bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Anti-Myeloma Effect of Bortezomib Is Associated with Osteoblastic Activity.
Authors: Zangari Maurizio , Najarian Kevin B. , Esseltine Dixie L. , Lee Choon-Kee , Barlogie Bart , Elice Francesca , Burns Michael J. , Yaccoby Shmuel , Richardson Paul , Sonneveld Pieter 
Published: April 2020

Abstract
Myeloma bone disease is characterized by osteolytic destruction associated with suppressed osteoblastic activity possibly through inhibition of the WNT signaling pathway. We already reported the association between increased alkaline phosphatase (ALP) and B but not dexamethasone response in myeloma patients (Zangari et al., BJH 2005, in press). To extend these observations, using data from the large APEX study, we have attempted to assess the relationship between quantitative alkaline phosphatase (ALP) changes during therapy with B and response and time to progression. We were interested in what threshold of increase in ALP (even within the normal laboratory range) might predict efficacy.
Methods: A group of patients with no biochemical evidence of liver disease and creatinine clearances &gt; 50 mL/minute treated with bortezomib 1.3 mg/m2 on days 1, 4, 8 and 11 for up to eight 3-week cycles followed by bortezomib 1.3 mg/m2 on days 1, 8, 15 and 22 for up to three 5-week cycles were studied. The percentage increase of ALP levels in responders and non-responders was analyzed at different thresholds and time points after baseline. Responses (the sum of CR plus PR) were based on EBMT criteria. The change from baseline ALP during therapy with B was compared in responders and non-responders.
Results: There were 229 patients with a median age of 60 years. 59% were male; median B2M was 3.2 mg/L, myeloma subtypes included IgG 64%, IgA 24%; median time from diagnosis was 41 months. The median ALP elevation from baseline for bortezomib responding patients was approximately 25% but even a 10% ALP increment at week 6 from baseline was strongly associated with response (P=0.0001). The most powerful predictor of response was a 25% ALP elevation at week 6.
Table 1 Parameter 25% ALP Elevation &lt;25% ALP Elevation p-value N 64 154 CR n (%) 10 (16) 12 (8) 0.0966 PR n (%) 35 (55) 42 (27) &lt;0.0001 CR/PR n (%) 45 (70) 54 (35) &lt;0.0001
Cox regression analysis showed that ALP elevation of 25% was not only associated with response rate but was also positively correlated with time to progression (P=0.0022).
For all bortezomib treated patients enrolled in the trial (n=333) regardless of their baseline (LFTs and creatinine), a 25% increase in ALP from the baseline to 6 week remained strongly associated with a very high response rate (p&lt;0.0001) and longer time to progression (206 vs 169 days) p=0.01.
Figure Figure
At the time of relapse after B therapy, the median ALP was 65.0 U/L and comparable to baseline level (65.5 U/L).
Conclusion:
During bortezomib treatment, markers of osteoblastic activation (such as ALP) may predict response and response duration in patients with multiple myeloma. In addition, this suggests that bone anabolism could affect myeloma growth. This observation needs to be confirmed prospectively.
Development of Retargeted CD38-Specific NK-92 Cell Line for Potential Anti-Myeloma Immunotherapy.
Authors: Yang Shihong , An Xin , Brown Ross D. , Ho Joy , Gibson John , Joshua Douglas E. , Wels Winfried , Sze Daniel M. 
Published: April 2020

Abstract
Multiple Myeloma (MM) is a B cell neoplasm in which malignant plasma cells accumulate in the bone marrow and produce excessive amounts of a monoclonal immunoglobulin. Among the large number of antigens expressed by plasma cells, CD38 is one of the highest expressed surface molecules, suggesting an alternative public tumour target. For instance, the myeloma cell line 8226 has a high surface expression of CD38. We plan to introduce genetically recombinant single-chain variable fragments (scFv) of anti-CD38 antibody, along with CD3 chain, into NK-92 a natural killer cell line which is highly cytotoxic against a variety of malignant cells (the only natural killer cell line being used in phase I/II clinical trials). It is likely that the retargeted NK-92 cells may specifically and efficiently kill CD38-expressing myeloma cells. By using a series of degenerate primer sets for mouse immunoglobulin variable region genes, VH and VL regions were isolated through reverse transcription PCR from OKT10 cell line. Further sequencing analysis provided novel information of the OKT10 immunoglobulin heavy and light chains. A PCR-based assembling strategy was employed to construct the scFv with FLAG sequence integrated as a tag. VH and VL domains were joined together with a hinge region of (Gly4Ser)3. The phagemid construct has been formed by subcloning the scFv fragment into the Sfi I/Not I cloning sites of the expression vector pHEN2, and subsequently transformed into the E. Coli HB2151. Five out of 7 transformed bacterial colonies screened did not contain the scFv fragment. For the remaining two clones with the correct-sized insert, we tested the specificity of the scFv fragments that were prepared through intracellular antibody capture as periplasmic proteins. We performed a competitive flow assay using periplasmic proteins with commercially available anti-CD38 PE conjugated antibodies on 8226 myeloma cell line. We found that one of these two clones (clone 6) showed a reduction in CD38 staining and presented as double peaks; while this pattern was not seen in the clone 7. Subsequent DNA sequencing revealed that there was a 100-bp truncation in the heavy chain region in clone 7; while the clone 6 has a complete productive heavy chain and light chain spanned by the linker region. We are going to transfect NK-92 cells with retrovial vector pL-scFv(clone 6) construct as the basis of an efficient approach for anti-myelome immunotherapy. (Dr. Daniel Sze is supported by the International Myeloma Foundation Senior Research Grant).
Chemosensitization of CDDP-Resistant Multiple Myeloma Cell Lines Following Treatment with Anti-TfR IgG3-Av.
Authors: Suzuki Eriko , Gan Xiao-Ho , Umezawa Kazuo , Helguera Gustavo , Schiller Gary , Penichet Manuel , Bonavida Benjamin 
Published: April 2020

Abstract
Multiple myeloma (MM) is an incurable proliferative disorder of monoclonal malignant plasma cells. Multiple agents conventional dose chemotherapy regimens have resulted in improved response rates, however, there have been no significant improvements in overall survival. Recently, Bortezomib has been introduced for the treatment of MM and has resulted in significant response rates. Nevertheless, additional strategies for direct treatment of MM patients are urgently needed. In general, cancer cells have been shown to express significantly higher levels of the transferrin receptor (TfR) and thus, this receptor is an attractive target for cancer therapy. We have generated an anti-TfR IgG3-avidin fusion protein (anti-TfR-IgG3-Av) that can deliver biotinylated molecules into cancer cells and exhibits intrinsic pro-apoptotic activity against hematopoietic malignant cells including myeloma (Ng P. et al., 2002, PNAS 99:10706–11). We have also reported that certain monoclonal antibodies (e.g., rituximab) can sensitize tumor cells to various chemotherapeutic drug-induced apoptosis (Jazirehi and Bonavida, 2005, Oncogene 65:264–76). Thus, this study investigated whether anti-TfR IgG3-Av sensitizes drug-resistant MM cell lines to drug-induced apoptosis. We initially examined the CDDP-resistant lymphoblastoid IM-9 and ARH-77 cell lines for chemosensitization using a suboptimal dose of anti-TfR IgG3-Av. Several methods were used to assess the cytotoxic activity, namely viable cell recovery and apoptosis by both PI and caspase-3 activation. The findings demonstrate that treatment of tumor cells with anti-TfR IgG3-Av for 18 h following treatment with CDDP for an additional 18 h resulted in significant cytotoxicity and apoptosis and synergy was achieved. The combined treatment also resulted in significant mitochondrial membrane depolarization, suggesting that apoptosis was achieved through a type-II pathway. In comparison with tumor cell treatment with anti-TfR-IgG3-Av, the anti-TfR IgG3 was a more potent chemosensitizer, based on both the concentration used and the extent of apoptosis achieved. These findings demonstrate that anti-TfRIgG3-Av is a potential therapeutic antibody that will reverse MM drug resistance when used in combination with chemotherapeutic drugs.
Sensing Iron: Reciprocal Regulation of Hepcidin mRNA by Soluble and Cell-Associated Hemojuvelin.
Authors: Lin Lan , Goldberg Y. Paul , Ganz Tomas 
Published: April 2020

Abstract
Human genetic studies identified HJV (also called HFE2) as the major cause for juvenile hemochromatosis (JH). Patients with HJV hemochromatosis have low urinary levels of hepcidin, the principal iron-regulatory hormone secreted by the liver. We attempted to establish the specific roles of HJV in iron metabolism, especially its relationship with hepcidin. Translation of the genomic sequence indicated a C-terminal GPI anchor for the protein product of HJV, hemojuvelin. This suggested that hemojuvelin may have either a soluble or a cell-associated form. In human hepatoma cell line Hep3B, knockdown of cellular HJV by siRNA decreased hepcidin expression, independently of the IL-6 pathway. Intriguingly, the addition of recombinant soluble hemojuvelin (rs-hemojuvelin) also suppressed hepcidin expression in primary human hepatocytes, in a log-linear dose-dependent manner, suggesting competition between soluble and cell-associated forms of hemojuvelin. Soluble hemojuvelin was found in human sera at concentrations similar to those required to suppress hepcidin mRNA in vitro. In cells engineered to express hemojuvelin, soluble hemojuvelin release was progressively inhibited by increasing iron or holotransferrin concentrations. Our study suggests that soluble and cell-associated hemojuvelin reciprocally regulate hepcidin mRNA levels, and that hemojuvelin may serve as a molecular messenger for iron homeostasis. Even in hepatocytes stimulated with IL-6, we observed strong suppression of hepcidin mRNA by rs-hemojuvelin. If rs-hemojuvelin or its active fragments also suppress hepcidin production in vivo, they could be used to alleviate anemia of inflammation.
High Response and Complete Remission Rates in Relapsed/Refractory Multiple Myeloma Treated with Bortezomib, Thalidomide, Dexamethasone and Zoledronic Acid (VTD-Z) Combination Therapy.
Authors: Teoh Gerrard , Tan Daryl , Hwang William , Koh Liang Piu , Chuah Charles , Ng Heng Joo 
Published: April 2020

Abstract
Purpose: Bortezomib (VELCADETM) is a novel proteasome inhibitor that has shown tremendous clinical efficacy either as a single agent or in combination with other cytotoxic agents in relapsed/refractory multiple myeloma (MM). In this study, we combined bortezomib (V) with thalidomide (Thal), dexamethasone (Dex) and zoledronic acid (Zol) to determine the efficacy and toxicity of this regimen (VTD-Z) in patients with relapsed/refractory MM.
Methods: Patients were treated for 2 to 11 three-weekly cycles of VTD-Z comprising: V 1.3 mg/m2 on days 1, 4, 8 and 11; Thal 50 mg ON; Dex 20 mg OM on days 1 to 4, 8 to 11, 15 to 18; and Zol 4 mg on day 1. Patients did not receive any prophylaxis for venous thrombosis. Response was graded according to Bladès criteria.
Results: Twelve patients (2 males, 10 females; median age 63.3 years) with relapsed/refractory MM were studied. Complex karyotypes were present in all (10 out of 10) patients who had karyotype studies performed at the time of diagnosis. Four patients had deletion chromosome 13 (del(13)). The overall response rate (RR) was 91.7% (11 out of 12), of which 50.0% (6) achieved complete remission (CR), 25.0% (3) achieved near-CRs (nCR), 16.7% (3) were partial responders (PR). There were no minimal responders (MR) and 1 (8.3%) non-responder (NR). Two patients who achieved CR had del(13). Transient grade 3 thrombocytopenia was the only severe adverse event and this was observed in 4 (33.3%) patients. Grade 2 peripheral neuropathy was observed in 4 (25.0%) of patients. None of the patients developed deep vein thrombosis or pulmonary embolism. The single NR was the only death that occurred during the period of study.
Conclusions: Our study demonstrates that the VTD-Z regimen is exceptionally effective and safe in patients with relapsed/refractory MM. The high CR/near-CR rates (total of 75.0%), attained after relatively few cycles of VTD-Z, suggest that these three drugs could be at least additive, if not, even synergistic. Longer term follow-up will be required to determine the durability of these responses.
Thalidomide Sensitizes 32D cl 3 Hematopoietic Progenitor Cells to Ionizing Irradiation.
Authors: Greenberger Emily E. , Epperly Michael W. , Franicola Darcy , Zhang Xichen , Jacobs Samuel , Greenberger Joel S. 
Published: April 2020

Abstract
Thalidomide (TL) has been used in the treatment of multiple myeloma and due to its anti-angiogenic effects has been postulated to be a potential radiosensitizer of tumors in vivo. We first investigated the effect of TL on growth of IL-3 dependent murine hematopoietic 32D cl 3 cells in semisolid methylcellulose containing medium also supplemented with TL ranging from 0 to 200 μM. The cells were incubated at 37°C for 7 days at which time colonies of greater than 50 cells were counted. TL increased cell colony formation 2-fold over the dose range of 10 to 100 μM. In serially decreasing concentrations of IL-3, TL (50 μM) was growth stimulatory of 32D cl 3 colony formation in 15% IL-3 and 10% IL-3 conditioned medium (146.2 ± 7.2 and 89.3 ± 2.4 colonies/500 cells plated in TL vs. 110.8 ± 11.4 and 58.0 ± 2.1 without TL, p = 0.0304 or 0.0006). To determine whether thalidomide was a radiosensitizer, cells were placed in: 0, 50, or 150 μM TL in three protocols:
1 hour before irradiation; 1 hour before irradiation plus also in methylcellulose following irradiation; or were placed in TL containing methylcellulose only following irradiation.
Cells were irradiated to doses ranging from 0 to 800 cGy and were plated in methylcellulose containing medium. Seven days later, colonies of greater than 50 cells were counted and analyzed by single hit, multi-target or linear quadratic models. Cells grown in 50 μM TL which stimulated cell growth resulted in an increased radiation resistance compared to control irradiated cells (D0 = 1.87 ± 0.02 vs. 1.30 ± 0.09 Gy, respectively, p = 0.0366). However, 150 μM TL (which did not stimulate cell growth) increased radiation sensitivity compared to control irradiated cells (D0 = 1.22 ± 0.5 vs. 1.64 ± 0.11 Gy, respectively, p = 0.0245). To determine whether thalidomide radiosensitized tumor cells in the therapeutic 150 μM range, SSC-VII murine squamous cell carcinoma cells were grown in TL at concentrations ranging from 0 to 200 μM and were assayed for colony formation. In contrast, SSC-VII cells showed no detectable stimulation or inhibition of cell growth for unirradiated cells in any concentration of TL. However, at 150 μM TL, SSC-VII cells were radiosensitive when incubated in TL before irradiation compared to control irradiated cells (D0 = 2.02 vs. 2.55 Gy, respectively). These results demonstrate that thalidomide may be a clinically valuable radiosensitizer.
Interim Analysis of a Phase II Trial of Bortezomib in Patients with Multiple Myeloma Who Failed To Achieve a Complete Response to Autologous Peripheral Blood Stem Cell Transplantation.
Authors: Rifkin Robert M. , Beveridge Roy A. , Brunvand Mark W. , Spitzer Gary , Kristi Boehm A. , Rauch Mary Ann , Zhan Feng , Asmar Lina 
Published: April 2020

Abstract
The majority of patients (pts) with multiple myeloma (MM) fail to achieve a complete response (CR) with standard induction therapy (ie, VAD, DVD, Thal-Dex). Bortezomib has been shown to induce a CR in ~10% of subjects with refractory/relapsed MM. This study was conducted to assess the safety and efficacy of bortezomib in converting pts, who failed to achieve a CR to high-dose chemotherapy, to complete responders.
A total of 68 pts are targeted for enrollment on the current study protocol. To date, 20 pts have enrolled and are the subject of this planned safety analysis. As of this submission, 18 pts have been treated and 16 pts have either completed or discontinued treatment. All pts have received at least one dose of bortezomib according to the following schedule. Bortezomib was administered at a dose of 1.3 mg/m2 via IV push on Days 1, 4, 8, and 11 every 21-days for 4 cycles. Patients who failed to demonstrate disease progression were eligible for 4 additional cycles of therapy administered according to the above schedule. The demographic data of the first 20 subjects enrolled are summarized in the table below.
Sixteen of 20 pts enrolled have now completed the study. Patients received a median of 4 cycles (range, 3–8). Reasons for discontinuation included: adverse events (n=3 neuropathy (± fatigue) and n=1 fatigue with constipation) and other (n=1 each, progressive disease, carpal tunnel, deemed ineligible). Six patients had dose(s) delayed and/or reduced due to adverse events (peripheral neuropathy; fatigue; neutropenia and herpes zoster; fatigue with neuropathy; nausea, vomiting, diarrhea, and sinus infection; hypertension, and fatigue with constipation).
Treatment-related Grade 3–4 toxicities have been infrequent and include: asthenia, peripheral neuropathy, and thrombocytopenia (11% each) and hypertension, abdominal pain, and neutropenia (5.6% each). Grade 4 toxicity was limited to 2 pts with thrombocytopenia. No life threatening infections or reactivation of herpes zoster have been observed. Bortezomib when administered in the post-transplant setting appears to be safe and well tolerated. Neuropathy and thrombocytopenia appear to be major factors in patient discontinuation. Following review of this planned interim analysis for safety, the study continues to accrue toward completion.
Patient characteristics Variable %Male:%Female 25:75 Mean age (range) 53 years (41–66) Number and percentage (%) of patients Myeloma Isotype IgA Kappa 3 (21.5) IgA Lambda 1 ( 7) IgG Kambda 4 (29) IgG Lambda 1 ( 7) Kappa light chain 3 (21.5) Lambda light chain 2 (14) Prior Chemotherapy 17 (85) Prior Radiation 5 (25) Type of Transplant Single 12 (60) Tandem 6 (30) Not Available 2 (10)
The Effects of Zoledronic Acid Dose and Infusion Rate on Pharmacokinetics (PK), Pharmacodynamics (PD), and Renal Function in Patients with Multiple Myeloma.
Authors: Berenson James R. , Major Pierre , Swift Regina A. , Vescio Robert , Rivera Christina , Bilic Sanela , Gauron Sonia , Denouel Jannick , Schran Horst 
Published: April 2020

Abstract
BACKGROUND: Zoledronic acid has demonstrated efficacy and safety in the treatment of bone lesions in patients with multiple myeloma. However, bisphosphonates are known to have dose-dependent and infusion rate-dependent effects on renal function and patients with multiple myeloma may have disease-associated compromise of renal function. The pharmacokinetics (PK), pharmacodynamics (PD), and safety of monthly intravenous (IV) infusions of zoledronic acid (4, 8, and 12 mg) were investigated in a subset of patients with multiple myeloma as part of an open-label, phase I, multiple-dose study. Doses higher than the approved 4-mg dose were infused over 1 or 2 hours based on an infusion-rate model that predicted a Cmax similar to that of the approved regimen (4 mg via 15-minute IV infusion). Moreover, levels of tumor markers were measured to assess potential clinical antitumor activity at various dose regimens.
METHODS: Patients received a total of 6 infusions of zoledronic acid in 28-day cycles: 4 mg via 15-minute infusion on day 1; 8 mg via 60-minute infusion on days 29 and 57; 12 mg via 120-minute infusion on days 85 and 113; and 4 mg via 15-minute infusion on day 141.
RESULTS: Ten patients with multiple myeloma were enrolled, and 7 completed all 6 infusions. Median age was 58 years, ECOG performance status was ≤ 1 in 9 patients and 2 in 1 patient; 80% had received prior antineoplastic therapy, and 100% had received prior radiation and/or surgery. Three patients discontinued because of abnormal laboratory values (n = 2) or withdrawal of consent (n = 1). The most common AEs were fatigue (60%), myalgia (50%), arthralgia (40%), and nausea (40%). Increased serum creatinine &gt; 0.3 mg/dL over baseline prompted a dose delay/reduction in 1 patient and discontinuation of treatment in 2 patients. In multiple myeloma patients, mean Cmax (± SD) was 329 (± 89) ng/mL after the first 4-mg dose infused over 15 minutes (1st infusion), 407 (± 105) ng/mL after the 2nd 8-mg dose infused over 60 minutes (3rd infusion), and 385 (± 114) ng/mL after the 2nd 12-mg dose infused over 120 minutes (5th infusion). Increasing doses produced proportional increases in drug exposure (AUC). Decreases from baseline were observed in serum markers of bone resorption; however, there were no consistent or meaningful changes in tumor marker levels (eg, serum IL-6).
CONCLUSIONS: This study indicates that by prolonging infusion times to 1 hour and 2 hours using 8-mg and 12-mg doses of zoledronic acid, respectively, Cmax can be maintained and is similar to that obtained with the approved 4-mg dose infused over 15 minutes.
Flow Cytometric Detection of the Expression of CXCR4 on CD34+ Cells and the Distribution of Lymphocyte Subsets in Allogeneic Hematological Grafts.
Authors: Lu Lulu , Song Yongping , Ma Baogen , Zhu Xiongpeng , Wei Xudong , Chen Zhizhe 
Published: April 2020

Abstract
Background and objectives: Normal human bone marrow (BM), cord blood (CB) and mobilized peripheral blood (MPB) are the most commonly used sources for allogeneic hematopoietic stem cell transplantation (HSCT). The aim of this study was to detect the expression of CXCR4 on CD34+ cells and to assess the distribution of lymphocyte subsets in each type allograft.
Methods: CD34+ cells were separated from BM (n=30), CB (n=30) and MPB (n=30) by the CD34 MultiSort Kit immunomagnetic bead system. The expression of CXCR4 on CD34+cells was assayed by double color flow cytometry. The lymphocyte subsets in each type of allograft were detected by three-color flow cytometry. The groups of monoclonal antibodies were used as the following: CXCR4-PE/CD34−Pecy5, CD8−FITC/CD4−R-PE/CD3−TC, CD45RA-FITC/CD45RO-PE/CD4−Pecy5, CD45RA-FITC/CD45RO-PE/CD8−Pecy5, and CD3−FITC/CD16+56-PE. Isotype-specific antibodies were used as controls.
Results:
The expression of CXCR4 of cord blood and mobilized peripheral blood CD34+ cells was lower than that of bone marrow cells (BM 40.21%±6.72%, CB 20.93%±3.96%, MPB 20.93%±3.96%, P &lt;0.05). The difference between cord blood and mobilized peripheral blood was not significant (P&gt;0.05). The CD3+CD8low and CD3+CD4−CD8low subsets were higher in BM than that of CB and MPB (BM 8.61%±1.40%, CB 3.31%±0.88%, MPB 5.11%±0.76%,P&lt;0.01). The relative frequencies of the naïve CD45RA+ CD45RO− phenotype among CD4+ and CD8high T cells were highest in CB, and it was higher in MPB than in BM grafts (BM 28.09%±4.52%, 41.86 %±3.31%; CB83.83%±12.24%, 86.69%±6.12%; MPB 43.58%±4.54%, 57.64%±4.77%, P&lt;0.01). Naïve T cells (CD45RA+ CD45RO−) were mobilized preferentially compared to memory T cells (CD45RA− CD45RO+)(P &lt;0.01); The relative frequencies of NKT (CD3+CD16+56+) among lymphocytes were lower in CB than that in BM and MPB (CB 0.77±0.19, BM4.15±1.10, MPB 4.13±0.84, P&lt;0.01).
Conclusion: BM, CB and MPB allografts differ widely in cellular makeup of CD34+ cells and lymphocyte subsets, which are associated with the distinct characteristics after allogeneic HSCT from different allogeneic hematological sources.