17 research outputs found
Association of human concentrative nucleoside transporter 3 gene expression and CYP2B6*6 Š°llele with the response to fludarabine plus cyclophosphamide in chronic lymphocytic leukemia patients
Hemioterapija na bazi fludarabina i ciklofosfamida predstavlja osnov leÄenja
pacijenata sa hroniÄnom limfocitnom leukemijom (HLL) mlaÄe životne dobi i bez znaÄajnih
komorbiditeta. Kod nekih pacijenata efekat terapije može biti uslovljen aberantnom
ekspresijom i/ili mutacijama gena koji utiÄu na farmakodinamiku i farmakokinetiku
antileukemijskih lekova. Sa druge strane, ustanovljeni su brojni kliniÄko-laboratorijski i
molekularno-genetiÄki markeri prognoze koji su znaÄajni u predikciji kliniÄkog toka,
prvenstveno vremena do prve terapije (time to first treatment, TTFT) i ukupnog
preživljavanja (overall survival, OS). Oni se poslednjih godina u cilju jaÄanja prognostiÄke
vrednosti kombinuju unutar razliÄitih prognostiÄkih modela, meÄu kojima su za predikciju
TTFT naroÄito 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 predviÄanje OS najznaÄajniji Internacionalni prognostiÄki indeks za HLL
(The International Prognostic Index for CLL, CLL-IPI). Cilj rada je da se ispita uticaj
farmakogenetike, taÄnije ekspresije gena SLC28A3 koji kodira hCNT3 protein, jedan od
transportera za fludarabin, i varijantnog alela CYP2B6*6 gena za citohrom P450 2B6, koji
uÄestvuje u metabolizmu ciklofosfamida, na ishod leÄenja FC protokolom kod pacijenata sa
HLL-om. TakoÄe, 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. KonaÄno, ispitaÄe se veza farmakogenetiÄkih karakteristika i
skorova.
Metodologija: U studiju je ukljuÄeno 57 pacijenata sa HLL-om koji su leÄeni FC
protokolom, veÄinom u prvoj liniji. Pacijenti su stratifikovani prema skorovima CLL-IPI,
PRS i MDACC 2011, nakon Äega je ispitana njihova prognostiÄka 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
HroniÄna limfocitna leukemija (HLL) izuzetno je heterogena bolest varijabilnog kliniÄkog toka. S jedne strane imamo, pacijente sa agresivnom i rezistentnom boleÅ”Äu od koje umiru svega par meseci nakon dijagnoze, dok s druge strane spektra postoje pacijenti sa indolentnom, sporo progredirajuÄom boleÅ”Äu koja ne zahteva leÄenje decenijama. Razlozi su samo delimiÄno poznati i veÄ su decenijama unazad tema mnogobrojnih nauÄnih istraživanja. Tako je razvijen koncept prognostiÄkih i prediktivnih faktora u HLL-u, koji imaju za cilj da predvide kliniÄki tok, odnosno terapijski ishod HLL-a. Liste prognostiÄkih i prediktivnih faktora su, sa boljim poznavanjem patofiziologije ove bolesti, svakom godinom sve duže, ali se i meÄusobno preklapaju. U ovom revijalnom radu izabrali smo aberacije TP53 gena i mutacioni status rearanžiranih IGHV (engl. immunoglobulin heavy variable) gena kao dva najznaÄajnija i najprouÄavanija faktora koji imaju i prognostiÄki i prediktivni znaÄaj.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 oļæ½ļæ½ļæ½ļæ½ļæ½ 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 ā„ 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 seļæ½ļæ½ļæ½ļæ½ļæ½ 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
aļæ½ļæ½ļæ½ļæ½ļæ½ er therapy initi ati on and to detect minimal residual disease aļæ½ļæ½ļæ½ļæ½ļæ½ 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-ĪŗB 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 patientsā 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-ĪŗB and PI3K signaling pathways, to evaluate
S100A4, S100A8, S100A9, S100A12 and NF-ĪŗB 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-ĪŗB 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 hroniÄne limfocitne leukemije (HLL) znaÄajno je unapreÄena u poslednje vreme. MeÄu nekoliko prognostiÄkih modela Äiji je cilj predviÄanje 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 prognostiÄki 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 znaÄaj pomenutih prognostiÄkih modela u pogledu predviÄanja TTFT i OS. Metode Analizirana kohorta je obuhvatila 57 neselektovanih bolesnika sa HLL Univerzitetskog kliniÄkog centra Srbije sa proseÄno agresivnijim profilom bolesti u odnosu na opÅ”tu populaciju de novo bolesnika sa HLL. Bolesnici su ocenjivani prema navedenim skorovima uz analizu TTFT i OS. Rezultati Bolesnici sa viÅ”im vrednostima CLL-IPI, PRS i MDACC 2011 primili su prvu terapiju znaÄajno ranije u poreÄenju 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 prognostiÄki znaÄaj u predikciji TTFT (p = 0,001, odnosno p = 0,018), dok je PRS ovaj znaÄaj izgubio. CLL-IPI je bio jedini znaÄajan prediktor OS u univarijantnoj (p = 0,005) i u multivarijantnoj analizi (p = 0,013). ZakljuÄak CLL-IPI, PRS i naroÄito MDACC 2011 su dobri prediktori TTFT Äak i u kohortama bolesnika sa agresivnijom boleÅ”Äu, dok je za predikciju OS od ova tri prognostiÄka modela CLL-IPI jedini primenljiv. Ovi rezultati pokazuju da bi prognostiÄke modele trebalo ispitati na bolesnicima sa HLL u razliÄitim fazama bolesti, kakvi se sreÄu u realnoj kliniÄkoj 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-Ī±) in HL, while with P-selectin (p<0.001), tumor growth factor-beta (TGF-Ī²) and factor VIII (p<0.05) in DLBCL and AML. Transendothelial migration of MNC was increased by TNF-Ī± (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-Ī² 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-ĪŗB 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 patientsā age was 56.2 years (range, 20ā87); 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 ā„1), bulky tumorous mass was present in 34.4% of patients, whereas 70.3% had extranodal localisation of lymphoma. MDA (6.66Ā±2.7 nmol/ml) was significantly increased (p<0.001, 2.6-fold), while PC (4.29Ā±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 (0.194Ā±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-1Ī² and TNF-Ī± 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-Ī± (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-Ī± (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-Ī² 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-Ī± (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-Ī² (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+: 7.49Ā±0.2, p<0.001) and activated monocytes (CD14+/CD16+: 3.75Ā±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-Ī±, 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.
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Razvoj Hodžkinovog limfoma u ileocekalnoj regiji pacijenta sa transplantiranim bubregom na dugoroÄnoj imunosupresivnoj terapiji - prikaz sluÄaja
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 poboljÅ”anje preživljavanja nakon transplantacije organa, brojne su komplikacije koje se mogu javiti nakon transplantacije, a razvoj maligniteta usled imunosupresije nastavlja da bude znaÄajan problem. Jedan od najÄeÅ”Äih tipova maligniteta koji se srecĢe kod ovakvih pacijenata je limfoproliferativni poremeÄaj koji se verovatno razvija kao posledica pomenute imunosupresije. Opisan je sluÄaj limfoma u ileocekalnoj regiji u 45-godiÅ”njeg muÅ”karca sa transplantiranim bubregom, koji je bio na imunosupresivnoj terapiji zasnovanoj na takrolimusu, tokom 10 godina. Literaturni podaci ukazuju na drugaÄiju lokalizaciju limfoma kao neželjene reakcije nakon dugoroÄnog koriÅ”Äenja takrolimusa, pa je prema naÅ”im saznanjima, ovo jedinstven sluÄaj razvoja limfoma u ileocekalnoj regiji. Ozbiljne neželjene reakcije i toksiÄni potencijal takrolimusa naglaÅ”avaju znaÄaj pronalaženja optimalnog balansa izmeÄu 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 patientsā 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 Ī²2-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