Association of human concentrative nucleoside transporter 3 gene expression and CYP2B6*6 аllele with the response to fludarabine plus cyclophosphamide in chronic lymphocytic leukemia patients

Hemioterapija na bazi fludarabina i ciklofosfamida predstavlja osnov lečenja pacijenata sa hroničnom limfocitnom leukemijom (HLL) mlađe životne dobi i bez značajnih komorbiditeta. Kod nekih pacijenata efekat terapije može biti uslovljen aberantnom ekspresijom i/ili mutacijama gena koji utiču na farmakodinamiku i farmakokinetiku antileukemijskih lekova. Sa druge strane, ustanovljeni su brojni kliničko-laboratorijski i molekularno-genetički markeri prognoze koji su značajni u predikciji kliničkog toka, prvenstveno vremena do prve terapije (time to first treatment, TTFT) i ukupnog preživljavanja (overall survival, OS). Oni se poslednjih godina u cilju jačanja prognostičke vrednosti kombinuju unutar različitih prognostičkih modela, među kojima su za predikciju TTFT naročito relevantni Skor rizika od progresije (Progression.Risk Score, PRS) i Skor MD Anderson centra za kancer iz 2011 (MD Anderson Cancer Center 2011 score, MDACC 2011), dok je za predviđanje OS najznačajniji Internacionalni prognostički indeks za HLL (The International Prognostic Index for CLL, CLL-IPI). Cilj rada je da se ispita uticaj farmakogenetike, tačnije ekspresije gena SLC28A3 koji kodira hCNT3 protein, jedan od transportera za fludarabin, i varijantnog alela CYP2B6*6 gena za citohrom P450 2B6, koji učestvuje u metabolizmu ciklofosfamida, na ishod lečenja FC protokolom kod pacijenata sa HLL-om. Takođe, cilj je da se u grupi ovih pacijenata utvrdi vrednost skorova CLL-IPI, PRS i MDACC 2011 u predikciji TTFT, OS, vremena bez progresije (progression free survival, PFS) i odgovora na terapiju. Konačno, ispitaće se veza farmakogenetičkih karakteristika i skorova. Metodologija: U studiju je uključeno 57 pacijenata sa HLL-om koji su lečeni FC protokolom, većinom u prvoj liniji. Pacijenti su stratifikovani prema skorovima CLL-IPI, PRS i MDACC 2011, nakon čega je ispitana njihova prognostička vrednost u pogledu TTFT, odgovora na terapiju, PFS i OS...Fludarabine plus cyclophosphamide (FC) chemotherapy is the basis of treatment protocols used in management of chronic lymphocytic leukemia (CLL). In some patients, response to therapy may be affected by aberrant function of genes involved in pharmacokinetics and pharmacodynamics of the drugs. On the other hand, numerous clinical, biological and genetic prognostic markers which predict the clinical course of CLL, primarily time to first treatment (TTFT) and overall survival (OS) have been established. In recent times, they have been combined to make different prognostic models in order to enhance their prognostic value. The most relevant prognostic models used for prediction of TTFT are the Progression-Risk Score (PRS), and the MD Anderson Cancer Center Score 2011 (MDACC 2011), while CLL-International Prognostic Index (CLL-IPI), although the most powerful for prediction of OS, is also being used to estimate TTFT. The aim of this research was to assess the impact of pharmacogenetic variability, namely expression of SLC28A3 gene and the presence of CYP2B6*6 variant allele, on the FC treatment efficacy. Also, one of the objectives was to investigate CLL-IPI, PRS, and MDACC 2011 prognostic values regarding TTFT, first line treatment response, progression-free survival (PFS) and OS. Finally, the association of pharmacogenetics and prognostic models was investigated. Methods: Fifty-seven CLL patients treated with FC, most of them in the first treatment line, were enrolled in this study. Patients were stratified according to the prognostic models CLLIPI, PRS and MDACC 2011 and their prognostic significance regarding TTFT, treatment response, PFS and OS was examined. Considering the fact that the loss of TP53 gene function is one of the main causes of chemoresistance, patients with mutated and/or deleted TP53 were excluded from pharmacogenetic analyses, thus eliminating the possibility of its influence on the treatment outcome..

Clinical significance of TP53 aberrations and IGHV mutational status in chronic lymphocytic leukemia

Hronična limfocitna leukemija (HLL) izuzetno je heterogena bolest varijabilnog kliničkog toka. S jedne strane imamo, pacijente sa agresivnom i rezistentnom bolešću od koje umiru svega par meseci nakon dijagnoze, dok s druge strane spektra postoje pacijenti sa indolentnom, sporo progredirajućom bolešću koja ne zahteva lečenje decenijama. Razlozi su samo delimično poznati i već su decenijama unazad tema mnogobrojnih naučnih istraživanja. Tako je razvijen koncept prognostičkih i prediktivnih faktora u HLL-u, koji imaju za cilj da predvide klinički tok, odnosno terapijski ishod HLL-a. Liste prognostičkih i prediktivnih faktora su, sa boljim poznavanjem patofiziologije ove bolesti, svakom godinom sve duže, ali se i međusobno preklapaju. U ovom revijalnom radu izabrali smo aberacije TP53 gena i mutacioni status rearanžiranih IGHV (engl. immunoglobulin heavy variable) gena kao dva najznačajnija i najproučavanija faktora koji imaju i prognostički i prediktivni značaj.Chronic lymphocytic leukemia (CLL) is a very heterogeneous disease with a variable clinical course. On the one side of the spectrum, there are patients with aggressive and resistant disease, of which they die only a few months after diagnosis while, on the other side, there are patients with an indolent, slowly progressive disease that does not require treatment for decades. The reasons for this are only partially known, but they have been the subject of numerous scientific studies during the last several decades. Consequently, the concept of prognostic and predictive factors in CLL have emerged, which aims to predict the clinical course and its therapeutic outcome. With the improvement of understanding the pathophysiology of this disease, the lists of prognostic and predictive factors are getting longer every year, but they also overlap. In this review, we singled out the aberrations of the TP53 gene and the IGHV (immunoglobulin heavy variable) gene mutational status as the two most important and most studied factors that have both prognostic and predictive significance

Signifi cance of molecular diagnosti cs in therapy of chronic lymphocytic leukemia

Chronic lymphocyti c leukemia (CLL) is a malignancy of mature CD5+ B lymphocytes that is characterized by excepti onal clinical and biological heterogeneity. The Rai and Binet staging systems, developed in the late 1970s to early 1980s, are used in clinical practi ce to strati fy CLL pati ents into risk categories and to help guide clinical follow-up opti ons: to treat or to watch and wait. However, in early-stage disease, these systems are unable to predict what pati ents will face the progression to a more aggressive disease. That means, a number of molecular markers with prognosti c and/ or predicti ve impact exist and their assessment is strongly recommended in all pati ents prior to treatment initi ati on. One of the fi rst recognized prognosti c genomic aberrati ons in CLL include those detected by fl uorescence in situ hybridizati on (FISH): del(17p), del(11q), trisomy 12 and del(13q), and the immunoglobulin heavy variable (IGHV) gene somati c hypermutati on (SHM) status. Moreover, the rapid development of genomics techniques greatly expanded the understanding of CLL at the molecular level in the past decade. This resulted in the discovery of many newer prognosti c markers based on chromosomal aberrati ons or gene mutati ons. For instance, next-generati on sequencing (NGS) studies have led to the discovery of recurrently mutated genes in CLL, such as NOTCH1, SF3B1, BIRC3, XPO1, POT1, NFKBIE and EGR2, that are associated with poor clinical outcome. Among all of these biomarkers, the disti ncti on between markers of prognosti c and predicti ve values should be made. Prognosti c markers refer to biomarkers that can provide informati on regarding the pati ent's outcome regardless of treatment. They are o����� en assessed before treatment to help guide decisions on to treat or not. Markers associated with overall survival (OS) or ti me to fi rst treatment (TTFT) represent such examples. On the other hand, predicti ve markers are related to therapeuti c interventi ons with the ability to predict treatment response to a drug. These markers are normally assessed when pati ents receive the parti cular therapy. Some markers can be both prognosti c and predicti ve. The Nati onal Comprehensive Cancer Network guideline recommends testi ng of TP53 geneti c alterati ons, IGHV mutati on status, and several well-established cytogeneti c markers for CLL prognosti cati on. Of these, TP53 mutati ons, IGHV unmutated status, del(17p), and del(11q), as well as complex karyotype (the presence of three or more unrelated clonal chromosomal abnormaliti es in a sample), are associated with poor prognosis. Normal karyotype and trisomy 12 are considered as intermediate prognosti c factors, whereas del(13q) is associated with a favorable prognosis. The higher frequencies of the previously menti oned unfavorable markers (except for IGHV) found in the treated populati on usually imply the clonal evoluti on during disease progression or change in clonal dynamics induced by therapies, especially chemotherapies. Diff erent molecular and genomic techniques are employed for detecti ng molecular biomarkers in CLL. For IGHV mutati on status, the preferred method is Sanger sequencing to detect mutati ons in genomic DNA or cDNA following PCR, and align the resulti ng sequences to the germline IGHV using the IMGT/V-QUEST analyti c tool, where ≥ 98% homology to the germ line is interpreted as unmutated, >2% nonhomology as mutated, and 97.0% to 97.9% is interpreted as borderline. Prognosti cally signifi cant chromosomal abnormaliti es are frequently detected using fl uorescence in situ hybridizati on, array comparati ve genomic hybridizati on or conventi onal karyotyping. Fluorescence in situ hybridizati on, although off ers a high sensiti vity and specifi city, requires prior knowledge of chromosomal lesions for the probe designs and are limited to the chosen panel genes. The technique has limitati ons in detecti ng possible complex cytogeneti c abnormaliti es, as well. On the other hand, karyotyping and array comparati ve genomic hybridizati on provide genome-wide coverage. Despite the fact that array comparati ve genomic hybridizati on does not eff ecti vely detect balanced chromosomal rearrangements, it uncovers more genomic abnormaliti es than karyotyping as the probe-based technology examines the chromosomal structure at a much higher resoluti on. Development in NGS technology in the past two decades, made the technique, especially targeted sequencing of gene panels, much less costly and accessible. Currently, in Serbia, geneti c techniques such as FISH, conventi onal karyotyping, Sanger sequencing and NGS are available for detecti on of CLL biomarkers. Advances in the understanding of CLL pathogenesis have consequently led to the development of several highly eff ecti ve targeted therapies, including Bruton tyrosine kinase (BTK), phosphati dylinositol 3-kinase, and BCL2 apoptosis regulator (BCL2) directed inhibitors. B-cell survival and proliferati on is regulated by the BCR signaling pathway. In normal B cells, BCR is triggered by anti gen ligati on, leading to acti vati on of a cascade of tyrosine kinases, including BTK. BCR signaling is aberrantly acti vated in many B-cell malignancies, including CLL. Ibruti nib has demonstrated high clinical effi cacy acti ng as an irreversible potent inhibitor of Bruton's tyrosine kinase and targets several key components of the BCR pathway. However, despite having 80% to 90% response rate, 10% to 15% of CLL pati ents, who respond initi ally, develop ibruti nib resistance and disease relapse in 2 to 3 years on ibruti nib treatment, mainly because of the acquisiti on of a BTK C481S mutati on. The mutati on prevents the drug from forming a covalent bond with the C481 residue that weakened the drug-BTK binding by 500-fold. As a result, BCR signaling and cell proliferati on were restored in the tumor cells. BTK mutati ons may be found in approximately 70% of CLL pati ents who progressed on ibruti nib treatment. Another resistance mechanism is through acquired acti vati ng mutati ons in PLCG2, which is found in approximately 10% of the cases. Given these evidences, the current Nati onal Comprehensive Cancer Network guideline recommends testi ng for BTK and PLCG2 mutati ons for CLL pati ents receiving ibruti nib who are suspected of having disease progression. NGS has become the opti mal method for detecti ng BTK or PLCG2 mutati ons in the se����� ng of ibruti nib treatment, as multi ple mutati ons in both genes may occur in the same specimen. Currently, approximately 20% of CLL pati ents who progressed on ibruti nib do not have either BTK or PLCG2 mutati ons; thus, with NGS, it is possible to uncover other less common but yet undefi ned drug-resistance mutati ons. In additi on to BTK and PLCG2 mutati ons known to confer ibruti nib resistance, other molecular markers have been associated with an upfront high risk of relapse on ibruti nib treatment. It has been reported that complex karyotype, del(17p)/TP53 mutati on, and del(18p) at baseline before ibruti nib treatment are strongly associated with disease relapse. Other approved targeted agents for CLL treatment include the phosphati dylinositol 3-kinase inhibitors idelalisib and duvelisib and BCL2 inhibitor venetoclax. For venetoclax, a novel BCL2-G101V mutati on was identi fi ed to prevent drug acti vity through drug-protein interacti on. Each pati ent with CLL may have several clinical and molecular markers of confl icti ng prognosti c signifi cance simultaneously, making the precise prognosti cati on challenging. Today is of the greatest importance to apply ultrasensiti ve techniques to reveal molecular relapses a����� er therapy initi ati on and to detect minimal residual disease a����� er pati ents achieve complete responses

Influence of inflammatory cytokines on S100A proteins expression in CLL patients

Background: S100A proteins possess a broad range of intra- and extracellular functions. The involvement of these proteins in inflammation-mediated responses is of particular interests, considering that inflammation represents one of the landmarks of malignancy. Processes such as inflammation and cellular stress trigger the release of S100A proteins to extracellular space, interacting with their receptors and activating numerous intracellular signaling pathways, for instance NF-κB and AP1. Through them, S100A proteins take part into regulation of some of the most essential cellular processes: cell differentiation, apoptosis, inflammation, proliferation, etc. Aims: The aim of the study is to assess the role of inflammation in activation of S100A proteins via proliferation and inflammation related signaling pathways. Methods: We observed 60 CLL patients’ samples to isolate mononuclear cells (MNC) and CD19+ cells. MNC of CLL patients were treated with pro-inflammatory IL-6 and anti-inflammatory IL-10 cytokines, and inhibitors of JAK1/2, NF-κB and PI3K signaling pathways, to evaluate S100A4, S100A8, S100A9, S100A12 and NF-κB protein expression by immunoblotting. Also, we used immunocytochemistry to analyse the number of the S100As immunopositive MNC of CLL patients.Results: S100A8 showed higher level of protein expression in MNC and CD19+cells in comparison to healthy control. The number of immunopositive S100A4 (p<0.05) and S100A9 cells (p<0.001) was signifi cantly decreased in CD19+cells and MNC, respectively of CLL patients in comparison to healthy control. In addition, S100A4 and S100A9 proteins expression had statistically significant lower level of expression in MNC. Also, IL-6 stimulated expression of S100A8 and S100A4 in MNC of CLL, while the expression of latter one was prevented by NF-κB and JAK1/2 inhibitors. IL-10 reduced expression of S100A8 and S100A12 in MNC of CLL.Summary/Conclusion: Pro-inflammatory IL-6 and anti-inflammatory IL-10 cytokines have opposite effect on inflammatory S100A8 protein in CLL, with a potential to be a prognostic marker

Comparative analysis of International Prognostic Index for chronic lymphocytic leukemia, progression-risk score, and MD Anderson Cancer Center 2011 score: A single center experience

Uvod/Cilj Prognoza hronične limfocitne leukemije (HLL) značajno je unapređena u poslednje vreme. Među nekoliko prognostičkih modela čiji je cilj predviđanje vremena do prve terapije (eng. TTFT) izdvajaju se skor rizika od progresije (eng. PRS) i skor Centra za rak MD Anderson iz 2011. God. (eng. MDACC 2011), dok se internacionalni prognostički indeks za HLL (eng. CLL-IPI), iako primarno ustanovljen za predikciju ukupnog preživljavanja (eng. OS), dobro pokazao i u predikciji TTFT. Cilj ovog rada je da se ispita značaj pomenutih prognostičkih modela u pogledu predviđanja TTFT i OS. Metode Analizirana kohorta je obuhvatila 57 neselektovanih bolesnika sa HLL Univerzitetskog kliničkog centra Srbije sa prosečno agresivnijim profilom bolesti u odnosu na opštu populaciju de novo bolesnika sa HLL. Bolesnici su ocenjivani prema navedenim skorovima uz analizu TTFT i OS. Rezultati Bolesnici sa višim vrednostima CLL-IPI, PRS i MDACC 2011 primili su prvu terapiju značajno ranije u poređenju sa bolesnicima sa nižim vrednostima ovih skorova (p = 0,002, p = 0,019 i p lt 0,001, redom). U multivarijantnoj analizi, MDACC 2011 i CLL-IPI su zadržali prognostički značaj u predikciji TTFT (p = 0,001, odnosno p = 0,018), dok je PRS ovaj značaj izgubio. CLL-IPI je bio jedini značajan prediktor OS u univarijantnoj (p = 0,005) i u multivarijantnoj analizi (p = 0,013). Zaključak CLL-IPI, PRS i naročito MDACC 2011 su dobri prediktori TTFT čak i u kohortama bolesnika sa agresivnijom bolešću, dok je za predikciju OS od ova tri prognostička modela CLL-IPI jedini primenljiv. Ovi rezultati pokazuju da bi prognostičke modele trebalo ispitati na bolesnicima sa HLL u različitim fazama bolesti, kakvi se sreću u realnoj kliničkoj praksi.Introduction/Objective Prognostication of chronic lymphocytic leukemia (CLL) has been substantially improved in recent times. Among several prognostic models (PMs) focused on the prediction of time to first treatment (TTFT), progression-risk score (PRS), and MD Anderson Cancer Center score 2011 (MDACC 2011) are the most relevant, while CLL-International Prognostic Index (CLL-IPI), although originally developed to predict overall survival (OS), is also being used to estimate TTFT. The aim of this study was to investigate CLL-IPI, PRS, and MDACC 2011 prognostic values regarding TTFT and OS. Methods The analyzed cohort included 57 unselected Serbian CLL patients from a single institution, with the basic characteristics reflecting more aggressive disease than in the general de novo CLL population. The eligible patients were assigned investigated PMs, and TTFT and OS analyses were performed. Results Patients with higher risk scores according to CLL-IPI, PRS, and MDACC 2011 underwent treatment significantly earlier than patients with lower risk scores (p = 0.002, p = 0.019, and p lt 0.001, respectively). In multivariate analysis, MDACC 2011 and CLL-IPI retained their significance regarding TTFT (p = 0.001 and p = 0.018, respectively), while PRS did not. CLL-IPI was the only significant predictor of OS both at the univariate (p = 0.005) and multivariate (p = 0.013) levels. Conclusion CLL-IPI, PRS, and particularly MDACC 2011 are able to predict TTFT even in cohorts with more advanced-disease patients, while for prediction of OS, CLL-IPI is the only applicable among the three PMs. These results imply that PMs should be investigated in more diverse CLL populations, as it is in real-life setting

Activation of coagulation factors and prothrombic properties of endothelium in hematological malignancies

Introduction: Patients with hematological malignancies have an increased risk of thrombotic complications, ranging from 3-5% in patients with lymphoma and acute myeloid leukemia (AML). The presented study observed the onset of thrombus formation to predict risk factors for thrombosis in lymphoid and myeloid malignancies. Methods: Coagulation factors, inflammatory signaling pathways and adhesion molecules have been observed in patients with Hodgkin lymphoma (HL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL) and AML. Their mononuclear cells (MNC) trans-endothelial migration through human microvascular endothelial cells (HMEC-1) monolayer is observed by Boyden chamber. Results: Thrombin was in positive correlation with tumor necrosis factor alpha (TNF-α) in HL, while with P-selectin (p<0.001), tumor growth factor-beta (TGF-β) and factor VIII (p<0.05) in DLBCL and AML. Transendothelial migration of MNC was increased by TNF-α (p<0.001) in DLBCL regardless of previous thrombosis. Regarding coagulation, factor VIII was increased in HL and AML (p<0.05), while tissue factor in non-Hodgkin lymphomas (DLBCL and FL, p<0.05). Tissue factor was in positive correlation with adhesion molecule P-selectin and factor VIII (p<0.05). P-selectin was increased in non-Hodgkin lymphomas (p<0.0001), while TGF-β only in FL (p<0.001). Fibrinolytic activity was decreased in plasma of patients with HL, DLBCL, and FL (p<0.05), but largely in AML (p<0.01) as measured by tissue-type plasminogen activator. Inflammatory NF-κB signaling has been activated in HL and DLBCL, while p38 signaling only in HL. Conclusion: Coagulation factors and inflammation are increased in hematological malignancies along with the interaction of the endothelium and circulating cells that predispose to thrombus formatio

Increased oxidative stress in diffuse large B-cell lymphoma

Background: Oxidative stress is caused by imbalance between excessive production of reactive oxygen species and decreased capabilities of antioxidant system, and it is recognized as a feature in cancerogenesis, as well in hematologic malignancies. Previous studies have shown increasing expression of oxidative stress markers and antioxidant enzymes in lymph nodes progressing to aggressive lymphomas. Aims: The aim of our study was to assess the clinical and prognostic significance of oxidative stress markers in patients with untreated diffuse large B-cell lymphoma (DLBCL). Methods: We analysed 64 patients diagnosed with DLBLC during 2018 and 2019, while 27 healthy volunteers (51.9% males) served as a control group. The plasma sample and laboratory analyses were obtained prior to initiation of specific hematologic treatment. After completion of the therapy, the patients were followed up for up to 4 years and for each of them progression free survival (PFS) and overall survival (OS) were calculated. Malondialdehyde (MDA) and protein carbonyl (PC) were used as markers of oxidative stress in plasma of patients and volunteers, while catalase is used as an antioxidant marker. Results: The mean patients’ age was 56.2 years (range, 20–87); 51.6% were males. Majority of patients were analysed before 1L therapy (n=61; 95,3%), and had following clinical stages: Ann Arbor stage I 23.4%, stage II 37.5%, stage III 15.6% and stage IV 23.4%. Majority of the patients had satisfactory performance status (73.5% had ECOG PS ≥1), bulky tumorous mass was present in 34.4% of patients, whereas 70.3% had extranodal localisation of lymphoma. MDA (6.66±2.7 nmol/ml) was significantly increased (p<0.001, 2.6-fold), while PC (4.29±2.7 nmol/mg) was also significantly increased (p=0.0027, 5-fold) in patients with DLBCL compared to healthy volunteers. In opposite, antioxidant catalase (0.194±0.06 IU/ml) was significantly reduced (p=0.0034, 1.9-fold) in patients with DLBCL. MDA was in significant (p<0.05) negative correlation with hemoglobin (r2=0.586) and LDH (r2=0.59) levels before chemotherapy. MDA was in significant (p<0.01) positive correlation with the age (r2=0.769) of patients with DLBCL at diagnosis. Moreover, MDA was in significant positive correlation with overall survival (p<0.01, r2=0.736) and progression-free survival (p<0.01, r2=0.736) of patients with DLBCL. PC was in negative correlation with the clinical stage (r2=0.103, p=0.146) and therapy response (r2=0.136, p=0.091) of DLBCL but did not reach significance. Summary/Conclusion: The elevated oxidative markers and reduced antioxidant support oxidative stress in patients with DLBCL, while MDA can be a prognostic marker of overall survival

Inflammation mediated thrombus formation in lymphomas

Background: Patients with lymphomas increased the risk of thrombotic complications, especially in diagnosis and during chemotherapy treatment, in the range of 2.9-4.2%. Aims: Our hypothesis is that inflammation and provoked immunity are responsible for generation of thrombus due to disturbed balance between coagulation and fibrinolysis. Methods: Quantification of neutrophil extracellular traps (NETs) from peripheral blood of 80 patients with Hodgkin lymphoma (HL), diffuse large B-cell lymphoma (DLBCL), and follicular lymphoma (FL) measuring circulating cell-free DNA (cfDNA) and myeloperoxidase (MPO) activity. The inflammatory cytokines, coagulation factors and chemokines are measured by enzyme-linked immunosorbent assay (ELISA) and flow cytometry in peripheral blood, while fibrinolytic activity by fluorescent tissue-type plasminogen activator (tPA) and urokinase plasminogen activator (uPA) assays. Using a Boyden chamber, trans-endothelial migration of mononuclear cells (MNC) across a monolayer of human microvascular endothelial cells (HMEC-1) will be observed. Results: The pro-inflammatory cytokines IL-1β and TNF-α were significantly increased in DLBCL and HL, but not the chemokines IL-8 and MCP-1. NETs were increased in the peripheral blood of patients with HL (p<0.05) as measured by cfDNA and MPO activity. In contrast, cfDNA was largely reduced in DLBCL with thrombosis (p<0.001). Trans-endothelial migration of MNC was decreased by IL-6, but increased by TNF-α (p<0.001) in DLBCL with thrombosis. In the absence of thrombosis, MNC of HL demonstrated increased trans-endothelial migration in the presence of pro-inflammatory IL-6 (p<0.01), while MNC of HL and DLBCL in the presence of TNF-α (p<0.05). Regarding coagulation, factor VIII was increased in HL (p<0.05), while tissue factor in non-Hodgkin lymphomas (DLBCL and FL, p<0.05). Adhesion molecule P-selectin was increased in lymphomas, mostly in non-Hodgkin lymphomas (p<0.0001), while TGF-β is only in FL (p<0.001). Fibrinogen was negatively correlated with cfDNA (p=0.021, r=-0.767) in HL, while in positive correlation with TNF-α (p=0.028, r=0.517), IL-8 (p=0.009, r=0.598) and MCP-1 (p=0.004, r=0.643) in FL and with TGF-β (p=0.007, r=0.748) in HL. In opposite to uPA, fibrinolytic activity was decreased in the plasma of patients with HL, DLBCL, and FL (p<0.05) as measured by tPA. The tPA was in negative correlation with MPO in HL (p=0.017, r=-0.783) and FL (p=0.006, r=-0.818), while positively correlated with cfDNA in DLBCL (p=0.034, r=0.402, Table 1). The uPA was in positive correlation with cfDNA (p=0.009, r=0.692) and fibrinogen (p=0.009, r=0.692) in FL. Tissue factor (CD142+) procoagulant microparticles derived from monocytes (CD14+: 7.49±0.2, p<0.001) and activated monocytes (CD14+/CD16+: 3.75±0.8%, p<0.05) were increased in DLBCL compared to healthy controls. Summary/Conclusion: Chronic inflammation is present in the examined lymphomas where TNF-α, as an activator of the immune response, is linked with the initiation of thrombus formation. Moreover, augmented innate immunity is accompanied by procoagulants that mutually support thrombosis. F

Razvoj Hodžkinovog limfoma u ileocekalnoj regiji pacijenta sa transplantiranim bubregom na dugoročnoj imunosupresivnoj terapiji - prikaz slučaja

Following solid organ transplantation, the patient's management includes the provision of immunosuppressive therapy to the recipient. All kidney transplant recipients require lifelong immunosuppression. Regardless the improving survival following solid organ transplantation, post-transplant complications such as the development of malignancy due to immunosuppression remain to be an issue. One of the most common malignancies encountered in the post-solid organ transplant is lymphoproliferative disorder likely developed as a consequence of immunosuppression. We report a case of plasma cell type lymphoma in the ileocecal region of a 45-year-old male kidney transplant patient who was on a tacrolimus-based regimen for ten years. Although literature data indicate differently localized lymphoma as an adverse reaction to the long term use of tacrolimus, to our knowledge, this is the first described case of lymphoma in the ileocecal region. Serious adverse drug reactions and potential toxicity of tacrolimus emphasize the importance of finding the optimal balance between effective drug concentration and the risk associated with its use.Nakon transplantacije organa, pacijenti primaoci organa se stavljaju na režim imunosupresivne terapije, a u primalaca bubrega ta terapija je doživotna. Bez obzira na poboljšanje preživljavanja nakon transplantacije organa, brojne su komplikacije koje se mogu javiti nakon transplantacije, a razvoj maligniteta usled imunosupresije nastavlja da bude značajan problem. Jedan od najčešćih tipova maligniteta koji se sreće kod ovakvih pacijenata je limfoproliferativni poremećaj koji se verovatno razvija kao posledica pomenute imunosupresije. Opisan je slučaj limfoma u ileocekalnoj regiji u 45-godišnjeg muškarca sa transplantiranim bubregom, koji je bio na imunosupresivnoj terapiji zasnovanoj na takrolimusu, tokom 10 godina. Literaturni podaci ukazuju na drugačiju lokalizaciju limfoma kao neželjene reakcije nakon dugoročnog korišćenja takrolimusa, pa je prema našim saznanjima, ovo jedinstven slučaj razvoja limfoma u ileocekalnoj regiji. Ozbiljne neželjene reakcije i toksični potencijal takrolimusa naglašavaju značaj pronalaženja optimalnog balansa između efektivne koncentracije leka i rizika vezanih za njegovu upotrebu

Expression Pattern and Prognostic Significance of the Long Non-Coding RNA Metastasis-Associated Lung Adenocarcinoma Transcript 1 in Chronic Lymphocytic Leukemia

Dysregulated expression of the long non-coding RNA MALAT1 has been implicated in the pathogenesis and progression of a variety of cancers, including hematological malignancies, but it has been poorly investigated in chronic lymphocytic leukemia (CLL). In this study, the expression of MALAT1 was measured using a quantitative reverse-transcriptase polymerase chain reaction in the peripheral blood mononuclear cells of 114 unselected, newly diagnosed CLL patients in order to analyze its association with clinical, laboratory, and molecular patients’ characteristics at diagnosis, as well as its prognostic relevance. MALAT1 was found to be upregulated in CLL patients in comparison to healthy controls, and expression levels were not related to age, leukocyte, lymphocyte and platelet count, serum β2-microglobulin, and IGHV somatic hypermutational status. On the other hand, high MALAT1 expression was associated with several favorable prognostic markers (high hemoglobin, low serum lactate dehydrogenase, earlier clinical stages, CD38-negative status), but also with unfavorable cytogenetics. Furthermore, an association between high MALAT1 levels and longer time to first treatment and overall survival in IGHV-unmutated CLL subtype was observed. In summary, our results imply that high MALAT1 expression at diagnosis may be a predictor of better prognosis and point to MALAT1 expression profiling as a candidate biomarker potentially useful in clinical practice