Clinical Significance of CD69 Expression In Chronic Lymphocytic Leukemia

Abstract Abstract 3574 Chronic lymphocytic leukemia (B-CLL) is a very heterogeneous disease with some patients experiencing rapid disease progression and others living for years without requiring treatment and therefore it is mandatory to find new prognostic markers. CD69 overexpression which resembles B cells at an earlier and greater state of activation (Damle, 2002 and 2007) and induces increased proliferation and survival of leukemic B-lymphocytes, may reflect an aggressive and progressive clinical outcome. The primary endpoints of our research were: 1) to determine progression free survival (PFS) and overall survival (OS) upon CD69 in univariate analysis; 2) to correlate CD69 with other clinical or biological prognostic factors such as age, Rai stages, lymphocyte doubling time, beta-2 microglobulin, CD38, CD49d, ZAP-70, cytogenetics by FISH and IgVH status and finally, 3) to confirm CD69 as an independent prognostic factor. We investigated 417 patients (pts), median age 66 years (range 33–89), 239 males and 178 females. With regard to modified Rai stages, 127 pts had a low stage, 272 an intermediate stage and 18 a high stage. CD69 was determined by multicolor flow cytometry, fixing the cut-off value at 30%. CD69+ pts were 111/417 (26.6%). CD69 <30% was significantly associated with low Rai stage (111/127; P<0.0001), lymphocyte doubling time >12 months (260/337; P=0.0006), beta-2 microglobulin <2.2 mg/dl (176/218; P=0.0005) and soluble CD23 <70 U/ml (199/245; P<0.0001). Significant associations were found between CD69 <30% and ZAP-70 <20% (189/243; P=0.01) or CD49d <30% (135/171; P=0.007). There were significant correlations between CD69 <30% and IgVH mutated status (323 total cases, 169/211; P=0.001). On the other hand, no significant correlation was found with FISH cytogenetics (337 studied cases). With regard to clinical outcome, interestingly, 79 (71%) of 111 of the CD69+ patients had received chemotherapy at the time of analysis (P<0.00001). Moreover, both shorter PFS and OS were observed in CD69+ patients (5% vs 40% at 16 years, P<0.0001 and 26% vs 76% at 20 years, P<0.0001). To further explore the prognostic impact of CD69, we investigated its expression within unmutated (112 pts) and mutated (211 pts) IgVH subsets. As a matter of fact, pts with CD69 <30% showed longer PFS and OS both within the unmutated subgroup (32% vs 10% at 5 years, P=0.01 [Figure] and 77% vs 38% at 12 years, P=0.04) and within the mutated subgroup (56% vs 22% at 12 years, P=0.0006 [Figure] and 94% vs 70% at 16 years, P=0.05). In multivariate analysis of PFS, FISH cytogenetics (P=0.00005), ZAP-70 (P=0.0001), CD69 (P=0.002), Rai stages (P=0.001) and IgVH status (P=0.004) were independent prognostic factors. With regard to OS, age > or <60 years (P=0.001), CD69 (P=0.004), ZAP-70 (P=0.01) and CD38 (P=0.03) were identified as significant. Noteworthy, here, we demonstrated that CD69 is able to improve the historical prognostic ability of the IgVH mutational status. Since the IgVH mutated subset represents a large and heterogeneous population with a variable progression, CD69 may better define prognosis within this subgroup. Therefore, CD69, determined by flow cytometry, should be considered a novel important prognostic parameter in B-CLL and has to be necessarily added in a new scoring prognostic system. In fact, its easy and rapid laboratory determination allows us to identify early progressive pts in order to take timely therapeutic decisions. Disclosures: No relevant conflicts of interest to declare.

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Lymphocyte Doubling Time As A Key Prognostic Factor To Predict Time To First Treatment In Early-Stage Chronic Lymphocytic Leukemia

Frontiers in Oncology ◽

10.3389/fonc.2021.684621 ◽

2021 ◽

Vol 11 ◽

Author(s):

Fortunato Morabito ◽

Giovanni Tripepi ◽

Riccardo Moia ◽

Anna Grazia Recchia ◽

Paola Boggione ◽

...

Keyword(s):

Prognostic Factors ◽

Chronic Lymphocytic Leukemia ◽

Doubling Time ◽

Lymphocytic Leukemia ◽

Prognostic Impact ◽

Newly Diagnosed ◽

Prognostic Role ◽

Mutational Status ◽

Binet Stage ◽

Time To First Treatment

The prognostic role of lymphocyte doubling time (LDT) in chronic lymphocytic leukemia (CLL) was recognized more than three decades ago when the neoplastic clone’s biology was almost unknown. LDT was defined as the time needed for the peripheral blood lymphocyte count to double the of the initial observed value. Herein, the LDT prognostic value for time to first treatment (TTFT) was explored in our prospective O-CLL cohort and validated in in two additional CLL cohorts. Specifically, newly diagnosed Binet stage A CLL patients from 40 Italian Institutions, representative of the whole country, were prospectively enrolled into the O-CLL1-GISL protocol (clinicaltrial.gov identifier: NCT00917540). Two independent cohorts of newly diagnosed CLL patients recruited respectively at the Division of Hematology in Novara, Italy, and at the Hospital Clinic in Barcelona, Spain, were utilized as validation cohorts. In the training cohort, TTFT of patients with LDT >12 months was significantly longer related to those with a shorter LDT. At Cox multivariate regression model, LDT ≤ 12 months maintained a significant independent relationship with shorter TTFT along with IGHV unmutated (IGHVunmut) status, 11q and 17p deletions, elevated β2M, Rai stage I-II, and NOTCH1 mutations. Based on these statistics, two regression models were constructed including the same prognostic factors with or without the LDT. The model with the LTD provided a significantly better data fitting (χ2 = 8.25, P=0.0041). The risk prediction developed including LDT had better prognostic accuracy than those without LDT. Moreover, the Harrell’C index for the scores including LDT were higher than those without LDT, although the accepted 0.70 threshold exceeded in both cases. These findings were also confirmed when the same analysis was carried out according to TTFT’s explained variation. When data were further analyzed based on the combination between LDT and IGHV mutational status in the training and validation cohorts, IGHVunmut and LDT>12months group showed a predominant prognostic role over IGHVmut LTD ≤ 12 months (P=0.006) in the O-CLL validation cohort. However, this predominance was of borden-line significance (P=0.06) in the Barcelona group, while the significant prognostic impact was definitely lost in the Novara group. Overall, in this study, we demonstrated that LDT could be re-utilized together with the more sophisticated prognostic factors to manage the follow-up plans for Binet stage A CLL patients.

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Clinical Significance of NOTCH1 mutations in Chronic Lymphocytic Leukemia.

Blood ◽

10.1182/blood.v120.21.2870.2870 ◽

2012 ◽

Vol 120 (21) ◽

pp. 2870-2870

Author(s):

Giovanni Del Poeta ◽

Michele Dal Bo ◽

Maria Ilaria Del Principe ◽

Federico Pozzo ◽

Dario Ragusa ◽

...

Keyword(s):

Prognostic Factor ◽

Chronic Lymphocytic Leukemia ◽

Clinical Outcome ◽

Prognostic Markers ◽

Univariate Analysis ◽

Independent Prognostic Factor ◽

Lymphocytic Leukemia ◽

Prognostic Impact ◽

Fish Cytogenetics ◽

Arms Pcr

Abstract Abstract 2870 Chronic lymphocytic leukemia (CLL) is a very heterogeneous disease ranging from rapid disease progression leading to death to a nearly normal life expectancy and therefore it is mandatory to find valid prognostic markers. Recent studies showed that activating mutations of NOTCH1 proto-oncogene occur in about 10% CLL at diagnosis and are associated with an unfavorable clinical outcome (Rossi et al, 2012). About 85% of NOTCH1 mutated CLL cases displayed a DCT7544–7545 frameshift deletion (hereafter NOTCH1 mutation), that has been demonstrated to predict NOTCH1 degradation impairment through the truncation of the C-terminal PEST domain. Given the possibility of targeting NOTCH1 with drugs currently under development, the primary endpoints of our research were: 1) to correlate NOTCH1 mutation with other clinical and biological prognostic factors; 2) to determine time to first treatment (TTFT) and overall survival (OS) upon NOTCH1 mutation in univariate analysis; 3) to validate NOTCH1 mutation as an independent prognostic factor. We investigated 463 pts, median age 65 years (range 33–89), 256 males and 207 females. With regard to modified Rai stages at diagnosis, 159 had a low stage, 290 an intermediate stage and 14 a high stage. NOTCH1 mutation was investigated by amplification refractory mutation system (ARMS) PCR at diagnosis or before any chemotherapeutic approach. The ARMS PCR approach was set up in order to identify NOTCH1 mutation when present in at least 10% of the alleles. Using this approach, NOTCH1 mutated pts were 45/463 (9.7%). Considering the association with markers of tumor burden and proliferation, NOTCH1 mutation correlated with intermediate/high Rai stages (37/45; P=0.002), multiple thoracic/abdominal lymphadenopathies and/or splenomegaly (26/45, P=0.003), beta-2 microglobulin >2.2 mg/ml (27/45; P=0.02), lymphocyte doubling time <12 months (19/45; P=0.0006) and soluble CD23>70 U/ml (26/39; P=0.00001). Significant associations were also found with the main biologic prognostic markers in CLL. In this regard, NOTCH1 mutation was associated with an unmutated IGHV status (available for 446 total cases, 30/43; P<0.0001), CD38>30% (26/45, P<0.0001), ZAP-70>20% (33/45; P<0.0001), and CD49d>30% (22/34; P=0.009). Finally, considering associations with specific chromosomal aberrations defined by FISH cytogenetics (available in 417 cases), significant correlations (P=0.003) were found between NOTCH1 mutation and trisomy 12 (14/41; 25%), and del11q (7/41;16% ), whereas only 2/43 NOTCH1 mutated cases presented 17p deletion. With regard to clinical outcome, 30/45 (67%) NOTCH1 mutated pts received chemotherapy vs 193/418 (46%) among NOTCH1 germ line CLL (P=0.01), with 15/45 (33%) vs 48/418 (11%) cases, belonging to the same subgroups, undergoing at least two lines of treatment (P=0.001). Moreover, both significant shorter TTFT and OS were observed in NOTCH1 mutated pts (7% vs 35% at 12 years, P=0.0006 and 34% vs 78% at 14 years, P<0.0001; Figures A, B). To further test the clinical value of NOTCH1 mutation in CLL, we investigated its prognostic impact in bivariate analyses with the main clinical/biological prognosticators. According to these analyses, pts with NOTCH1 mutation showed shorter OS both within the ZAP-70 positive (>20%, 188 pts) and unmutated IGHV (<2%, 144 pts) subsets (24% vs 50% at 14 years; P=0.006, and 18% vs 52% at 14 years, P=0.04, respectively; Figures C, D). In multivariate analysis of OS, NOTCH1 mutation (P=0.02) together with age (P=0.001), FISH cytogenetics (P=0.001) and CD38 (P=0.002) was confirmed to be an independent prognostic factor, after correcting for colinearity with IGHV status. Therefore, NOTCH1 mutation, as determined by ARMS PCR, is a novel important prognostic parameter in CLL to be considered in drawing prognostic scores. In addition, NOTCH1 might represent a commendable therapeutic target for specific inhibitors to be employed especially in NOTCH1 mutated CLL. Disclosures: No relevant conflicts of interest to declare.

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Beta-2-Microglobulin Is a Reliable Tumor Marker in Chronic Lymphocytic Leukemia

Acta Haematologica ◽

10.1159/000205558 ◽

1989 ◽

Vol 81 (4) ◽

pp. 181-185 ◽

Cited By ~ 33

Author(s):

Samuele Di Giovanni ◽

Gaetano Valentini ◽

Paolo Carducci ◽

Paolo Giallonardo

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Tumor Marker ◽

Lymphocytic Leukemia ◽

Beta 2 ◽

Beta 2 Microglobulin

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Predicting Time to Initial Treatment in Early Stage CLL - Integration of Classical and Novel Prognostic Factors.

Blood ◽

10.1182/blood.v112.11.2093.2093 ◽

2008 ◽

Vol 112 (11) ◽

pp. 2093-2093

Author(s):

Michael J. Keating ◽

Susan Lerner ◽

Xuemei Wang ◽

Laura Z. Rassenti ◽

Susan O’Brien ◽

...

Keyword(s):

Prognostic Factors ◽

Lymph Node ◽

Initial Treatment ◽

Early Stage ◽

Tumor Burden ◽

Fish Cytogenetics ◽

Watch And Wait ◽

Mutation Status ◽

Beta 2 ◽

Beta 2 Microglobulin

Abstract CLL patients (pts) who are not eligible for initiation of treatment by the NCI Working Group criteria (1996) are traditionally monitored on a “watch and wait” approach. There is increasing interest in initiating therapy earlier in pts who are considered high risk. The most important advice pts require in the “watch and wait” category is whether they will need treatment and when. There are a number of traditional prognostic factors that potentially identify those “watch and wait” (W&W) pts most likely to need treatment. These have recently been amplified by the discovery of new prognostic factors such as the immunoglobulin (IgVH) mutation status, FISH cytogenetics, ZAP-70, and CD38 expression on CLL cells. Most reports have emphasized the new prognostic factors without consideration of traditional characteristics such as tumor burden, stage, etc. Since January 2004 FISH, IgVH mutation status, ZAP-70, and CD38 were characterized for most pts presenting to M. D. Anderson Cancer Center. We identified an early stage low tumor burden group of 826 pts (W&W) defined with as having lymph nodes < 3 cm in the longest dimension, splenomegaly < 5 cm, and Rai stage 0 – II. Twenty-four percent of these pts have received treatment in the first two years of follow-up. From the traditional prognostic factors, number of lymph node sites (cervical, axillary, and inguinal), absolute lymphocyte count (ALC), serum beta-2-microglobulin (B2M) levels were highly correlated with time-to-initial treatment (P<.001). The predictive power of these characteristics was also confirmed in an earlier historic patient population. When these three characteristics are evaluated in multivariate analysis, IgVH mutation status, ZAP-70, and FISH cytogenetics all add independent prognostic power. When these three novel characteristics were evaluated in multivariate model with the traditional characteristics, the most important were the number of node sites, ALC, B2M, FISH, IgVH mutation status, and then ZAP-70. Clinical only +Mutation Status +ZAP 70 +FISH + All 3 No. Lymph Node Sites 1 1 1 1 1 ALC 2 2 2 2 2 Beta-2-microglobulin 3 4 4 4 4 Mutation Status -- 3 -- -- 5 ZAP-70 -- -- 3 -- 6 FISH -- -- -- 3 3 Integration of novel prognostic factors into readily clinically available characteristics will enable us to develop a nomogram for predicting probability of each patient requiring therapy at one year, two years, and eventually five years. This nomogram will be presented when completed and then will be available for validation in independent patient populations studied by other groups.

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The Prognostic Significance of Positive Direct Antiglobulin Test in Chinese Patients with Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood.v112.11.4183.4183 ◽

2008 ◽

Vol 112 (11) ◽

pp. 4183-4183

Author(s):

Wei Xu ◽

Jianyong Li ◽

Xin Cao ◽

DAN-Xia Zhu ◽

Lin Yao ◽

...

Keyword(s):

Prognostic Factors ◽

Chronic Lymphocytic Leukemia ◽

Prognostic Significance ◽

Prognostic Indicator ◽

Lymphocytic Leukemia ◽

Direct Antiglobulin Test ◽

Chinese Patients ◽

Prognostic Impact ◽

Antiglobulin Test ◽

Positive Direct

Abstract Chronic lymphocytic leukemia (CLL) is the most common type of adult leukemias in the Western countries, however, infrequent in the Eastern. Autoimmune hemolytic anemia (AHA) is a complication in chronic lymphocytic leukemia (CLL). The direct antiglobulin test (DAT) may be positive at some time during the disease course in up to 35% of cases, but overt AHA occurs less frequently. The aim of the study was to explore the prognostic impact of positive DAT in Chinese patients with CLL and its correlation with other prognostic factors, including Binet stages, lymphocyte count in peripheral blood, lactate dehydrogenase (LDH), β2-microglobulin (β2-MG), IgVH mutation status, ZAP-70, CD38 and cytogenetic abnormalities. Out of the 80 Chinese patients with CLL, positive DAT was found in 21 (30.6%) cases. The incidence of positive was 12.5% in Binet A, 23.8% and 44.4% in Binet B and C, respectively. The incidence of positive DAT was significantly increased at Binet C, compared with Binet A (P=0.006), and the presence of higher LDH and β2-MG levels correlated strongly with positive DAT (P=0.006 and P=0.004, respectively). Patients with unmutated IgVH genes had higher incidence of positive DAT than did patients with IgVH mutations (P=0.042), and positive DAT was also associated with higher level of ZAP-70 and CD38 (P=0.004 and P<0.001, respectively). We also analyzed positive DAT in different cytogenetic subgroups. Higher incidence of positive DAT was found in patients with unfavorable cytogenetic aberrations (deletion in 17p13 or 11q22) in contrast to lower level in good risk cytogenetics (deletion in 13q as the sole abnormality) (P = 0.002). Positive DAT was associated with poor outcome. Survival analysis showed that the patients with positive DAT had significantly shorter OS (mean, 106.3 months) (95% CI, 74.7 to 137.8 months) than the patients negative DAT (mean, 151.5 months) (95% CI, 122.3 to 180.6 months) (P=0.024). Patients treated with fludarabine were not likely to remain DAT positive and to change from negative to positive (P=0.209). In conclusion, DAT status provides a new prognostic indicator and correlates with other clinical or laboratory prognostic factors, and might be applied for the assessment of prognosis in patients with CLL.

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Clinical Significance of Apoptosis in Chronic Lymphocytic Leukemia (CLL)

Blood ◽

10.1182/blood.v124.21.834.834 ◽

2014 ◽

Vol 124 (21) ◽

pp. 834-834

Author(s):

Giovanni Del Poeta ◽

Maria Ilaria Del Principe ◽

Michele Dal Bo ◽

Francesco Buccisano ◽

Dario Ragusa ◽

...

Keyword(s):

Flow Cytometry ◽

Prognostic Factor ◽

Conflicts Of Interest ◽

Intermediate Stage ◽

Independent Prognostic Factor ◽

Lymphocytic Leukemia ◽

Prognostic Impact ◽

Development Of Resistance ◽

Beta 2 Microglobulin ◽

Very High

Abstract Impaired programmed cell death is an important factor both in the pathogenesis of CLL (Berndt et al, Nat Genet 2013) and in the development of resistance to chemoimmunotherapy (Fegan et al, Adv Exp Med Biol 2013). It has become increasingly clear that in CLL the balance between the pro- (Bax) and anti-apoptotic members (Bcl-2) of the Bcl-2 family determines the chemotherapy sensitivity and ultimately progression free (PFS) and overall survival (OS). Moreover, the today availability in clinical use of novel potent oral pro-apoptotic BH3 peptidomimetics such as ABT 199 (Seymour et al, ASH 2013) emphasized the importance of Bcl-2 family-targeted therapy, prompting us to analyze the real impact of Bax/Bcl-2 ratio on CLL prognosis. The primary aims of our research were: 1) to correlate Bax/Bcl-2 ratio with other clinical and biological prognostic factors; 2) to determine PFS and OS upon Bax/Bcl-2 ratio; 3) to confirm Bax/Bcl-2 as an independent prognostic factor. Therefore we investigated 502 pts, median age 65 years (range 33-89), 279 males and 223 females. With regard to modified Rai stages at diagnosis, 170 patients had a low stage, 318 an intermediate stage and 14 a high stage. Bax/bcl-2 ratio was calculated by flow cytometry, dividing mean fluorescence intensity (MFI) of bax by MFI of bcl-2 on CD19+CD5+ CLL cells. The threshold was set at the median value >1.5 (range 0.27-6.10). Two hundred sixty- six patients were Bax/Bcl-2 ratio positive (266/502; 53%). Higher Bax/Bcl-2 ratio was significantly associated with low Rai stage, lymphocyte doubling time >12 months, beta-2 microglobulin <2.2 mg/dl and soluble CD23<70 U/ml (P<0.0001). Moreover, higher Bax/Bcl-2 was greatly represented within the low risk (normal or del13q) cytogenetics (221/341; P<0.0001). Noteworthy, significant correlations were found between lower Bax/Bcl-2 ratio and IGHV unmutated status (130/168; P<0.0001) or NOTCH1 (49/58; P<0.0001) or TP53 mutations (29/37; P=0.00007). With regard to clinical outcome, significant shorter PFS and OS were observed in patients with lower Bax/Bcl-2 ratio (10% vs 52% at 16 years; P<0.0001 and 46% vs 79% at 16 years; P<0.0001, respectively). To further explore the prognostic impact of Bax/Bcl-2 ratio, we investigated its expression within IGHV unmutated (168 pts) and TP53 mutated subgroups (37 pts), notoriously at worst prognosis. As a matter of fact, higher Bax/Bcl-2 ratio identified patients with a significant longer PFS (43% vs 10% and 50% vs 10% at 7 years; P=0.00002 and P=0.039, respectively, Figure), so suggesting its very high prognostic impact. In multivariate analysis of PFS (489 patients), Bax/Bcl-2 ratio (P<0.0001) together with modified Rai stages (P<0.0001), cytogenetics (P=0.0001), IGHV status (P<0.0001) and TP53 (P=0.001) was confirmed to be an independent prognostic factor. Therefore, the apoptotic index Bax/Bcl-2, performed by flow cytometry, is a powerful prognostic marker being able to identify patients at different prognosis also within IGHV unmutated and TP53 mutated subsets which are at very high risk. The modern strategies to downregulate Bcl-2 and shift the balance toward cellular demise, such as the BH3 mimetic ABT 199, could be carefully and precisely monitored by using this simple but powerful flow cytometric approach. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

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