Targeting cytohesin-1 suppresses acute myeloid leukemia progression and overcomes resistance to ABT-199.

Adhesion molecules play essential roles in the homeostatic regulation and malignant transformation of hematopoietic cells. The dysregulated expression of adhesion molecules in leukemic cells accelerates disease progression and the development of drug resistance. Thus, targeting adhesion molecules represents an attractive anti-leukemic therapeutic strategy. In this study, we investigated the prognostic role and functional significance of cytohesin-1 (CYTH1) in acute myeloid leukemia (AML). Analysis of AML patient data from the GEPIA and BloodSpot databases revealed that CYTH1 was significantly overexpressed in AML and independently correlated with prognosis. Functional assays using AML cell lines and an AML xenograft mouse model confirmed that CYTH1 depletion significantly inhibited the adhesion, migration, homing, and engraftment of leukemic cells, delaying disease progression and prolonging animal survival. The CYTH1 inhibitor SecinH3 exerted in vitro and in vivo anti-leukemic effects by disrupting leukemic adhesion and survival programs. In line with the CYTH1 knockdown results, targeting CYTH1 by SecinH3 suppressed integrin-associated adhesion signaling by reducing ITGB2 expression. SecinH3 treatment efficiently induced the apoptosis and inhibited the growth of a panel of AML cell lines (MOLM-13, MV4-11 and THP-1) with mixed-lineage leukemia gene rearrangement, partly by reducing the expression of the anti-apoptotic protein MCL1. Moreover, we showed that SecinH3 synergized with the BCL2-selective inhibitor ABT-199 (venetoclax) to inhibit the proliferation and promote the apoptosis of ABT-199-resistant leukemic cells. Taken together, our results not only shed light on the role of CYTH1 in cell-adhesion-mediated leukemogenesis but also propose a novel combination treatment strategy for AML.

Orelabrutinib for the treatment of relapsed or refractory MCL: a phase 1/2, open-label, multicenter, single-arm study.

Relapsed or refractory (r/r) mantle cell lymphoma (MCL) is an aggressive B-cell malignancy with a poor prognosis. Bruton tyrosine kinase (BTK) is a mediator of B-cell receptor signaling and is associated with the development of B-cell lymphomas. Patients with r/r MCL were enrolled in this phase 1/2 study and treated with orelabrutinib, a novel, highly selective BTK inhibitor. The median number of prior regimens was 2 (range, 1-4). The median age was 62 years (range, 37-73 years). Eligible patients received oral orelabrutinib 150 mg once daily (n = 86) or 100 mg twice daily (n = 20) until disease progression or unacceptable toxicity. A dose of 150 mg once daily was chosen as the preferred recommended phase 2 dose. After a median follow-up duration of 23.8 months, the overall response rate was 81.1%, with 27.4% achieving a complete response and 53.8% achieving a partial response. The median duration of response and progression-free survival were 22.9 and 22.0 months, respectively. The median overall survival (OS) was not reached, and the rate of OS at 24 months was 74.3%. Adverse events (AEs) occurring in >20% of patients were thrombocytopenia (34.0%), upper respiratory tract infection (27.4%), and neutropenia (24.5%). Grade ≥3 AEs were infrequent and most commonly included thrombocytopenia (13.2%), neutropenia (8.5%), and anemia (7.5%). Three patients discontinued treatment because of treatment-related adverse events (TRAEs), but no fatal TRAEs were reported. Orelabrutinib showed substantial efficacy and was well tolerated in patients with r/r MCL. This trial was registered at www.clinicaltrials.gov as #NCT03494179.

Low incidence of hepatitis B virus reactivation in patients with hematological malignancies receiving novel anticancer drugs: A report from a high epidemic area and literature review.

BACKGROUND: More and more novel anticancer drugs have been approved for patients with hematological malignancies in recent years, but HBV reactivation (HBV-R) data in this population is very scarce. This study aimed to evaluated HBV-R risk in patients with hematological malignancies receiving novel anticancer drugs. METHODS: HBV markers and serum HBV DNA levels of patients with hematological malignancies receiving novel anticancer drugs in a tertiary cancer hospital were retrospectively collected. HBV-R risk in the whole cohort and subgroups was described. The relevant literature was reviewed to make a pooled analysis. RESULTS: Of 845 patients receiving novel anticancer drugs, 258 (30.5%) were considered at risk for HBV-R. The median duration of exposure to novel drugs was 5.6 (0.1-67.6) months. The incidence of HBV-R was 2.1% in patients with past HBV infection without prophylactic antiviral treatment (PAT) and 1.2% in all patients at risk of HBV-R. In a pooled analysis of 11 studies with 464 patients, the incidence of HBV-R was 2.4% (95% CI: 1.3-4.2) in all at-risk patients receiving novel anticancer drugs and 0.6% (95% CI: 0.03-3.5) in patients with anticancer drugs plus PAT. The incidence of death due to HBV-R was 0.4% (95% CI: 0.1-1.6) in all at-risk patients and 18.2% (95% CI: 3.2-47.7) in patients with HBV-R. CONCLUSION: Most episodes of HBV-R are preventable, and most cases with HBV-R are manageable. We recommend that novel anticancer drugs should not be intentionally avoided when treating cancer patients with HBV infection.

[Regulation of TRIP13 on Proliferation and Apoptosis of B-Cell Lymphoma Cells and Its Mechanism].

OBJECTIVE: To explore the regulatory effect of TRIP13 on the proliferation and apoptosis of B-cell lymphoma cells and its possible molecular mechanism by knocking down/overexpressing TRIP13 on the cell lines Granta-519 and JVM-2. METHODS: Lentiviral transfection technology was used to construct Granta-519 and JVM-2 cells with knocked down or overexpressed TRIP13 and their control cells. The efficiency of transfection was determined by fluorescence microscopy. The efficiency of knockdown and overexpression was evaluated by real-time quantitative PCR and Western blot. The proliferation was detected by CCK-8 assay. The apoptosis was detected by the Annexin V-APC single staining. The cell cycle was detected by the PI staining. The expression levels of P53, MDM4, and BCL-2 were evaluated by Western blot. RESULTS: After TRIP13 was knocked down, the proliferation ability of Granta-519 and JVM-2 cells was significantly reduced, and the apoptosis rate significantly increased. After TRIP13 was overexpressed, the proliferation ability of Granta-519 and JVM-2 cells was significantly enhanced, and the apoptosis was significantly reduced. After TRIP13 was knocked down, Granta-519 cells had obvious G1 phase arrest, and JVM-2 cells had obvious G1 and G2/M phase arrest. After TRIP13 was knocked down in Granta-519 cells, the expression of BCL-2 protein decreased, while MDM4 protein increased. After TRIP13 was overexpressed, the expression of MDM4 protein decreased. After TRIP13 was overexpressed in JVM-2 cells, the expression of BCL-2 protein increased. CONCLUSION: TRIP13 promotes the proliferation of B-cell lymphoma cells, inhibits their apoptosis, and affects their proliferation and apoptosis by participating in the regulation of the cell cycle. TRIP13 promotes the expression of BCL-2 proteins and inhibits the expression of MDM4 protein in B-cell lymphoma cells.

The MAGNOLIA Trial: Zanubrutinib, a Next-Generation Bruton Tyrosine Kinase Inhibitor, Demonstrates Safety and Efficacy in Relapsed/Refractory Marginal Zone Lymphoma.

PURPOSE: Marginal zone lymphoma (MZL) is an uncommon non-Hodgkin lymphoma with malignant cells that exhibit a consistent dependency on B-cell receptor signaling. We evaluated the efficacy and safety of zanubrutinib, a next-generation selective Bruton tyrosine kinase inhibitor, in patients with relapsed/refractory (R/R) MZL. PATIENTS AND METHODS: Patients with R/R MZL were enrolled in the phase II MAGNOLIA (BGB-3111-214) study. The primary endpoint was overall response rate (ORR) as determined by an independent review committee (IRC) based on the Lugano 2014 classification. RESULTS: Sixty-eight patients were enrolled. After a median follow-up of 15.7 months (range, 1.6 to 21.9 months), the IRC-assessed ORR was 68.2% and complete response (CR) was 25.8%. The ORR by investigator assessment was 74.2%, and the CR rate was 25.8%. The median duration of response (DOR) and median progression-free survival (PFS) by independent review was not reached. The IRC-assessed DOR rate at 12 months was 93.0%, and IRC-assessed PFS rate was 82.5% at both 12 and 15 months. Treatment was well tolerated with the majority of adverse events (AE) being grade 1 or 2. The most common AEs were diarrhea (22.1%), contusion (20.6%), and constipation (14.7%). Atrial fibrillation/flutter was reported in 2 patients; 1 patient had grade 3 hypertension. No patient experienced major hemorrhage. In total, 4 patients discontinued treatment due to AEs, none of which were considered treatment-related by the investigators. CONCLUSIONS: Zanubrutinib demonstrated high ORR and CR rate with durable disease control and a favorable safety profile in patients with R/R MZL.

[Application of lenalidomide in chronic lymphocytic leukemia].

In recent years, the incidence of chronic lymphocytic leukemia (CLL) is increasing. Microenvironment and immune system play a key role in the pathogenesis of CLL. The immune system is aggravated by the use of chemotherapeutic agents, such as fludarabine and cyclophosphamide with rituximab(FCR) which are the current standards in frontline therapy. This leads to an increase of infection incidence in patients, resulting in a poor prognosis. The present situation was changed by lenalidomide. Recent studies indicated that lenalidomide monotherapy in treatment of refractory or relapsed CLL patients, the overall response rate(ORR) reached about 32%-47%, CR roughly was 7%-13%; when lenalidomide and rituximab were combined for treatment of refractory or relapsed CLL patients, the ORR reached about 53%-66%, CR about 12%-13%. Moreover, when lenalidomide and ofatumumab were combined, the efficacy is improved significantly and the adverse reactions are greatly reduced. The adverse reactions are neutrophilic granulocytopenia, thrombocytopenia, anemia, tumor lysis syndrome(TLS), tumor flare reaction(TFR) and venous thromboembolism(VTE). This review focuses on the related studies and the latest progress about lenalidomide in CLL.

[Expression and prognostic significance of TOSO in CD19+ B cells from Chinese CLL patients].

OBJECTIVE: To investigate the expression and its prognostic significance of TOSO in CD19(+) B cells from Chinese chronic lymphocytic leukemia (CLL) patients. METHODS: The expression of TOSO was detected by absolute quantitative reverse transcription-polymerase chain reaction (RT-PCR) in a cohort of 85 untreated CLL patients from March 2006 to September 2010. Their status of immunoglobulin heavy chain variable (IGVH) somatic mutation, ZAP70 and CD38 were analyzed. RESULTS: The expression of TOSO was significantly elevated in CLL patients versus the healthy population (8.30 ± 2.99 vs 6.63 ± 1.22, P = 0.036) and other B cell lymphoproliferative diseases (8.30 ± 2.99 vs 7.12 ± 1.13, P = 0.023). The expression of TOSO was elevated in the IGVH non-mutated group versus the mutated group (9.87 ± 1.08 vs 7.61 ± 3.03, P = 0.000). The expression of TOSO was significantly elevated in Binet stage C patients versus in Binet stage B and Binet stage A patients (9.91 ± 3.03 vs 8.73 ± 1.86 vs 7.27 ± 2.83, P = 0.000). The expression of TOSO rose to (9.37 ± 2.12) in the chemotherapy group. And it was significantly higher than that in the observation group (7.19 ± 3.23, P = 0.001). At the cut-off point of 55 years old, the younger patients had a higher expression of TOSO than the elders (9.10 ± 2.06 vs 7.95 ± 3.22, P = 0.049). The expression of TOSO in ZAP70 and CD38-positive groups had no difference with those in the negative groups. CONCLUSION: The over-expressed TOSO in CLL cells is associated with a progressive disease. It may be an important prognostic factor for CLL.

[Overrepresentation of specific gene segments of expressed immunoglobulin heavy chain variable region among unmutated and mutated patients with chronic lymphocytic leukemia].

OBJECTIVE: To investigate the overrepresentation of specific gene segments of immunoglobulin heavy chain variable region (IgVH) among unmutated and mutated chronic lymphocytic leukemia (CLL) patients and its prognostic implication. METHODS: Multiplex PCR was used to identify the expression of IgVH segment and its mutation status in CLL. RESULTS: Analyses were successfully performed in 80 of 85 samples. Marked skewed IgVH families were disclosed. The most commonly used VH was VH3 (40.0%), followed by VH4 (30.0%), VHI (13.8%), VH2 (10.0%) and VH5, VH7 (2.5%). Fifty-six patients (70.0%) had mutated VH, 24 (30.0%) unmutated VH. Nine cases (11.3%) were with 100% germline sequence. Fifteen cases (15/24, 62.5%) in VH4, 29 (29/32, 90.7%) in VH3, and 4 (4/11, 36.3%) in VH1 had mutated VH. The most frequently used IgVH gene was VH4-39 (13.8%), and VH4-34 (8.8%). J4 (36/66, 54.5%) and D3 (25/66, 37.8%) were the most frequently used in J and D genes. The progression-free survival (PFS) was 82 and 17 months (P = 0.000), and the overall survival (OS) was 90 and 41 months (P = 0.009), respectively, for mutated and unmutated cases. Recurrent CDR3 sequences were found in our patients and 2 patients with VH1-69 had CDR3 sequences highly similar to those reported in literature. CONCLUSION: There is difference in IgVH gene segment usage and mutational status in different area CLL patients. Recurrent CDR3 sequences were found in specific IgVH gene segments, which highlights the importance of immunoglobulin mediated stimulation in the development of CLL.

[The expression of microRNA-155 and microRNA-146a and its' clinical value in chronic lymphoproliferative disorders].

OBJECTIVE: To investigate the expression of microRNA-155 and microRNA-146a in the CD19(+) B cells of chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), splenic marginal zone lymphoma (SMZL), and to analyze its clinical significance. METHODS: Peripheral blood (PB) (78 cases) and bone marrow (BM) samples (9 cases) from 53 CLL patients, 13 MCL patients, 19 SMZL patients, and 12 healthy donors were collected. Mononuclear cells were isolated and B cells were purified with a CD19(+) magnetic-bead system. Total RNA was extracted from purified CD19(+) cells and microRNAs expression were measured using the TaqMan microRNA quantitative PCR. The results combined with the clinic data of patients were analysed. RESULTS: (1) The expression of microRNA-155 in CLL (4.49 ± 0.83) was significantly higher than in MCL (3.83 ± 0.45) and SMZL (3.80 ± 0.61) (P < 0.05); (2) The level of microRNA-146a in SMZL (3.81 ± 0.59) was significantly higher than in CLL (2.58 ± 0.90) and MCL (2.27 ± 0.88) (P < 0.01); (3) The level of microRNA-155 was significantly higher in IgVH unmutated patients than in mutated patients in CLL (P = 0.012); (4) The microRNAs expression had no statistical difference between two prognostic groups in CLL. CONCLUSION: (1) The expression of microRNA-155 and microRNA-146a is different in malignant lymphoproliferative disorders (LPD); (2) Deregulation of the microRNAs expression might play a critical role in the pathogenesis and prognosis in the LPD.

[Expression of microRNA-223 and its clinical value in B lymphoproliferative disorders].

OBJECTIVE: To detect the expression of microRNA-223 and analyze its clinical value in B lymphoproliferative disorders. METHODS: Peripheral blood samples (n = 78) and bone marrow samples (n = 9) were collected from patients with chronic lymphocytic leukemia (CLL, n = 53), mantle cell lymphoma (MCL, n = 13), splenic marginal zone lymphoma (SMZL, n = 9) and healthy donors (n = 12) at our hospital from 2003 to 2010. Mononuclear cells were isolated and B cells purified with a CD19(+) magnetic-bead system. Total RNA was extracted from purified CD19(+) cells and the expression of microRNA-223 measured by TaqMan microRNA quantitative polymerase chain reaction (PCR). The clinical data of these patients were collected and their outcomes analyzed with SPSS 16.0 software. RESULTS: (1) The levels of microRNA-223 in CLL, MCL and SMZL were 4.58 ± 0.62, 4.03 ± 0.54 and 4.63 ± 0.57 respectively. And they were significantly lower than that in normal B cells (5.69 ± 0.60, P < 0.01). The expression of microRNA-223 decreased significantly in MCL versus CLL and SMZL (P < 0.05). There was no statistical difference between CLL and SMZL (P > 0.05). (2) The down-regulation of microRNA-223 was associated with disease aggressiveness in CLL. Patients with unmutated immunoglobulin heavy chain variable region (IgVH) expressed significantly a lower level of microRNA-223 (4.05 ± 0.69 vs 4.67 ± 0.51, P = 0.003). In 13q-negative patients, the expression of microRNA-223 decreased more significantly than that in 13q-positive patients (4.25 ± 0.67 vs 4.76 ± 0.45, P = 0.044). (3) Using receiver operating characteristic (ROC) curve analysis, the microRNA-223 cutoffs were defined according to the IgVH mutational status. The patients were divided into the positive and negative subgroups. The median progression-free survival (PFS) of microRNA-223 positive patient subgroup was 48 months. It was significantly longer than the negative subgroup (P = 0.001). In the microRNA-223 positive subgroup, no patient died at the end of follow-up. CONCLUSIONS: MicroRNA-223 may play an important role in the pathogenesis of B lymphoproliferative disorders. The down-regulation of microRNA-223 is associated with disease aggressiveness and poor prognostic factors in CLL. It may become a new reliable prognostic predictor.

[Expression of stereotyped B-cell receptor and its clinical value in chronic lymphocytic leukemia].

OBJECTIVE: To investigate the expression pattern of stereotyped B-cell receptor (BCR) and its prognostic significance in Chinese chronic lymphocytic leukemia (CLL) patients and evaluate the relationship to other prognostic markers. METHODS: Multiplex polymerase chain reaction (PCR) was used to identify the immunoglobulin variable heavy-chain (IGHV) segment and its mutation status in 116 CLL patients from April 1992 to April 2010. For CDR3-driven clustering, all in-frame IGHV-D-J rearrangements were aligned by the multiple sequence alignment software ClustalW2. RESULTS: There were 102 of 116 samples from newly diagnosed CLL were successfully analyzed. IGHV CDR3 genes were identified in 73/102 cases. Fourteen patients (19.2%) carried stereotyped BCR. A high percentage of carried stereotyped BCR was observed in IGHV non-mutated group versus mutated group (40.9% vs 11.8%, P = 0.005). Patients with stereotyped BCR had a higher frequency of deletion (17q) (33.3% vs 10.7%, P = 0.045). The median progression-free survival (PFS) in patients with stereotyped BCR was much shorter than other patients (39 vs 84 months, P = 0.002). CONCLUSION: The patients with stereotyped BCR have a poor prognosis. It highlights the importance of immunoglobulin mediated stimulation in the development of CLL.

[Analysis of efficacy and related factors in 85 chronic myeloid leukemia patients treated with imatinib mesylate].

The objective of this study was to evaluate the efficacy of Imatinib on patients with chronic myeloid leukemia (CML) in chronic phase and to analyze its influencing factors. 85 patients received Imatinib mesylate at a dose of 300-600 mg orally per day, and were evaluated for hematologic, cytogenetic, and molecular responses. The results showed that the median follow-up was 21 (range 9 - 78) months. Cumulative complete hematological remission (CHR) rate was 100%, major cytogenetic remission (MCyR) rate was 80%, complete cytogenetic remission (CCyR) rate was 67.1% and complete molecular remission (CMoR) rate was 36.4%. The median time to complete hematological remission (CHR) was 1 (range 1 - 3) month, to complete cytogenetic remission (CCyR) was 6 (range 1 - 24) months. The estimated overall survival rates for patients who received Imatinib for 1, 2, 3 years were (98.7 +/- 1.3)%, (96.5 +/- 2.5)% and (90.1 +/- 6.6)% respectively. The estimated progression-free survival rates at 1, 2, 3 years were (97.6 +/- 1.6)%, (96.1 +/- 2.2)% and (90.0 +/- 1.4)% respectively. The CHR, MCyR and CCyR between low risk, intermediate risk and high risk groups according to the Sokal scoring system and between primarily treated and retreated groups all had no difference. The overall survival of patients who achieved MCyR or CCyR was better than that in patients only achieved hematologic remission (p = 0.026), but there was no significant difference in progression-free survival between them. Univariate analysis for efficacy of Imatinib mesylate revealed that WBC count < 100 x 10(9)/L (p = 0.024), Hb level > or = 130 g/L (p = 0.036), and peripheral basophil count < or = 0.05 (p = 0.024) before therapy were independent favourable factors for achieving MCyR or CCyR. It is concluded that the patients with CML in chronic phase treated with Imatinib can achieve the best hematologic remission and higher cytogenetic remission, it should be considered that the imatinib is a drug of the first-line therapy for untreated and treated patients with CML.