bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cyclooxygenase-2 (COX-2) in Multiple Myeloma: Prognostic Marker or Therapeutic Target?.
Authors: Owen Roger G. , Fan Im , O’Connor Sheila J.M. , Davies Faith E. , Rollett Rebecca A. , Child J. Anthony , Rawstron Andy C. 
Published: April 2020

Abstract
Cyclooxygenase-2 (COX-2) is the key enzyme involved in prostaglandin synthesis. It appears to support the growth of a number of solid tumours including colon, breast, ovary, lung and uterine cervix and may be an important therapeutic target in at least some of these tumours. COX-2 expression has recently been evaluated (by immunohistochemistry using polyclonal anti-COX-2 antibodies) in multiple myeloma (MM) where expression was documented in 33–57% of patients. COX-2 expression in these studies was strongly associated with an adverse outcome. In addition there is some emerging data to suggest that the use of aspirin in MM may improve survival rates. In order to further evaluate this we have used a monoclonal antibody (Clone SP21, Labvision, Fremont, Ca) to assess COX-2 expression in both normal and neoplastic plasma cells. 52 specimens were assessed using standard streptavidin-biotin immunoperoxidase techniques using a known COX-2+ colon cancer as a positive control. Strong uniform COX-2 expression was seen in 32/33 (97%) of myeloma patients assessed and was also documented in all patients with MGUS (n=10). COX-2 expression was also documented in reactive plasmacytic lesions (oral mucosa, skin and lymph node, n=6) as well as normal bone marrow plasma cells (n=6). Megakaryocytes stained positively in all bone marrow biopsies examined and provided a useful positive internal control while erythroid, myeloid and lymphoid cells were consistently negative. We would conclude that COX-2 is strongly expressed by both normal and neoplastic plasma cells suggesting that COX-2 is a potential therapeutic target in MM. The apparent increase in the proportion of myeloma patients expressing COX-2 in the present study reflects the use of a monoclonal antibody in our immunohistology studies. The fact that polyclonal antibodies identify a lower proportion of patients who appear to have an inferior outcome suggests that the level of expression is of prognostic significance rather than its presence or absence. This is worthy of further study using more appropriate techniques such as RQ-PCR.
Detection of FGFR3 by Immunohistochemistry in Multiple Myeloma Is Highly Predictive of t(4;14) by FISH and Is Associated with CD56 (NCAM1) Expression.
Authors: Bahlis Nizar J. , Roland Birgitte , Maggliocco Toni , Auer Ivona , Stewart Douglas A. , Mansoor Adnan 
Published: April 2020

Abstract
Recent studies have demonstrated that neural cell adhesion molecule CD56 (NCAM1) is involved in multiple adhesive interactions with several different classes of ligands on the cell surface and in the extracellular matrix. One of these ligands is fibroblast growth factor receptor (FGFR) family. While a direct interaction between CD56 (NCAM1) and FGFR3 is yet to be reported we have speculated that such interaction might exist in myeloma cells with aberrant FGFR3 expression. In order to test this hypothesis we have retrospectively examined the presence of t(4;14) in 127 multiple myeloma samples and its correlation with the presence of deletion 13 by FISH; FGFR3 and cyclin D1 expression by immunohistochemistry and CD56 expression as determined by flow cytometry. FISH detected t(4;14) in 32 (25.1%) of 127 MM patient specimens. 71.4% of t(4;14) positive samples were found to have deletion of chromosome 13, in contrast to only 34% in t(4;14) negative samples. Aberrant expression of FGFR3 was detected by immunohistochemistry in 14/21 or 66.7% of t(4;14) positive samples. No expression of FGFR3 was detected in 10 t(4;14) negative samples. As expected low Cyclin D1 expression by IHC was detected in t(4;14) positive samples (5/21 or 23.8%). More importantly all samples positive for FGFR3 by IHC were CD56 positive (14/14 or 100%) and in 21/22 (95.5%) of t(4;14) positive in contrast to 62% in t(4;14) samples. Further work is currently being conducted in order to examine the downstream signaling pathways that might be activated as a result of such possible interaction.
Antiproliferative and Proapoptotic Effect of Vitamin D in Bortezomib (B) Resistant Human Myeloma Cell Line (HMCL).
Authors: Chanan-Khan Asher Alban , Romano Michelle , Pera Paula , Ersing Noreen , Muindi Josephia , Trump Donald L. , Johnson Candace 
Published: April 2020

Abstract
-Introduction: B is a proteasome inhibitor approved for treatment of multiple myeloma (MM) patients. Despite initial clinical efficacy all patients eventually become resistant to bortezomib and die of progressive disease. Development of newer agents with novel antitumor mechanisms is thus warranted. In vitro antiproliferative and proapoptotic effects of vitamin D are reported in various tumor models including MM. This is mediated through vitamin D receptor (VDR) which is also expressed on MM cells. Effect of calcitriol (D3) or its analog paricalcitol (D2) in HMCL resistant to B have not been previously examined. We investigated the antitumor potential of D3 and D2 in HMCL sensitive or resistant to B.
Method & Results: HMCL sensitive (OPM-2) or resistant to B (OPM-2/BR) were treated with various concentration of D2 or D3. Significant growth inhibition and induction of apoptosis was noted with D2 and D3 that was maximal at 3 days (p&lt;0.01). This antitumor response was noted despite resistance to B. Treatment of MM cells with vitamin D (D2 or D3) was associated with a loss of full-length PARP, decrease in p21 and induction of p27. Levels of the pro-survival molecules P-Akt and P-Erk were also reduced while total Akt and Erk remain unchanged.
Conclusion: We demonstrate that the antiproliferative and proapoptotitc effect of vitamin D in HMCL are mediated through interruption of the Akt pathway and were independent of resistance to B. We have initiated a phase I clinical trial to investigate the clinical utility of paricalcitol in relapsed or refractory MM patients.
OSU03012, a Novel Celecoxib Derivative, Induces Apoptosis in Multiple Myeloma Cells Independently of Caspase Activation.
Authors: Zhang Shuhong , White Valerie L. , Johnson Amy , Chen Ching-Shih , Farag Sherif S. 
Published: April 2020

Abstract
Multiple myeloma (MM) is a clonal disorder affecting terminally differentiated B cells, with the accumulation of plasma cells in the bone marrow. Previous studies showed that OSU03012 is a novel celecoxib derivative lacking cyclooxygenase-2 inhibitory activity that induces apoptosis in various types of cancer cells and is being developed as an anti-cancer therapy in the NCI Rapid Access to Intervention Therapy (RAID). Here, we examined the in vitro effect of OSU03012 in MM cell lines (U266, ARH-77, IM-9 and RPMI8226). Cytotoxicity data indicated that mean LC50 (lethal concentration 50%) of OSU03012 was 6.25±0.86 μM at 24 hours and 4.23±0.87 μM at 72 hours in these four cell lines. Using annexin V/PI (propidium iodide) flow cytometry assay, OSU03012 was shown to induce apoptosis in MM cells. OSU03012 activated caspases-8, -9, and -3, induced PARP (POLY ADP-RIBOSE Polymerase) cleavage, and reduced survivin and XIAP expression after 6 and 24 hour exposure. Although the caspase inhibitor Q-VD-OPH treatment strongly blocked OSU03012-induced PARP cleavage, it did not inhibit OSU03012-induced apoptosis of MM cells. The pan-caspase inhibitor z-VAD-fmk did not prevent OSU03012 mediated cell death. Cell death with OSU03012 treatment was associated with significant down-regulation of phospho-Akt. Several substrates of AKT, including phospho-GSK-3 beta (Ser9), phospho-FoxO1a (Ser256) and phospho-MDM2 (Ser166) were also down-regulated by OSU03012 drug. OSU03012 triggered both early (6h) and late (24h) down-regulation of cyclin D1 expression, but cyclin A and B1 expression was down-regulated only at 24h. There was no induction of p21 or p27 protein levels by OSU03012. After 24-hour exposure, low concentration (1–5 μM) OSU03012 arrested MM cell lines in the G1 phase of the cell cycle while high concentration (10 μM) OSU03012 induced G2 phase arrested. OSU03012 decreased both phospho-Stat3 (Ser727) and Stat3 expression. OSU03012 has on effect on phosphorylated MAP kinase kinase1/2 (pMEK1/2) but it decreased MEK1/2 expression at 24h. The expression levels of Bcl-2 family proteins, Bcl-2, Mcl-1, BAX, and BIM did not alter with OSU03012 treatment suggesting that Bcl-2 members may not play direct or significant roles in inducing cell death. Taken together, we conclude that OSU03012 is potently active against MM cells by predominantly caspase-independent mechanisms, and may involve downstream pathways consequent to phopho-Akt down-regulation. These studies provide preclinical rationale for investigating OSU03012 in the treatment of MM.
OSU-HDAC42, a Novel Histone Deacetylase Inhibitor, Induces Apoptosis in a Caspase-Dependent Manner and Induces p21WAF1/CIP1 and p16 Expression in Multiple Myeloma Cell Lines.
Authors: White Valerie L. , Zhang Shuhong , Lucas David , Chen Ching-Shih , Farag Sherif S. 
Published: April 2020

Abstract
Multiple myeloma (MM) is a neoplastic disorder characterized by accumulation of slowly-proliferating clonal plasma cells. OSU-HDAC42 [a.k.a. (S)-HDAC-42] is a novel histone deacetylase inhibitor that induces apoptosis in various types of cancer cells and is being developed as an anti-cancer therapy in the NCI Rapid Access to Intervention Therapy (RAID) program. In this study, we tested the in vitro activity of OSU-HDAC42 against human MM cells.
OSU-HDAC42 induced myeloma cell death, with an LC50 of less than 1.6μM after 48 hours in the four cell lines tested - U266, IM-9, RPMI 8226 and ARH-77 using the MTT assay. OSU-HDAC42 induced cleavage of caspases 3, 8 and 9, as well as polyADP-ribose polymerase (PARP). Addition of the pan-caspase inhibitor Q-VD-OPH before exposure to the drug prevented apoptosis at 48 hours, as determined by Annexin V/propidium iodide staining. These results indicate that OSU-HDAC42 induced apoptosis by a mainly caspase-dependent manner. Bax expression was up-regulated at 24 and 48 hours, while Bcl-2 remains relatively constant. Mcl-1 showed increasing cleavage at increasing doses of OSU-HDAC42. These findings support a mitochondrial pathway of apoptosis. Cell cycle suppressor proteins p21WAF1/CIP1 and p16 were also significantly induced after treatment with the drug, suggesting that OSU-HDAC42 may also acts on pathways to halt cell cycle progression.
In addition, the gp130 (signal-transducing) subunit of the IL-6 receptor was down-regulated by OSU-HDAC42 exposure. The tyrosine-phosphorylated form of STAT3, which is phosphorylated by dimerized gp130, was also dramatically reduced following incubation with OSU-HDAC42, supporting the finding that gp130 expression is diminished. As IL-6 is an important growth and survival factor for MM cells, down-regulation of gp130 may be an important mechanism for the activity of OSU-HDAC42 against MM cells. TRAIL, FasL, XIAP, and p53 expression were not affected by OSU-HDAC42. While other HDAC inhibitors have been shown to activate the death receptor pathway or down-regulate XIAP, this was not observed with OSU-HDAC42 in myeloma cells.
In conclusion, OSU-HDAC42 has in vitro activity against myeloma cells and acts via activation of caspases, inducing the cell cycle suppressors p21WAF1/CIP1 and p16, as well as interfering with the IL-6 signal transduction pathway.
Upregulation of MMP-2 & 9 by Co-Culture of Human Myeloma Cell Lines and Endothelial Cells May Be Responsible for Protection Against Cytotoxic Drugs.
Authors: Paneesha Shankaranarayana , Adya Raghu , Khanji Hemali , Leung Ed , Vijayasekar C. , Pratt Guy , Borg Anton , Rose Peter , Morris Alan 
Published: April 2020

Abstract
Multiple myeloma is a clonal lymphoproliferative disorder characterised by the proliferation of plasma cells in the bone marrow. Inspite of good initial response, it is associated with universal relapse. We hypothesise this is due to sanctuary provided to myeloma cells by the endothelium. Matrix metalloproteinases (MMPs) are shown play a role in cell growth, invasion, angiogenesis, metastasis and bone degradation. We show here the protection offered by endothelial cells to human myeloma cell lines in in-vitro co-culture with upregulation of MMP-2 & 9 and the role of GM6001 MMP inhibitor (Ilomastat) in overcoming this protection.
Human myeloma cell lines (H929, RPMI 8226, U266 & JJN3) with or without endothelial cells (human umbilical vein endothelial cells and EaHy 926 cell line) in-vitro co-culture were treated with melphalan, dexamethasone, arsenic trioxide and Ilomastat. Cytotoxicity/proliferation were assessed by the alamarBlue™ assay (Serotec) and validated by Annexin V-FITC apoptosis detection Kit (Calbiochem) and BrDU proliferation assay (BD Pharmingen™). Gelatin Zymography was used to demonstrate activity of MMP-2 & 9 in the supernatant. MMP-2 and 9 mRNA expression was quantified by Real Time Quantitative PCR (ROCHE).
Co-culture of human myeloma cell lines with endothelial cells lead to increase in the proliferation of myeloma cell lines and also protected them from the cytotoxicity of chemotherapeutic agents. MMP-2 & 9 activity was upregulated by the co-culture. MMP-2 mRNA expression in human myeloma cell lines increased following 4 hr co-culture. Treatments with Ilomastat lead to the suppression of proliferation in co-culture in a dose dependent manner, associated with a reduction of MMP-2 and 9 activity. Our study shows endothelial cells offer protection to human myeloma cell lines in the presence of cytotoxic agents. This may result in the sanctuary of myeloma cells in bone marrow leading to ultimate relapse of disease. Our study also demonstrates the upregulation of MMP-2 and 9 by co-culture and increased cytotoxicity achieved by the inhibition of MMPs. Further studies are needed to determine the exact role of MMPs in myeloma biology as MMP inhibition may be an interesting therapeutic target and help in averting relapse in multiple myeloma.
PAD Is a Highly Effective Regimen for the Treatment of Patients with Relapsed Refractory Multiple Myeloma.
Authors: Delforge Michel , Janssens Ann , Dierickx Daan , Maertens Johan , Theunissen Koen , Vandenberghe Peter , Verhoef Gregor 
Published: April 2020

Abstract
Background and aim of the study:
Bortezomib (Velcade®) is a first-in-class potent and reversible proteasome inhibitor that has been approved for the treatment of relapsed and/or refractory multiple myeloma (MM). With single agent bortezomib, complete (CR) and partial (PR) response rates have been reported in 27% of patients with relapsed refractory (Richardson 2003), and in 38% of patients with relapsed MM (Richardson 2005). Addition of dexamethasone (dex) to bortezomib can increase the response rates (Jagannath, 2005), and a synergistic anti-myeloma effect has been described when bortezomib is combined with cytotoxic drugs like anthracyclins and melphalan (Ma, 2003). With the combination of PAD (bortezomib, adriamycin and dex) a 95% CR+PR rate in untreated myeloma patients has been reported (Oakervee, 2005), but data on the effectiveness of PAD in the relapsed, refractory setting are currently lacking.
Patients and Treatment:
We have studied the anti-myeloma effect of PAD in 8 unselected relapsed refractory MM patients (M/F:6/2; mean age 61 y, range:50–75). The mean time since diagnosis was 6 years (range:2–11). Previous treatment consisted of a median of 6 treatment lines including autologous (n = 6) and allogeneic (n = 2) transplantation, various forms of combination chemotherapy (n = 8) including anthracyclins (n = 7), pulse dose dex (n = 7), thalidomide −/+ dex (n = 7), bortezomib (n = 8) without (n = 3) or with dex (n = 5). PAD was administered as previously published (Oakervee, 2005), but with 12 days of dex administration also during during the odd cycles.
Results:
A median of 4 cycles of PAD were administered (range: 2–6). The CR+ PR rate was 88% according to the EBMT criteria: one patient achieved a CR (IF+), one a VGPR, and 5 patients a PR. One patient had a limited amount of paraprotein secretion but multiple extramedullary plasmacytomas. After 2 cycles of PAD she had &gt; 50% reduction in tumormass on PET-CT imaging. On average, the best response occurred already after 2 cycles of PAD. 4/8 patients relapsed after a mean interval of 5 months and 3 patients are still in remission at + 5 months. Two patients developed septicaemia with one toxic death. Hematological grade III/IV toxicities included: neutropenia (4/8), and thrombocytopenia (4/8). Neuropathy pre-existed in 4/8 patients and was not aggravated with appropriated dose reductions of bortezomib.
Conclusion:
Using a combination of bortezomib, adriamycin and dexamethasone (PAD), we could obtain at least a PR in 8/8 unselected highly refractory myeloma patients who had, amongst many other agents, previously been exposed to bortezomib and/or adriamycin and/or dexamethasone. Major toxicities of PAD were hematological with one toxic death because of septicaemia. We believe that our observation warrants further studies of the PAD regimen in relapsed MM.
Reduced Intensity Conditioning Regimen for Allografting Following Autografting Has Strong Anti-Myeloma Activity and Delays Disease Progression Compared to Tandem Autografting.
Authors: Carella A.M. , Lerma E. , Nati S. , Congiu A. , Rossi E. , Spriano M. , Vimercati R. 
Published: April 2020

Abstract
Multiple Myeloma (MM) is still an incurable disorder despite the numerous attempts to exploit new therapy approaches for it. The better understanding of its molecular pathogenesis has led to the development of effective, novel therapeutic agents like thalidomide and the proteasome inhibitor bortezomib. High-dose therapy with stem cell transplantation and novel targeted therapies represent two approaches to overcome resistance of multiple myeloma cells to conventional treatments. Autografting (AutoSCT) has been limited by high-relapse rates and conventional allografting (AlloSCT) by excessive TRM and toxicity. Reduced intensity conditioning for transplant (RICT), a less toxic procedure for AlloSCT that aims to exploit graft versus tumor effect, has been shown to achieve remissions in MM. High-dose therapy/AutoSCT followed shortly thereafter by RICT might improve outcomes in MM as compared to AutoSCT or conventional AlloSCT used alone. We compared two retrospective cohort of patients who underwent either tandem AutoSCT (HDT consisted of Melphalan 200 mg/m2) or AutoSCT followed closely by related RICT (patients with HLA-matched siblings). The two groups were matched for pre-transplant therapy, disease status at transplant, time from diagnosis to transplant. GVHD prophylaxis for RICT patients consisted of CyA/MTX. The major results are summarized in the Table. In the AutoSCT/RICT group the complete remission rate was higher (p=0.004) and the risk of disease progression after transplant was significantly reduced. All patients who reached CR responded after full chimerism and GVHD developed. This finding confirms the existence of a graft-versus-myeloma effect. Since the first clinical signs of response in remitters patients were noted between 70 and 120 days and maximum response between 160 and 200 days after RICT (after DLI in one patient), these responses should be considered immunological responses. Our data demonstrated that RICT following an AutoSCT can mediate a potentially curative graft-versus-myeloma effect and reduces the incidence of disease progression.
Tandem ASCT (N=35) ASCT+RICT (n=20) Age, median 56 (range, 38–66) 51 (range, 34–63 Median number of prior cycles of Chemoth. 4 (range, 3–6) 4 (range, 3–6) Time from Dx to 1st AutoSCT (median mo.) 6 (range, 5–60) 9 (range, 7–42) Conditioning Regimen for AutsoSCT Melphalan (200mg/m2) Melphalan (200mg/m2) Conditioning regimen for RICT --- TBI/Fludarabine Median days from AutoSCT to RICT --- 80 Complete Remission rate 14% 50% Disease-free Survival at 4 yrs 14% 45% Overall Survival at 4 yrs 28% 40% Transplant-Related Mortality 0% 0%
Clinical CMRF-56 Blood Dendritic Cell Preparations for Immunotherapy of Multiple Myeloma.
Authors: Vari Frank , Rossetti Tony , Freeman Jennifer , Hart Derek 
Published: April 2020

Abstract
Aim: Despite advances in therapy, such as stem cell transplantation and new drugs such as the proteosome inhibitors, multiple myeloma (MM) remains incurable for most patients. Immune therapy, which uses dendritic cells (DC) to generate cytotoxic T lymphocyte (CTL) responses targeting residual disease is a promising, potentially curative strategy, compatible with other therapies. We have shown that blood DC can be isolated from MM patients and that they induce CTL responses (J. Immunotherapy.2005; 28:322). The CMRF-56 monoclonal antibody (mAb) detects an antigen expressed on blood DC after a short period of in vitro culture. To create a GMP compatible DC purification platform for clinical investigation of DC immunotherapy, we developed a method for single step CMRF-56 mAb positive immunoselection of blood DC using magnetic beads.
Methods and Results: CMRF-56 mAb immunoselection produced a blood DC enriched population containing 33.2±3.5% CD11c+ blood DC (including CD1c+, CD141+ and CD16+ subsets) and 0.6±0.2% CD123hi plasmacytoid blood DC (n=6). Additional CMRF-56 positive B cells and monocytes comprised 14.7±5.8% and 32.4±3.4% of the preparations respectively (n=6). CMRF-56 selected cells stimulated CTL responses against defined tumour associated antigens, including MART-1 peptide and the MM associated antigen, HM1.24. Furthermore they generated strong MM specific CTL responses after incubation with MM cell lysates. Brief DC activation with GM-CSF enhanced these responses, improving MM-specific CTL activity (Cr release) in cultures primed with MM lysate loaded CMRF-56 selected cells from 18.9±7.4% to 44.1±8.0% (n=3, p=0.051). We have now developed and validated a procedure for isolation of CMRF-56+ blood DC using the CliniMACS device. Apheresed mononuclear cell preparations were cultured, labelled with antibody and blood DC immunoselected in a closed system, free of human or animal serum supplements, in a manner compatible with regulatory requirements for cell therapy. Simultaneously, our team has developed a bioprocess for production of the murine biotinylated CMRF-56 mAb, suitable for the large scale synthesis required for multi-centre clinical trials. It incorporates components that remove contaminating DNA, host cell proteins, viral particles and endotoxin. The first batch of clinical grade CMRF-56 mAb has been produced and used for successful laboratory scale blood DC isolation. We have also completed a number of clinical scale blood DC preparations from healthy donors (n=10) and MM patients (n=2). There were sufficient blood DC isolated from both the healthy donors and MM patients for the planned vaccination schedule. The average blood DC purity was 38% in healthy donors and 29% in MM patients, while the average yields were 35 and 44%, respectively.
Conclusions: When additional studies on MM patients are completed these studies will provide the necessary pre-clinical validation data to commence a Phase I Clinical Trial.