Expression Profiles of Long Non-Coding RNA GAS5 and MicroRNA-222 in Younger AML Patients

Acute myeloid leukemia (AML) is a heterogeneous malignant disease both on clinical and genetic levels. AML has poor prognosis and, therefore, there is a constant need to find new prognostic markers, as well as markers that can be used as targets for innovative therapeutics. Recently, the search for new biomarkers has turned researchers' attention towards non-coding RNAs, especially long non-coding RNAs (lncRNAs) and micro RNAs (miRNAs). We investigated the expression level of growth arrest-specific transcript 5 (GAS5) lncRNA in 94 younger AML patients, and also the expression level of miR-222 in a cohort of 39 AML patients with normal karyotype (AML-NK), in order to examine their prognostic potential. Our results showed that GAS5 expression level in AML patients was lower compared to healthy controls. Lower GAS5 expression on diagnosis was related to an adverse prognosis. In the AML-NK group patients had higher expression of miR-222 compared to healthy controls. A synergistic effect of GAS5(low)/miR-222(high) status on disease prognosis was not established. This is the first study focused on examining the GAS5 and miR-222 expression pattern in AML patients. Its initial findings indicate the need for further investigation of these two non-coding RNAs, their potential roles in leukemogenesis, and the prognosis of AML patients

Експресиони образац дуге некодирајуће РНК GAS5 и микроРНК- 222 код млађих пацијената оболелих од АМЛ

Akutna mijeloidna leukemija (AML) je heterogeno maligno oboljenje kako na kliničkom tako i na genetičkom nivou. AML ima lošu prognozu, i zato postoji konstantna potreba za novim prognostičkim markerima, kao i markerima koji mogu biti mete za inovativne terapeutike. U skorije vreme potraga za novim biomarkerima je naučnicima skrenula pažnju ka nekodirajućim RNK, naročito dugim nekodirajućim RNK (dnkRNK) i mikroRNK (miRNK). U ovom radu ispitali smo nivo ekspresije „growth arrest-specific transcript 5“ (GAS5) dnkRNK kod 94 mlađih osoba obolelih od AML, kao i nivo ekspresije miR-222 u grupi od 39 osoba obolelih od AML sa normalnim kariotipom (AML-NK), u cilju da ispitamo njihov prognostički potencijal. Naši rezultati su pokazali da je nivo ekspresije GAS5 kod obolelih od AML niži u poređenju sa zdravim kontrolama, i češći je u grupi sa nepovoljnom prognozom. U AML-NK grupi pacijenti su imali povišenu ekspresiju miR-222 u poređenju sa zdravim kontrolama. Sinergistički efekat „GAS5low/miR-222 high“ nije utvrđen. Ovo je prva studija koja je ispitala interakciju ekspresije GAS5 i miR-222 kod obolelih od AML. Naši rezultati indikuju potrebu za daljim ispitivanjem ove dve nekodirajuće RNK i njihove potencijalne uloge u leukemogenezi i prognostici AML pacijenata.Акутна мијелоидна леукемија (АМЛ) је хетерогено малигно обољење како на клиничком тако и на генетичком нивоу. АМЛ има лошу прогнозу, и зато постоји константна потреба за новим прогностичким маркерима, као и маркерима који могу бити мете за иновативне терапеутике. У скорије време потрага за новим биомаркерима је научницима скренула пажњу ка некодирајућим РНК, нарочито дугим некодирајућим РНК (днкРНК) и микроРНК (миРНК). У овом раду испитали смо ниво експресије „growth arrest-specific transcript 5“ (GAS5) днкРНК код 94 млађих особа оболелих од АМЛ, као и ниво експресије miR-222 у групи од 39 особа оболелих од АМЛ са нормалним кариотипом (АМЛ-НК), у циљу да испитамо њихов прогностички потенцијал. Наши резултати су показали да је ниво експресије GAS5 код оболелих од АМЛ нижи у поређењу са здравим контролама, и чешћи је у групи са неповољном прогнозом. У АМЛ-НК групи пацијенти су имали повишену експресију miR-222 у поређењу са здравим контролама. Синергистички ефекат „GAS5low/miR-222 high“ није утврђен. Ово је прва студија која је испитала интеракцију експресије GAS5 и miR-222 код оболелих од АМЛ. Наши резултати индикују потребу за даљим испитивањем ове две некодирајуће РНК и њихове потенцијалне улоге у леукемогенези и прогностици АМЛ пацијената.Knjiga sažetaka: Treći Kongres biologa Srbije, Zlatibor, Srbija 21 - 25. 9. 2022

The influence of BCL2, BAX, and ABCB1 gene expression on prognosis of adult de novo acute myeloid leukemia with normal karyotype patients

Background: Deregulation of the apoptotic process underlies the pathogenesis of many cancers, including leukemia, but is also very important for the success of chemotherapy treatment. Therefore, the gene expression profile of main apoptotic factors, such as anti-apoptotic BCL2 (B-cell lymphoma protein 2) and pro-apoptotic BAX (BCL2-associated X), as well as genes involved in the multi-drug resistance (ABCB1), could have significant impact on the prognosis and could be used as targets for specific therapy.Patients and methodsWe analyzed the expression of BCL2, BAX, and ABCB1 in bone-marrow samples collected at diagnosis from 51 adult patients with acute myeloid leukemia with normal karyotype (AML-NK) using real-time polymerase chain reaction method, and examined their prognostic potential.ResultsIncreased expression of BCL2 (BCL2+) was associated with the presence of chemoresistance (p = 0.024), while patients with low BAX expression were more prone to relapse (p = 0.047). Analysis of the combined effect of BCL2 and BAX expression showed that 87% of patients with BAX/BCL2low status were resistant to therapy (p = 0.044). High expression of ABCB1 was associated with BCL2+ status (p < 0.001), and with absence FLT3-ITD mutations (p = 0.019).ConclusionsThe present analysis of BCL2, BAX, and ABCB1 gene expression profiles is the first study focusing solel

Expression levels of BCL2, BAX and MDR1 as pharmacotranscriptomic and prognostic markers of prognosis in acute myeloid leukemia

Acute myeloid leukemia (AML) is a malignancy of hematopoetic tissue which occurs due to a halt in differentiation, loss of proliferation control and dysregulated apoptosis of myeloid progenitor cells. In many cancers, as well as AML, dysregulation of apoptosis constitutes the basis of pathogenesis and this phenomenon is important for chemotherapy success. Pharmacotranscriptomic markers of AML prognosis could be targets of specific therapy. The anti-apoptotic gene BCL2 (B-cell lymphoma protein 2), the pro-apoptotic BAX (BCL2-associated X) and genes involved in drug resistance, like MDR1 could have a significant impact on AML prognosis and therapy response. Bone-marrow samples at diagnosis were collected from 51 adult patients with AML-NK. Expressions of BCL2, BAX and MDR1 were analysed using the real-time polymerase chain reaction method. Statistical evaluation was performed. The presence of chemoresistance was found to be associated with overexpression of BCL2 (BCL2+) (p=0.018), while underexpression of BAX in patients has shown a greater affinity towards relapse (p=0.034). Evaluating the expressions of BCL2 and BAX in a combined effect has shown that 87% of patients with BAX/BCL2low status were resistant to therapy (p=0.024). BCL2+ status was associated with high expression of MDR1 (p<0.001). Likewise, high expression of MDR1 was associated with the absence of NPM1 and FLT3-ITD mutations (p=0.048 and p=0.010, respectively). This is the first study that focused only on AML-NK patients, when it comes to analysis of BCL2, BAX and MDR1 gene expression profiles. The results of this preliminary study have shown that high BCL2 expression would likely lead to resistance from chemotherapy, making anti-BCL2 treatment a viable option in patients with this expression profile. A study on a larger group of patients could clarify the prognostic importance of the studied genes in adult AML-NK patients and improve the precision medicine approach in the field of hematology.Book of abstracts: International Conference of Biochemists and Molecular Biologists in Bosnia and Herzegovina - ABMBBIH May, 202

The Role of the Spleen and the Place of Splenectomy in Autoimmune Hemolytic Anemia—A Review of Current Knowledge

Autoimmune hemolytic anemia (AIHA) is a rare, very heterogeneous, and sometimes life-threatening acquired hematologic disease characterized by increased red blood cell (RBC) destruction by autoantibodies (autoAbs), either with or without complement involvement. Recent studies have shown that the involvement of T- and B-cell dysregulation and an imbalance of T-helper 2 (Th2) and Th17 phenotypes play major roles in the pathogenesis of AIHA. AIHA can be primary (idiopathic) but is more often secondary, triggered by infections or drug use or as a part of other diseases. As the location of origin of autoAbs and the location of autoAb-mediated RBC clearance, as well as the location of extramedullary hematopoiesis, the spleen is crucially involved in all the steps of AIHA pathobiology. Splenectomy, which was the established second-line therapeutic option in corticosteroid-resistant AIHA patients for decades, has become less common due to increasing knowledge of immunopathogenesis and the introduction of targeted therapy. This article provides a comprehensive overview of current knowledge regarding the place of the spleen in the immunological background of AIHA and the rapidly growing spectrum of novel therapeutic approaches. Furthermore, this review emphasizes the still-existing expediency of laparoscopic splenectomy with appropriate perioperative thromboprophylaxis and the prevention of infection as a safe and reliable therapeutic option in the context of the limited availability of rituximab and other novel therapies

Can pharmacogenetics impact the therapeutic effect of cytarabine and anthracyclines in adult acute myeloid leukaemia patients?: A Serbian experience

Background: Cytarabine-anthracycline-based induction chemotherapy remains the standard of care for remission induction among patients with newly diagnosed acute myeloid leukaemia (AML). There are remarkable differences in therapy response among AML patients. This fact could be partly explained by the patients' genetic variability related to the metabolic paths of cytarabine and anthracyclines. This study aims to evaluate the effect of variants in pharmacogenes SLC29A1, DCK, ABCB1, GSTM1, and GSTT1, as well as laboratory and AML-related parameters on clinical outcomes in adult AML patients. Methods: A total of 100 AML patients were included in the study. Pharmacogenetic variants SLC29A1 rs9394992, DCK rs12648166, ABCB1 rs2032582, and GSTM1 and GSTT1 gene deletions were detected by methodology based on PCR, fragment analysis and direct sequencing. The methods of descriptive and analytic statistics were used. Survival analysis was done using the Kaplan-Meier method using the Log-Rank test. Results: This is the first study of adult AML pharmacogenetics in the Serbian population. Clinical outcomes in our cohort of AML patients were not impacted by analysed variants in SLC29A1, DCK, ABCB1 and GSTT1, and GSTM1 genes, independently or in combinations. Achievement of complete remission was identified as an independent prognostic indicator of clinical outcome. Conclusions: The population-specific genomic profile has to be considered in pharmacogenetics. Since the data on AML pharmacogenetics in European populations is limited, our results contribute to knowledge in this field and strongly indicate that a high-throughput approach must be applied to find particular pharmacogenetic markers of AML in the European population