scholarly journals Recurrent Chromosomal Abnormalities in Tumoral Lesions of Small Lymphocytic Lymphoma/Chronic Lymphocytic Leukemia: A Large-Scale Fluorescent in-Situ Hybridization Study on Tissue Biopsy Sections

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 4282-4282
Author(s):  
Pedro Horna ◽  
Kathryn E. Pearce ◽  
Rhett P. Ketterling ◽  
Jess Peterson
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Chromosomal Abnormalities ◽  
Lymphocytic Leukemia ◽  
Small Lymphocytic Lymphoma ◽  
Tissue Sections ◽  
Tissue Biopsy ◽  
Trisomy 12

Background: Small lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL) is a lymphoproliferative disorder of small mature B-cells, most commonly presenting with peripheral blood and bone marrow involvement. Tumoral lesions are variably encountered, resulting from either lymph node involvement or, less commonly, infiltration into virtually any extramedullary site. The prognostic assessment of patients with SLL/CLL relies strongly on the identification of recurrent chromosomal abnormalities, which are routinely tested by fluorescence in situ hybridization (FISH) on peripheral blood or bone marrow specimens. However, the incidence of such recurrent chromosomal abnormalities on tumoral lesions has only been studied on small series, and its concordance with peripheral blood and/or bone marrow testing remains unknown. We hereby report a large series of chromosomal abnormalities detected on tumoral SLL/CLL lesions, based on a validated FISH panel for formalin-fixed/paraffin-embedded (FFPE) biopsy sections. Methods: FISH was performed at Mayo Clinic on FFPE tissue sections from biopsies obtained from May 2014 to November 2018, as clinically indicated. Pathology reports and H&E-stained sections were reviewed and were consistent with a diagnosis of SLL/CLL in all cases. Probes were validated to detect previously described chromosomal abnormalities in SLL/CLL, including trisomy 12 (+12) and deletions of 6q23 (6q-), 11q22.3 (11q-), 13q14.3 (13q-) and 17p13.1 (17p-). Available FISH results performed on peripheral blood or bone marrow samples from these patients were also reviewed for comparison. Results: Tissue biopsies involved by SLL/CLL from 346 patients were evaluated by FISH. The majority of specimens were either lymph nodes (65%) or soft tissue masses (29%). The median age was 66.9 years (range: 36 to 91), and the male to female ratio was 2.1:1. FISH abnormalities were identified in 60% of evaluated tissue sections. The most frequently detected aberration was +12 (35%), followed by 13q- (24%), 11q- (15%), 17p- (6%) and 6q- (2%) (Figure A). In particular, the incidence of +12 was significantly higher, and the incidence of 13q- significantly lower compared to frequencies previously reported on peripheral blood or bone marrow specimens (p<0.01 for both abnormalities, in comparison to a recently published cohort of 1585 patients: Br J Haematol 2016;173:105). Most cases had 1 abnormality (55%), with fewer patients having 2 abnormalities (13%), and only 1 case showing 3 abnormalities. Of 47 patients with 2 or more chromosomal aberrations, most (72%) had 13q- in combination with either 11q- (7 cases), +12 (10 cases) or 17p- (7 cases) (Figure 1B). Of 29 patients with positive blood or bone marrow FISH results within 12 months of tissue biopsy, 7 patients (24%) had discrepant results, all of which were limited to discordant +12 and/or 13q- detection. In 14 additional patients with positive blood or bone marrow FISH results more than 12 months before or after tissue biopsy, discordant results were found in 3 cases (21%), all of which indicated gains of 17p- (2 cases) or 11q- (1 case) on the follow-up analysis. Conclusions: We hereby report the largest documented series (to the best of our knowledge) of FISH results performed on tumoral lesions of SLL/CLL. Trisomy 12 was overrepresented in tumoral lesions, as compared to the reported predominance of 13q- in leukemic involvement. Discrepant results between concurrent tumoral and leukemic FISH testing were limited to differences in +12 and 13q- in our series. We also document the acquisition of aberrancies of unfavorable prognosis (17p- and 11q-) on follow up FISH analysis in a small subset of patients, which should be taken into account when considering repeat FISH studies. Figure Disclosures Horna: MorphoSys AG: Research Funding.

Detection of Chromosomal Abnormalities in Chronic Lymphocytic Leukemia Increased by Interphase Fluorescence In Situ Hybridization in TPA-Stimulated Peripheral Blood Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4927-4927
Author(s):  
Anna Aventin ◽  
Jana Sanchez
Keyword(s):  
In Situ Hybridization ◽  
Chronic Lymphocytic Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Chromosomal Abnormalities ◽  
Lymphocytic Leukemia ◽  
Peripheral Blood Cells ◽  
Trisomy 12

Abstract Chromosomal abnormalities, namely deletion 11q-,13q-, 17p- and trisomy 12, have prognostic significance for patients with chronic lymphocytic leukemia (CLL). Several studies have demonstrated that the interphase fluorescence in situ hybridization technique (I-FISH) in CLL identifies such genomic aberrations in a higher frequency than classical karyotyping, including stimulated cultures using B-cell specific mitogens. However, there appears to be no information in the literature comparing I-FISH on non-cultured and cultured cells in CLL. A total of 56 samples from 49 patients with CLL were studied using commercially available probes for chromosomes 11q22.3(ATM), 13q14(13S272), 17p13(p53) and 12 centromere(D12Z3). We compared the results obtained by I-FISH-PBMC and those by interphase fluorescence in situ hybridization on TPA-stimulated peripheral blood cells (I-FISH-TPA) used for conventional cytogenetics in order to evaluate the usefulness of I-FISH-TPA. The proportion of abnormal nuclei obtained with the I-FISH-TPA was higher than that found with I-FISH-PBMC (P<0.001). Consequently, 15 cases with a negative or borderline result by I-FISH-PBMC became positive by I-FISH-TPA for deletion 11q- (n=2), 13q- (n= 9) and trisomy 12 (n=4). In all but one of these, chromosomal abnormalities were reconfirmed by either metaphase-FISH or conventional G-banding. Disease detection thus increased from 51% with I-FISH-PBMC to 78% with I-FISH-TPA. Interestingly, all 15 cases which reached the diagnostic thresholds for deletion 11q-,13q- and trisomy 12 had a slight lymphocytosis. An absolute lymphocyte count <8.7×109/l was found to be the critical threshold (P=0.037) below which I-FISH-TPA should be performed rather than I-FISH-PBMC. We have shown that I-FISH-TPA can not only detect a higher proportion of abnormal interphase nuclei but can also identify abnormal CLL cases which may be overlooked by I-FISH-PBMC, especially those with low absolute lymphocyte counts. I-FISH-TPA is thus a reliable technique for clinical diagnostics in CLL.

Interphase Fluorescence In Situ Hybridization Detection of Cytogenetic Abnormalities in B-Cell Chronic Lymphocytic Leukemia.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4992-4992
Author(s):  
Wei Xu ◽  
Jianyong Li ◽  
Jinlan Pan ◽  
Li Li ◽  
Hairong Qiu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Situ Hybridization ◽  
B Cell ◽  
Fluorescence In Situ Hybridization ◽  
Peripheral Blood ◽  
Chromosome Aberrations ◽  
Lymphocytic Leukemia ◽  
Chromosome 12 ◽  
Molecular Cytogenetic

Abstract The most frequent chromosomal abnormalities in B-cell chronic lymphocytic leukaemia (B-CLL) are deletions on 13q14 and 17p13, trisomy 12 and 14q32 rearrangement. Conventional metaphase cytogenetic analysis underestimates the frequency of specific chromosome aberrations in B-CLL due to the low rate of spontaneous mitoses and poor response to mitogen stimulation. The aim of this study was to investigate the incidence of chromosomal changes in bone marrow or peripheral blood cells (or both) of B-CLL patients using a molecular cytogenetic method, interphase fluorescence in situ hybridization (I-FISH). Probes for 13q14 (D13S319), 17p13 (P53 gene), the centromere of chromosome 12 (D12Z3) and 14q32 (Ig10 and Y6) were applied to detect chromosomal aberrations on bone marrow and peripheral blood smears from 83 B-CLL patients (60 male, 23 female,). Molecular cytogenetic aberrations were found in 60 (72.3%) cases, and 8 (9.6%) patients showed two kinds of abnormalities. The most frequent abnormalities detected in our patients was deletions of 13q14 in 34 cases (41.0%), followed by trisomy of chromosome 12 in 16 patients (19.3%), deletions of 17p13 in 10 patients (12%) and 14q32 rearrangement in 8 patients (9.6%). Statistical analyses were performed to correlate the molecular cytogenetic findings with Binet stages. No apparent differences in distribution were noted for anomalies del(13q14), del(17p13), +12 or 14q32 rearrangement among patients with various Binet stages. FISH was found to be a more rapid, exact and sensitive technique for the analysis of chromosome aberrations in CLL. FISH could provide accurate information of molecular cytogenetics for CLL.

Cyclin D1 mRNA Expression In CLL: a Pilot Study.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4619-4619
Author(s):  
Heidi Mocikova ◽  
Sona Pekova ◽  
Lenka Zejskova ◽  
Martin Spacek ◽  
Tomas Kozak
Keyword(s):  
Bone Marrow ◽  
Overall Survival ◽  
Prognostic Factors ◽  
Cyclin D1 ◽  
Mrna Expression ◽  
Peripheral Blood ◽  
Newly Diagnosed ◽  
Rt Pcr ◽  
Trisomy 12

Abstract Abstract 4619 Background. Expression of cyclin D1 demonstrated by immunohistochemistry is seen in 95% of mantle cell lymphomas and in other lymphoproliferative diseases including 10% of chronic lymphocytic leukemias (CLL). This study analyzed the impact of cyclin D1 positivity on time to treatment (TTT) and overall survival (OS) measured by RT PCR in newly diagnosed CLL patients and correlation with reported prognostic factors. Patients and methods. Level of cyclin D1 (quantitative real-time RT-PCR with a specific TaqMan flurescent hybridization probe) mRNA expression was analysed in 72 samples (57 peripheral blood and 15 bone marrow) from patients with newly diagnosed CLL. Cyclin D1 expression (cut-off according to ROC curve >3 over the threshold expression in healthy donors) was reported as positive. Fisher's exact test was used to analyze the relationship of cyclin D1 positivity and prognostic factors: del17p, del11q, unmutated IgVH, trisomy 12, ZAP 70 and CD38 positivity, elevated B2microglobulin, elevated LDH and lymphocyte doubling time (LDT) <6 months. The comparison of time to treatment (TTT) and prognostic factors with cyclin D1 positivity was calculated via Spearman correlation coefficient. Survival curves were calculated by Kaplan-Meier survival analysis and comparison between subgroups was performed by the log-rank test. Results. Cyclin D1 was positive in 29 (40%) CLL patients. Although cyclin D1 was not statistically significant for TTT (P=0,145), a trend was observed, suggesting a negative prognostic impact of cyclin D1 overexpression in CLL. Following variables correlated significantly with TTT: del17p (P=0.037), del11q (P=0.003), unmutated IgVH (P=0.004), trisomy 12(P=0.024), positive CD38 (P=0.014), elevated B2microglobulin (P<0.001), elevated LDH (P<0.001) and LDT <6 months (P<0.001). Del 17p, del 11q, trisomy 12 and elevated B2microglobulin were independent factors for TTT in the multivariate analysis. None of these factors were significant for overall survival due to the short follow-up. Conclusion. Cyclin D1 measured by RT-PCR from peripheral blood or bone marrow has no statistically singificant impact on TTT or OS, though a trend pointing to cyclin D1 overexpression in CLL as a negative prognostic marker can be suggested. This data should be confirmed on a larger cohort of patients with a longer follow-up. Disclosures: No relevant conflicts of interest to declare.

The Use of Tetradenoylphorbol Acetate Stimulated Peripheral Blood Cells for Interphase Fluorescence In-Situ Hybridization Analysis May Enhance Its Prognostic Value for Patients with Chronic Lymphocytic Leukemia

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4152-4152
Author(s):  
Julio Delgado ◽  
Anna Aventin ◽  
Javier Briones ◽  
Jana Sanchez ◽  
Josep Nomdedeu ◽  
...  
Keyword(s):  
Overall Survival ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Prognostic Value ◽  
Mononuclear Cells ◽  
Lymphocytic Leukemia ◽  
Peripheral Blood Cells ◽  
Peripheral Blood Mononuclear ◽  
Trisomy 12

Abstract Chronic lymphocytic leukemia (CLL) is a mature B-cell malignancy characterized by a variable clinical course. Genomic aberrations, as identified by interphase fluorescent in situ hybridization (I-FISH), have a remarkable predictive power in terms of response to therapy, time to first treatment and overall survival of these patients. I-FISH studies can be performed either on unstimulated peripheral blood mononuclear cells (I-FISH-PBMC) or on tetradecanoylphorbol acetate (TPA) stimulated peripheral blood cells (I-FISH-TPA). We have previously observed that I-FISH-TPA could identify genomic abnormaties that might be overlooked with I-FISH-PBMC. The aim of the study was to evaluate whether this increased detection of genomic abnormalities, as identified by I-FISH-TPA, was clinically relevant for a group of consecutive CLL patients. Blood samples from 47 CLL patients were stimulated with TPA and cultured in RPMI medium supplemented with fetal calf serum. Peripheral blood mononuclear cells were isolated using density-gradient centrifugation and fixed according to standard methods. I-FISH was performed on both TPA stimulated and unstimulated cells using 11q22.3 (ATM), 13q14 (13S272), 17p13.1 (p53), and centromeric 12 (D12Z3) probes. 200 nuclei were evaluated for each probe, and cut-off points were set at 6%, 7%, 5% and 2% for del(11q), del(13q), del(17p) and trisomy 12, respectively. Metaphase FISH and conventional cytogenetics were also performed in selected cases. For all patients, chemotherapy was initiated according to Cheson criteria and treatment-free and overall survival curves were plotted using SPSS software. Following a modified version of Dohner’s hierarchical model, patients were divided in those with del(17p) and/or del(11q) and those with other or no genomic abnormalities. Fourteen cases with negative or bordeline results with I-FISH-PBMC became positive with I-FISH-TPA for del(11q) (2 cases), del(13q) (9 cases) and trisomy 12 (3 cases). In all but one patient, either conventional karyotyping or metaphase FISH confirmed these abnormalities. I-FISH-TPA provided a better prediction of treatment-free interval compared to I-FISH-PBMC (P= 0.002 vs 0.019, see Figures 1 and 2). In particular, two patients with no cytogenetic abnormalities detected by I-FISH-PBMC required chemotherapy 3 and 11 months after diagnosis, more in keeping with the presence of del(11q) found using I-FISHTPA. Furthermore, I-FISH-TPA also improved the overall survival prediction compared to I-FISH-PBMC (P= 0.036 vs 0.042). In summary, I-FISH-TPA increased the detection rate and had an improved prognostic value compared to I-FISH-PBMC. Further studies with larger numbers of patients are warranted. Figure Figure Figure Figure

Detailed Safety Analysis of Venetoclax Combined with Rituximab in Patients with Relapsed/Refractory Chronic Lymphocytic Leukemia

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 2033-2033 ◽  
Author(s):  
Danielle M. Brander ◽  
Michael Y. Choi ◽  
Andrew W. Roberts ◽  
Shuo Ma ◽  
L. Leanne Lash ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chronic Lymphocytic Leukemia ◽  
Board Of Directors ◽  
Peripheral Blood ◽  
Research Funding ◽  
Lymphocytic Leukemia ◽  
Advisory Committees ◽  
Grade 3 ◽  
Over Time

Abstract Background: Venetoclax (VEN) is a selective, potent, orally bioavailable BCL-2 inhibitor FDA-approved for patients with del(17p) chronic lymphocytic leukemia (CLL) and who have received ≥1 prior therapy. Based on preclinical evidence of synergy, VEN plus rituximab is being assessed in an ongoing Phase 1b study. Methods: Patients with relapsed/refractory (R/R) CLL received daily VEN with stepwise ramp-up over 3-4 weeks to reach daily doses of 200-600mg. After 1 week at the target dose, monthly rituximab was added for 6 doses. Responses and progression were assessed by iwCLL criteria with CT scan and bone marrow biopsy. Bone marrow assessments were done at screening, completion of combination therapy (month 7), and 2 months after clinical/radiologic criteria of iwCLL response were met. Minimal residual disease (MRD) was assessed in peripheral blood and marrow aspirates using ≥4 color flow cytometry (min sensitivity: 0.01%). Data cutoff was 04March2016, with analysis focusing on updated safety of cytopenias experienced on the course of treatment. Results: Forty-ninepatients enrolled (48 CLL/1 SLL). Patients had received a median of 2 prior therapies (range: 1-5) and disease in 25 (51%) was considered refractory to the most recent therapy. Median time on study was 28 (<1-42) months, with 31 patients active on study. Eighteen patients discontinued: 11 due to disease progression, 3 due to toxicity (peripheral neuropathy [1], MDS [1], and death due to TLS [1]), 3 withdrew consent, and 1 was lost to follow up. Across all doses, the most common AEs of any grade were diarrhea (57%), neutropenia (55%), upper respiratory tract infection (55%), and nausea (51%). Peripheral blood cytopenias were the most common Grade 3/4 AEs (neutropenia [53%], thrombocytopenia [16%], anemia [14%], febrile neutropenia [12%], and leukopenia [12%]). Twenty-seven (55%) patients had a history of neutropenia, of whom 6 were receiving G-CSF support prior to starting VEN. Overall, in the first month of therapy, 15 (31%) experienced an AE of neutropenia (any grade). Thereafter, the rate of new AEs of neutropenia decreased over time. While there was individual patient variability, mean ANC was stable over time. Overall, 26 (53%) patients had Grade 3/4 neutropenia. Neutropenia was generally well tolerated and managed by G-CSF support in 24 patients, in addition to ≥1 dose modification in 11 of the 24 patients. Of 8 (16%) patients who experienced grade 3 infections, 2 were while neutropenic. There were no grade 4 infections. Among the 11 (22%) patients who developed any-grade thrombocytopenia, none occurred within 2 weeks of a reported bleeding-related AE. One patient had thrombocytopenia overlapping with disease progression on therapy. Objective response rate for all patients was 86% (n=42), with 51% (n=25) who had complete response (CR/CRi; 12 achieved CR/CRi by month 7). At the completion of combination therapy (month 7), 39 patients had evaluable bone marrow assessments. Thirty (77%) had no histologic evidence of CLL in the bone marrow and 22 patients (56%) had attained bone marrow MRD-negativity. In longer follow up at any point during treatment for all 49 patients, 37 (75%) patients achieved complete marrow clearance and 28 (57%) achieved marrow MRD-negativity. Conclusions: Transient manageable neutropenia was the most common AE, with first onset usually seen within the first month of treatment and the onset of new neutropenia AEs decreased over time. No patients discontinued the study due to cytopenias. Patients were able to continue on study and high rates of response to treatment were observed. VEN given with rituximab achieved rapid and profound reductions in disease burden in peripheral blood and bone marrow. 77% of evaluable patients achieved morphologic clearance by month 7, and 57% were MRD-negative at any point on study. Figure 1 Figure 1. Disclosures Brander: TG Therapeutics: Research Funding; Gilead: Honoraria. Roberts:AbbVie: Research Funding; Servier: Research Funding; Janssen: Research Funding; Genentech: Research Funding; Genentech: Patents & Royalties: Employee of Walter and Eliza Hall Institute of Medical Research which receives milestone payments related to venetoclax. Ma:Pharmacyclics, LLC, an AbbVie Company: Consultancy, Honoraria, Research Funding, Speakers Bureau; Janssen: Consultancy, Honoraria, Speakers Bureau; Gilead: Consultancy, Honoraria, Research Funding, Speakers Bureau; Genentech: Consultancy, Honoraria, Speakers Bureau; Novartis: Research Funding; Xeme: Research Funding; AbbVie: Research Funding. Lash:AbbVie: Employment. Verdugo:AbbVie: Employment, Other: may own stock. Zhu:AbbVie Inc.: Employment, Other: may own stock. Kim:AbbVie: Employment. Seymour:Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Gilead: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Genentech: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; AbbVie Inc.: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

scholarly journals Fluorescent in situ hybridization and cytogenetic studies of trisomy 12 in chronic lymphocytic leukemia

Blood ◽  
1993 ◽  
Vol 81 (10) ◽  
pp. 2702-2707 ◽  
Author(s):  
SM Escudier ◽  
JM Pereira-Leahy ◽  
JW Drach ◽  
HU Weier ◽  
AM Goodacre ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chronic Lymphocytic Leukemia ◽  
Median Survival ◽  
Chromosomal Abnormalities ◽  
Residual Disease ◽  
Clonal Evolution ◽  
Lymphocytic Leukemia ◽  
Cytogenetic Studies ◽  
Trisomy 12

Abstract Cytogenetic studies (CG) of 475 chronic lymphocytic leukemia (CLL) cases showed trisomy 12 in 6.1% or 26% of patients with abnormal karyotypes. Fluorescence in situ hybridization (FISH) detected trisomy 12 in 35% of 117 CLL patients. Only 34.6% of cases detected by FISH were detected by CG. Twelve patients had low levels of trisomic cells (4% to 11%) relative to clonal B cells (47.5% to 86%), suggestive of clonal evolution. Untreated patients with trisomy 12 were predominantly male (P < .05) and had an increased incidence of splenomegaly (P < .03). Patients with trisomy 12 were more likely to be previously treated and had advanced Binet stage compared with those without trisomy 12. The median survival was shorter in patients with trisomy 12 (7.8 years) and patients with other chromosomal abnormalities without trisomy 12 by FISH (5.5 years) than in patients with diploid karyotypes (14.4 years). The response to fludarabine was similar to that of patients with diploid karyotypes, but there was a trend for earlier disease progression. FISH detected residual disease in all patients with trisomy 12 in complete (n = 6) or partial remission (n = 4). As few as 1 trisomic cell in 5,000 was detected by performing FISH on fluorescence-activated cell sorter-sorted cells. Trisomy 12 was absent in T cells in patients with trisomy 12. We conclude that FISH identifies trisomy 12 approximately 2.6 times more often than CG, readily identifies minimal residual disease, and predicts for a shorter median survival.

Array Comparative Genomic Hybridization: A Better Alternative for Genomic Profiling and Prognosis of Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4572-4572
Author(s):  
Annette Leon ◽  
Deborah Sevilla ◽  
Joseph M Marino ◽  
Kenneth D Moreno ◽  
Rev Obrera ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chronic Lymphocytic Leukemia ◽  
Comparative Genomic Hybridization ◽  
Peripheral Blood ◽  
Array Comparative Genomic Hybridization ◽  
Genetic Material ◽  
Lymphocytic Leukemia ◽  
Comparative Genomic ◽  
Small Lymphocytic Lymphoma ◽  
Genomic Hybridization

Abstract Abstract 4572 Traditional cytogenetic techniques such as chromosome analysis and fluorescence in situ hybridization (FISH) have provided valuable genetic information in the evaluation of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). However, many genomic abnormalities remain undetected due to the limitations intrinsic to these techniques. We designed, validated, and clinically applied a combined targeted-whole genome custom oligonucleotide microarray for the evaluation of hematological malignancies. Incorporation of array comparative genomic hybridization (aCGH) analysis has enabled us to identify previously undetected copy number changes with an increased resolution and sensitivity and more precisely determine genomic breakpoints and gene content with diagnostic and prognostic relevance in CLL/SLL. A cohort of 300 patients diagnosed with CLL/SLL was studied using concurrent cytogenetics, FISH and aCGH analyses on either peripheral blood or bone marrow aspirate samples. Array CGH identified clinically significant genomic alterations in 75% of the cases compared to 57% and 47% by cytogenetics and FISH analyses, respectively. There were no statistically significant differences detected between peripheral blood and bone marrow samples. Recurrent alterations such as deletions in chromosomes 11q (ATM, adverse prognosis), 13q (RB1, DLEU1, 2, and 7, MIR15A and 16–1, favorable prognosis in general), 17p (TP53, adverse prognosis) and gain of one copy of the entire chromosome 12 (ETV6, KLRK1, CD27, KRAS, MDM2, HIP1R, KITLG, intermediate prognosis) were identified in 55% of cases by aCGH. Of these cases, chromosome and/or FISH analyses did not detect these abnormalities in 42% and 27% of cases respectively. In addition, aCGH study more fully characterized the interstitial deletion in chromosome 13q recently suggested to have different outcomes based on size and gene content. Important findings exclusively by aCGH analysis in 7% of cases include loss of genetic material in chromosomes 3p (KAT2B, FHIT, clonal evolution), 6q (MAP3K7, PRDM1, AIM1, TNFAIP3, intermediate prognosis), 8p (TNFRSF10D, disease progression), 9q (JAK2, MLLT3, CDKN2A and B, DAPK1) and gain of genetic material in chromosome 2p (ACP1, MYCN, ALK, REL, BCL11A) associated with unmutated IGHV status, advanced disease stage and adverse prognosis. The results obtained in this study demonstrate the superior resolution and detection rate of aCGH technology as well as the importance of incorporating this methodology into current algorithms for the diagnosis and prognosis of CLL/SLL. Disclosures: No relevant conflicts of interest to declare.

Bone Marrow May Be More Informative Than Peripheral Blood To Evaluate Minimal Residual Disease During 1st and 2nd Year Follow Up After First-Line Fludarabine, Cyclophosphamide, and Rituximab For Chronic Lymphocytic Leukemia

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1621-1621
Author(s):  
Paolo Strati ◽  
Michael J. Keating ◽  
Susan O'Brien ◽  
Jan A. Burger ◽  
Alessandra Ferrajoli ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chronic Lymphocytic Leukemia ◽  
Peripheral Blood ◽  
Residual Disease ◽  
Response Assessment ◽  
Lymphocytic Leukemia ◽  
First Line ◽  
Final Response ◽  
Minimal Residual

Abstract Background Minimal Residual Disease (MRD) status at end of first-line chemoimmunotherapy is an independent prognostic factor for patients (pts) with chronic lymphocytic leukemia (CLL). In the CLL8 trial of the German CLL Study Group, peripheral blood (PB) was monitored for MRD during follow up. Because the microenvironment is important for CLL cell growth and survival and typically it is the last site to eliminate residual disease with chemoimmunotherapy, bone marrow (BM) might be a more reliable site to monitor MRD. Methods Two-hundred thirty-seven pts with CLL and an indication for therapy (IWCLL-WG 2008) received first-line fludarabine, cyclophosphamide, and rituximab (FCR) on protocol between 09/2008 and 09/2012. MRD was prospectively assessed in BM and/or PB by flow cytometry using the highly sensitive international standardized approach, 2 months after the last course of treatment (final response assessment) and every 3-6 months thereafter. Kaplan-Meier estimates were compared using the log-rank test. Results Sixty-one percent of pts were male, 21% were >65 years old, 40% had Rai stage III-IV, 41% had beta2-microglobulin (B2M) ≥4 mg/L, 61% had unmutated IGHV, and 21% had FISH analysis positive for deletion 11q and 7% for deletion 17p. Seventy-five percent of pts received ≥3 total courses of FCR. The complete remission (CR) and overall response (OR) rates were 65 and 97%, respectively. BM MRD negativity was achieved in 59% of pts at final response assessment. For monitoring, BM MRD was assessed in 121 pts during the 1st year and in 30 pts during the 2nd year after completion of treatment with FCR; all samples were serial. PB MRD was assessed in 106 pts during the 1st year and in 57 during the 2nd year of follow up; again all samples were serial. BM MRD negativity was observed in 63 (52%) pts during the 1st year of follow up and in 15 (50%) pts during the 2nd year. PB MRD negativity was observed at the same staging times in 81 (76%) and 29 (51%) pts, respectively. Concurrent BM and PB samples were taken during the 1st year in 51 pts, and in 6 pts during the 2nd year of follow up. We evaluated the association between MRD negativity during the 1st and 2nd year of follow-up and progression-free survival (PFS). BM MRD positive status was associated with shorter PFS when assessed during both the 1st and 2nd year of follow up (p<0.001 and p=0.001, respectively; Figure). In contrast, PB MRD positive status did not correlate with PFS for either time (p=0.15 and p=0.79, respectively; Figure). Conclusions After first-line FCR for pts with CLL, positive BM MRD may identify pts at higher risk for progression. Based on this finding, BM may be preferred to assess MRD status and pts with positive BM MRD could be considered for maintenance or consolidation strategies. Additional studies confirming these findings are warranted. Disclosures: No relevant conflicts of interest to declare.

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