Aplicació de la citogenètica convencional i la hibridació in situ a l'estudi de síndromes limfoproliferatives cròniques B

Descripció del recurs: 26 febrer 2002Titól obtingut de la portada digitalitzadaLes síndromes limfoproliferatives cròniques B (SLPC-B) són un grup divers de malalties que tenen en comú la proliferació neoplàsica i acumulació de cèl·lules limfoides de tipus B de tipus madur. Dins d'aquest grup, distingim bàsicament les leucèmies i els limfomes. El diagnòstic de les SLPC-B amb expressió leucèmica es basa en l'estudi integrat de la morfologia i l'immunofenotip de les cèl·lules leucèmiques, així com de les característiques citogenètiques i moleculars. Existeixen una sèrie d'entitats que poden ser confoses fàcilment i requereixen un estudi exhaustiu que permeti la seva classificació correcta. En el present treball s'analitzen tres d'aquestes entitats, la Leucèmia Limfàtica Crònica B atípica (LLC-Ba), el Limfoma de Cèl·lules del Mantell (LCM) i el Limfoma Esplènic de la Zona Marginal (LEZM) amb la intenció de trobar marcadors citogenètics que permetin separar-les, de manera que sigui possible establir un diagnòstic diferencial amb certesa. També es fa esment de la limfocitosi B Policlonal Persistent (LBPP). S'han estudiat 27 LLC-Ba, 13 LCM, 19 LEZM i 1 LBPP amb tècniques de citogenètica convencional i hibridació in situ, amb sondes centromèriques pels cromosomes 3, 12, 17 i 18, de locus específic per les regions 13q14 i 17p13 i de pintat cromosòmic. Entre les LLC-Ba, un 37% han presentat alteracions citogenètiques i l'alteració citogenètica més freqüent ha estat la trisomia 12 (63%), mentre que les delecions de 13q14 i 17p13 s'han detectat en % molt baix. Respecte els LCM, el 77% han presentat alteracions citogenètiques i dins d'aquests, en un 70% s'ha detectat la t(11;14)(q13;q32), alteració citogenètica característica d'aquesta entitat. Altres cromosomes implicats han estat: 1, 2, 9, 13 i 17 en forma d'alteracions estructurals principalment. No s'ha detectat la presència de trisomia 12 en cap pacient. En els LEZM, un 58% han presentat un cariotip alterat. Els cromosomes més freqüentment implicats han estat: 1, 3, 7, 8 i 14. Dels estudis inclosos en la tesi i d'altres posteriors s'han pogut definir la trisomia parcial del braç llarg del cromosoma 3 (+3q) i la delecioó del braç llarg del cromosoma 7 (del(7q)) com a alteracions més freqüents. No s'ha detectat presència de t(11;14) en cap dels casos, i la trisomia 12, així com la deleció de 17p13 només s'ha detectat en un pacient. D'aquest estudi es conclueix que les alteracions citogenètiques dels tres grups són clarament diferents i que el seu coneixement permet establir un millor diagnòstic diferencial d'aquestes malalties. La LBPP és una entitat que encara no se sap si és una neoplàsia de progressió lenta, o un fenòmen reactiu. S'ha inclòs en l'estudi amb la intenció de fer esment de la seva existència i de la possibilitat de ser confosa amb alguna de les altres entitats.B-cell lymphoproliferative disorders (BCLPD) constitute a group of pathologies in which mature B cells proliferate and accumulate in different tissues. Among BCLPD, leukaemic and lymphomatous diseases can be distinguished. In the present work, BCLPD with leukaemic expression are studied, taking into account at the same time morphologic, immunophenotypic, cytogenetic and molecular data. It is remarkable that the diagnosis of some BCLPD with leukaemic expression can be difficult because of some similar characteristics. In these cases, an exhaustive study will be required to a correct classification. In this thesis, three different entities would be analyzed: atypical B-cell lymphocytic leukaemia (aCLL), mantle cell lymphoma (MCL) and splenic marginal zone B-cell lymphoma (SMZBCL). The aim of the present study was to study the above mentioned entities by means of conventional cytogenetics and in situ hybridization techniques to analyze if different cytogenetic markers could be identified to establish a differential diagnosis among them. In addition, persistent polyclonal B-cell lymphocytosis (PPBL) was included in the study. Twenty seven aCLL, 13 MCL, 19 SMZBCL and 1 (PPBL) were studied cytogenetically and using in situ hybridization probes for centromeric regions of chromosomes 3, 12, 17 and 18, and for specific locus 13q14 (Rb) and 17p13 (p53). Among aCLL, 37% presented cytogenetic aberrations, being trisomy 12 the most frequent abnormality (63%), besides, 13q14 and 17p13 deletions were found in low percentage. Regarding MCL, chromosomal aberrations were found in 77% of patients, and among them, 70% presented a t(11;14)(q13;q32). Other affected chromosomes were 1, 2, 9, 13 and 17. No patient presented trisomy 12. In SMZBCL group, an abnormal karyotype was found in 58% of patients. The most frequently involved chromosomes were 1, 3, 7, 8 and 14. The most frequent structural abnormalities associated to SMZBCL were +3q and del(7q). No case presented +12, nor t(11;14)(q13;q32), and only one case showed 17p13 deletion. In conclusion, cytogenetic abnormalities seem to be different in each studied group, and its knowledge could be useful to establish a differential diagnosis among these pathologies. PPBL still is a controversial entity, because it is not clear if it represents a neoplastic disease with a very slow progression rate or if it represents a reactive phenomena

Oligonucleotide array-CGH identifies genomic subgroups and prognostic markers for tumor stage mycosis fungoides

Mycosis fungoide (MF) patients who develop tumors or extracutaneous involvement usually have a poor prognosis with no curative therapy available so far. In the present European Organization for Research and Treatment of Cancer (EORTC) multicenter study, the genomic profile of 41 skin biopsies from tumor stage MF (MFt) was analyzed using a high-resolution oligo-array comparative genomic hybridization platform. Seventy-six percent of cases showed genomic aberrations. The most common imbalances were gains of 7q33.3q35 followed by 17q21.1, 8q24.21, 9q34qter, and 10p14 and losses of 9p21.3 followed by 9q31.2, 17p13.1, 13q14.11, 6q21.3, 10p11.22, 16q23.2, and 16q24.3. Three specific chromosomal regions, 9p21.3, 8q24.21, and 10q26qter, were defined as prognostic markers showing a significant correlation with overall survival (OS) (P=0.042, 0.017, and 0.022, respectively). Moreover, we have established two MFt genomic subgroups distinguishing a stable group (0-5 DNA aberrations) and an unstable group (>5 DNA aberrations), showing that the genomic unstable group had a shorter OS (P=0.05). We therefore conclude that specific chromosomal abnormalities, such as gains of 8q24.21 (MYC) and losses of 9p21.3 (CDKN2A, CDKN2B, and MTAP) and 10q26qter (MGMT and EBF3) may have an important role in prognosis. In addition, we describe the MFt genomic instability profile, which, to our knowledge, has not been reported earlier

Reduced expansion of CD94/NKG2C + NK cells in chronic lymphocytic leukemia and CLL-like monoclonal B-cell lymphocytosis is not related to increased human cytomegalovirus seronegativity or NKG2C deletions

Data de publicació electrònica: 22-02-2021Introduction: Dysregulated NK cell-mediated immune responses contribute to tumor evasion in chronic lymphocytic leukemia (CLL), although the NK cell compartment in CLL-like monoclonal B-cell lymphocytosis (MBL) is poorly understood. In healthy individuals, human cytomegalovirus (HCMV) induces the expansion of NK cells expressing high levels of CD94/NKG2C NK cell receptor (NKR) specific for HLA-E. Methods: We analyzed the expression of NKG2A, NKG2C, ILT2, KIR, CD161, and CD57 in 24 MBL and 37 CLL. NKG2C was genotyped in these patients and in 81 additional MBL/CLL, while NKG2C gene expression was assessed in 26 cases. In 8 CLL patients with increased lymphocytosis (≥20 × 109 /L), tumor HLA-E and HLA-G expression was evaluated. Results: NKR distribution did not significantly differ between MBL and CLL patients, although they exhibited reduced NKG2C+ NK cells compared with a non-CLL group (4.6% vs 12.2%, P = .012). HCMV+ patients showed increased percentages of NKG2C+ NK cells compared with HCMV- (7.3% vs 2.9%, P = .176). Frequencies of NKG2C deletions in MBL/CLL were similar to those of the general population. Low/undetectable NKG2C expression was found among NKG2C+/- (45%) and NKG2C+/+ (12%) patients. CLL cases with increased lymphocytosis displayed especially reduced NKG2C expression (1.8% vs 8.1%, P = .029) and tumor cells with high HLA-E (>98%) and variable HLA-G expression (12.4%, range: 0.5-56.4). CLL patients with low NKG2C expression (<7%) showed shorter time to first treatment (P = .037). Conclusion: Reduced percentages of CD94/NKG2C+ NK cells were observed in CLL and MBL patients independently of HCMV serostatus and NKG2C zygosity, particularly in CLL patients with increased lymphocytosis, which could potentially be related to the exposure to tumor cells

Trisomy 8, a cytogenetic abnormality in Myelodysplastic Syndromes, is constitutional or not?

Isolated trisomy 8 is not considered presumptive evidence of myelodysplastic syndrome (MDS) in cases without minimal morphological criteria. One reason given is that trisomy 8 (+8) can be found as a constitutional mosaicism (cT8M). We tried to clarify the incidence of cT8M in myeloid neoplasms, specifically in MDS, and the diagnostic value of isolated +8 in MDS. Twenty-two MDS and 10 other myeloid neoplasms carrying +8 were studied. Trisomy 8 was determined in peripheral blood by conventional cytogenetics (CC) and on granulocytes, CD3+ lymphocytes and oral mucosa cells by fluorescence in situ hybridization (FISH). In peripheral blood CC, +8 was seen in 4/32 patients. By FISH, only one patient with chronic myelomonocytic leukemia showed +8 in all cell samples and was interpreted as a cT8M. In our series +8 was acquired in all MDS. Probably, once discarded cT8M by FISH from CD3+ lymphocytes and non-hematological cells, +8 should be considered with enough evidence to MDS

Patients with chronic lymphocytic leukemia and complex karyotype show an adverse outcome even in absence of TP53/ATM FISH deletions

Genomic complexity identified by chromosome banding analysis (CBA) predicts a worse clinical outcome in CLL patients treated either with standard or new treatments. Herein, we analyzed the clinical impact of complex karyotypes (CK) with or without high-risk FISH deletions (ATM and/or TP53, HR-FISH) in a cohort of 1045 untreated MBL/CLL patients. In all, 99/1045 (9.5%) patients displayed a CK. Despite ATM and TP53 deletions were more common in CK (25% vs 7%; P < 0.001; 40% vs 5%; P < 0.001, respectively), only 44% (40/90) patients with TP53 deletions showed a CK. CK group showed a significant higher two-year cumulative incidence of treatment (48% vs 20%; P < 0.001), as well as a shorter overall survival (OS) (79 mo vs not reached; P < 0.001). When patients were categorized regarding CK and HR-FISH, those with both characteristics showed the worst median OS (52 mo) being clearly distinct from those non-CK and non-HR-FISH (median not reached), but no significant differences were detected between cases with only CK or HR-FISH. Both CK and TP53 deletion remained statistically significant in the multivariate analysis for OS. In conclusion, CK group is globally associated with advanced disease and poor prognostic markers. Further investigation in larger cohorts with CK lacking HR-FISH is needed to elucidate which mechanisms underlie the poor outcome of this subgrou

Oligonucleotide array-CGH identifies genomic subgroups and prognostic markers for tumor stage mycosis fungoides

Mycosis fungoide (MF) patients who develop tumors or extracutaneous involvement usually have a poor prognosis with no curative therapy available so far. In the present European Organization for Research and Treatment of Cancer (EORTC) multicenter study, the genomic profile of 41 skin biopsies from tumor stage MF (MFt) was analyzed using a high-resolution oligo-array comparative genomic hybridization platform. Seventy-six percent of cases showed genomic aberrations. The most common imbalances were gains of 7q33.3q35 followed by 17q21.1, 8q24.21, 9q34qter, and 10p14 and losses of 9p21.3 followed by 9q31.2, 17p13.1, 13q14.11, 6q21.3, 10p11.22, 16q23.2, and 16q24.3. Three specific chromosomal regions, 9p21.3, 8q24.21, and 10q26qter, were defined as prognostic markers showing a significant correlation with overall survival (OS) (P=0.042, 0.017, and 0.022, respectively). Moreover, we have established two MFt genomic subgroups distinguishing a stable group (0-5 DNA aberrations) and an unstable group (>5 DNA aberrations), showing that the genomic unstable group had a shorter OS (P=0.05). We therefore conclude that specific chromosomal abnormalities, such as gains of 8q24.21 (MYC) and losses of 9p21.3 (CDKN2A, CDKN2B, and MTAP) and 10q26qter (MGMT and EBF3) may have an important role in prognosis. In addition, we describe the MFt genomic instability profile, which, to our knowledge, has not been reported earlier

Oligonucleotide array-CGH identifies genomic subgroups and prognostic markers for tumor stage mycosis fungoides

Mycosis fungoide (MF) patients who develop tumors or extracutaneous involvement usually have a poor prognosis with no curative therapy available so far. In the present European Organization for Research and Treatment of Cancer (EORTC) multicenter study, the genomic profile of 41 skin biopsies from tumor stage MF (MFt) was analyzed using a high-resolution oligo-array comparative genomic hybridization platform. Seventy-six percent of cases showed genomic aberrations. The most common imbalances were gains of 7q33.3q35 followed by 17q21.1, 8q24.21, 9q34qter, and 10p14 and losses of 9p21.3 followed by 9q31.2, 17p13.1, 13q14.11, 6q21.3, 10p11.22, 16q23.2, and 16q24.3. Three specific chromosomal regions, 9p21.3, 8q24.21, and 10q26qter, were defined as prognostic markers showing a significant correlation with overall survival (OS) (P=0.042, 0.017, and 0.022, respectively). Moreover, we have established two MFt genomic subgroups distinguishing a stable group (0-5 DNA aberrations) and an unstable group (>5 DNA aberrations), showing that the genomic unstable group had a shorter OS (P=0.05). We therefore conclude that specific chromosomal abnormalities, such as gains of 8q24.21 (MYC) and losses of 9p21.3 (CDKN2A, CDKN2B, and MTAP) and 10q26qter (MGMT and EBF3) may have an important role in prognosis. In addition, we describe the MFt genomic instability profile, which, to our knowledge, has not been reported earlier

Patients with chronic lymphocytic leukemia and complex karyotype show an adverse outcome even in absence of TP53/ATM FISH deletions

Genomic complexity identified by chromosome banding analysis (CBA) predicts a worse clinical outcome in CLL patients treated either with standard or new treatments. Herein, we analyzed the clinical impact of complex karyotypes (CK) with or without high-risk FISH deletions (ATM and/or TP53, HR-FISH) in a cohort of 1045 untreated MBL/CLL patients. In all, 99/1045 (9.5%) patients displayed a CK. Despite ATM and TP53 deletions were more common in CK (25% vs 7%; P < 0.001; 40% vs 5%; P < 0.001, respectively), only 44% (40/90) patients with TP53 deletions showed a CK. CK group showed a significant higher two-year cumulative incidence of treatment (48% vs 20%; P < 0.001), as well as a shorter overall survival (OS) (79 mo vs not reached; P < 0.001). When patients were categorized regarding CK and HR-FISH, those with both characteristics showed the worst median OS (52 mo) being clearly distinct from those non-CK and non-HR-FISH (median not reached), but no significant differences were detected between cases with only CK or HR-FISH. Both CK and TP53 deletion remained statistically significant in the multivariate analysis for OS. In conclusion, CK group is globally associated with advanced disease and poor prognostic markers. Further investigation in larger cohorts with CK lacking HR-FISH is needed to elucidate which mechanisms underlie the poor outcome of this subgroup

Increased risk of revision of cementless stemmed total hip arthroplasty with metal-on-metal bearings

<div><p><b>Background and purpose —</b> Data from the national joint registries in Australia and England and Wales have revealed inferior medium-term survivorship for metal-on-metal (MoM) total hip arthroplasty (THA) than for metal-on-polyethylene (MoP) THA. Based on data from the Nordic Arthroplasty Register Association (NARA), we compared the revision risk of cementless stemmed THA with MoM and MoP bearings and we also compared MoM THA to each other.</p><p><b>Patients and methods —</b> We identified 32,678 patients who were operated from 2002 through 2010 with cementless stemmed THA with either MoM bearings (11,567 patients, 35%) or MoP bearings (21,111 patients, 65%). The patients were followed until revision, death, emigration, or the end of the study period (December 31, 2011), and median follow-up was 3.6 (interquartile range (IQR): 2.4–4.8) years for MoM bearings and 3.4 (IQR: 2.0–5.8) years for MoP bearings. Multivariable regression in the presence of competing risk of death was used to assess the relative risk (RR) of revision for any reason (with 95% confidence interval (CI)).</p><p><b>Results —</b> The cumulative incidence of revision at 8 years of follow-up was 7.0% (CI: 6.0–8.1) for MoM bearings and 5.1% (CI: 4.7–5.6) for MoP bearings. At 6 years of follow-up, the RR of revision for any reason was 1.5 (CI: 1.3–1.7) for MoM bearings compared to MoP bearings. The RR of revision for any reason was higher for the ASR (adjusted RR = 6.4, CI: 5.0–8.1), the Conserve Plus (adjusted RR = 1.7, CI: 1.1–2.5) and “other” acetabular components (adjusted RR = 2.4, CI: 1.5–3.9) than for MoP THA at 6 years of follow-up.</p><p><b>Interpretation —</b> At medium-term follow-up, the survivorship for cementless stemmed MoM THA was inferior to that for MoP THA, and metal-related problems may cause higher revision rates for MoM bearings with longer follow-up.</p></div