ROS compared to NBM GMP. The 3 MDS with Ringed Sideroblasts (RS) showed more elevated ROS in HSC and GMP compared to the not RS low/intermediate-1 MDS; the 2 monosomy 7 patients displayed higher ROS levels in each subpopulation compared to the normal karyotype MDS; the only del(5q) patient did not show relevant differences in ROS levels compared to the median of the normal karyotype MDS ROS. The 9 high transfusion burden patients exhibited higher ROS in HSC and GMP compared to NBM HSC and GMP. These data were not confirmed in low transfusion burden (n:2) and non-transfused patients (n:26). In low/intermediate-1 MDS, a direct correlation between ferritin values and ROS levels in progenitors, but not in HSC, was detected. Interestingly, low/intermediate-1 risk patients that are no longer responding to recombinant human erythropoietin (rh-EPO) showed higher ROS levels in GMP and HSC. Our data showed that ROS can play a role in the pathogenesis and maintenance of low and intermediate-1 risk MDS clone; ROS status can be influenced by several clinical factors as ferritin levels and rh-EPO treatment. In this scenario, high ROS levels can contribute to genetic instability and influence progression to AML. Further biological studies are needed to elucidate ROS role in MDS pathogenesis and analyze the possible benefit of antioxidant drugs added to the standard MDS treatments.

Lymphocytic Infiltrate and p53 Protein Expression as Predictive Markers of Response and Outcome in Myelodysplastic Syndromes Treated with Azacitidine.

High-risk Myelodysplastic syndromes (MDS) represent therapeutical challenges and are usually managed with hypomethylating agents such as azacitidine. Given the lack of data in the literature concerning azacitidine effects on bone marrow, we retrospectively analyzed 57 high-risk MDS cases in order to identify any changes induced by azacitidine therapy or relevant correlations between therapy response and pre- or post-treatment features. Azacitidine treatment had no significant impact on bone marrow cellularity or morphological dysplastic features. On the contrary, although not statistically significant, we observed a slight decrease in CD34+ and CD117+ blasts and p53+ precursors after treatment. Moreover, pre-treatment IPSS-R cytogenetic score (p = 0.004), lymphocytic infiltrate (p = 0.017) and p53+ elements (p = 0.001) correlated with AML progression; pre-treatment lymphocytic infiltrate was also linked to better response to therapy (p = 0.004), suggesting an anti-tumoral role of bone marrow microenvironment. Post-treatment blast count impacted negatively on overall survival (p = 0.035) and risk of leukemic progression (p = 0.04), while both post-treatment lymphocytic infiltrate and p53+ elements showed significant correlation with treatment response (p = 0.004 and p = 0.003 respectively). Higher post-treatment p53+ elements correlated also with risk of leukemic progression (p = 0.013). Our results suggest the possible role of lymphocytic infiltrate and p53+ elements as predictive markers in MDS treated with azacitidine, disclosing new chapters in the understanding of MDS evolution and treatment.

Genomic variations in patients with myelodysplastic syndrome and karyotypes without numerical or structural changes

Myelodysplastic syndrome (MDS) is an onco-hematologic disease with distinct levels of peripheral blood cytopenias, dysplasias in cell differentiation and various forms of chromosomal and cytogenomic alterations. In this study, the Chromosomal Microarray Analysis (CMA) was performed in patients with primary MDS without numerical and/or structural chromosomal alterations in karyotypes. A total of 17 patients was evaluated by GTG banding and eight patients showed no numerical and/or structural alterations. Then, the CMA was carried out and identified gains and losses CNVs and long continuous stretches of homozygosity (LCSHs). They were mapped on chromosomes 1, 2, 3, 4, 5, 6, 7, 9, 10, 12, 14, 16, 17, 18, 19, 20, 21, X, and Y. Ninety-one genes that have already been implicated in molecular pathways important for cell viability were selected and in-silico expression analyses demonstrated 28 genes differentially expressed in mesenchymal stromal cells of patients. Alterations in these genes may be related to the inactivation of suppressor genes or the activation of oncogenes contributing to the evolution and malignization of MDS. CMA provided additional information in patients without visible changes in the karyotype and our findings could contribute with additional information to improve the prognostic and personalized stratification for patients.

Screening for myeloid mutations in patients with myelodysplastic syndromes and AML with myelodysplasia-related changes

IntroductionOne of the most critical complications in myelodysplastic syndromes (MDS) is the progression to acute myeloid leukemia (AML). The dynamics of clonal evolution in MDS and how acquired mutations can be used as biomarkers to track disease progression remains under investigation. Objective and methodHerein, we investigated the frequency of common myeloid clonal mutations (FLT3, NPM1, JAK2, IDH1 and IDH2) in 88 patients with MDS and 35 AML patients with myelodysplasia-related changes, followed at a single reference center in northeastern Brazil. ResultsOverall, 9/88 (10%) of the MDS patients and 9/35 (26%) of the secondary AML patients had at least one mutation. While the JAK2 V617F mutation was the most frequent in the MDS patients, the FLT3, NPM1, IDH1 and IDH2 mutations were more frequently found in the secondary AML group. Furthermore, there was a higher frequency of FLT3, NPM1, IDH1 and IDH2 mutations in MDS patients classified as high-risk subtypes than in those of lower risk. ConclusionDespite the limited sample size, our data suggest that mutations in FLT3, NPM1, IDH1 and IDH2 genes could be potential biomarkers to detect early disease progression in MDS.

Acute myeloid leukemia M1 FAB secondary to myelodysplastic syndrome with excess blasts type 2

Acute myeloid leukemia occurs naturally in the evolution of myelodysplastic syndrome (MDS), typically in MDS with excess blasts type 2. The risk of progression is around 33%. We present the case of an elderly patient who was diagnosed with MDS single dysplasia in 2016. He was treated with erythropoietin. In 2021, the disease evolved into MDS with excess blast type 2 and then in acute myeloid leu­ke­mia in a short period of time.

Intracellular ROS profile in hematopoietic progenitors of MDS patients: association with blast count and iron overload

ABSTRACT Objectives Reactive oxygen species (ROS) are under scrutiny as a participant in the pathophysiology of myelodysplastic syndrome (MDS) and the progression of MDS to acute myeloid leukemia (AML). Measurement of intracellular ROS (iROS) is particularly important since iROS is a direct indicator of cellular health and integrity. Methods We developed a technique to measure standardize iROS (siROS) level in lymphocytes and bone marrow (BM) CD34+ hematopoietic progenitors using the fluorescent probe dichlorofluorescein (DCF). We then quantified the siROS in 38 consecutive BM specimens from 27 MDS patients over the course of 10 months. Disease outcome of these patients were also assessed. Results High serum ferritin, high blast count and poor IPSS were associated with inferior survival and AML progression in this cohort. High blast MDS patients had lower siROS in their BM CD34+ cells than those of low blast patients, consistent with increased reliance on glycolysis and enhanced ROS defense in high blast MDS. We also observed narrower siROS distribution in the BM CD34+ cells of high blast patients, suggesting that loss of heterogeneity in ROS content accompanies the clonal evolution of MDS. Furthermore, we observed a strong correlation between CD34+ cells siROS and serum ferritin level in high blast patients. In one case, iron chelation therapy (ICT) resulted in parallel decreases in serum ferritin and CD34+ cells siROS. Conclusion Our findings established the siROS profile in early hematopoietic cells of MDS patients and its relationship with blast count and iron overload.

Molecular Pathogenesis and Treatment of Myelodysplastic Syndromes.

Myelodysplastic syndromes (MDS) are clonal hematological disorders arising from hematopoietic stem cells that have accumulated various genetic abnormalities. MDS are heterogeneous in nature but uniformly characterized by chronic and progressive cytopenia from ineffective hematopoiesis, dysplasia in single or multiple lineages, and transformation to acute leukemia in a subset of patients. The genomic landscape revealed by next-generation sequencing has provided a comprehensive picture of the molecular pathways involved in MDS pathogenesis. Recurrent mutational targets in MDS are the genes involved in RNA splicing, DNA methylation, histone modification, transcription, signal transduction, cohesin complex and DNA repair. Sequential acquisition of mutations in these sets of genes serves as a driver for the initiation, clonal evolution and progression of MDS. Based on these findings, novel agents targeting driver mutations of MDS are currently under development and expected to improve the clinical outcome of MDS in the coming decades.

Predisposition to transformation from myelodysplastic syndrome into acute myeloid leukemia – case presentation

Predisposition to transformation from myelodysplastic syndrome into acute myeloid leukemia – case presentation

Aberrant Bone Marrow Microenvironment in Therapy Related Myeloid Neoplasm (t-MN)

Introduction: Therapy-related myeloid neoplasm (t-MN) is a lethal second hematological malignancy following chemotherapy (CT) and radiotherapy (RT) for primary cancers. It accounts for 15-20% of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). AML and MDS are considered to be hematopoietic stem cell (HSC)-autonomous disorders, in which initiation and progression are mainly driven by HSC-intrinsic genetic events. However, emerging data suggest that bone marrow (BM) microenvironment plays critical role in initiation and evolution of MDS and AML (Raaijmakers et al., 2010). Malignant clones can also shape the BM-microenvironment conducive for its survival and proliferation (Medyouf et al., Cell Stem Cell 2014). Although CT/RT can damage BM-microenvironment, very limited studies assessed role of BM-microenvironment in t-MN pathogenesis. Aim: To assess BM-microenvironment changes induced by malignant HSC and changes induced by previous genotoxic stress on BM-microenvironment, we compared BM-mesenchymal stromal cells (MSC) from t-MN patients, with BM-MSC from 1) patients with two unrelated cancers, one of the cancer being MDS/AML, without prior exposure to CT/RT (Double cancers, DC), 2) primary MDS patients (pMDS) and 3) age matched healthy controls (HC). Methods: We characterized BM-MSC from t-MN (n=10), DC (n=8), pMDS (n=6) and age-matched healthy controls (n=10). Morphology (Fei et al., 2014), clonogenic potential (Geyh et al., 2013), proliferation (Prata et al., 2010) and cellular senescence was assessed using previously described methodology. Differentiation potential was assessed by the respective lineage cytochemical staining and quantification of positive cells (mineral quantification for osteogenic cells, and Nile red for adipocytes). DNA damage response was determined by assessing H2AX phosphorylation in the MSC. Results: Only 70% of BM-MSC from t-MN cohort could be expanded to passage 6 compared to 100% of MSC cultures from pMDS, DC and HC. Proliferation rate, assessed by population doubling time, and clonogenic potential was significantly reduced in t-MN patients compared to p-MDS, DC and HC (Fig 1Ai-ii). This was further substantiated by higher senescence rates, assessed by β-galactosidase positive cells at passage 3 (HC 7%±1.9%; pMDS 39%±6%; DC 27%±1%; t-MN 68%±4%) (Fig 1B). Together, it demonstrates that MSC from t-MN have significantly impaired proliferation capacity and higher senescence rate compared to HC, pMDS and DC patients. Interestingly, proliferation capacity and senescence rate was not significantly different between MSC from DC and pMDS patients. We also compared DNA damage repair, following sub-lethal dose of RT, in MSC from t-MN, pMDS and HC. DNA damage repair in t-MN MSC was significantly impaired compared to pMDS and HC (Fig. 1F). Impaired DNA repair could be due to pathogenic germline mutation in DNA repair pathways in some t-MN patients (Singhal et al., ASH 2018). Although MSC from pMDS and DC appeared disorganized, they maintain fibroblast-like morphology similar to HC-MSC. Whereas, most of the MSCs from t-MN cases lost spindle shape morphology and were significantly larger than DC, pMDS and HC (p&lt;0.0001; Fig 1C). Further characterization of the MSC's phenotype revealed that ability to form a mineralized matrix was significantly increased in t-MN MSC compared to HC, pMDS and DC (p=0.04; Fig 1Di-ii). In contrast, quantification of lipid-laden Nile-red-stained adipocytes in t-MN MSCs showed a 5-fold decrease in adipocytes formation compared to HC and ~2-fold compare to pMDS and DC (Fig 1Ei-ii). Conclusions: Our data demonstrate that BM-MSCs from patients with myeloid malignancies are significantly abnormal as compared to age-matched healthy controls. BM-MSCs from T-MN patients have significantly reduced proliferative, clonogenic and DNA repair capacity and have higher senescence rate as compared to BM-MSCs from patients with double cancers. The critical difference between t-MN and DC is previous exposure to CT/RT, providing evidence that prior CT/RT leads to long-term damage to BM-microenvironment, which could be contributing t-MN pathogenesis. Disclosures No relevant conflicts of interest to declare.