bluuebunny/crossref_metadata_2025_split huggingface dataset:
Durable Remission with Decreased HTLV-I Proviral Load after Reduced-Intensity Stem Cell Transplantation (RIST) in Patients with Adult T-Cell Leukemia/Lymphoma (ATL).
Authors: Tanosaki Ryuji , Nosaka Kisato , Mineishi Shin , Mori Shin-ichiro , Kim Sung-Won , Kami Masahiro , Kannagi Mari , Sonoda Shunro , Okamura Jun , Fukuda Takahiro , Tobinai Kensei , Takaue Yoichi 
Published: April 2020

Abstract
Background. ATL is an HTLV-I-associated hematological malignancy. The majority of ATL patients are not curable using current chemotherapy with median survival time of 13 months. Recently, patients undergoing allo-SCT with conventional conditioning regimen were reported to have a 3-year overall survival (OS) rate of 45%, however, with high transplant-related mortality (TRM) rate as much as 40% (Fukushima T. Leukemia19:829, 2005). Since most ATL patients are over 50 years old with various complications, limited numbers of patients are eligible for conventional transplant. We have already conducted a phase I multi-center trial testing the feasibility of RIST in ATL patients, where 2-year OS was 33% and some differences in the outcome were suggested among participating institutes. Hence, we investigated the efficacy of RIST performed exclusively in our single institute (NCCH, Japan).
Patients and Methods. Between Oct 2000 and Jun 2005, 16 ATL patients underwent RIST from a related donor. Since 3 have been enrolled into an on-going multi-center trial, 13 patients were analyzed in this retrospective study, including 3 who were enrolled into the above-mentioned multi-center trial (Okamura J. Blood105;4143, 2005). Median age was 51 years old (range, 44–61), 8 males and 5 females, and 8 were of acute type and 5 of lymphoma. The disease status at transplant was 3 CR, 7 PR and 3 primary refractory. Three donors were HTLV-I healthy carriers. Eleven were HLA 6/6, 2 were 5/6, 12 were PBSCT and 1 was BMT. Conditioning regimen was fludarabine 30 mg/m2 x6, busulfan 4 mg/kg x2 with (n=5) or without (n=8) rabbit ATG 2.5 mg/kg x2. Prophylaxis of GVHD was CSP alone.
Results. Engraftment was rapid (median, 11 days) in all cases, and there was no TRM. Acute GVHD of grade II–IV developed in 7, and chronic extensive GVHD in 5. Eleven patients achieved CR after RIST, and 6 relapsed or progressed, among whom 2 achieved CR after cessation of CSP, donor lymphocyte infusion and local irradiation, and have been disease-free for more than 1 year. Ten patients are alive, 9 without disease, with median follow-up time of 26 mos (range, 4–45 mos). HTLV-I proviral titers determined using real time PCR decreased after RIST and became undetectable in 8 out of 13 patients. In 3, including one transplanted from an HTLV-I carrier donor, anti-HTLV-1 antibody transiently decreased and became nearly undetectable.
Conclusions. RIST is feasible and safe in ATL patients, and it has not only an anti-ATL but also an anti-HTLV-I activity. This is the first report documenting the efficacy of RIST for ATL in terms of survival and remission durability.
Figure Figure
Potent Cytotoxicity Against Multiple Myeloma and Acute Myeloid Leukemia Cells by a New Histone Deacetylase Inhibitor ITF2357.
Authors: Rambaldi Alessandro , Micò Caterina , Cuppini Lucia , Domenghini Marcello , Borleri Gian Maria , Barbui Anna Maria , Leoni Flavio , Mascagni Paolo , Salvi Anna , Golay Josee , Introna Martino 
Published: April 2020

Abstract
ITF2357 (Italfarmaco, Italia) is a novel hydroxamic acid-based HDAC inhibitor (HDACi) that has shown reduced toxicity in phase I studies. We have investigated the cytotoxic and anti-proliferative activities of ITF2357 in multiple myeloma (MM) and acute myelogenous leukaemia cells (AML). ITF2357 had a strong cytotoxic activity in 8/9 MM and 6/6 AML cell lines, with a mean IC50 of about 0.2 μM, a concentration largely attained following oral administration of safe doses of ITF2357 to healthy individuals. In contrast SAHA, the prototypic hydroxamic HDACi, showed an IC50 of about 1 μM or above in all cases. The cytotoxic activity of ITF2357 was due to induction of apoptosis, as documented by detection of annexin V and cleaved caspase 3. The ITF2357 induced hyperacetylation of histones in cell lines resistant or sensitive to the cytoytoxic activity of the drug is under investigation in order to further define its mechanism of action. ITF2357 had also more potent cytotoxic activity compared to SAHA against freshly isolated CD138+ purified MM cells and AML samples, with an IC50 of about 0.1 μM in 3/3 MM and 13/15 AML cases. Sensitive AML cases included five cases of FAB M1, five M2 and three M4. Three of the sensitive cases carried a t(8;21) translocation, 2 an inv(16), 6 had a normal and 2 a complex karyotype. Furthermore four of the sensitive AML also carried a flt3 internal tandem duplication and 3 a type A mutation in the nucleophosmin 1 gene (NPM1). The 2 more resistant AML cases (one M1 and one M5, both with normal karyotype) showed nonetheless a response to the drug with an IC50 of about 0.5 μM. We have also developed a culture system to grow freshly isolated AML cells for at least 3 weeks in vitro on human mesenchymal cells (MSC). Interestingly, ITF2357 was cytotoxic for primary AML cells stimulated to grow in optimal conditions on MSCs, at the same dose as in standard short term cultures. In contrast ITF2357 was not cytotoxic for MSCs even at the 1 μM concentration. The strong cytotoxic activity of ITF2357 on MM and AML has provided the framework for ongoing phase I studies of ITF2357 in these malignancies.
Raf Inhibitor BAY 43-9006 Induces Bim Dephosphorylation and Activates the Intracellular Apoptotic Pathway in AML.
Authors: Zhang Weiguo , Konopleva Marina , McQueen Teresa , Cortes Jorje , McCubrey James , Andreeff Michael 
Published: April 2020

Abstract
We have previously demonstrated constitutive activation of MAPK signaling in 70% of primary AML samples (Millela et al, JCI108:851–859, 2001), suggesting that upstream kinases (Raf and MEK) may play a role in the leukemic transformation of myeloid cells. BAY 43-9006 is a small molecule Raf kinase inhibitor that has demonstrated potent anti-tumor activity against solid human tumors in xenograft models. In this study, we tested the hypothesis that BAY 43-9006 inhibits leukemia cell growth and/or induces apoptosis by suppressing the activity of the MAPK pathway. In the in vitro kinase assay, BAY 43-9006 inhibited both Raf-1 and B-Raf-mediated MEK1 phosphorylation in a dose-dependent manner, with Raf-1 kinase being more sensitive to the inhibitory effects of BAY 43-9006 (IC50Raf-1, 1.37 μM vs. IC50B-Raf, 4.64 μM). BAY 43-9006 suppressed MEK1/2 and ERK phosphorylation in the AML cell lines OCI-AML3, HL-60, U937 and KG-1 in a dose-dependent manner after 24 hr treatment. Unexpectedly, BAY 43-9006 also inhibited AKT phosphorylation on Ser473 (after 4.5 hrs). BAY 43-9006 inhibited growth of AML cells in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50) of BAY 43-9006 was 0.39, 1.14, 2.86 and 2.80 μM, respectively in OCI-AML3, HL-60, U937 and KG-1 cells after 72 hrs. This growth-inhibitory effect was mediated by a dose-dependent induction of cell cycle arrest in G1 mediated by the down-regulation of the cell cycle-related proteins cyclin E, cdk2 and cdc2, followed by induction of apoptosis after 72 hrs. In primary AML patient samples, BAY 43-9006 not only inhibited cell growth and induced apoptosis after 48–72 hrs in vitro, but also preferentially inhibited colony formation of AML progenitor cells compared to normal bone marrow cells [IC50: 2.33 μM vs. 9.34μM (CFU-GM), 5.69 μM (Erythroid) and 3.75 μM (Mixed), respectively]. Time-course analyses demonstrated that BAY 43-9006 suppressed phosphorylation of the pro-apoptotic protein Bim (at 4.5 hrs), caused loss of the mitochondrial membrane potential and cytochrome c release (at 6 hrs) followed by cleavage of caspases-3 and -9 but not of caspase-8, suggesting primary involvement of the intrinsic mitochondrial pathway. Furthermore, the pro-apoptotic proteins Bim and Bax were up-regulated after 48 hrs of BAY 43-9006 treatment, and the level of the inhibitor-of-apoptosis protein Survivin was down-regulated after 48 hrs. In summary, our data demonstrates that BAY 43-9006 inhibits Raf-MEK-ERK signaling and induces apoptosis in AML via Bim de-phosphorylation and activation of the intrinsic apoptotic pathway. The potential of BAY 43-9006 in the therapy of AML patients will be tested in a Phase I clinical trial.
Acquired Bortezomib (Velcade®)-Resistance Is Accompanied by Downmodulation of Proteasome Activity and Upregulation of Tripeptidyl Peptidase II (TPPII) and a Ubiquitin-Specific Protease.
Authors: Driessen Christoph , Gogel Jeanette , Kraus Marianne , Ovaa Huib , Beck Alexander , Rückrich Thomas 
Published: April 2020

Abstract
Because cell lines can be adapted to proteasome inhibition in vitro (Pfeifer G. et al., Science 283: 978–981, 1999.), secondary resistance of malignant cells towards Bortezomib (Velcade®) in vivo is a likely scenario. We have succeeded to adapt the human AML cell line HL-60 to Bortezomib in vitro, so that the adapted subline (HL-60a) shows normal viability and growth rate at 40 nM Velcade®, while cell death is induced above 10nM in the parental line. We hypothesized that alternative proteolytic pathways might allow the continued proliferation and survival of Bortezomib-adapted cells, and assessed the activity-profiles of proteasomal subunits, ubiquitin-specific proteases and lysosomal cysteine proteases in a functional proteomics approach, using recently developed synthetic affinity probes. These tools for the first time allow semiquantitative visualization of the individual members of these protease families, based on their activity, in contrast to western blot which lacks activity information or to the turnover of fluorogenic substrates, which is not truly protease-specific. After 72h of culture in Bortezomib-free medium (wash out phase), HL-60a cells contained significantly reduced levels of active proteasomal β1,β2, and β5 subunits, compared to the parental line, as confirmed by a reduced turnover of the β5-selective fluorogenic substrate Suc-LLVY-AMC. A panel of 7 different ubiquitin-specific proteases (USP) was visualized in both types of cells, using the affinity probe HAUbVS. Of these, a 97 kD USP that we have identified as USP14 by mass spectrometry-based sequencing, was significantly upregulated in HL-60a cells. By contrast, the activity of lysosomal cysteine proteases remained unchanged in HL-60a cells. Because the cytosolic protease tripeptidyl peptidase II (TPPII) can partially substitute for proteasome activity in lactacystine-treated cell lines, we assessed TPPII-activity using a fluorogenic substrate. We observed a significant upregulation of TPPII-activity in HL-60a cells, compared to non-adapted controls. Inhibition of TPPII, however, was not sufficient to restore Bortezomib-sensitivity in HL-60a cells. Interestingly, the combination of Bortezomib with the HIV protease inhibitor Ritonavir induced synergistic cytotoxicity both in HL-60 and HL-60a cells. Thus, Bortezomib-resistance is accompabied by upregulation of protease activities in alternative pathways. Combining different protease inhibitors like Bortezomib and Ritonavir might be a promising option to overcome primary or secondary Bortezomib resistance.
The Ellipticine Derivative NSC 338258 Has Anti-Myeloma Activity.
Authors: Tian Erming , Stephens Owen , Holbeck Susan , Barlogie Bart , Shaughnessy John 
Published: April 2020

Abstract
We recently discovered that amplification and overexpression of the CKS1B gene, which regulates the ubiquitination and proteasomal degradation of the cyclin-dependent kinase inhibitor p27Kip1, is linked to a poor prognosis in myeloma. We used the COMPARE algorithm (NCI, Developmental Therapeutics Program) to identify correlations between the expression of CKS1B in the NCI 60 cell line panel and the GI50 of nearly 50,000 anticancer compounds. This analysis revealed a strong correlation between CKS1B expression and the anticancer activity of Ellipticine and multiple derivatives of it. Ellipticine (5,11-Dimethyl-6H-Pyrido[4,3]Carbazole, MW=246.3), an alkaloid isolated from Apocyanaceae, is a topoisomerase II poison that induces topoisomerase II-dependent DNA cleavages. Previous studies have shown that the anti-neoplastic mechanism of Ellipticine is to form covalent DNA adducts mediated by human cytochromes P450. We obtained Ellipticine and twelve other CKS1B-correlated antineoplastic compounds from the NCI-DTP. In vitro analyses were carried out with 12 myeloma cell lines. Cell viability was detected using CellTiter-Glo Luminescent Assay (Promega, Co.). We found that one of the Ellipticine derivatives, NSC 338258 (EPED3) showed significant cell kill activity not observed with Ellipticine or the other compounds in all cell lines tested. In a dose response analysis of cell kill using 0.2, 0.02, and 0.002 uM EPED3, 12 out of 12 myeloma cell lines showed at least 50% reduction in cell viability within 24 hrs at 0.2 uM compared to no treatment controls. By 48 hours essentially all cell growth was inhibited. Only weak to moderate cell kill was noted at 0.02 and 0.002 uM at all time points tested. We next tested the activity of the topoisomerase II inhibitor VP-16-213 at the same concentrations and time periods and found only modest reduction of cell viability at day 5 at 0.2uM. Essentially no effect on cell viability was observed at 0.02 and 0.002 uM at any of the time points tested. We next tested whether EPED3 had a comparable anti-myeloma activity as Adriamycin at the same concentrations. Twelve myeloma cell lines were exposed to EPED3, VP-16-213, and Adriamycin at 0.2mM for a total of 5 days. Results of the mean relative luminescence showed that EPED3 promoted an immediate cell proliferation arrest within the first 24 hours of culture, with almost complete cell kill within 3 days. The other drugs were less effective with Adriamycin inducing a &gt; 90% reduction only after 4 days and VP-16 a &gt; 70% reduction at day 5. These data suggests that EPED3 may represent a potential new therapeutic for myeloma. Studies are currently underway to test the efficacy of EPED3 in our in-vivo SCID-Hu mouse model of primary myeloma and results of these studies will be presented.
Baicalein Inhibits IL-6-Mediated Multiple Signaling Cascades through Jak Family Kinase in Human Myeloma Cells.
Authors: Ma Zi , Liu Shangqin , Otsuyama Ken-ichiro , Kawano Michio M. 
Published: April 2020

Abstract
Chemoresistance is a major problem in the treatment of patients with multiple myeloma(MM). Interleukin 6(IL-6), the most important growth factor for myeloma cells, mediates multiple signals which play a central role not only in the cell survival and the proliferation but also in the chemoresistance. The agents that suppress the IL-6-mediated signaling have the potential for the treatment of MM, such as PS-341 and curcumin. We have previously shown that baicalein from Scutellaria radix inhibited the proliferation and induced apoptosis of human myeloma cells, partly due to the suppression of IkB phosphorylation, resulting in the downregulation of NF-kB activity (Blood105:3312,2005). In order to clarify whether baicalein could inhibit IL-6-triggered signaling cascades in myeloma cells, we first demonstrated that baicalein inhibited not only the activation of MAPK, but also the phosphorylation of Stat3 and Stat1 induced by IL-6. Baicalein could also inhibit the phosphorylation of Akt triggered by IL-6, but not IGF-1. Since gp130 is phosphorylated by Jak family kinase after IL-6 stimulation, we examined whether Jak kinases could be involved in the baicalein-induced blockade of IL-6-induced signaling mediating the survival and growth of myeloma cells and the drug-resistance. Furthermore, we showed by phosphoprotein immunoblotting analysis that baicalein inhibited the activity of Jak2 and Tyk2 in myeloma cell lines (U266 and NOP-2 cells). Therefore, our results indicate that baicalein can also inhibit IL-6-induced signaling through Jak family kinase in myeloma cells, and combined with its suppressive effect on NF-kB activity baicalein can lead to more effective suppression of the proliferation and survival of myeloma cells.
Melphalan Resistance in Multiple Myeloma Can Be Overcome by Proteasome Inhibition or Histone Deacetylation: Lack of Syngergism of the Combination.
Authors: Sonneveld Pieter , Kamst Eric , de Knegt Yvonne , Klarenbeek Naomi , Schoester Martijn 
Published: April 2020

Abstract
Multiple Myeloma (MM) is a disease of monoclonal plasma cells in the bone marrow which has a transient response to classic chemotherapy. At diagnosis, induction chemotherapy followed by high-dose melphalan (HDM) with stem cell support is used in most patients to achieve a clinical response. Because all patients will ultimately relapse, the treatment of melphalan-refractory disease represents a major clinical challenge and new agents are needed to overcome melphalan resistance. We have investigated the anti-myeloma efficacy of two new classes of targeted agents, i.e. proteasome inhibition and histone deacetylation inhibition alone or in combination in the melphalan sensitive MM1S and the Melphalan refractory MM1MEL2000 cell lines. The IC50 values of Bortezomib (B), Melphalan (M) and LAQ824 (L) in MM1S were 2.1 nM, 1.9 uM and 1.7 nM, respectively and in MM1MEL2000 3.9 nM, 50 uM and 4.0 nM. Using isobologram analysis a synergysm between B and L was observed in the sensitive, however not in the melphalan refractory cell line. These data indicate that B proteasome inhibition and histone deacetylation inhibition may be effective ways to overcome melphalan resistance. However, the previously reported synergism between these drugs does not seem to occur in melphalan resistant cells. The gene expression profiles of these cell lines were analysed using the Affymetrix U133plus 2.0 gene chip before and after treatment with melfaphalan or the proteasome inhibitor B or the histone deacetylation inhibitor L or the combination of B and L.
Genes that were highly expressed in the melphalan refractory derivate cell line MM1MEL2000 as compared with wild-type MM1S included GP M6B, ADAM23 and HTPAP. Following melphalan exposure, TMF1, a CEBp glucocorticoid interaction factor, WHSC1L1, a MMSET homologue with EGF like domain and several transcription factors had highly increased expression as compared to MM1S. With exposure to B combined with L, increased expression in MM1MEL2000 over MM1S was observed for GTP exchange factor TIAM1 which interacts with RAS and JNK, and the lymphoid enhancer factor, a notch transcription factor. It is concluded that Bortezomib and the histone deacetylase inhibitor LAQ824 are effective agents to overcome melphalan resistance in multiple myeloma. However, the combination fails to show the synergism observed in melphalan sensitive cells. Gene analysis sofar does not provide a clear explanation for this lack of synergism. A comprehensive summary of the observed shifts of gene expression profiles in melphalan resistant cells following exposure to these agents, will be presented.
Overexpression and Preferential Nuclear Translocation of the Transcription Factor Yin Yang 1 (YY1) in Human Bone Marrow-Derived Multiple Myeloma.
Authors: Huerta-Yepez Sara , Baritaki Stravoula , Hernandez-Cuecto Angeles , Anguino-Hernandez Yu-Mei , Neshat Mehran , Chen Haiming , Campbell Richard A. , Gordon Melinda S. , Berenson James R. , Bonavida Benjamin 
Published: April 2020

Abstract
Yin Yang 1 (YY1) is a ubiquitously expressed zinc-finger transcriptional regulator that can function as an activator, a repressor, or an initiator binding protein. YY1 has been implicated in the control of cell growth, development, differentiation and tumor suppression. We have recently reported that YY1 regulates the transcription of death receptors in carcinoma and lymphoma and thus, YY1 regulates tumor sensitivity to immune-mediated apoptosis (Huerta-Yepez, et al., 2004, Oncogene23: 4993–5003; Vega, M., et al., 2005, Journal of Immunology175: 2174–2183). In addition, we have also reported that human prostate tumor tissues overexpress YY1 (Seligson, et al., 2005, International Journal of Oncology 27:131). This study investigated the expression of YY1 in multiple myeloma (MM) with the objective of determining whether YY1 plays a role in the progression and drug- resistance of MM. We have initiated these studies by examining nine bone marrow (BM) samples derived from patients with MM. Immunohistochemical studies were performed for the detection and cytoplasmic or nuclear localization of YY1 in the MM cells and also in adjacent normal mature and immature cells. The intensity of staining by the anti-YY1 antibody was scored and the relative intensity was calculated. Two slides from each patient were analyzed and 200 cells per slide were scored. Mean intensities of all samples were calculated and the data were subjected to statistical analysis. YY1 expression in normal BM was low and primarily of cytoplasmic origin. In contrast, YY1 was significantly overexpressed in MM cells. The mean intensity in the MM was approximately three-four fold higher than that of the normal cells and was primarily of nuclear origin (p-value &lt; 0.05). The signals that control shuttling YY1 are undefined. The expression of YY1 in normal mature and immature cells was low and there was comparable staining in the cytoplasm and the nucleus. Analysis of the cell distribution expressing YY1 by flow cytometry revealed that greater than 50% of the cells in the CD38+ subset expressed YY1. In addition, the MM tumor cells also expressed high level of pleiotrophin (PTN) and the patients had high levels of circulating PTN. PTN is a growth factor for MM and PTN transcription is regulated by an initiator element and could thus be a target of YY1-mediated transcriptional control. These findings suggest that YY1 overexpression may be involved in the pathogenesis of MM and is a potential target for therapeutic intervention. Current studies are aimed at examining the prognostic and diagnostic roles of YY1 and PTN in MM and molecular mechanisms that underlie their overexpression in MM.