bluuebunny/crossref_metadata_2025 huggingface dataset:
Treatment of CD 20+ Hodgkin Lymphoma (Lymphocyte Predominant) with Monoclonal Antibody Rituximab after First-Line Treatment Is Effective and Might Improve the Outcome
Authors: Genadieva-Stavrik Sonja , Pivkova Alexandra , Ivkovski Ljube , Cevreska Lidija
Published: April 2020

Lymphocyte-predominant Hodgkin disease (LPHD) presents as early-stage disease with slow disease progression and excellent initial response to conventional chemotherapy. Although 96% of LPHD patients experience a complete remission (CR) upon first line treatment, many relapses occur. Residual tumor cells in Hodgkin’s patients can survive standard therapy and cause relapse. Thus, the elimination of minimal residual disease after first line treatment might improve the outcome in Hodgkin disease. CD20+ is strongly expressed by malignant cells in LPHD, but so far there is limited information regarding the efficacy of Rituximab in this patient population. A 66 year-old woman presented with bilateral inguinal lymphadenopathy in 2004 and complained on malaise and weight loss. Lymph node biopsy confirmed a diagnosis of LPHD and immunohistochemistry confirmed that CD20 antigen was expressed on more than 30% of malignant cells. Staging revealed II B disease. There were lymph node enlargements in the pelvis on CT. Her blood count, erythrocyte sedimentation rate, and albumin were normal with elevation of and lactate dehydrogenase and alkaline phoshatase. She was treated with ABVD regiment 8 cycles and CR was achieved. After standard chemotherapy patient with CD20+ LPHD received four weekly doses of Rituximab at 375 mg/m2 and maintenance therapy with additional four doses of Rituximab every three months for the next year. Treatment was well tolerated without any complications. Evaluation of disease assessment included physical examination and computed tomography, bone marrow biopsy and routine analyses, after the end of treatment and every 3 months after for the first two years, and on six months for the following year. There were no abnormalities on CT after the end of the treatment. Patient was in complete remission. CT after 36 months showed no detectible tumor mass. Patient remains still in complete remission three years after chemotherapy without detectible tumor mass.

Our case report suggests that Rituximab is both safe and effective in patients with CD20+ LPHD. The optimal treatment for patients with LPHD remains uncertain. Current research aims to identify alternative treatment approaches that possess significant activity but less toxicity. Radiotherapy for early-stage patients and chemotherapy or combined modality treatment for advanced-stage patients remains to be standard of care. Initial observed benefit of Rituximab treatment in this group of patients suggests that there is a role for monoclonal antibodies in optimal treatment approach, especially in improving the outcome. Further studies with Rituximab are warranted in this patient population.
Targeting Early Events of B-Cell Receptor Signaling in Chronic Lymphocytic Leukemia: Suppressed Syk and PLCγ2 Activities Predict Apoptotic Response of Leukemic Cells to Dasatinib
Authors: Wang Y. Lynn , Song Zibo , Lu Pin , Leonard John P. , Coleman Morton , Martin Peter , Wu Chaowei , Lee Francis Y. , Knowles Daniel M. , Furman Richard
Published: April 2020

B cell receptor (BCR) signaling plays an essential role in the pathogenesis of chronic lymphocytic leukemia. In a subset of patients with a poor clinical outcome, BCR ligation leads to increased cell metabolism and cell survival (Cancer Research66, 7158–66, 2006). Based on these findings, we tested whether targeting BCR signaling with dasatinib, an inhibitor of Src kinase, would interfere with the signaling cascade and cause death of CLL B cells. CLL leukemic cells were isolated from 34 patients and were incubated with or without dasatinib at a low dose of 128 nM. Among 34 cases, viability of leukemic cells was reduced by 2% to 90%, with an average of ~50% reduction on day 4 of ex vivo culture. Further study showed that CLL B cells undergo death by apoptosis via the intrinsic pathway which involves the generation of reactive oxygen species. Analysis of the Src family kinases showed that phosphorylation of Src, Lyn and Hck was inhibited by dasatinib not only in those cases that responded to dasatinib with apoptosis, but also in those that did not respond well (<20% apoptosis). Further analysis revealed that suppressed activity of two downstream molecules, Syk and PLC

Statistical analysis showed a significant correlation between CLL dasatinib response and their IgVH mutation and ZAP70 status. Cases with worse prognoses by these criteria have a better response to the kinase inhibitor. Lastly, we have also found that ZAP70 positive cases showed a greater degree of PLC
Design and Synthesis of Novel Derivatives of ATRA Demonstrate the Combined Importance of Acid Moiety and Conjugated Double Bonds in Its Binding to PMLRAR-Alpha Oncogene in Acute Promyelocytic Leukemia
Authors: Goel Swati , Schinke Carolina , Bhagat Tushar , Mo Yongkai , Zhou Li , Heuck Christoph , Alencar Christina , Pahanish Perry , Anguiano Jaime , Kabalka George , Platanias Leonidas C , Verma Amit , Das Bhaskar
Published: April 2020

All trans retinoic acid (ATRA) is the treatment of choice for Acute Promyelocytic Leukemia (APL) and binds to the transcriptional repressor PML-RAR-alpha oncogene. The binding of ATRA to RAR-alpha relieves transcriptional repression and leads to induction of genes important for myeloid differentiation. ATRA consists of an acid moiety at position 15 and is also characterized by conjugated double alkene bonds (Fig1A). The exact contributions of these structural components of ATRA in its clinically important binding to RAR-alpha receptor are not very well studied. In an effort to improve the therapeutic index and to study its binding mechanism, we synthesized novel retinoic acid analogs (retinoids). Our objective was to construct retinoids to test these structure functional correlates. 3 novel retinoids created were synthesized as follows:

Compound 13a (Fig1B): consisting of modifying conjugated alkene backbone while keeping acid moiety intact.

Compound 13b (Fig1C): consisting of modifying conjugated alkene backbone and converting acid to ester moiety.

Compound 13c (Fig1D): consisting of modifying conjugated alkene and cycloalkene ring backbone and keeping acid moiety intact.

We tested the ability of these newer retinoids to bind to RAR-alpha as well as inhibit the proliferation of PML-RAR-alpha containing acute promyelocytic leukemic cell line NB4. Reporter assays using a RAR- luciferase demonstrated that Compound 13a successfully relieved transcriptional repression by RAR-alpha and led to downstream gene induction. This was significantly less compared to wild type ATRA demonstrating the importance of C9–C10 double bonds in this binding. Compounds 13b and 13c could not relieve RAR mediated repression of gene induction demonstrating that the acid moiety plays a important role in this binding. Functional studies were consistent with these observations and showed that both ATRA and 13a were able to inhibit the proliferation of APL cells while 13b, 13c were not. Flow cytometry showed that 13a led to partial differentiation of NB4 cells, as evident from CD11b expression after five days of culture. 13b and 13c did not lead to any differentiation, while ATRA led to the greatest expression of CD11b.

Our results demonstrate that both the acid moiety and conjugated double bonds are important in binding to RAR-alpha and the subsequent transcriptional repression by retinoic acid. These insights will be important in future efforts to improve the stability and efficacy of ATRA and also have implication in efforts focused on treating ATRA resistant cases of APL.

Figure Figure
Effects of Imatinib on the Pharmacokinetics of Acetaminophen in Korean Patients with Chronic Myelogenous Leukemia (CML)
Authors: Kim Dong-Wook , Park Sa-Hee , Jin Yu , Hayes Michael , Wang Yanfeng
Published: April 2020

Background: Drug-drug interactions between imatinib (IM) and drugs which are substrates of CYP2C9, CYP2D6- and CYP3A4/5 isozymes have been characterized in clinical studies as IM has demonstrated an inhibitory effect on these isozymes in vitro in human liver microsomes [Gleevec® Prescribing Information]. IM also exhibited a weak in vitro inhibitory effect on UGTs-mediated O-glucuronidation using acetaminophen as a probe substrate (Ki 58.5 μM). The present study evaluates the clinical significance of PK interactions between IM 400 mg/d and acetaminophen, an often used analgesic/antipyretic medicine, in CML patients.

Methods: Newly diagnosed CML patients (n=12) treated at St Mary’s Hospital, the Catholic University of Korea first received a single oral dose of acetaminophen 1000 mg on day 1 after an overnight fasting. On days 2–8, IM 400 mg was administered daily (qd). On day 8, another 1000 mg dose of acetaminophen was administered together with IM 400 mg. Blood and urine samples over 24 hours were collected following the morning dose on days 1 and 8 to determine the plasma PK and renal excretion of acetaminophen and its glucuronide (G) and sulfate (S) conjugate metabolites (two major metabolites of acetaminophen). Additional blood samples were collected on day 8 to measure the plasma exposure of IM and its major metabolite CGP74588 (CGP) in patients.

Results: Preliminary results from 6 patients showed that the plasma Cmax and AUC values of acetaminophen and the AUC ratios of conjugate G and S to acetaminophen were all similar between day 1 (Control) and day 8 (with IM) (Table 1). The renal clearance values for acetaminophen and its two conjugate metabolites were also similar between days 1 and 8. Following 7 days treatment at IM 400 mg qd, the IM plasma Cmax and AUC values were 2040±477.7 ng/mL and 31.8±8.68 μg*hr/mL, respectively. The oral clearance of IM in these Korean patients, 13.4±3.6 L/hr, was comparable to that reported in Caucasian CML patients, 10–14 L/hr [Schmidli 2005]. PK samples from an additional six patients have been collected, and results will be available for reporting by December 2008.

Acetaminophen (A) Cmax (μg/mL) AUC (μg*hr/mL) G/A AUC ratio S/A AUC ratio Day 1 (n=6) 15.6±8.3 52.4±24 2.66±0.75 0.77±0.11 Day 8 (n=6) 12.0±5.0 49.0±19.6 2.70±0.96 0.85±0.17 Day 8/Day 1 ratio 0.92±0.42 0.95±0.08 1.0±0.11 1.1±0.15 Imatinib (IM) Cmax (ng/mL) AUCt (μg*hr/mL) Cmin (ng/mL) CGP/IM AUC ratio Day 8 (n=6) 2040±477.7 31.8±8.68 947±323 0.15±0.035

Conclusion: Preliminary results showed that coadministration of imatinib 400 mg/d had no effect on the PK and metabolism of acetaminophen 1000 mg in newly diagnosed CML patients.
Research of Apoptosis Induction by the Application of Magnetic Nanoparticle of Fe3O4 Loaded with Daunorubicin and 5-Bromotetrandrine in K562/A02 Leukemic Cells
Authors: Chen Bao-An , Shen Ming-fang , Cheng Jian , Gao Feng , Xu Wen-lin , Shen Hui-lin , Ding Jia-hua , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Sun Xin-chen , Cheng Hong-yan , Deng Yu-xia , Li Guo-hong , Liu Li-jie , Chen Wen-ji , Wang Jue-qiong , Yuan Peng , Pei Xiao-ping , Chen Ning-na , Wang Xue-mei
Published: April 2020

Objective: To observe the apoptosis of K562/A02 cells induced by application of Fe3O4-Magnetic Nanoparticle (Fe3O4-MNPs) loaded with Daunorubicin(DNR) and 5-Bromotetrandrine (BrTet) and to explore its possible molecular mechanism.

Methods: We detected the apoptosis of K562/A02 cells after treatment with Fe3O4-MNPs and BrTet with DNR alone or their combination by flow cytometry(FCM),Wright staining and DNA ladder. The expression of Bax and Bcl-2 protein was measured by Western blot.

Results: The result showed apoptotic characteristics after treated with this composite for 48 hours. Flow cytometry assay showed the percentage of apoptotic cells which were treated with Fe3O4-MNPs and BrTet with DNR alone or their combination for 48 hours increased from (7.9%±0.35)% to (15.5%±0.87)%, (19.7%±1.04)% and(40.8%±1.74)%, (P<0.05. After treated with this composite, the typical apoptotic morphological features appeared: condensation of cells and nuclear, disintegration of nuclear chromatin, and apoptotic body could be observed under microscope; DNA agarose gel electrophoresis showed ladder bands; the expression level of Bax protein increased (P < 0.01) and Bcl-2 protein decreased markedly (P < 0.01).

Conclusion: Our results suggest that Fe3O4-MNPs or BrTet with DNR induces apoptosis in K562/ADM cells, and when used together,they have distinct synergism. The up-regulation of Bax protein and the down-regulation of Bcl-2 protein may play important roles.
Analysis on Characteristics of Dysplasia in 345 Patients with Myelodysplastic Syndrome
Authors: Xu Xiaoping , Ji Meirong , Chen Bobin , Lin Guowei
Published: April 2020

Objective To investigate the characteristics of dysplasia in myelodysplastic syndrome (MDS).

Methods Collect 716 samples of adult patients with abnormal blood routine but unclear cause between July 04, 2003 and March 14, 2007. Based on the gold standard of WHO MDS classification, all cases were detected on cytomorphological observation, cytochemical stain, bone marrow pathological study, cytogenetics, flow cytometry, and ect. The bone marrow cytological study on some abnormal hematopoietic cells has a diagnostic value to determine clonal or non-clonal diseases and assess sensitivity and specificity.

Results In the complicated various dysplasia of hematopoietic cells, the following characteristics can be the main basis of cytomorphological diagnosis:

One of granular Auer bodies, micronucleus (MN), or nuclear budding; Erythroid nuclear budding; Megakaryocytes presented in peripheral blood; Myeloblast or prorubricyte exhibited in peripheral blood; ringed sideroblasts>1%.

The subordinate basis of cytomorphological diagnosis was as follows:

Granular pseudo Pelger-H≥et anomaly, hard nucleus segmentation, unsynchronous development of nuclei, ring-shaped nuclei, and aggregation of nuclear chromatin. Erythroid multi-nuclei, odd nucleus, mother-daughter nucleus, nuclear fragmentation, vacuole, and anisocytosis; micromegakaryocytes.

Conclusion Cytomorphologic is the base for the diagnosis of MDS, however, it presents certain limit, especially cytomorphological change does not possess specificity for early MDS, hereby, it requires to combine other detection methods.
The Multiple Myeloma Genomics Portal
Authors: Liefeld Ted , Gao Qing , Chapman Michael A , Tonon Giovanni , Auclair Daniel , Reich Michael , Golub Todd R.
Published: April 2020

Current integrative genomics projects are enabling new discoveries in cancer research through the ability to combine multiple modalities of data, e.g. gene expression, copy number, RNAi, exon resequencing, and epigenetics. However, the tools to access and analyze this data have traditionally been out of the reach of clinicians and research biologists, due to the widely distributed nature of the available data and the significant learning curve required to use the analytical tools. This problem is particularly relevant to the multiple myeloma research community because of the heterogeneity of genomic aberrations underlying this disease and lack of a central repository of multi-modal multiple myeloma data. To address these problems, the Broad Institute has created a pilot Multiple Myeloma Genomics Portal, http://www.broad.mit.edu/mmgp, which serves as an interface between biologists (and clinical investigators), analytical tools, and multiple myeloma datasets. The Portal provides access to a number of advanced gene expression, gene set enrichment, and copy number analyses and visualizations within an easy to use Web interface. These analyses can be performed on the datasets hosted on the Portal, which include previously published curated, high quality genomic multiple myeloma datasets as well as a new reference collection of multiple myeloma samples. The Portal is continuously updated with new datasets, data types, and analytical capabilities as they become available. The Portal’s accessibility allows it to serve as a significant venue for investigators from other fields, engaging a broader range of investigators in exploring these data, and its design is readily adaptable to integrative genomics studies of other cancer types.
Flow Cytometric Detection of Minimal Residual Disease in Multiple Myeloma Patients
Authors: Kovarova Lucie , Buresova Ivana , Pour Ludek , Michalek Jaroslav , Hajek Roman
Published: April 2020

Background: Multiple myeloma (MM) is a hematological malignancy characterized by expansion of neoplastic plasma cells (PCs). Despite of advances in treatment, achieved in past ten years, many patients relapse due to minimal residual disease (MRD) existence.

Aim: Monitoring of number and phenotype of plasma cells in transplanted MM patients by multiparametric flow cytometry.

Patients and Methods: Bone marrow of 24 MM patients (13 male, 11 female) after autologous stem cells transplantation was used. Analyses were done in 1st month and 1st year after transplantation. Relapsed patients were analyzed earlier. Plasma cells were identified as CD138+CD38+ cells, CD19 and CD56 markers were used for discrimination of normal polyclonal (N-PCs) and abnormal monoclonal (A-PCs) plasma cells, respectively.

Results: Median number of PCs in whole group of patients in 1st month after transplantation was 0.1% (range 0.0–2.5%), median number of CD19+ N-PC 39.6% (5.0–79.2%), CD56+ A-PC 31.7% (3.5–90.9%) and N-PC/A-PC ratio was 1.1 (0.1–22.6). There were identified 8 relapsed patients during 1st year from transplantation, all with detectable monoclonal immunoglobulin, and with median number of PCs 10.6% (range 0.1–76.6%), N-PC 0.2% (0.0–56.2%), A-PC 60.1% (0.8–99.9%) and N-PC/A-PC ratio 0.0 (0.0–3.0). In a group of non-relapsed patients in 1st year after transplantation was median number of PCs 0.2% (range 0.0–1.9%), N-PC 64.2% (6.3–90.0%), A-PC 21.7% (3.0–93.2%) and N-PC/A-PC ratio 3.4 (0.1–25.0). There is an evidence that relapse is relate to loss of CD19 expression on PCs and to higher number of CD56+ PC.

Conclusion: Flow cytometric analysis of PCs phenotype profile is relatively quick and sufficiently sensitive method that can be used for monitoring of MRD. Detailed analysis of higher number of patients and comparison of their data with other parameters (immunofixation etc.) are necessary for verification of this method and for its application in routine diagnostics.
Anti-Heparanase Therapy in Combination with Conventional Chemotherapy Potently Inhibits Multiple Myeloma Growth in Vivo
Authors: Ritchie Joseph P. , Naggi Annamaria , Torri Giangiacomo , Casu Benito , Pisano Claudio , Carminati Paolo , Vlodavsky Israel , Sanderson Ralph D. , Yang Yang
Published: April 2020

Targeted agents directed at specific intracellular signaling pathways and the tumor microenvironment have improved the management of many cancers, including multiple myeloma. Work by our lab has demonstrated that heparanase, an endo-β-D-glucuronidase which cleaves the glycosidic bond of heparan sulfate chains at only a few sites, is expressed by a subpopulation of myeloma cells. Expression of heparanase promotes myeloma growth and spontaneous metastasis to bone, correlates with increased microvessel density and is an indicator of poor patient prognosis. This makes heparanase a viable target for myeloma therapy. Current approaches for treating multiple myeloma are ineffective, with relapse occurring frequently. Recently, a modified non-anticoagulant species of heparin, designated SST0001, was identified as a specific inhibitor of heparanase. SST0001, when tested in multiple in vivo models of myeloma, potently inhibited both angiogenesis and tumor growth in a dose dependent manner. However, in vitro, it did not have direct tumor cell cytotoxicity. Therefore, because SST0001 apparently targets the tumor microenvironment and conventional chemotherapy directly targets the tumor cell, we hypothesized that combination therapy would synergistically decrease tumor burden and provide an alternative biological approach to treating myeloma. In this study, using a syngeneic subcutaneous model of myeloma, we screened three potential novel combinations; SST0001 + dexamethasone, SST0001 + doxorubicin, and SST0001 + bortezomib. While all three combinations were effective in blocking tumor growth, SST0001 + dexamethasone was identified as being the most potent and was chosen for further testing. In two separate SCID-subcutaneous models using the CAG and MM1.R dexamethasone resistant human myeloma cell lines, SST0001 + dexamethasone also proved to be a potent inhibitor of tumor growth in vivo. These results identify an effective combination therapy, which is not cell line specific, for further development and translation into clinical trials.
PKC412 Shows Strong Anti-myeloma effects in in-Vitro-Studies
Authors: Elmaagacli Ahmet H , Koldehoff Michael , Steckel Nina K , Beelen Dietrich
Published: April 2020

Background. The protein kinase C (PKC) inhibitor PKC412 (N-benzylstaurosporine) is a derivate of the naturally occurring alkaloid staurosproine and has been shown to inhibit the conventional isoforms of PKC (alfa, beta1, beta2 and gamma). PKC412 has been shown to have an antitumor effect on non-small cell lung cancer and acute leukemia with FLT3 mutations, but little is known about its effect on multiple myeloma up to date.

Methods. Since PKC is also an inhibitor of a tyrosin kinase which is associated with VEGF, and inhibits the release of Interleukin-6, TNF alfa, and that of growth factor dependent C-FOS, we postulated that PKC412 might have also strong anti-myeloma features. Here we evaluated the anti-myeloma effect of PKC412 in the multiple myeloma cell lines INA-6, OPM-2 and RPMI 8226 by measuring its effect on their proliferation rate, the apoptosis rate and the Interleukin-6 mRNA expression.

Results. PKC412 showed strong anti-myeloma effects in all three celllines. 50nM of PKC412 was enough to drop the proliferation rate in all three cell lines under 10% compared to untreated cells(p<0.01). The apoptosis rate increased in INA cell line up to 2,5 times and in RPMI cell line up to 3 times (p<0.05), whereas only a moderate increase was observed in the OPM2 cell line with 500nM of PKC412. As expected, the IL-6 mRNA expression decreased after PKC412 treatment in all three cell lines more than 50%. The addition of Bevacizumab to PKC412 in RPMI and OPM-2 cell lines did not increased the apoptosis rate significantly, whereas the addition of short-interference RNA (RNAi) against VEGF increased the apoptosis rate in RPMI 8226 cells about 20% (p<0.05) and in OPM-2 cells up to 30% (p<0.01) compared to PKC412 alone, which was also associated concordantly with a further reduction of the proliferation rate in RPMI and OPM-2 cells up to 30%.

Conclusions. PKC412 shows strong anti-myeloma effects and might be effective also in the treatment of patients with multiple myeloma. These in-vitro studies might encourage to initiate clinical trials with PKC412 in patients with multiple myeloma.
Bortezomib, Doxorubicin and Dexamethasone (PAD) Combination Therapy Followed by Thalidomide and Dexamethasone (TD) as a Salvage Treatment for Relapsed Multiple Myeloma (MM): Analysis of Efficacy and Safety
Authors: Lee Jae Hoon , Suh Cheolwon , Lee Sung Sook
Published: April 2020

Introduction: PAD was reported to be highly effective regimen as an induction therapy before high dose therapy. TD is an another effective regimen with no cross resistance. We conducted a phase II study with PAD followed by TD in relapsed MM to test effectiveness of this combination.

Method: Patients were planned to receive 6 cycles of PAD, (bortezomib 1.3 mg/m2 days 1, 4, 8 and 11, doxorubicin 4.5 mg/m2 days 1–4, dexamethasone 40 mg days 1–4, every 21 days). Responders following 6 cylces of PAD received 12 cycles of TD (thalidomide 100 mg days 1–28 and dexamethasone 40 mg days 1–4, every 28 days). In patients with progression during PAD therapy, regimen was changed to 12 cycles of thalidomide 200 mg days 1–28 and dexamethasone 40 mg days 1–4, every 28 days. Response was assessed by EBMT criteria, with additional categories of nCR and VGPR. Adverse events were graded by the NCI-CTC, Version 3.0.

Result: This study aimed to enroll 40 patients till Oct 2007 and we are reporting the result with 40 patients. Efficacy could be assessed in 34 patients. After two cycles of PAD, 34 patients showed response with 6 CR. Overall response rate to 6 cylces of PAD was 90.9% with 36.4% CR. Seven of total 24 patients with TD showed further improvement of response status with 7 additional CR. Overall response to PAD followed by TD was 94%: CR 57.6%, nCR 15.2%, VGPR 6%, PR 15.2%, SD 3%, PD 3%. There was no prognostic factor for CR+nCR achieving in the univariate analysis. The median follow-up was 16.2 months with 2 year OS 54.5%. The median PFS time from study treatment was 18 mon (95% CI, 11.0~24.9 mon), median OS was not reached. The median OS time from first diagnosis was 112.9 mon (95% CI, 24.6~201.1 mon). One hundred seventy-eight PAD cycles (median 6, range 1–6) in 38 patients were assessable for safety. The most common hematologic toxicity was thrombocytopenia, with grade 3/4 in 39.5%. Grade 3/4 neutropenia was observed in 10.5%. Sensory neuropathy occurred with grade 2 in 26.3% and grade 3 in 13.2%. The median dose intensity was 1.35 mg/m2/week for bortezomib and 5.21 mg/m2/week for doxorubicin, which correspond 78% and 86.8% of the planned dose intensities, respectively. A total of 185 TD treatment cycles (median 2, range 0–12 cycles) was administered. One patient die by the pneumonia.

Conclusion: PAD followed by TD in patients with relapsed multiple myeloma is very active and tolerable.
Ritonavir Sensitizes Multiple Myeloma Cells to Bortezomib-Induced Apoptosis
Authors: Shibata Robert , Bassermann Florian , Chandra Pranil , Resemann Henrike , Mazumder Amitabha , Pagano Michele , Ibrahim Sherif
Published: April 2020

The ubiquitin-proteasome system plays a central role in the regulation of cell growth and cell proliferation by controlling the abundance of key cell cycle proteins. Increasing evidence indicates that unscheduled proteolysis of many cell cycle regulators contributes significantly to tumourigenesis and is indeed found in many types of human cancers, including colon and renal cell cancer, non hodgkin lymphomas, and multiple myelomas. Protein degradation pathways are also targets for cancer therapy, as shown by the successful introduction of bortezomib (B), an inhibitor of the 26S proteasome, for the therapy of multiple myeloma. Recently, protease inhibitors conventional in HIV therapy (e.g., ritonavir (R)) have been found to exert an antiproliferative effect on different tumor types, including multiple myeloma cells. More recently, ritonavir has been shown to induce endoplasmic reticulum stress and to sensitize sarcoma cells towards bortezomib induced apoptosis.

In this study, we show that the combination of therapeutic concentrations of bortezomib and ritonavir acts synergistically on cultured multiple myeloma cell lines (U266, RPMI 8226, ARH-77 (2 variants-One is more resistant)). Cell proliferation was significantly reduced in an overadditive fashion as compared to individual treatments (proliferation as % of DMSO control after 48hrs: CRL-1621 (ARH-77): B: 40%, R: 50%, R&B: 7%, CCL-155 (RPMI 8226): B: 37%, R: 50%, B+R: 6%; n=3). Moreover, we found proteasome inhibition by bortezomib to be associated with low levels of expression of Skp2 and consequent stabilization of its target p27Kip1, a negative cell cycle regulator at the G1/S cell cycle transition. Accordingly, bortezomib induced cell cycle arrest at the G1/S transition followed by caspase dependent apoptosis. In contrast, only mild induction of apoptosis and no activation of Caspase 3 were observed in myeloma cells treated with ritonavir, although growth arrest was present. Interestingly, growth arrest in ritonavir-treated cells was associated with an accumulation in G2 phase.

In a limited clinical study, we treated two patients with stage III relapsed and refractory myeloma with a combination of bortezomib at standard doses and ritonavir at 600 mg twice a day. Both patients had MR or greater lasting for more than 2 months. Only grade 2 GI toxicity was seen.

Taken together, our findings suggest different mechanisms of action for bortezomib and ritonavir on myeloma cells and suggest that the combination of the two drugs may be a valid therapeutic strategy.
Identification of Key Factors That Are Associated with the Clinical Anti-Tumor Effect of Zoledronic Acid in Multiple Myeloma
Authors: Teoh Gerrard , Koh Liang Piu
Published: April 2020

Despite compelling preclinical data, only anecdotal reports have shown that zoledronic acid (ZOMETA, Zol) exerts an anti-multiple myeloma (MM) effect in humans. We postulate that this could be related to the dose-intensity of Zol, since anti-MM effects were clearly observed in mice given a dose equivalent of 8.0 mg (8.0 EQmg) weekly; but not in humans who received only 4.0 mg monthly. We describe 3 patients, who were treated with 2- to 3-fold higher dose-intensity of Zol; who demonstrated brisk clinical improvements. When the equivalent dose of Zol was normalized to the patient’s body surface area, the maximum EQmg rate of Zol administered was between 11.4 EQmgmax and 12.0 EQmgmax per 28 days. Since the potency of Zol is measured by the degree of inhibition of differentiation of osteoclast precursors (pOC) (i.e. monocytes) to osteoclasts (OC) in vitro, we next investigated whether high-dose rate Zol had an in vivo inhibitory effect on this renewable pOC pool. We demonstrate that there was a linear relationship between the absolute monocyte count (AMCO) and the stage of MM at presentation. Specifically, patients with International Staging System (ISS) stage I disease had the lowest AMCO levels, whereas ISS stage III patients had the highest. We also show that treatment with Zol resulted in progressive decreases in AMCO levels, and these changes correlated with response to therapy. By contrast, AMCO levels increased in patients who either failed to respond to treatment or who had progressive disease. Finally, since Zol is known to induce the gamma delta T cells in vitro, we studied the recovery patterns of various T cell subsets following treatment using a high dose-intensity Zol-based regimen called “dtZ”. To our surprise, there was no difference in gamma delta T cell numbers, but instead bone marrow CD4+ CD25+ regulatory T cells were significantly decreased in patients who achieved CR, as compared to those that did not. These data suggest that high dose-intensity Zol has a very desirable anti-tumor profile in MM that could be related to inhibition of both myelomagenic processes as well as the immunosuppressive elements of host anti-tumor defense.
Analysis of JAK2V617F and MPLW515L/K Mutation in 30 Patients with Early Stage Myeloproliferative Disease
Authors: Zhang Ri , Fan Zhen , Zhang Xuhui , Fei Hairong
Published: April 2020

The classic Bcr-Abl negative myeloproliferative disorders (MPDs) comprise polycythemia vera (PV), essential thrombocythemia (ET), and idiopathic myelofibrosis (IMF). They are considered clonal stem cell disorders. Recently two acquired somatic mutations JAK2V617F and MPLW515L/K have been identified to play an important role in pathogenisis of MPDs. JAK2V617F was detected in almost all PV and nearly 50% in ET and IMF, and the MPLW515L/K mutation occured in only 1% and 4% in ET and IMF respectively. However it is interesting that there are some patients with slightly increased blood cells did not meet the criteria of PV and ET clinically. These early prefibrotic stages of MPDs are overlooked by both the Polycythemia Vera Study Group (PVSG) and the 2001 WHO criteria for the diagnosis of PV, ET and IMF. In order to investigate the prevalence of JAK2V617F and MPLW515L/K mutation in patients with early MPDs without a secondary cause, genomic DNA from bone marrow or blood mononuclear cells of 30 patients with early MPDs was screened by allele specific polymerase chain reaction (AS-PCR) for JAK2V617F and MPLW515L/K mutations and the history of thrombosis was assessed retrospectively by using patient files. Results showed that 14 out of 30 patients (46.7%) were positive for the JAK2V617F mutation, and none of them had the MPLW515L/K. Of the 14 patients with JAK2V617F mutation, a history of thrombosis was comfirmed in 5 patients(35.7%), while none of 16 patients with WT JAK2 had the thrombosis history. The average age of patients with the JAK2V617F mutation exceeded the patients with wild type JAK2(P<0.05), whereas there is no statistical significance on sexual Athe counts of white cell and the occurance of complications (thrombosis) between the patients with and without the JAK2V617F mutation. All the patients have been followed up since onset and 22 patients have available results. Among them, 12 patients with the burden of JAK2V617F turned out to be MPDs, however only 1 out of 10 patients without this mutation also evolved to MPDs. Our results suggest that JAK2V617F mutation can occur in a certain percentage of these early stage of MPDs patients and has help to diagnose early MPDs. Because these patients might have a high rate of complications, proper treatment of the MPD including aspirin and/or cytoreductive therapy is recommended in latent MPDs patients associated with the JAK2V617F mutation.
The Clinical and Histological Characterization of Malignant Lymphoma, Kikuchi-Fujimoto Disease and L-Forms Acid-Fast Bacillus Infection
Authors: He Jie , Li Rui-ping , Wang Jian-hua , Chen Bao-An
Published: April 2020

Objective: Malignant lymphoma (ML), Kikuchi-Fuljimoto disease (KFD) and L-forms Acid-fast bacillus infection (L-forms TB) usually affect the cervical lymph nodes. KFD and L-form TB are often confused with ML and take misdiagnosis and inappropriate treatment. The diagnosed of these diseases is on basis of an excision biopsy of affected lymph nodes.

Methods: We analyzed 63 patients (including case diagnosed in our hospital) by clinical characteristic, laboratory, pathological morphology, histochemistry and immunohistochemistry.

Results: Of 63 cases, 43 were ML, 17 were KFD and 3 were L-forms TB. In ML disease, 46% were male and 54% were female, mean age was 54 (11–82). 9% cases have persistent fever and associated with hepatosplenomegaly. In KFD, 47% were male and 53% were female, mean age was 26 (17–45). 59% cases with cervical hymphadenopath and no fever, 41% had painful, accompanied by fever. hepathosplenomegaly, occasionally leucopenia and lypadenopathy especially involving cervical region not responsive to antibiotic treatment. 29% lymph tissues were morphologically, and characterized by a necrosis in the cortical or paracortical areas of enlarged lymph node around the necrotic area. T lymphocytes were found and absence of granulocytes. 30% cases were mistaken to ML. L-forms TB disease, there were fever, clinical and histological were similar KFD and ML. 3 cases were misdiagnosis to ML or KFD. Laboratory analyses: PAM and PAS were positive, and anti acid test to found L-forms bacterium in tissues.

Conclusions: Kikuchi-Fujimoto disease and L-forms Acid-Fast bacillus infection are easily confused histological and clinically with malignant lymphoma. Clinicians and pathologists must be aware of this condition. Although Malignant lymphoma, Kikuchi-Fujimoto disease and L-forms Acid-fast bacillus infection can be found any age, but should be considered for malignant lymphoma in older patients. Kikuchi-Fujimoto disease is predominantly a self-limiting disease of the young and most cases with fever, nodal biopsy showing fragmentation necrosis and karyorrhexis, not responsive to antibiotic treatment. In the L-forms Acid-fast bacillus infection, the PAM and PAS are positive, and have an effective antibiotic treatment.
Tumor Necrosis Factor Alpha (TNF-a) Blockade and Risk of Lymphoma: Systematic Review and Analysis of Literature
Authors: Ramanarayanan Jeyanthi , Pahuja Shalu , Czuczman Myron S , Hernandez-Ilizaliturri Francisco J.
Published: April 2020

BACKGROUND: TNF-α plays a critical role in the regulation of cytokine mediated cancer immunosurveillance. Evidence suggests that depletion of TNF-α may result in abrogated anti-tumor immunity and increase in the risk of malignancies. Anti-TNF-α therapy (infliximab, adalimumab and etanercept) is known to halt the progression of certain inflammatory disorders and has been approved for the treatment of at least five autoimmune/inflammatory conditions. Given the widespread use of these biological agents, it is vital to explore the associated increased risk of malignancies especially lymphoma.

OBJECTIVE: Risk of lymphoma associated with anti-TNF-α therapy is controversial in rheumatoid arthritis (RA) and has not been extensively studied in other inflammatory disorders. With the expanding role of anti-TNF-α therapy, we evaluated the risk of lymphoma among all approved indications for which they are currently in use.

METHODOLOGY: We conducted a systematic electronic search of MEDLINE using the terms TNF-α, infliximab, adalimumab, etanercept, rheumatoid, psoriasis, arthritis, inflammatory bowel disease, ankylosing spondylitis from January 1998 to December 2007. Studies analyzed were restricted to those in English language, full text published articles, randomized controlled trials (RCT), meta-analysis (MT), review articles, extension studies, reports from national databases (ND) and postmarketing surveillance studies. Risk of lymphoma from ND was obtained as odds ratio with 95% confidence interval. Incidence of lymphoma reported from RCT was analyzed as percentage fraction of the total number of patients enrolled in the study and follow up.

RESULTS: Overall 51 studies (2 MT, n=10,370; 4 ND, n=29,099 and 45 RCT, n=54,637) that included anti-TNF-α therapy in inflammatory disorders were analyzed. Treatment/follow up period were variable from 12 weeks to 8 years. Ten patients (0.09%) from MT treated with anti-TNF-α therapy developed lymphoma vs 0 in the non-anti-TNF-α group. Report from one of the 3 ND showed an increased risk (11.5 RR 95% CI 3.7–26.9) of lymphoma among the anti-TN-α exposed group. Among the RCT, 16 patients (0.02%) in the anti-TNF-α group developed lymphoma vs 3 (0.005%) in the control group.

CONCLUSION: Existing data suggests that anti-TNF-α therapy in rheumatoid arthritis is associated with an increased risk of lymphoma but this may be attributed to the severity of the disease itself. Among other inflammatory disorders, a rise in lymphoma risk could not be established. Whether anti-TNF-α therapy in the long term can in fact decrease the incidence of lymphoma by altering the disease severity remains to be determined.
MYST3/NCOA2-Induced Acute Myeloid Leukemia in Transgenic Fish
Authors: Zhuravleva Julia , Paggetti Jérôme , Martin Laurent , Hammann Arlette , Solary Eric , Bastie Jean-Noël , Delva Laurent
Published: April 2020

The MYST3/NCOA2 (MOZ/TIF2) fusion gene generated by the inv(8)(p11q13) chromosomal abnormality was described in a specific subgroup of acute myeloid leukemias (AML) that represents less than 5% of AML4/5. This abnormality fuses MYST3 (MOZ), a member of the MYST family of histone acetyl-transferases (HAT) to NCOA2 (TIF2), a member of the p160 HAT family. The transforming properties of MYST3/NCOA2 were demonstrated in mouse committed myeloid progenitors in vitro and in vivo. Hematopoiesis is very similar in zebrafish and in higher vertebrates. Homologues of a large number of genes involved in mammalian myelopoiesis were identified in this animal model. We have recently shown that ncoa2 (tif2) played a role in zebrafish primitive hematopoiesis. This animal also represents a model for investigating leukemogenesis. Transgenic expression of rag2-EGFP-mMyc or rag2-ICN1-EGFP induces a T-cell acute lymphoblastic leukemia (ALL) whereas transgenic expression of the ETV6/RUNX1 fusion gene induces a B-cell type ALL. We generated a transgenic zebrafish in which the MYST3/NCOA2 fusion gene was expressed under control of the spi1 (pu.1) promoter. An AML developed in two of 180 MYST3/NCOA2-EGFP-expressing embryos, 14 and 26 months after injection of the fusion gene in a one cell embryo, respectively. This leukemia was characterized by an extensive invasion of kidneys by myeloid blast cells. This model, which is the first zebrafish model of acute myeloid leukemia, demonstrates the oncogenic potency of MYST3/NCOA2 fusion gene.
Aberrant PDGFRA Signalling Is Sustained by An Autocrine Loop in Peripheral T-Cell Lymphoma Not Otherwise Specified
Authors: Piccaluga Pier Paolo , Rossi Maura , Esposito Aurora , Tazzari PierLuigi , Ricci Francesca , Agostinelli Claudio , Righi Simona , Gazzola Anna , Went Philip , Alviano Francesco , Bagnara Gian Paolo , Zinzani P.L. , Pileri Stefano A.
Published: April 2020

Background. Peripheral T-cell lymphoma not otherwise specified (PTCL/NOS) is the most common T-cell lymphoma; however, it remains a complex entity showing great variety regarding morphology, immunophenotype and clinical behaviour. Recently, gene expression profiling (GEP) revealed PDGFRA as possible PTCL-associated molecule, by nominating it as potential therapeutic target.

Aims and Methods. We investigated the possible determinants of PDGFRA activity in PTCL/NOS. We performed GEP of 28 PTCLs/NOS and 20 samples of normal T-cell subpopulations, by using the Affymetrix HG-U133 2.0 plus microarray. Moreover, we built tissue-microarrays (TMAs) including 145 PTCLs/NOS for protein analyses. The PDGFRA locus (4q1.1–4q1.3) was studied by FISH on those TMAs, while direct sequencing of all PDGFRA exon and introns as well as of the promoter region was performed in 90 cases. Immunohistochemistry (IHC) and ELISA were adopted in order to study the expression of PDGF-A, PDGF-B and PDGF-C on tissue sections and in supernatants from PTCL/NOS cell cultures, respectively. Finally, the expression of PDGFRA and its activated (phosphorilated) form, p-PDGFRA, was assessed by IHC on TMAs, and by flow-citometry in PTCL/NOS cultured cells before and after the exposure to an anti-PDGF ligand neutralizing antibody (R&B System).

Results. First, GEP showed over-expression of PDGFRA in all tested PTCLs/NOS, while IHC confirmed the expression of PDGFRA and p-PDGFRA. FISH, SNPs analysis and direct sequencing showed preserved integrity of PDGRA locus. We then studied PDGF-A, PDGF-B and PDGF-C, which turned out to be expressed and secreted by PTCL/NOS cells. In order to verify the hypothesis of an autocrine loop, we tested whether the remotion of PDGF ligands from the supernatants of cultured PTCL/NOS cells determined PDGFRA de-phosphorilation. Notably, 48h after the exposure to 20 mg of anti-PDGF antibody, we already appreciated a significant effect with reduction of PDGFRA phosphorilation up to 75%.

Conclusions. Taken together, our data demonstrate that PDGFRA activity is sustained by an autocrine loop in PTCL/NOS.
Oral Contraceptive-Associated Thromboembolism. A Retrospective Analysis of Thrombophilic Work-out and Long Term Follow up in 57 Cases
Authors: Papadakis Emmanouil , Efremidou Smaragda , Kartsios Haris , Mpraimi Margarita , Kokoviadou Kiriaki , Loukidis Konstantinos , Papaioannou Georgia , Korantzis Ioannis
Published: April 2020

Introduction: The increased risk of venous thrombosis in women taking oral contraceptives (OCs) has been recognized since the early 1960s. Coexistence of hereditary risk factors appears to have an additive effect. Women under OCs that carry the factor V Leiden mutation have a 35-fold increased risk of thromboembolic events compared to women without the mutation who are not on OCs. Evaluation of family and personal history is the mainstay of prophylaxis prior to OC administration, but often family thrombophilia or thromboembolic (TE) events are not reported prior to OCs prescription.

Patients-Methods: Fifty-seven women with a median age of 28 (21–48) years, which suffered OC-associated TE, were studied. The median period of OC therapy prior to TE event was 2 months (0.5–60). Fifty-five of them experienced VTE while 2 suffered stroke. Leg thrombosis was the most common clinical finding [37/55 (67,2%) patients] Apart from personal and family history, Thrombophilia investigation included measurement of : serum Homocysteine, Antithrombin, Protein C and S, Lipoprotein (a), Activated Protein C (APC) resistance, antiphospholipid antibodies and lupus anticoagulant. In addition the presence of FV Leiden, FII 20210 GA mutations and MTHFR 677 CT polymorphism were determined.

Results: A high prevalence of the factor V Leiden mutation was detected in the study group; 50% had APC-resistance test positive, 26 (45%) patients were found to be heterozygous and 3 (5,2%) homozygous for the FV Leiden mutation. Lp(a) elevation was observed in 19,3% and Homocysteine elevation in 15,8% of patients. In 9 women (15,8%) both family history and thrombophilic profile were negative. Serious VTE events (2 abdominal and 6 CNS thromboses) were observed only in the Leiden subgroup. During the follow up period ranging from months to 18 years, 3 women (6,25%) experienced a miscarriage and 14 suffered additional VTE events (25%) and they are currently on permanent anticoagulation.

Conclusions : Universal thrombophilia screening of women prior to prescription of OCs is not advisable as it does not appear to be cost effective. However, screening certain subgroups, such as women with a known personal or family history, may be of great value. If a full thrombophilic profile can’t be performed, a mere activated protein C resistance test, that reflects the presence of the factor V Leiden mutation, may provide an easy and cheap way of identifying and consulting properly women at higher risk for VTE prior to OC use. Women with OC-associated VTE and thrombophilia carry a substantial recurrence risk that persists for years.
Retinal Artery Occlusion and Other Arterial Thrombotic Events in Women Using Yasmin
Authors: Shum Merrill K , Nacopoulos Dimitrios A
Published: April 2020

The third-generation combined oral contraceptive pill, Yasmin®, which contains 30 ug ethinyl estradiol and 3 mg of drospirenone, has been marketed as safe and effective, but the risks of arterial thrombotic events in women using Yasmin® are unknown. We report a case of retinal artery occlusion in a woman using Yasmin®, and review the literature for other cases of arterial thrombotic events in women taking Yasmin®. An extensive search using Pubmed, Ovid, and Medline, revealed three previously reported cases of arterial thrombotic events, in women using Yasmin® (see table below). Similar to the three prior cases, our case study had no known acquired or hereditary thombophilic conditions. Unlike the previous three cases, our subject did not smoke and developed her arterial thrombus late, approximately three years after initiating Yasmin®. It appears that the risk for arterial thrombotic events may occur at anytime. Large scale studies determining relative incidence, as well as post-marketing adverse effects are needed to determine risk and incidence while using Yasmin®.

Studies reporting arterial thrombotic events in women using Yasmin®

Author Year Age of patient Type of thrombosis Days on OC before thrombosis Associated risk factors Comments Vaya, et al. (1) 2003 21 TIA 15 days Smoking (15/day) Apparent recovery Cabou, et al. (2) 2006 33 MI 15 days Smoking (6/day), surgery for ectopic pregnancy 15 days before MI Pt had taken 2nd generation OCP in the past without complications, recovered Girolami, et al. (3) 2007 18 Central retinal artery occlusion 10 days Smoking (8/day) No recovery, two years after event Shum, et al. 2008 21 Central retinal artery occlusion 3 years None Pt had taken 1st generation OCP for 2 years without complications, not yet recovered after 4 months