Higher percentage of in vitro apoptotic cells at time of diagnosis in patients with chronic lymphocytic leukemia indicate earlier treatment requirement: Ten years follow up

Introduction. Chronic lymphocytic leukemia (CLL) has an extremely variable clinical course. Biological reasons for that wide variation in clinical course and survival rates in CLL patients are not fully understood. Objective. The aim of the study was to evaluate the value of spontaneous apoptosis of CLL cells in vitro determined at presentation of disease, in prediction of treatment requirements and evolution of the CLL. Methods. Malignant B cells were isolated from the whole blood of 30 newly diagnosed CLL patients and cultured for 24 hours in RPMI-1640 medium supplemented with 10% of serum obtained from the same CLL patient. Cells were later fixed and processed for embedding in Epon, or cell smears were prepared and stained with TUNEL technique. Results. Ten-year follow-up revealed that patients with lower percentage of cells in apoptosis at presentation of disease had significant longer time treatment initiation (log rank test p<0.05). On the contrary, apoptosis of CLL cells was not shown to have significant impact on survival of patients (Kaplan Meier log rank test p>0.05). Conclusion. The results of this study emphasize the importance of apoptosis of CLL cells at the time of the initial diagnosis in pathobiology of this disease.

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High Resolution Array-CGH Provides New Insights Into the Prognosis of Chronic Lymphocytic Leukemia (CLL): Is 8p Loss Worse Than 17p Loss?.

Blood ◽

10.1182/blood.v114.22.2339.2339 ◽

2009 ◽

Vol 114 (22) ◽

pp. 2339-2339

Author(s):

Andrea Rinaldi ◽

Michael Mian ◽

Davide Rossi ◽

Francesco Forconi ◽

Clara Deambrogi ◽

...

Keyword(s):

Cox Regression ◽

Clinical Course ◽

Conflicts Of Interest ◽

Univariate Analysis ◽

Lymphocytic Leukemia ◽

Rank Test ◽

Western World ◽

Clinical Parameters ◽

Log Rank Test ◽

The Impact

Abstract Abstract 2339 Poster Board II-316 BACKGROUND: CLL, the most common adult-onset leukemia in the Western world, has a heterogeneous clinical course. Many advances have led to a better understanding of its pathogenesis and to improvements in treatment strategies, but striking solutions are still missing. We conducted a study to evaluate the impact of genomic aberrations on the clinical course. METHODS: From January 1980 to May 2008, 395 frozen samples of CLL patients, were prospectively collected in four centers. Extracted DNA was analyzed with Affymetrix Human Mapping 6.0 arrays. Normal matched DNA was analyzed for one fourth of the cases. Correlations between minimal common regions (MCR) and clinical parameters were evaluated with the Fisherôs-exact test and their impact on OS with the log-rank test. A p-value after Bonferroni multiple test correction (MTC) (p-adj.) <0.05 was considered as statistically significant. Up to now 266 samples have been analyzed. RESULTS: Analysis of the clinical parameters (CPs) and known risk factors (Rai/Binet, age, doubling time, LDH, beta2, IGVH status, p53 mutations, telomere length, CD38, 11q, 17p) was consistent to previous published series. ZAP70 did not affect the clinical course, likely due inter-laboratories variability. After a median follow up of 53 months, 143/239 (60%) of the patients have started therapy and 63/261 (24%) died. 5-yr OS was 82%. Fisher test between the MCRs and CPs revealed an inverse relation between the presence of trisomy 12 by FISH and del13q14.3, an association between del17p and losses of 8p regions and between CD38 and 12q gain. Before MTC, 46 MCRs had a significant impact on OS and 67. After MTC, 3 regions maintained their role: 8p22 loss (38/248, 15%, p-adj.=0.002, median OS: 26 months vs. 48), 17p13.3-11.2 loss (20/248, 8%, p-adj.=0.001; median OS: 10 months vs. 48). In univariate analysis, the log-rank test among pts with 8p-/17p- (8/248, 3%), 8p- (30/248, 12%), 17p- (12/248, 5%), wild type (198/248, 80%) was statistically significant (p<0.001; see figure). Importantly, none of the analyzed clinical and biological parameters was associated with this aberration. CONCLUSIONS: Loss of 8p22 designated a CLL subgroup with a worse outcome among all patients and in the subset with 17p loss. Our data suggested that this aberration might constitute an independent prognostic factor to be evaluated in independent studies. Results, including a Cox regression model, will be presented on all 395 cases. Disclosures: No relevant conflicts of interest to declare.

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Demographics, Treatment Patterns, Safety, and Real-World Effectiveness in Patients ≥70 Years of Age with Chronic Lymphocytic Leukemia Receiving Bendamustine with or without Rituximab,

Blood ◽

10.1182/blood.v118.21.3914.3914 ◽

2011 ◽

Vol 118 (21) ◽

pp. 3914-3914

Author(s):

Kathryn S. Kolibaba ◽

Avani D. Joshi ◽

James A. Sterchele ◽

Michael Forsyth ◽

Erin Alwon ◽

...

Keyword(s):

Clinical Trials ◽

Chronic Lymphocytic Leukemia ◽

Partial Response ◽

Real World ◽

Progressive Disease ◽

Lymphocytic Leukemia ◽

Social Security Administration ◽

Grade 3

Abstract Abstract 3914 Background Bendamustine is a unique, well-established alkylating agent with multifaceted actions leading to cancer cell death in several hematologic malignancies. In a phase 3 trial in treatment-naïve patients with chronic lymphocytic leukemia (CLL), rates of response and progression-free survival (PFS) were significantly superior to those for chlorambucil [Knauf WU et al. J Clin Oncol 2009;27:4378–84]. In vitro studies have found that the cytotoxic activity of bendamustine against CLL-derived cell lines is synergized by rituximab, an anti-CD20 monoclonal antibody [Demidem A et al. Cancer Biother Radiopharm 1997;12:177–86]. Older patients may demonstrate lower tolerance to chemoimmunotherapy [Foon KA & Hallek MJ. Leukemia 2010;24:500–11], and published clinical data on bendamustine-rituximab in CLL are scarce. Thus, this retrospective study sought to characterize a population of adults ≥70 years old with CLL receiving bendamustine with or without rituximab, describe patterns of care, assess data on real-world effectiveness outside of the controlled environment of clinical trials [Waldthaler C et al. Wien Klin Wochenschr 2011;123:269–275], and assess safety. Methods Records were extracted from US Oncology iKnowMed (iKM) record databases for all outpatients ≥70 years old with CLL (but no other tumor) and more than 1 visit recorded (but not enrolled in clinical trials) who received bendamustine between March 2008 and May 2010. Patients were classified as treatment-naïve or relapsed (including ≥ second-line therapy). To ascertain mortality, the iKM data were supplemented with vital-status data from the Social Security Administration Death Index. The overall response rate (ORR) included complete response (CR), nodal partial response (nPR), and partial response (PR). PFS was time from first bendamustine dose to progressive disease (change in line of therapy), relapse, or death from any cause. Data from patients who did not die, or had no progression and were lost to follow-up were censored. Results Among 91 patients, the mean (SD) initial age at beginning of first therapy was 77.4 (5.6) years, age at diagnosis was 70.3 (6.5) years, and 63.7% were male. Of the 16 (17.6%) treatment-naïve patients, 10 had received bendamustine monotherapy and 6 received bendamustine-rituximab. Of the 75 (82.4%) relapsed patients, 20 had received bendamustine monotherapy and 55 received bendamustine-rituximab. The observed ORR for treatment-naïve patients was 56.3% (n=9; 18.8% CR, 37.5% PR, and 0 nPR); 6.3% had progressive disease. For relapsed patients, the ORR was 58.7% (n=44; 13.3% CR, 44.0% PR, and 1.3% nPR); 24.0% had progressive disease. Among patients with data, median PFS for 16 treatment-naïve patients has not been reached (median follow-up 15.1 months); for 73 relapsed patients, PFS was 18.4 months. Kaplan-Meier estimates for PFS over time for each group are shown in the Figure. No unexpected toxicities were seen. The overall rate of blood/bone marrow toxicities (all grades) was 40.7%; grade 3/4 rates were 18.8% for treatment-naïve patients and 25.3% for relapsed patients. Other grade 3/4 adverse events (AEs) included upper respiratory infection and abdominal pain in the relapsed group as well as rash and sepsis in both groups (n=1 each). The most frequent nonhematologic AEs (≥5%, any grade) were fatigue (33.0%), weight loss (11.0%), infection (9.9%; herpes zoster [n=2]; cryptococcal sepsis, Klebsiella sepsis, and pneumonia [n=1 each]), gastrointestinal (8.8%), fever (8.8%), pulmonary (6.6%), and rash (5.5%). o = data censored. Conclusions In this retrospective chart review of patients ≥70 years old with CLL, bendamustine, either alone or with rituximab, provided meaningful response rates and was generally well tolerated. The length of PFS of both treatment-naïve and relapsed patients was clinically meaningful. This research was sponsored by and conducted in collaboration with Cephalon, Inc., Frazer, PA. Disclosures: Sterchele: Cephalon, Inc.: Employment. Beygi:Cephalon, Inc.: Employment. Kennealey:Cephalon, Inc.: Employment.

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Early Results Of a Phase II Study Of Lenalidomide and Rituximab As Frontline Treatment Of Patients With Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood.v122.21.4183.4183 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4183-4183

Author(s):

Paolo Strati ◽

Michael J. Keating ◽

Christina Hinojosa ◽

Stefan Faderl ◽

Susan C. Smith ◽

...

Keyword(s):

Pancreatic Cancer ◽

Chronic Lymphocytic Leukemia ◽

Lymphocytic Leukemia ◽

Metastatic Pancreatic Cancer ◽

Categorical Variables ◽

Rank Test ◽

Early Results ◽

Baseline Characteristics ◽

Grade 3

Abstract Introduction The combination of lenalidomide and rituximab is an active treatment for patients (pts) with relapsed Chronic Lymphocytic Leukemia (CLL), with 66% Overall Response Rate (ORR) and a median Progression Free Survival (PFS) of 17.4 months (JCO 2013). We therefore sought to evaluate its efficacy and tolerability in untreated pts. Methods Twenty-five pts with untreated CLL have been enrolled. Rituximab (375 mg/m2 intravenously) was administered weekly during cycle 1 and on day 1 of cycles 3 to 12. Lenalidomide was started on day 9 of cycle 1 at 10 mg orally and administered daily continuously. Each cycle was 28 days. Rituximab was administered for 12 cycles; lenalidomide could continue indefinitely. Response was assessed every 3 cycles by 2008 NCI-WG criteria. Treatment-related toxicity was assessed using Common Terminology Criteria for Adverse Events (version 3.0). The primary end point of this study was ORR. Secondary objectives included PFS and Overall Survival (OS). PFS and OS were calculated using Kaplan-Meier estimates and compared using the log-rank test. Categorical variables were compared using Fisher's exact test (two tailed). Differences were considered significant if p≤.05. Results Baseline characteristics of the 25 pts are shown in Table 1. Twenty-four pts are evaluable for toxicity (one pt was taken off study after only 7 days due to the diagnosis of widely metastatic pancreatic cancer) and 20 for response (2 pts too early, 3 pts early discontinuation). So far 5 pts received 1 cycle, 1 pt received 2 cycles, 6 pts received 3 cycles, 5 received up to 6 cycles and 8 received 12 or more cycles of therapy. ORR by 2008 NCI-WG criteria is 85%, including 2 (10%) complete remission (CR) and 15 (75%) partial remissions (PR). No significant association between ORR and baseline characteristics was observed. At the latest follow-up, the median dose of lenalidomide was 5 (2.5-10) mg and 14 (56%) pts needed at least one interruption because of toxicity. Grade 3-4 toxicities are summarized in Table 2. No episodes of grade 3-4 tumor flare were observed and 7 (29%) pts experienced a grade 1 or 2 tumor flare. Eight (32%) pts have so far discontinued treatment: 1 because of metastatic pancreatic cancer diagnosed after only 1 week on study, 2 because of progressive disease (after 4 months and 13 months), and 5 because of toxicity (skin rash in 3 pts, a deep venous thrombosis in 1 pt, and persistent neutropenia in 1 pt). Two of the 5 pts who discontinued treatment have required subsequent therapy. Twenty-four pts are alive, 1 pt died of metastatic pancreatic cancer after treatment discontinuation. At a median follow-up of 9 (1-17) months, median PFS and median OS have not been reached. Conclusions Our initial experience with the combination of lenalidomide and rituximab as front-line treatment of CLL indicates that this combination is tolerated by the majority of pts and responses are seen in 85% of them. Enrolment in this study is ongoing. Disclosures: O'Brien: CELGENE: Consultancy. Ferrajoli:CELGENE: Research Funding.

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The Combination of Complex Karyotypes' Subtypes and IGHV Mutational Status Provides Prognostic and Predictive Information in Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood-2018-99-114721 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 1844-1844

Author(s):

Andrea Visentin ◽

Laura Bonaldi ◽

Gian Matteo Rigolin ◽

Francesca Romana Mauro ◽

Annalisa Martines ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Research Funding ◽

Board Member ◽

Advisory Board ◽

Lymphocytic Leukemia ◽

Rank Test ◽

Predictive Information ◽

Excellent Outcome ◽

Mutational Status

Abstract INTRODUCTION. Complex karyotype (CK), defined by the presence of at least 3 chromosomal abnormalities, is a heterogeneous cytogenetic category associated with adverse prognosis in several hematologic malignancies. Recently, Rigolin et al. provided evidence that CK with major structural abnormalities (CK2) at chronic lymphocytic leukemia (CLL) diagnosis negatively impact on the time to first treatment (TTFT) and overall survival (OS) (Rigolin GM, BJH 2018). However, it is unknown whether the prognostic strength of CK could be implemented when combined with stable markers such as the IGHV mutational status. In the present study, we assessed the prognostic and predictive role of the combination of CK subtypes and IGHV status in a large CLL series. METHODS. Stimulated cytogenetics with CpG+IL2 was performed in 736 CLL patients in 3 referenced Italian hematological centers. According to Rigolin et al, CK2 cases included unbalanced translocations, addition, insertion, derivative and marker chromosomes. All other CK were classified as type 1 (CK1). An IGHV gene sequence homology >98% was considered as unmutated (U-IGHV), as opposed to mutated (M-IGHV). Treatment was initiated according to the iwCLL guidelines. TTFT and OS were calculated from diagnosis to first treatment or death, respectively, or last known follow-up. Survival curves were compared with the log-rank test and p<.05 was considered as significant. Harrell concordance index (c-index) was used to compare our prognostic model with Dohner's and FISH-IGHV models. RESULTS. We focused on 520 out of the 736 patients with cytogenetic and IGHV status assessed within 12 months from diagnosis. The median age at diagnosis was 63 years, 322 (62%) were males, 68% at Binet A stage, 45% U-IGHV, 48 harbored TP53 abnormalities, 99 a CK (28 CK1 and 71 CK2), 232 received at least one line of therapy (31% FCR, 16% BR, 8% ibrutinib, 5% chlorambucil-antiCD20, 40% other treatments) and 80 died over a median follow-up of 5.8 years. 71 (14%) harbored CK2, 214 (41%) CK1 or U-IGHV and 235 (45%) M-IGHV without CK2. The former group were characterized by a higher prevalence TP53 (38% vs 8% vs 3%, p<0.0001) and cytogenetics abnormalities but lower cases with low-risk FISH (i.e. 13q or normal; 38% vs 54% vs 91%, p<0.0001) as compared with others two groups. We observed that subjects with CK2 had a shorter TTFT (median years 1.97, 3.40 and 19.1, p<0.0001) and 5 years OS (67%, 85%, 93%, p<0.0001) compared to cases with CK1/U-IGHV, or M-IGHV without CK. These data were confirmed in multivariate analysis. The worse prognosis of CK2 patients was independent of TP53 status (p values 0.0770 and 0.8122 for TTFT and OS, respectively). The c-indexes for our model were 69% and 68% for TTFT and OS, respectively, and were not inferior to those calculated with Dohner's (64% and 61%) and FISH-IGHV (69% and 63%) models. The combination of these two markers also provides predictive information after first-line therapy (p<0.0001 for both TTFT and OS). In particular, among 107 patients treated with FCR or BR just one of the M-IGHV cases relapsed but none died after a median follow-up of 43 months as compared with the other two subgroups (3-year PFS 92%, 69% and 23%, p<0.0001; 3-year OS: 100%, 94%, 62%, p<0.0001). CONCLUSIONS. In this study, we demonstrated that the combination of CK subtypes and IGHV status provides important prognostic and predictive data in CLL. Moreover, our model was not inferior to other commonly used prognostic scores. While patients with M-IGHV without any subtypes of CK showed an excellent outcome with chemoimmunotherapy, new alternative therapies should be explored for patients with CK2. Disclosures Visentin: janssen: Consultancy, Honoraria. Rigolin:Gilead: Research Funding. Mauro:abbvie: Other: board member; janssen: Other: board member. Foà:JANSSEN: Other: ADVISORY BOARD, Speakers Bureau; NOVARTIS: Speakers Bureau; INCYTE: Other: ADVISORY BOARD; CELTRION: Other: ADVISORY BOARD; ABBVIE: Other: ADVISORY BOARD, Speakers Bureau; AMGEN: Other: ADVISORY BOARD; GILEAD: Speakers Bureau; CELGENE: Other: ADVISORY BOARD, Speakers Bureau; ROCHE: Other: ADVISORY BOARD, Speakers Bureau. Cuneo:Roche: Other: advisory board, Speakers Bureau; Gilead: Other: advisory board, Speakers Bureau; Abbvie: Other: advisory board, Speakers Bureau; janssen: Other: advisory board, Speakers Bureau. Trentin:Gilead: Research Funding; Janssen: Research Funding; Abbvie: Honoraria; Roche: Membership on an entity's Board of Directors or advisory committees.

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Effect of Aggressive Haemoperfiusion on the Clinical Course of Patients with Paraquat Poisonin

Human & Experimental Toxicology ◽

10.1177/096032719301200411 ◽

1993 ◽

Vol 12 (4) ◽

pp. 323-327 ◽

Cited By ~ 34

Author(s):

Kouichiro Suzuki ◽

Nobukatsu Takasu ◽

Toru Okabe ◽

Shinichi Ishimatsu ◽

Akinori Ueda ◽

...

Keyword(s):

Survival Time ◽

Clinical Course ◽

Survival Rates ◽

Urine Volume ◽

Rank Test ◽

Log Rank Test ◽

Paraquat Poisoning ◽

Initial Plasma ◽

Number Of Patients ◽

The Difference

The effect of aggressive haemoperfusion; i.e. haemoperfusion of 10 h or more during the first 24 h after ingestion, on the clinical course of paraquat poisoning was studied. Among 40 patients admitted within 15 h after ingestion of paraquat with an SIPP of less than 100 (h x μg ml-1), 21 received aggressive haemoperfusion and 19 received conventional haemoperfusion; i.e. haemoperfusion of less than 10 h during the same period. Survival rates of patients with severity between an SIPP of 100 and Proudfoof's curve in the two groups were compared by the log-rank test. Aggressive haemoperfusion did not improve the outcome but did improve the survival rates; that is, the number of patients surviving at particulalr points in time (P<0,05). The length of haemoperfusion for the aggressive haemoperfusion group was longer than that for the conventional group on the first day (P<0.001 ), but the difference was insignificant during the following two days. Neither the time from ingestion to haemoperfusion, urine volume from the first to third day, nor initial plasma-paraquat concentrations and SIPP were significant between groups. These findings imply that aggressive haemoperfusion reduces the severity of paraquat poisoning and elongates survival time. We, therefore, propose that the efficacy of more aggressive haemoperfusion, such as the 'continuous haemoperfusion' proposed by Okonek et al., should be further studied.

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Risk Factor Analysis of Intravesical Recurrence After Laparoscopic Nephroureterectomy for Upper Tract Urothelial Carcinoma

10.21203/rs.3.rs-885355/v1 ◽

2021 ◽

Author(s):

Masato Yanagi ◽

Tsutomu Hamasaki ◽

JunJun Akatsuka ◽

Yuki Endo ◽

Hayato Takeda ◽

...

Keyword(s):

Urothelial Carcinoma ◽

Survival Rates ◽

Urine Cytology ◽

Rank Test ◽

Free Survival ◽

Log Rank Test ◽

Kaplan Meier ◽

Positive Urine ◽

Laparoscopic Nephroureterectomy

Abstract Background: One of the major concerns of patients with upper tract urothelial carcinoma (UTUC) treated with nephroureterectomy is intravesical recurrence (IVR). The purpose of the present study was to investigate the predictive risk factors for IVR after laparoscopic nephroureterectomy (LNU) for UTUC.Methods: Clinicopathological and surgical information were collected from the medical records of 73 patients treated with LNU for non-metastatic UTUC, without a history of or concomitant bladder cancer. The association between IVR after LNU and clinicopathological and surgery-related factors, including preoperative urine cytology and pneumoperitoneum time, was analyzed using Cox proportional hazards regression models and the Kaplan–Meier method with log-rank test.Results: During the median follow-up time of 39.1 months, 18 (24.7%) patients had subsequent IVR after LNU. The 3- and 5-year IVR-free survival rates were 76.5% and 74.3%, respectively. In the multivariate Cox regression analysis, positive preoperative urine cytology (hazard ratio [HR]: 3.55; 95% confidence interval [CI]: 1.326–11.327; p=0.011) and prolonged pneumoperitoneum time of ≥ 210 min (HR: 3.40; 95% CI: 1.271–10.692; p=0.014) were independent prognostic factors for IVR-free survival. In patients with positive urine cytology, the Kaplan–Meier method with log-rank test revealed that the 3-year and 5-years IVR free survival rates were 46.3% and 39.7%, respectively, in patients with a prolonged pneumoperitoneum time of ≥ 210 min, which was significantly lower than that in their counterparts (76% and 76%, respectively, p=0.041).Conclusions: In UTUC patients with positive urine cytology, the occurrence of IVR is highly probable when the pneumoperitoneum time of LNU is prolonged (≥ 210 min). Strict follow-up after LNU is highly recommended for these patients.

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Durable Responses to Single-Agent Ibrutinib in Monoallelic—but Not in Biallelic—TP53 Aberrated Patients with Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood-2018-99-114472 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 3111-3111

Author(s):

Christian Brieghel ◽

Inhye E. Ahn ◽

Sarah E. M. Herman ◽

Adrian Wiestner ◽

Carsten Utoft Niemann

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Cumulative Incidence ◽

Research Funding ◽

Limit Of Detection ◽

Single Agent ◽

Lymphocytic Leukemia ◽

Rank Test ◽

Excellent Outcome ◽

Synonymous Mutations

Abstract Introduction In chronic lymphocytic leukemia (CLL), TP53 aberration (TP53ab: del(17p) and/or TP53 mutation) remains the only biomarker directly influencing clinical practice (Blood. 2018;131(25):2745-2760). Further, TP53abs are early events that can lead to acquisition of additional somatic mutations, which suggest genomic instability caused by loss of p53, especially upon therapy. Single-agent ibrutinib is an effective therapy for both treatment-naïve (TN) and relapsed or refractory (R/R) CLL patients including those with del(17p). However, long-term progression-free survival (PFS) and overall survival are inferior for ibrutinib treated patients with TP53abs and R/R CLL due to early (<2 years) development of Richter's transformation or late (≥2 years) acquired resistance mutations in BTK and PLCG2. Aim To assess PFS and time to progression (TTP) in CLL patients treated with ibrutinib based on number of TP53abs. Methods Twenty-nine TP53 aberrated CLL patients from a phase II study of single-agent ibrutinib were included (Lancet Onc. 2015;16(2):169-176, www.clinicaltrials.gov #NCT01500733): 21 TN and 8 with R/R CLL. All patients received ibrutinib 420 mg once daily until progressive disease (PD) or intolerable side effects. The study was approved by the Institutional Review Board and conducted in accordance with the Declaration of Helsinki. The TP53 gene (exons 2-10) was assessed from baseline patient samples by deep targeted Next Generation Sequencing. Library preparation was performed according to manufacturer's protocol (Nimblegen) and pooled libraries were sequenced as paired-end on a NextSeq (2x150 base PE, Illumina). A bioinformatic pipeline was developed in CLC Biomedical Genomics Workbench 3.0 (Qiagen) including a validated dilution step resulting in a limit of detection of 0.2% variant allelic frequency (VAF). Synonymous mutations and single nucleotide polymorphisms were excluded. Patients were followed from treatment initiation until death, PD or end of follow-up, whichever came first. Pairwise log-rank test was applied for PFS, while Aalen-Johansen estimates of cumulative incidence rates were applied for TTP considering death as a competing risk. Analyses downstream of CLC were performed with R version 3.4.1. Results Twenty-four of 29 (83%) patients harbored 124 TP53 mutations (TP53muts) with a median of 0.7% VAF (IQR 0.3-3.1%): 22 high burden (VAF >10%) and 102 low burden TP53muts (VAF ≤10%) including 67 minor TP53muts (VAF <1%). Missense mutations accounted for 73%, while 14% of mutations were located to splice sites and another 14% were indels including 8 frameshift, 7 nonsense, and 2 in-frame mutations. Ninety-four percent of mutations were located within exons 5-9. Twenty-two patients had both del(17p) and TP53mut, 5 had del(17p) only, and 2 patients had only TP53muts of whom 1 had a single TP53:c.847C>T, which was predicted to encode functional p53 (http://p53.iarc.fr). Based on the number of TP53abs, 6 patients had only 1 TP53ab and 23 had >1 TP53ab. All patients with >1 TP53ab had biallelic TP53 disruption including 1 patient harboring a total of 38 TP53muts without concomitant del(17p). With a median follow-up time of 5.0 years (IQR: 4.9-6.0), PFS was 100% for patients with only 1 TP53ab. Among the 23 patients with >1 TP53ab, 9 patients had PD and 1 patient had died without progression (Figure 1A). There was no difference in PFS between patients with 2 TP53ab and patients with >2 TP53ab (Figure 1B). The cumulative incidence of PD for patients with >1 TP53ab was 39%, while no patients with only 1 TP53ab had progressed. Even though 5/6 patients with only 1 TP53ab were also TN, having >1 TP53ab impacted PFS negatively among both patients with TN (4 with PD and 1 death) and R/R CLL (5 with PD). Conclusion Patients with monoallelic TP53 aberration demonstrate excellent outcome on single-agent ibrutinib regardless of prior treatment status. Biallelic TP53 aberration identifies patients who have inferior outcome with ibrutinib monotherapy and may benefit from combination therapy. Among patients with biallelic TP53 disruption, having more than two TP53 aberrations does not portend a worse prognosis. The prognostic role of biallelic TP53 aberration calls for validation in independent, prospective studies and in clinical trials using targeted combination regimens. Disclosures Brieghel: Rigshospitalet, Denmark: Research Funding; Arvid Nilson's Fund: Research Funding. Wiestner:Pharmacyclics LLC, an AbbVie Company: Research Funding. Niemann:Janssen: Consultancy, Research Funding; Novo Nordisk Foundation: Research Funding; Gilead: Consultancy; CSL Behring: Consultancy; Danish Cancer Society: Research Funding; Roche: Consultancy; AstraZeneca: Consultancy; Abbvie: Consultancy, Research Funding; Novartis: Consultancy.

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ZAP-70 Expression, VH-Mutation Status, Genomic Aberrations and Prognosis in Chronic Lymphocytic Leukemia.

Blood ◽

10.1182/blood.v104.11.1920.1920 ◽

2004 ◽

Vol 104 (11) ◽

pp. 1920-1920 ◽

Cited By ~ 1

Author(s):

Alexander Kröber ◽

Dirk Kienle ◽

Dirk Winkler ◽

Andreas Bühler ◽

Till Seiler ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Gene Rearrangement ◽

Progressive Disease ◽

Clinical Course ◽

Surrogate Marker ◽

Lymphocytic Leukemia ◽

Mutation Status ◽

Free Survival ◽

Genomic Aberrations

Abstract The VH status is a strong prognostic marker in chronic lymphocytic leukemia (CLL). ZAP-70, a zeta associated tyrosine kinase physiologically expressed by T-cells, is overexpressed in VH unmutated CLL and could therefore serve as a surrogate marker for the VH status. We analyzed ZAP-70 expression (n=96), the VH status (n=75) and genomic aberrations (n=84) in a single center CLL cohort to study associations among these parameters and to assess their relative prognostic value. ZAP-70 expression was measured by 4-colour flow cytometry (CD5, CD19, CD3/56, ZAP-70) applying an unconjugated anti-ZAP-70-antibody (Upstate, clone 2F3.2) according to Crespo et al., NEJM 2003. ZAP-70 expression was positive (cut-off 20%) in 67% and negative in 33% of cases. VH was mutated in 33% and unmutated in 67% of cases. Unfavorable genomic aberrations (17p−, 11q−) were more frequently observed in cases with unmutated VH (46 vs. 9%) and in ZAP-70 positive cases (39 vs. 20%), while favorable genomic aberrations (13q− as single aberration) occurred more frequently in VH mutated (48 vs. 17%) and ZAP-70 negative subgroups (50 vs. 18%). ZAP-70 expression predicted the VH status in 84% of cases. At a median follow up time of 47 months (m), the median treatment free survival (TFS) of ZAP-70 positive and negative cases was 31 and 86 m (p=.057). The median TFS of the VH unmutated and VH mutated subgroups were 24 and 172 m (p<.001). Within the follow up time 10 deaths occurred. Of these, 8 cases exhibited high ZAP-70 expression and an unmutated VH, whereas 2 cases showed discordant results. Overall, discordant results for ZAP-70 expression and VH status were identified in 12 cases (ZAP-70 positive/VH mutated, 8 cases; ZAP-70 negative/VH unmutated, 4 cases). Of the 8 VH mutated cases with high ZAP-70 expression, only 1 case exhibited unfavorable genomic aberrations, 4 remained in stable disease, 4 developed progressive disease, 3 patients required therapy, and 1 of these 3 died within follow up time. Two of the 3 patients who required therapy, including the patient who died, showed a mutated V3-21 gene rearrangement, associated with an unfavorable outcome. Among the 4 cases with an unmutated VH and low ZAP-70 expression, 2 cases exhibited unfavorable genomic aberrations, 3 cases required therapy, 1 of these 3 died, and for one patient no clinical data were available. In summary, the imbalanced distribution of high risk genomic aberrations was similar when comparing the subgroups according to ZAP-70 expression and VH status. In our series an unmutated VH status predicted for shorter TFS, whereas high ZAP-70 expression did not reach significance. ZAP-70 expression was associated with unmutated VH, but a substantial number of cases showed discordant results for ZAP-70 expression and VH status. The pattern of genomic aberrations and the clinical course of the discordant cases were typical for their respective VH status. Compared to ZAP-70 expression the VH status appeared to be more informative in the prediction of the clinical course in our series of CLL patients.

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Evaluation of Integrated CLL Scoring System (ICSS) in 420 Patients with Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood.v128.22.5563.5563 ◽

2016 ◽

Vol 128 (22) ◽

pp. 5563-5563 ◽

Cited By ~ 1

Author(s):

Andrea Visentin ◽

Federica Frezzato ◽

Silvia Imbergamo ◽

Valentina Trimarco ◽

Veronica Martini ◽

...

Keyword(s):

High Risk ◽

Predictive Accuracy ◽

Lymphocytic Leukemia ◽

Low Risk ◽

Rank Test ◽

High Risk Patients ◽

Log Rank Test ◽

Risk Patients ◽

Normal Fish

Abstract BACKGROUND Chronic Lymphocytic Leukemia (CLL) is one of the most common hematological malignancies in Western countries. The disease is characterized by heterogeneous clinical course and outcome. During the last 15 years several clinical, biological and molecular prognostic factors have been identified, validated and some of them are currently used in patients' and treatment management. To improve the predictive accuracy of these markers, they have been combined into prognostic indexes (W. Wierda, JCO 2011, D. Rossi, Blood 2012, J. Bahlo, Haematologica 2015). Werecently proposed the Integrated CLL Scoring System (ICSS) based on cytogenetic abnormalities by FISH, IGHV mutational status and CD38 expression from 212 patients (A. Visentin et al, Clin Lymph Myeloma & Leuk 2015). The aim of this study was to validate the prognostic power of our index into a larger series of 420 CLL patients. METHODS 420 CLL patients referred to the Hematology Unit of Padua University Hospital from 1989 to 2015 were recruited in this study. According to ICSS, patients were classified as: low-risk, those patients with 13q deletion or normal FISH, IGVH mutated and CD38<30%; high-risk, subjects with 17p or 11q deletion and/or IGVH unmutated and CD38>30%; intermediate-risk, all remaining patients. Treatment free survival (TFS) was calculated as time from diagnosis to treatment (event), death or last known follow-up (censored). Overall survival (OS) was calculated from the date of diagnosis to death for any cause (event) or last known follow-up (censored). TFS and OS were compared with log-rank test and plotted using Kaplan-Meier method. The predictive accuracy of ICSS was evaluated by the Harrel's concordance index (c-index); a value >0.5 implies a good predictive ability. RESULTS The median age of our cohort was 62 years; 64% were male and 85% were Binet stage A at diagnosis. Cytogenetic analysis by FISH showed that 41 patients harbored 17p deletion, 50 11q deletion, 236 13q deletion, 44 trisomy 12 and 49 had normal FISH. 236 (56%) patients had IGHV gene homology >98% (i.e. mutated IGHV) and 103 (25%) expressed more then 30% of CD38. According to ICSS 202 (48%) subjects were classified as low-risk, 83 (20%) intermediate-risk and 135 (32%) high-risk. After a median follow-up of 81 months, the median TFS for ICSS classes of risk were 211, 70 and 27 months (log-rank test, p<0.0001, Figure 1A). The estimated 10-year TFS were 61%, 37% and 10% for low, intermediate and high-risk patients. The median OS were 213 and 136 months for intermediate and high-risk, while it was not reached for low-risk patients (Log-rank test, p<0.0001, Figure 1B). After 10 years from diagnosis the estimated OS were 88%, 79% and 57%, respectively. These data were confirmed by a multivariate analyses. In fact, high-risk patients had 5.3 and 4.0 times risk of start treatment and death than low-risk subjects, respectively (p<0.0001). This model was statistically internally validated, showing c-indexed of 0.712 and 0.693 for TFS and OS, respectively. In multivariate analyses, variables confirmed to predict adverse prognosis were male gender (p=0.0183), age>65years (p<0.0001), Rai III-IV (p=0.0025), Binet C (p=0.0002), 17p deletion (p=0.0002), TP53 abnormalities (p=0.0051), unmutated IGVH (p<0.0001), CD38>30% (p=0.0044) and high-risk ICSS (p<0.0001). CONCLUSIONS We herein provide evidence of the prognostic power and feasibility of ICSS into a large population of CLL patients. The use of this prognostic index could help physician into follow-up schedule, since high-risk patients should be monitored more often given the estimated increased risk of progression. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

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Benign monoclonal B cell lymphocytosis--a benign variant of CLL: clinical, immunologic, phenotypic, and cytogenetic studies in 20 patients

Blood ◽

10.1182/blood.v64.1.244.244 ◽

1984 ◽

Vol 64 (1) ◽

pp. 244-252 ◽

Cited By ~ 60

Author(s):

T Han ◽

H Ozer ◽

M Gavigan ◽

R Gajera ◽

J Minowada ◽

...

Keyword(s):

B Cell ◽

Clinical Course ◽

Normal Karyotype ◽

Lymphocytic Leukemia ◽

Thymidine Uptake ◽

Humoral And Cellular Immunity ◽

Essentially Normal ◽

Cell Counts

Abstract From 1951 through 1978, we have seen 20 cases of stage O chronic lymphocytic leukemia (CLL) without disease progression for 6.5–24 years. The cohort included 7 males and 13 females, aged 48–77 years at the time of diagnosis. None presented with anemia, thrombocytopenia, or neutropenia nor developed cytopenias during follow-up. Mean total lymphocyte count in these patients was 20,100/microL, with ranges from 10,000 to 43,700 at the time of diagnosis, and was 20,600, with ranges from 1,000 to 47,200, at last follow-up. Of 12 patients studied, 8 and 4 were phenotyped as heavy chain mu delta- and mu-type, respectively, with 7 kappa- and 4 gamma-type (no light chain was detectable in one patient). Of 13 patients studied, one had a slightly elevated IgG level and two had slightly depressed serum IgA and IgM levels. All patients had positive delayed hypersensitivity responses to at least one of five skin test antigens. Each of seven patients studied for an in vitro leukocyte thymidine uptake had a low level of [3H]thymidine incorporation. Nine of 12 patients studied had elevated total T cells, and the remaining 3 had normal T cell counts. In vitro unseparated lymphocyte response to phytohemagglutinin showed normal kinetics of DNA synthesis, with a peak response on day 3 or 4 of culture in 4 and slightly or moderately depressed and/or delayed kinetics in 8 patients studied. Cytogenetic analyses by Q- or G-banding techniques of polyclonal B cell mitogen-stimulated lymphocytes in all six patients studied showed normal karyotypes. These data are consistent with a previously undescribed syndrome involving a monoclonal B cell lymphocytosis, a prolonged asymptomatic or benign clinical course, and essentially normal humoral and cellular immunity and normal karyotype. Our observations indicate that these 20 patients with stage O CLL have a benign clinical course and that they may also be designated as benign monoclonal B cell lymphocytosis ( BMBL ), a benign variant of CLL.

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