Driver Mutations and Single Copy Number Abnormalities Identify Binet Stage A Patients with Chronic Lymphocytic Leukemia with Aggressive Progression

The correlation between progression and the genetic characteristics of Binet stage A patients with chronic lymphocytic leukemia (CLL) detected by whole exome sequencing (WES) was analyzed in 55 patients. The median follow-up for the patients was 102 months. During the follow-up, 24 patients (43%) progressed. Univariate Cox analysis showed that the presence of driver mutations, the accumulation of two or more mutations, the presence of adverse mutations, immunoglobulin heavy chain genes (IGHV) mutation status and unfavorable single copy number abnormalities (SCNAs) were associated with a higher risk of progression. Particularly, the occurrence of an adverse mutation and unfavorable SCNAs increased the risk of progression nine-fold and five-fold, respectively. Nevertheless, only the occurrence of adverse mutations retained statistical significance in the multivariate analysis. All patients carrying an unfavorable mutation progressed with a median progression-free survival (PFS) of 29 months. The accumulation of two or more mutations also increased the risk of progression with a median PFS of 29 months. The median PFS of patients with unfavorable SCNAs was 38 months. Combining mutations and SCNAs, patients may be stratified into three groups with different prognostic outcomes: adverse (17% probability of five-year PFS), protective (86% probability of five-year PFS) and neither (62% probability of five-year PFS, p < 0.001). Overall, the analysis of the mutational status of patients with CLL at an early stage of the disease may allow the identification of patients with a high risk of progression. The feasibility of an early therapeutic intervention in these particular patients requires further investigation.

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Risk Factors Associated with Richter's Transformation in Patients with Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood-2018-99-112442 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 1697-1697 ◽

Cited By ~ 1

Author(s):

Yasmin Ben-Dali ◽

Mariam Hussein Hleuhel ◽

Michael Asger Andersen ◽

Christian Brieghel ◽

Erik Clasen-Linde ◽

...

Keyword(s):

Risk Factors ◽

Chronic Lymphocytic Leukemia ◽

Incidence Rate ◽

Cumulative Incidence ◽

Research Funding ◽

Univariate Analysis ◽

Lymphocytic Leukemia ◽

Increased Risk ◽

Binet Stage

Abstract Background Richter's transformation (RT) refers to the development of an aggressive lymphoma in patients with chronic lymphocytic leukemia (CLL) or small lymphocytic leukemia (SLL). Roughly, 2-10 % of patients with CLL develop RT most often as diffuse large B-cell lymphoma (DLBCL) or Hodgkin lymphoma (HL). Aim This study aimed to assess the incidence rate and risk factors for RT for patients with CLL in a nationwide cohort. Furthermore, we want to assess prognostic risk factors for patients with RT. Methods All patients diagnosed with CLL in Denmark between 2008 and 2016 were included in this study. Clinical data was retrieved from the Danish National CLL Registry (DCLLR), whereas all histologically verified DLBCL, HL and/or transformation diagnoses for patients with CLL were retrieved from the Danish National Pathology Registry. Patients were followed from date of CLL diagnosis until date of RT, death or end of follow-up, whichever came first. The time to RT was estimated as cumulative incidence considering death as a competing risk. Stepwise Cox analysis with backward elimination was applied to identify independent risk factors for RT in patients with CLL. Results A total of 3771 CLL patients were identified, and followed for 14165 person-years. With a median follow-up of 4.3 (IQR (2.4;6.6)) years, 120 (3%) CLL patients had a transformation diagnosis, of which 4 patients were excluded due to misdiagnosis. DLBCL accounted for 78/116 (67%) cases, HL for 15/116 (13%) cases and one patient presented with both DLBCL and HL. In the remaining 22/116 (19%) cases the subtype of the transformation was either unspecified or unclassified RT. The median time to RT was 3.4 (IQR (1.8;5.7)) years from CLL diagnosis and the median overall survival (OS) after development of RT was 4.9 (IQR (0.7;8.4)) years. The cumulative incidence of RT, calculated by Aalen-Johansen estimator, at 5 and 8 years post-CLL diagnosis were 3.3% and 7.9% respectively (Figure 1). The annual crude incidence rate of RT was approximately 0.7% per year for all CLL patients. In all, 918 (24%) patients received CLL-related treatment, of whom 59 (6.4%) patients developed RT, resulting in a cumulative incidence of RT of 7% after 5 years and 11% after 8 years. At the time of CLL diagnosis, patients treated for CLL prior to RT diagnosis had a worse median OS (1.49 years) compared to RT patients who were untreated for CLL (6.16 years). In the univariate analysis, RT was significantly associated with male gender, advanced Binet stage (B or C), unmutated IGHV status (CLL-U), elevated beta-2-microglobulin (>3.5 mg/L) and elevated lactate dehydrogenase (>205 U/L). Of cytogenic aberration, deletion 13q (del(13q)) had a protective effect on the risk of RT, whereas deletion 11q (del(11q)) and deletion 17p (del(17p)) increased the risk. In the multivariable model, advanced Binet stage (HR 2.86 (1.82;4.51), p<0.001), del(17p) ((HR 3.74 (2.12;6.61), p<0.001) and CLL-U ((HR 2.30 (1.46;3.63), p<0.001) showed an independent correlation with development of RT. ZAP70 and CD38 were excluded from statistical analyses due to incomplete data and high inter-laboratory variation. Among RT patients, CLL-U, trisomy 12 and del(17p) at CLL diagnosis as well as ECOG Performance Status (PS) (i.e. PS≥1) at time of RT diagnosis correlated with poor OS in univariate analysis. Both del(17p) and PS≥1 were independently associated with an increased risk of death in a multivariable analysis (HR 2.9, (1.1;7.7), p=0.04 and HR 3.0, (1.0;3.1), p=0.05, respectively). Conclusions To the best of our knowledge, we here report the largest study on RT assessing nationwide data of consecutive patients diagnosed with CLL. The incidence of RT in this unselected population was 3.3% after 5 years while the median OS for patients from time of RT was 4.9 years. Advanced Binet stage, del(17p) and CLL-U were significantly and independently associated with an increased risk of RT. Del(17p) at CLL diagnosis and PS≥1 at RT diagnosis were significant predictors for death for patients with RT. For patients diagnosed with RT prior to any CLL treatment, a less severe disease course with a median OS of 6.16 years was demonstrated. Contrary, the median OS for patients receiving prior CLL treatment was 1.49 years. Thus, assessment of different treatment options for patients developing RT based on whether they have received prior CLL treatment or not is warranted. Figure 1. Figure 1. Disclosures Ben-Dali: Rigshospitalet: Research Funding. Hleuhel:Rigshospitalet: Research Funding. Brieghel:Arvid Nilson's Fund: Research Funding; Rigshospitalet, Denmark: Research Funding. Niemann:Danish Cancer Society: Research Funding; Novo Nordisk Foundation: Research Funding; Janssen: Consultancy, Research Funding; Abbvie: Consultancy, Research Funding; Novartis: Consultancy; Roche: Consultancy; Gilead: Consultancy; AstraZeneca: Consultancy; CSL Behring: Consultancy.

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MicroRNA-29c and 223 Are Powerful Prognostic Factors for Chronic Lymphocytic Leukemia and Improve Risk Stratification When Combined with ZAP70 and LPL in a qPCR Score.

Blood ◽

10.1182/blood.v112.11.1066.1066 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1066-1066

Author(s):

Basile Stamatopoulos ◽

Nathalie Meuleman ◽

Dominique Bron ◽

Benjamin Haibe-Kains ◽

Pascale Saussoy ◽

...

Keyword(s):

Prognostic Factors ◽

Chronic Lymphocytic Leukemia ◽

Noncoding Rna ◽

Poor Prognosis ◽

Lymphocytic Leukemia ◽

Early Therapy ◽

Mutational Status ◽

Igvh Mutational Status ◽

Binet Stage

Abstract Background: MicroRNAs (or miR) are a novel class of small noncoding RNA involved in gene regulation. Aberrant microRNA expression has been recently associated with chronic lymphocytic leukemia (CLL) outcome. Currently, the heterogeneous evolution of this disease can be predicted by several prognostic factors. Nevertheless, a better individualization of the outcome in a given patient is still of utmost interest. Methods: In the current study, we investigated the expression of two microRNAs, miR-29c and miR-223, compared them to other biological or clinical markers and proposed a quantitative real-time PCR (qPCR) score to better assess CLL outcome. All cut-offs were calculated by ROC curve analysis maximising the correlation with the immunoglobulin variable heavy chain (IgVH) mutational status; statistical differences were evaluated by Mann Whitney test or Kruskal-Wallis test ; treatment-free (TFS) and overall (OS) survival differences were investigated by log-rank test or Cox proportional hazard ratio (HR). Results: miR-29c and miR-223 expression decreased significantly with progression along Binet Stage A to C (P=0.0010 and P=0.0183, respectively), and were significantly lower in poor prognosis subgroups defined by cytogenetic abnormalities, IgVH mutational status, lymphocyte doubling time, solubleCD23, β2-microglobulin, ζ-associated protein 70 (ZAP70), lipoprotein lipase (LPL) and CD38 expression. Furthermore, miR-29c and miR-223 could predict TFS (n=110, P=0.0015 and P<0.0001, respectively) and OS (n=110, P=0.0234 and P=0.0008, respectively). Regarding all these results, we developed a qPCR score (from 0 to 4 poor prognostic markers) combining miR-29c, miR-223, ZAP70 and LPL in order to stratify treatment and death risk in a 110 patient cohort with a median follow-up of 72 months (range, 2–312). Patients with a score of 0/4, 1/4, 2/4, 3/4, and 4/4 had a median TFS of >312, 129, 80, 36 and 19 months, respectively (HR=17.00, P<0.0001). Patient with a score of 0–1/4, 2–3/4 and 4/4 had a median OS of >312, 183 and 106 months, respectively (HR=13.69, P=0.0001). Interestingly, during the first 50 months after diagnosis, only 10% of patients with a 0/4 score required a treatment, when compared to 100% of the 4/4. Furthermore, during the total follow-up (312 months), patients with a 4/4 score had a 27-fold higher risk to be treated and a 31-fold higher risk to die comparing to patients with a 0/4 score. This score was validated by a 10-fold cross-validation (prediction accuracy of 82%). Finally, in Binet stage A patients (n=77), this score remained relevant and significant for TFS and OS prediction (HR=18.56, P<0.0001 and HR=12.5, P=0.0068, respectively). Conclusions: we showed that (i) miR-29c and miR-223 levels were decreased in poor prognosis patients regarding several well-known prognostic factors; (ii) a low level of these two microRNAs is thus associated to disease aggressiveness, tumor burden and poor clinical evolution; (iii) we also showed that these two microRNAs could predict TFS and OS; (iv) we proposed a qPCR score to better individualize evolution of a particular CLL patient. This score will help to identify patients who will need early therapy and require thus a closer follow-up.

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Preliminary Results of a Phase II Open-Label, Randomized Study of the BH3 Mimetic Protein Navitoclax (ABT-263) with or without Rituximab for Treatment of Previously Untreated B-Cell Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood.v120.21.190.190 ◽

2012 ◽

Vol 120 (21) ◽

pp. 190-190 ◽

Cited By ~ 3

Author(s):

Herbert Eradat ◽

Sebastian Grosicki ◽

John Catalono ◽

Walter Cosolo ◽

Irina Dyagil ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Phase Ii ◽

Research Funding ◽

Response Rate ◽

Response Assessment ◽

Lymphocytic Leukemia ◽

Preliminary Results ◽

Binet Stage ◽

Bh3 Mimetic

Abstract Abstract 190 Introduction: Overexpression of Bcl-2 in Chronic Lymphocytic Leukemia (CLL) is associated with enhanced CLL-cell resistance to spontaneous or chemotherapy-induced apoptosis. The BH3 mimetic protein navitoclax (ABT-263) specifically inhibits Bcl-2, and related proteins Bcl-xL and Bcl-w, and can induce apoptosis of CLL cells in vitro. Phase I evaluation in relapsed/refractory CLL patients demonstrated 35% overall response rate (Roberts, 2012). Dose-limiting thrombocytopenia due to Bcl-xL inhibition was mitigated using a lead-in dosing schedule to allow the bone marrow to achieve a compensatory increase in platelets prior to dose escalation to the MTD of 250 mg. Based on the promising single-agent data, a Phase II trial randomized trial compared the safety, pharmacokinetics, and biologic activity of treatment with navitoclax and rituximab (RTX) versus RTX alone. Methods: Patients with CLL who required initial treatment according to iwCLL criteria (Hallek et al, 2008) were stratified by Binet stage and high-risk cytogenetic features (17p deletion and/or 11q deletion), and randomized 1:1:1 to receive RTX weekly for 8 wks (375 mg/m2 wk 1, 500 mg/m2 wks 2–8) (Arm A), or RTX for 8 wks plus navitoclax daily for 12 wks (250 mg/day following a 7–14 day lead-in period of 100 mg/day) (Arm B), or RTX for 8 wks plus navitoclax daily as in Arm B, but continued treatment with navitoclax until disease progression, relapse, or unacceptable toxicity (Arm C). Arm A to Arm B crossover was permitted. Response rate was assessed by iwCLL CLL response criteria at week 12, and every 12 weeks during follow-up. The study was stopped after the last patient had completed ≥ 12 weeks of treatment and week-12 response assessment. Results: Baseline characteristics and prognostic factors for the 118 randomized patients were generally balanced among the three treatment arms. Median age was 63 years (range 38–94), and 55% were Binet stage B+C. Median baseline lymphocyte count was 53,000 mm3 (range 7,000–552,000/mm3). FISH analyses identified higher than expected rates of deletion of 11q or 17p in the CLL cells of 32% or 28% of patients, respectively. Median time on study was 32 weeks overall (24 wks for Arm A, 33 wks Arm B, and 44 wks Arm C). AEs of Grade 3–4 that were more common (> 5% greater) in a navitoclax-treated arm compared with the RTX arm included thrombocytopenia, neutropenia, leukopenia, anemia, GI symptoms (diarrhea, abdominal pain), chills, fatigue, ALT/AST/bilirubin elevations, and infusion-related reactions (to RTX). Thrombocytopenia, neutropenia, and hepatic enzyme elevations were generally reversible when navitoclax was stopped and/or dose-reduced; however, 12 patients (15%) discontinued navitoclax due to laboratory abnormalities (9 due to ALT elevations). Neutropenia responded to growth factors. One serious event of epistaxis occurred related to the thrombocytopenia. Two deaths occurred on study, one on the RTX-only arm due to a pulmonary embolus and one on Arm B due to hypotension and dyspnea related to a severe RTX infusion reaction. Investigator-assessed objective response (CR and PR) rate was 35% for Arm A, 55% for Arm B (p=0.19 vs A), and 70% for Arm C (p=0.0034 vs A). All responses were PRs except for 2 CRs in Arm C. All responses were confirmed by CT (and BM for CR) ≥ 8 wks after clinical response assessment. While the presence of 17p deletion appeared to result in a lower response rate to RTX alone (Arm A, ORR 18%, 2/11 pts), it did not appear to affect the response to ABT-263 and RTX (Arm B, ORR 73%, 8/11 pts); Arm C, ORR 50%, 5/10 pts. Limited PFS results appeared consistent with the responses by arm, with a longer PFS associated with the longer duration of ABT-263 treatment on Arm C; however, the magnitude of PFS differences could not be precisely quantified due to the limited follow-up and patient number. Preliminary pharmacokinetic analysis did not detect any drug interaction between navitoclax and RTX. Conclusions: Navitoclax in combination with RTX weekly × 8 was generally well-tolerated as initial therapy for CLL patients and demonstrated greater clinical activity than treatment with RTX alone as well as responses in patients with 17p deletion. The preliminary results of this study indicate that a BH3-mimetic inhibitor of Bcl-2 could be highly effective when used in combination with RTX for treatment of patients with CLL. Disclosures: Eradat: Genentech: Research Funding. Off Label Use: BH3 Mimetic Protein Navitoclax (ABT-263). Catalono:Genentech: Consultancy. Kipps:Genentech: Research Funding. Zheng:Genentech: Employment. Yalamanchili:Genentech: Employment. Sahasranaman:Genentech: Employment. Hurst:Genentech: Employment. Ho:Genentech: Employment.

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BIRC3 disruption and Copy Number Aberrations in Chronic Lymphocytic Leukemia (CLL) Patients with 11q Deletion

Blood ◽

10.1182/blood.v124.21.3295.3295 ◽

2014 ◽

Vol 124 (21) ◽

pp. 3295-3295

Author(s):

Ilaria Del Giudice ◽

Silvia Bonina ◽

Marilisa Marinelli ◽

Luciana Cafforio ◽

Sara Raponi ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Copy Number ◽

Poor Prognosis ◽

Conflicts Of Interest ◽

Progression Free Survival ◽

Chromosome Analysis ◽

Lymphocytic Leukemia ◽

The Other ◽

11Q Deletion

Abstract Introduction. Chronic lymphocytic leukemia (CLL) with 11q deletion has been associated to a poor prognosis, but the clinical course of patients carrying this lesion is variable. This aberration, most often monoallelic, is present in 10-17% of newly diagnosed CLL and in 20-30% of patients with progressive or chemorefractory disease. The minimal deleted region (MDR) (2-3 Mbp) is located on the 11q22.3-q23.1 region and includes ATM. Moreover, 30-40% of 11q- CLL have also an inactivating ATM mutation on the other allele. The deleted region on 11q can also include BIRC3, a gene that is often deleted or mutated in advanced/chemorefractory stages of the disease. Although BIRC3 disruption has been associated to a poor prognosis, its prognostic implications in addition to ATM deletion are not well defined. The aim of this study was to perform a copy number aberration (CNA) and gene sequencing analyses on a cohort of CLL patients with 11q- in order to identify subgroups with potential prognostic relevance based on: i) the inclusion of BIRC3 in the deleted region; ii) the presence of BIRC3 mutation; iii) the presence of other CNAs. Methods. The study has included 55 untreated CLL patients followed at our Institution or enrolled in GIMEMA clinical trials (2003-2013). Genomic DNA was extracted from peripheral blood samples. CNA analysis was performed by genomic hybridization on the CytoScan HD array (Affymetrix), which contains more than 2.6 x 106 markers for copy number analysis and 750.000 SNPs. Data were analyzed using both Partek Genomics Suite and ChAS (Chromosome Analysis Suite, Affymetrix) software. The resulting CNAs were verified by visual examination of the plotted copy number profiles. BIRC3 mutations (exons 6-9) were evaluated by Sanger sequencing. Time to first treatment (TFT) was calculated from the date of diagnosis to the date of first therapy or last follow-up; progression-free survival (PFS) from the date of first therapy to the date of progression, death or last follow-up. Results. Baseline characteristics of the 55 cases were as follows: median age at diagnosis 59 years (range 39-84), male gender in 81.8% of patients, progressive disease in 62%. All patients showed 11q- by FISH (median 80%, range 25-99% of nuclei); germline IGHV were present in 96.4% of cases; TP53 deletion in 1 case and TP53 mutation in none; NOTCH1 mutation in 4/40 cases; SF3B1 mutation in 5/40 (all mutually exclusive with only 1 case having both SF3B1 and BIRC3 mutations). By CytoScan HD array, the size of 11q- was very variable, ranging from 0.36 Mbp to 65.14 Mbp; the MDR was located on 11q22.3 region, encompassing 4 genes (ACAT1, ATM, CUL5, NPAT). BIRC3 was included in the deleted region in 45/55 cases (81.8%) and was mutated in 4/54 (7.5%), being always deleted on the other allele. Beside 11q-, 51 cases (92.7%) showed several additional CNAs (average 4.9, range 1-14 per patient), with 5 recurrent lesions: 2p gain in 11 cases, del4(p15.2) in 6, del19(p13.3) in 6, 8q gain in 5 and del4(q22.1) in 4. BIRC3 deletion was not associated to the number of additional CNAs nor to specific CNA. After a follow-up of 59.6 months (range 7.4-229.7), 40 of 47 evaluable patients have received treatment (median TFT 15.8 months, range 0-167). BIRC3 deleted cases (n=37) showed a TFT not significantly different from WT cases (n=10). Conversely, BIRC3 mutation was associated to a shorter TFT (p <0.0001), 0.5 vs 19.3 months of WT cases. Notably, 3/4 cases with the biallelic BIRC3 disruption had a marked hyperleukocytosis at diagnosis (212.4-302.8 x 109L). The presence of additional CNAs was associated to a shorter, though not significant, TFT considering either >3 CNAs larger than 5 Mb (n=14) or >10 CNAs (n=5), or the presence of 2p gain, del4(p15.2), del19(p13.3) or 8q gain. So far, 22 patients have been evaluated for PFS after first-line therapy (median 28.7 months). BIRC3 deleted cases (n=17) were not associated to a shorter PFS compared to WT cases (n=5), in line with the results from Rose-Zerilli et al (Haematologica 2014). Conclusions. Among CLL with 11q-: 1) BIRC3 deletion involves more than 80% of cases, whilst the mutation is rare (7.5%); 2) BIRC3 deletion is not associated to a higher genomic complexity; 3) BIRC3 deletion does not seem to influence TFT or PFS of 11q- CLL; 4) BIRC3 mutation is strongly associated to a short TFT; 5) BIRC3 biallelic lesions can be associated to a marked hyperleucocytosis at diagnosis and immediate need of treatment. Disclosures No relevant conflicts of interest to declare.

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Monoclonal B-cell lymphocytosis and early-stage chronic lymphocytic leukemia: diagnosis, natural history, and risk stratification

Blood ◽

10.1182/blood-2015-02-585059 ◽

2015 ◽

Vol 126 (4) ◽

pp. 454-462 ◽

Cited By ~ 96

Author(s):

Paolo Strati ◽

Tait D. Shanafelt

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Risk Stratification ◽

B Cell ◽

Cell Count ◽

Early Stage ◽

Lymphocytic Leukemia ◽

High Count ◽

Genetic Characteristics ◽

Increased Risk ◽

Risk Of Progression

Abstract Monoclonal B lymphocytosis (MBL) is defined as the presence of a clonal B-cell population in the peripheral blood with fewer than 5 × 109/L B-cells and no other signs of a lymphoproliferative disorder. The majority of cases of MBL have the immunophenotype of chronic lymphocytic leukemia (CLL). MBL can be categorized as either low count or high count based on whether the B-cell count is above or below 0.5 × 109/L. Low-count MBL can be detected in ∼5% of adults over the age of 40 years when assessed using standard-sensitivity flow cytometry assays. A number of biological and genetic characteristics distinguish low-count from high-count MBL. Whereas low-count MBL rarely progresses to CLL, high-count MBL progresses to CLL requiring therapy at a rate of 1% to 2% per year. High-count MBL is distinguished from Rai 0 CLL based on whether the B-cell count is above or below 5 × 109/L. Although individuals with both high-count MBL and CLL Rai stage 0 are at increased risk of infections and second cancers, the risk of progression requiring treatment and the potential to shorten life expectancy are greater for CLL. This review highlights challenging questions regarding the classification, risk stratification, management, and supportive care of patients with MBL and CLL.

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Accessing Genomic Alternations in Chronic Lymphocytic Leukemia Using an NGS-Based Comprehensive Genomic Profiling Assay

Blood ◽

10.1182/blood-2020-141690 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 8-8

Author(s):

Segun C Jung ◽

Maya Thangavelu ◽

Hyunjun Nam ◽

Ryan Bender ◽

Sally Agersborg ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Copy Number ◽

Patient Management ◽

Structural Changes ◽

Sex Chromosome ◽

Single Gene ◽

Hematologic Malignancies ◽

Lymphocytic Leukemia ◽

Driver Mutations ◽

Loss Of Function

Background: Next generation sequencing (NGS) is an integral component in the characterization of hematologic malignancies, including chronic lymphocytic leukemia (CLL). Fluorescence in situ hybridization (FISH) and conventional cytogenetics (CC) are cost effective and are currently the gold standard for detecting copy number abnormalities (CNAs) in hematologic malignancies. NGS is emerging as a comprehensive assay that can detect CNAs while surveying the whole genome for single nucleotide variants and loss of heterozygosity (CN-LOH). Identifying CNA events in addition to mutations and RNA fusions may help identify and characterize the highly complex genetic landscape of hematologic malignancies. Methods: A custom total nucleic acid (TNA) NGS panel was designed which consists of mutation profiles of 297 genes, transcriptome profile of 213 genes, and genomic backbones of 14 chromosomes to identify unbalanced abnormalities. Two-hundred seventy CLL patients were included in the study (abnormalities detected in 236 cases in total: 61 cases by CC; 230 cases by FISH; and 53 cases by both CC and FISH, and no abnormalities detected in 34 cases by both FISH and CC). Mutation profiles including SNVs, indels, and structural changes were interrogated with a custom bioinformatic pipeline which utilized PureCN and CNVkit algorithms to identify structural changes. NGS results were compared to results of CC and FISH. CNA detection of sex chromosome and balanced rearrangement including translocation and inversion was excluded from the analysis Results: CNAs were detected by NGS in 56 of 61 cases (91%) reported by CC and in 178 of 230 cases (77%) detected by FISH. Seventy-seven CNAs detected by CC and 202 CNAs detected by FISH were identified by NGS. NGS failed to detect 13q deletion, detected by FISH in 48 cases. Abnormalities not detected by neither cytogenetics nor FISH were detected by NGS in 108 (gain) and 32 (loss) cases. In addition, we observed abnormalities in 9 of 34 cases by NGS reported as normal by both FISH and cytogenetics. CN-LOH was detected in 9% of cases predominantly on 13q, 17p and 22q. In addition to trisomy 12, gains of 20p and 20q were observed in each 72 (30%) and 43 (18%) cases. CN gains of 7p, 8q, and 17q were also observed in 12%, 12%, and 7% of cases, respectively. Oncogenic driver mutations in KRAS (p.G12D) and (p.G13D) were observed in four and five cases with CN gains, respectively. IKZF3, a recurrent hotspot pathogenic mutation in CLL and a potential prognostic marker that may positively regulate MYC, was detected in five patients with CN gains. CN loss of 11q, 2q, 13q, 3p, 17p, 21q, and 6q were among the most common chromosomes with CN loss (Figure 1). Notably, LOH of RB1, DLEU7, COG3, and FOX1 genes on 13q, of TP53, WRAP53, SLC52A1, CTC1, and ABR genes on 17p and of PRDM1, EPHA7, and CASP8AP2 genes on 6q were observed. Identifying cases with 13q14 deletions that include RB1 could change the CLL patient management due to the aggressive clinical course. Recurrent loss of function mutations in KMT2C (p.E2798Gfs*11), NOTCH1 (p.P2514Rfs*4), and TP53 (p.H179R) in 7q, 9q, and 17p were observed. Identifying both CN loss combined with loss of function mutations in tumor suppressors could help improve patient care. Conclusions: Abnormalities detected by cytogenetics were mostly detected by NGS, but NGS offers a higher resolution including CN events of various length, LOH events, and single gene mutations. CNAs detected at higher resolution is useful in identifying patients with 13q14 loss that include/exclude RB1 which may affect patient management. However, an accurate detection of the CNA could be affected in part by a baseline established by a panel of normal and the depth of coverage. Differences in sensitivity of methodologies can also be attributed to in vitro proliferation and tissue culture conditions utilized for CC analysis. CC and FISH can identify clones with multiple abnormalities as well as clonal evolution. Comprehensive genomic profile including high resolution copy number changes and mutational profiles, detectable by NGS, may provide better profiling for a patient for clinical management. Disclosures Jung: NeoGenomics: Current Employment. Thangavelu:NeoGenomics: Current Employment. Nam:NeoGenomics: Current Employment. Bender:NeoGenomics: Current Employment. Agersborg:NeoGenomics: Current Employment. Weiss:Bayer: Other: speaker; Genentech: Other: Speaker; Merck: Other: Speaker; NeoGenomics: Current Employment. Funari:NeoGenomics: Current Employment.

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The complex karyotype landscape in chronic lymphocytic leukemia allows to refine the risk of Richter syndrome transformation

Haematologica ◽

10.3324/haematol.2021.278304 ◽

2021 ◽

pp. 0-0

Author(s):

Andrea Visentin ◽

Laura Bonaldi ◽

Gian Matteo Rigolin ◽

Francesca Romana Mauro ◽

Annalisa Martines ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Lymphocytic Leukemia ◽

Prognostic Impact ◽

Complex Karyotype ◽

Biological Variables ◽

Richter Syndrome ◽

Binet Stage ◽

The Impact

Complex karyotype (CK) at chronic lymphocytic leukemia (CLL) diagnosis is a negative biomarker of adverse outcome. Since the impact of CK and its subtypes, namely type-2 CK (CK with major structural abnormalities) or high-CK (CK with C5 chromosome abnormalities), on the risk of developing Richter syndrome (RS) is unknown, we carried out a multicenter reallife retrospective study to test its prognostic impact. Among 540 CLL patients, 107 harbored a CK at CLL diagnosis, 78 were classified as CK2 and 52 as high-CK. Twenty-eight patients developed RS during a median follow-up of 6.7 years. At the time of CLL diagnosis, CK2 and high-CK were more common and predicted the highest risk of RS transformation, together with advanced Binet stage, unmutated (U)-IGHV, 11q-, TP53 abnormalities. We integrated these variables into a hierarchical model: high-CK and/or CK2 patients showed a 10-year time to RS (TTRS) of 31%; U-IGHV/11q-/TP53 abnormalities/Binet stage B-C patients had a 10-year TTRS of 12%; while mutated (M)-IGHV without CK and TP53 disruption a 10-year TTRS of 3% (p<0.0001). We herein demonstrated that CK landscape at CLL diagnosis allows to refine the risk of RS transformation and we recapitulated clinico-biological variables into a prognostic model.

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microRNA-29c and microRNA-223 down-regulation has in vivo significance in chronic lymphocytic leukemia and improves disease risk stratification

Blood ◽

10.1182/blood-2008-11-189407 ◽

2009 ◽

Vol 113 (21) ◽

pp. 5237-5245 ◽

Cited By ~ 165

Author(s):

Basile Stamatopoulos ◽

Nathalie Meuleman ◽

Benjamin Haibe-Kains ◽

Pascale Saussoy ◽

Eric Van Den Neste ◽

...

Keyword(s):

Prognostic Factors ◽

Chronic Lymphocytic Leukemia ◽

Disease Risk ◽

Lymphocytic Leukemia ◽

Aberrant Expression ◽

Early Therapy ◽

Disease Evolution ◽

Binet Stage

Abstract Aberrant expression of microRNAs has been recently associated with chronic lymphocytic leukemia (CLL) outcome. Although disease evolution can be predicted by several prognostic factors, a better outcome individualization in a given patient is still of utmost interest. Here, we showed that miR-29c and miR-223 expression levels decreased significantly with progression from Binet stage A to C were significantly lower in poor prognostic subgroups (defined by several prognostic factors) and could significantly predict treatment-free survival (TFS) and overall survival (OS). Furthermore, we developed a quantitative real-time polymerase chain reaction (qPCR) score combining miR-29c, miR-223, ZAP70, and LPL (from 0 to 4 poor prognostic markers) to stratify treatment and death risk in a cohort of 110 patients with a median follow-up of 72 months (range, 2-312). Patients with a score of 0/4, 1/4, 2/4, 3/4, and 4/4 had a median TFS of greater than 312, of 129, 80, 36, and 19 months, respectively (hazard ratio, HR0/4 < 1/4 < 2/4 < 3/4 < 4/4 = 17.00, P < .001). Patients with a score of 0-1/4, 2-3/4, and 4/4 had a median OS of greater than 312, of 183 and 106 months, respectively (HR0/4 < 1/4 < 2/4 < 3/4 < 4/4 = 13.69, P = .001). This score will help to identify, among the good and poor prognosis subgroups, patients who will need early therapy and thus will require a closer follow-up.

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Rituximab Added to Cyclophosphamide, Vincristine and Prednisolone in Heavily Pretreated Chronic Lymphocytic Leukemia.

Blood ◽

10.1182/blood.v106.11.5023.5023 ◽

2005 ◽

Vol 106 (11) ◽

pp. 5023-5023

Author(s):

Henrique Coelho ◽

Cristina Gonçalves ◽

Claudia Casais ◽

Marisol Guerra ◽

Alexandra Mota ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Disease Progression ◽

Lymphocytic Leukemia ◽

Clinical Activity ◽

Front Line ◽

Pulmonary Aspergillosis ◽

Heavily Pretreated ◽

Treatment Complication ◽

Binet Stage

Abstract Purpose: Although patients with chronic lymphocytic leukemia (CLL) typically respond to available front-line chemotherapy, the response is often temporary, and patients frequently undergo relapse. The authors report the beneficial clinical activity and tolerability of an immunochemotherapeutic regimen consisting of rituximab added to cyclophosphamide, vincristine and prednisolone (R-CVP), in patients with heavily pretreated CLL. Methods: The therapeutic regimen consisted of: cyclophosphamide 750 mg/m2 iv on day one, vincristine 1.4 mg/m2 iv on day one and prednisolone 100 mg po for five consecutive days every 4 weeks, combined with rituximab at a dose of 375 mg/m2 on day 1 of each cycle. Patients were evaluated after three cycles, with responding patients receiving up to eight cycles. No infectious prophylaxis was administered. Results: Five patients have been treated with R-CVP: median age 65 years (range 52–68), advanced Binet stage (four stage C and one stage B) and heavily pretreated (range 1–3 prior therapies). All patients received at least three cycles of R-CVP. Clinical and hematological response was achieved in two patients. Three patients had stable disease. One patient was re-induced once after disease progression. Data from a median follow-up of 112 months (range 52–128 months) showed that three patients remained free of disease progression 8 months after the first cycle of R-CVP (range 2–14 months). The principle treatment complication was pulmonary aspergillosis. Conclusion: These promising results suggest salvage therapy with R-CVP is beneficial in patients with heavily pretreated CLL.

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Prognosis of Binet stage A chronic lymphocytic leukemia patients: the strength of routine parameters

Blood ◽

10.1182/blood-2010-06-288274 ◽

2010 ◽

Vol 116 (22) ◽

pp. 4588-4590 ◽

Cited By ~ 30

Author(s):

Rémi Letestu ◽

Vincent Lévy ◽

Virginie Eclache ◽

Fanny Baran-Marszak ◽

Dominique Vaur ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Prognostic Index ◽

Progression Free Survival ◽

Cost Effective ◽

Lymphocytic Leukemia ◽

Free Survival ◽

Cd38 Expression ◽

Recent Developments ◽

Binet Stage

Recent developments in the management of chronic lymphocytic leukemia (CLL) patients have made necessary the availability of dependable prognostic factors. We have developed a prognostic index derived from the multivariate analysis of 339 stage A patients at diagnosis, exhaustively studied for classical and recent predictive markers. Only 4 biologic parameters were found to be independent predictors of progression-free survival (PFS): serum thymidine kinase (sTK), lymphocytosis, β2-microglobulin, and CD38 expression. Two groups were distinguishable: cases with no or 1 risk factor (among whom 85% did not progress after 7 years), and cases with 2 or more factors showing a median PFS of 20 months. Finally, we propose an easy, fast, cost-effective strategy for a trustworthy prognostication in stage A patients, who currently represent more than 80% of the CLL population, allowing physicians to adapt follow-up individually.

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