bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transdifferentiation of Human Myeloma Cells and Their Dedifferentiation by IL-6.
Authors: Kawano Michio M. , Liu Shangqin , Otsuyama Kenichiro , Asaoku Hideki 
Published: April 2020

Abstract
Human primary myeloma cells as well as myeloma cell lines lack the expression of Pax-5 gene and CD19 expression on their surface, and myeloma cells are well known to be heterogeneous with regard to their morphology and surface phenotype. The expression of translineage markers, such as CD33, CD7, CD56, CD4 or CD86, has been reported to be observed in most cases of multiple myeloma, although their positive % in the bone marrow mononuclear cells are variable case by case. We found that the expression of CD33 in Liu01, a subclone fromU266 cells and vitamin D3-treated ILKM3 cells, correlated to the monocytoid morphology with convoluted nuclei and the increased expression of C/EBPα. The expression of CD56 in myeloma cells is also well known, but its meaning remains to be clarified. We examined whether CD56(+) myeloma cells also express either CD7, the NK lineage marker, or neuron-specific enolase (NSE), the neuronal cell marker. The expression of NSE was found in CD56(+) myeloma cell lines (NOP-2 and Liu01 cells) as well as primary myeloma cells both by RT-PCR and immunostaining. The expression of CD7 in Liu01 and forskolin-stimulated U266 cells was observed and compatible with large granules in the cytoplasm, and showed an increase expression of perforin mRNA. However, these findings were not observed in Pax-5(+) B cell lines. Interestingly, IL-6 could downregulate the expression of CD33, CD7 or CD56 in primary myeloma cells as well as Liu01 cells. Therefore, these data suggest that Pax-5(−) myeloma cells, but not Pax-5(+) B cells, are capable of multi-lineage differentiation, and IL-6 can induce their dedifferentiation. We propose here that the myeloma cells dropping out of their lineage (B cell linage) can become stem cell-like cells; we call them “stone cells”(stem cell-like cells dropping out of the lineage).
Expression of Human Amyloidogenic Immunoglobulin Light Chains in Mice Leads to Organ Toxicity.
Authors: Ward Jennifer E. , Brenner Daniel , Cui Lei , Liao Ronglih , Connors Lawreen H. , Hoo Pam Soo , Eberhard Jeremy , Christensen Thomas , O’Hara Carl , Skinner Martha , Seldin David C. 
Published: April 2020

Abstract
Recent evidence from the study of different amyloidogenic proteins challenges the dogma that tissue damage is solely the result of amyloid fibril deposition. To examine whether amyloidogenic human immunoglobulin light chains (LCs) may cause acute toxic effects prior to the development of fibrillar tissue deposits in vivo, we have generated amyloidogenic LC-expressing cell lines and transplanted them into mice. A full length lambda-6 light chain was cloned from cDNA prepared from bone marrow of a patient with aggressive multi-organ AL amyloidosis. The LC was subcloned into an expression vector with a CMV promoter and transfected into SP2/0 plasmacytoma cells. Stably transfected cells were injected into syngeneic Balb/c and RAG−/− mice. Four-six weeks later, echocardiograms were performed and the mice were euthanized and serum, urine, and tissues were collected. Mice injected with LC-producing cells, but not control untransfected SP2/0 cells, had detectable circulating human LC in their serum, and 6 of 9 RAG−/− mice excreted LC and albumin in the urine. These mice had evidence of bradycardia by echocardiography, with 4 of 12 mice having heart rates lower than 500 bpm while no controls had heart rates that low, and upregulation of markers of cell stress in the heart. In the kidney, there was evidence of amorphous protein deposits and early glomerulopathy by electron microscopy in two mice examined, but no fibril deposition. Thus, short-term expression of human amyloidogenic LC in mice in vivo produces alterations in heart and kidney function prior to the development of fibrillar deposits.
Natural Killer Cells with NKDG2D Cytotoxicity but NKp30-Dull Phenotype Predominate in Myelodysplastic Syndrome (MDS).
Authors: Bai Fanqi , Painter Jeffrey S. , Alan Cantor , JianXiang Zou , Wei Sheng , Djeu Julie Y. , List Alan F. , Epling-Burnette P.K. 
Published: April 2020

Abstract
Natural Killer (NK) function in patients with MDS as measured by non-MHC-restricted cytotoxicity and activation-dependent cell cytotoxicity (ADCC) are reduced in patients with MDS, however, the mechanisms of the functional impairment are not known. Tumor cytolysis occurs through orchestrated control by inhibitory NK receptors (NKRs) and activating NKRs, which control signaling events that lead to polarized movement of perforin-containing granules toward the NK-tumor contact area. We found that NK cells from 23 out of 35 patients with MDS (66%) displayed reduced lysis of K562 tumor cells compared to age-matched normal controls (p&lt;0.01). To better characterize this defect, we evaluated patient NK function against differential tumor targets including the MDS1 cell line established from an MDS patients. We found that MDS1 incited non-MHC-restricted lysis. Unactivated PBMCs, unactivated NK cells, NK cell lines (NK92 and NKL) but not purified unactivated T cells from normal donors killed MDS1 in 4-hr 51Cr-release assays. Normal NK cells and NK cell lines were also found to rapidly redistrubute perforin granules after exposure to MDS1suggesting that a perforin-dependent lytic pathway was activated. We then performed simultaneous cytolytic assays with K562, MDS1, and the 721.221 B cell lymphoma cell line as target cells. We found that NK cells from MDS patients had greater lytic activity against MDS1 (average 24% vs. average 8% at 50:1 Effector:Target ratio, respectively, p&lt;0.01) Antibody-blocking experiments demonstrated that the NKL cell line and PBMCs from 8 out of 10 MDS patients predominantly used the NKG2D activating receptor to kill MDS1. Consistent with this finding, we showed that MDS1 cells express the major human stress-inducible endogenous proteins MICA and MICB, which are NKG2D ligands. In contast, lysis by NK92 cells and normal PBMCs was not appreciably reduced by NKG2D blocking antibodies suggesting that other unidentified NKR(s) also mediate lysis. To identify the NKRs expressed in MDS patients, we performed immunophenotyping for both the activating NKRs and inhibitory NKRs compared to age-matched normal controls. We found that two activating receptors, NKp30 and CD244 (2B4), were significantly reduced on NK cells from all MDS patients regardless of their ability to lyse NK targets. Inhibitory NKR expression and function were normal. Interestingly, NKG2D expression correlated with reduced cytolytic function. Similar to studies on normal NK cells with low NKp30 and NKp46 (NCRdull) phenotypes, these results suggest that low NKp30 expression leads to predominant NKG2D utilization for tumor cell lysis, which is reduced in MDS patients with defective NK function. Our findings provide critical information about potential importance for immunosurviellance through NKG2D-NKG2D ligands.
CpG in the Immunotherapy of B-Cell Lymphoma.
Authors: Li Jiali , Song Wenru , Levy Ronald 
Published: April 2020

Abstract
CpG is an immune stimulator that can activate both innate and adaptive immunity. By binding to its receptor, TLR9, CpG can activate dendritic cells and B cells by inducing the secretion of cytokines, and increasing the expression of costimulatory molecules, such as CD80 and CD86. In this study, CpG1826, a B type CpG was given intratumorally to BALB/c mice bearing a A20 transplantable B cell lymphoma as a s.c local tumor with accompanying metastatic disease. In this model, CpG alone delayed the growth of tumor, however, the majority of the treated tumor-bearing mice still died. We therefore tested a new therapeutic strategy, a combination of cyclophosphomide (CTX), followed by subsequent intratumoral injection of CpG. The rationale for this strategy is that the danger signals generated by chemotherapy attract the migration of immature DC to the tumor site, and these DCs are activated by subsequent injection of CpG. Our animal studies showed that the combined maneuver can completely regress well-established tumor (Fig.1). The combination of CTX with CpG could cure well-established large size (&gt;1.2cm in diameter) tumors, and the mice remained tumor-free for 5-6 months after the initial treatment. Conversely, the majority of mice that received CTX or CpG alone died of lymphoma. We also showed that this combined effect was CpG injection site dependent, as CTX combined with intratumoral injection of CpG had a much better therapeutic effect (100% survival), as compared to injection of CpG into a distant normal site (20% survival). Our studies also indicated that this combined effect is CD8+T cell dependent, as systemic effects were lost when we repeated this experiment in a CD8+ T cell knockout mouse model. We also showed that CD4+ T-cells and B cells are not required in this system by depleting CD4+ T cells and by using a B cell knockout mouse model. Most importantly, we demonstrated the systemic effect of this combination strategy in mice bearing tumors at two different sites. As shown in Fig. 2, in combination with CTX, injection of one site with CpG controlled the growth of the second site tumor as opposed to a transient anti-tumor effect from chemotherapy or CpG alone. The systemic effect induced by injecting a single tumor site led to significant prolongation of overall survival (p&lt;0.01). In summary, this combined chemoimmunetherapeutic strategy has showed both local and systemic anti-tumor effects, which may be a potent anti-tumor strategy in the clinic.
Fig. 1 Fig. 1. Fig. 2 Fig. 2.
Synergistic Activity of the Proteasome Inhibitor PS-341 with Non-Myeloablative 153-Sm-EDTMP in a Syngeneic, Orthotopic Model of Multiple Myeloma.
Authors: Goel Apollina , Dispenzieri Angela , Geyer Susan M. , Greiner Suzanne , Russell Stephen J. 
Published: April 2020

Abstract
Multiple myeloma is a highly radiosensitive malignancy but, at the present time, radionuclide-based interventions have no proven place in disease management. Bone-seeking radionuclides such as 153-Sm-EDTMP and 166-Ho-DOTMP are promising agents for systemic delivery of ionizing radiation to sites of myeloma disease activity, but they are associated with significant myelosuppression at therapeutically effective doses and have therefore been used only in the context of myeloma stem cell transplantation protocols. We previously reported that the proteasome inhibitor PS-341 potently and selectively sensitizes myeloma cell lines and primary myeloma cells to the lethal effects of ionizing X-irradiation (Goel et al, Exp Hematol. 33, 784, 2005). To determine whether PS-341 is equally effective in sensitizing myeloma cells to ionizing beta-radiation and to extend our initial observations to an in vivo model, we combined PS-341 with the bone-seeking radionuclide 153-Sm-EDTMP. In vitro clonogenic assays were performed using a panel of myeloma cell lines and demonstrated synergistic killing following co-treatment with PS-341 and 153-Sm-EDTMP. Using the orthotopic, syngeneic 5TGM1 myeloma model, the median survivals of mice treated with saline, two doses of PS-341 (0.5 mg/kg), or a single non-myeloablative dose of 153-Sm-EDTMP (22.5 MBq) were 21, 22 and 28 days, respectively. In contrast, mice treated with combination therapy comprising two doses of PS-341 (0.5 mg/kg), one day prior to and one day following 153-Sm-EDTMP (22.5 MBq) showed a greatly prolonged median survival of 49 days. Correlative studies indicated that, compared to single-agent therapy, combination treatment with PS-341 and 153-Sm-EDTMP rapidly reduced the clonogenicity of bone-marrow resident 5TGM1 cells, slowed the elevation of serum myeloma-associated paraprotein levels, and was associated with longer term preservation of bone mineral density. The myelotoxicity of single agent and combination therapies was evaluated by comparing peripheral blood cell counts in each of the treatment groups, and by clonogenicity assays of hematopoietic progenitors isolated form normal mice receiving identical treatment regimens. Treatment with 153-Sm-EDTMP led to significant, but transient, myelosuppression which did not differ between animals treated with 153-Sm-EDTMP alone versus those treated with the combination of PS-341 plus 153-Sm-EDTMP. In summary, PS-341 is a potent in vivo radiosensitizer that greatly enhances the therapeutic potency, without increasing myelotoxicity, of skeletal targeted radiotherapy in the syngeneic, orthotopic 5TGM1 myeloma model. Based on these findings, we propose to conduct a phase I clinical trial at Mayo Clinic to evaluate the combination of PS-341 plus non-myeloablative skeletal targeted radiotherapy (using 153-Sm-EDTMP) in patients with advanced or treatment-refractory multiple myeloma.
Identification of Target Antigens Recognized in MGUS for Immunotherapeutic Approaches.
Authors: Blotta Simona , Bellucci Roberto , Tassone Pierfrancesco , Prabhala Rao , Neri Paola , Shammas Masood , Batchu Ramesh R. , Bae Jooeun E. , Li Cheng , Venuta Salvatore , Ritz Jerome , Anderson Kenneth C. , Munshi Nikhil C. 
Published: April 2020

Abstract
Monoclonal gammopathy of undetermined significance (MGUS) is a benign condition with 30% of patient developing symptomatic multiple myeloma (MM). The events leading to the transformation from MGUS to MM remain unclear, however several evidences have supported the idea that the immune system, in patients with MGUS, may play a role in controlling the progression to MM; and the identification of antigenic targets could open the way for future immunotherapeutic approaches to delay or prevent such progression. To identify target antigens that are recognized by the immune system in MGUS, we have screened a cDNA expression library, produced from purified CD138+ bone marrow myeloma cells, with high dilution (1:500) serum from 3 MGUS patients with stable disease for 1 to 4 years. Using this approach we were able to identify a panel of 11 gene products to which an antibody response was observed in serum from individuals with MGUS. Responses appeared to be directed against intracellular proteins involved in variety of cellular functions such as apoptosis (SON, Hip1), DNA and RNA binding (KIAA0530, GPATC4), signal transduction regulators (AKAP11), transcriptional co-repressors (IRF2BP2), developmental proteins (OFD1) and proteins involved in the ubiquitin-proteosome pathway (PSMC1). Interestingly PSMC1, the 26S subunit of proteosome, has recently emerged as a potential target for antimyeloma therapies and AKAP11 located on the 13q14.11, a locus frequently involved in the pathogenesis of aggressive MM. Importantly, two of these identified genes with antibody response have been previously identified by SEREX screenings; KIAA0530 has been reported to induce a strong antibody response in solid tumors such as head and neck, testicular, breast and colorectal cancers and SON, a protein involved in apoptosis with similarities to oncoproteins such as myc and mos, has been previously reported by us to induce an antibody response associated with a complete response after donor lymphocyte infusion in patients with MM. Our studies are now focused on testing these proteins in a series of MGUS serum at different stages of disease as well as in early stage myeloma. Although preliminary, these data open the possibility to identify target antigens that are important in the disease process of MGUS and may allow us to design future vaccines and immunotherapeutic approaches targeting these antigens in MGUS as well as in MM.
DKK-1 Is a Widely Expressed, Potent Tumor-Associated Antigen in Multiple Myeloma Recognized by Cytotoxic T Lymphocytes.
Authors: Yi Qing , Qian Jianfei , Yang Jing , Wang Siqing , Zhan Fenghuang , Shaughnessy John D. , Wezeman Michele 
Published: April 2020

Abstract
Idiotype proteins have been used for immunotherapy in multiple myeloma (MM); and the results thus far have been disappointing. Therefore, the identification and use of novel and more potent tumor-associated antigens are urgently needed to improve the efficacy of the treatment. Dickkopf-1 (DKK1), a secreted protein and Wnt signaling pathway inhibitor, is highly expressed by the tumor cells of most myeloma patients and may be responsible for suppressed osteoblast formation. Based on these data and the fact that DKK1 is not expressed in normal tissues except placenta, prostate, and mesenchymal stem cells, we hypothesized that DKK1 is a novel tumor-associated antigen in MM. In this study we examined the capacity of DKK1 to trigger the induction of antimyeloma cytotoxic T lymphocyte (CTL) responses. Using DKK-1 peptide-pulsed dendritic cells (DCs), we successfully generated DKK-1 peptide (DKK120-29 and DKK166-74V)-specific CTL lines and clones from HLA-A2+ MM patients and healthy individuals. In our experiments, mature DCs obtained from cultures of blood monocytes were pulsed with HLA-A2-restricted DKK1 peptides. Autologous T cells were then stimulated weekly with these DCs, and cytotoxicity was examined against DKK1 peptide-pulsed T2 cells, myeloma cell lines, and primary myeloma cells isolated from patients. After 4 cycles of in vitro stimulation and subsequent T cell cloning and expansion, specific CD8+ CTL lines and clones were obtained. These CTLs not only had cytolytic activity against DKK1 peptide-pulsed T2 cells, but also significantly lysed HLA-A2+ MM cell line U266 and primary tumor cells in vitro. No killing was observed against HLA-A2+ normal lymphocytes including B cells or HLA-A2− myeloma cell lines or primary myeloma cells from patients. The CTL response could be inhibited by anti-HLA-A2 antibody indicating that the response was indeed restricted by HLA-A2 molecules. IFN-γ, but not IL-4, was secreted by the T cells indicating that the effector cells were type-1 CD8+ T cells. Further functional studies are underway. Thus, these results demonstrate that DKK1-specific CTLs are able to lyse myeloma tumor cells including primary myeloma cells from patients, and identify DKK1 as a potentially important antigenic target for antimyeloma immunotherapeutic strategies
Anti-CD20 Monoclonal Antibody (mAb) with Enhanced Affinity for CD16 Activates NK Cells at Lower Concentrations and More Effectively Than Rituximab ®.
Authors: Weiner George J. , Bowles Julie A. , Link Brian K. , Campbell Mary A. , Wooldridge James E. , Breitmeyer James B. 
Published: April 2020

Abstract
Growing evidence indicates the affinity of mAb for CD16 (FcgRIII) plays a central role in the ability of the mAb to mediate anti-tumor activity. Both polymorphisms in CD16 and structure of mAb Fc can impact on the affinity between CD16 and mAb. Little is known about how affinity between mAb and CD16 impacts on the ability of mAb to activate NK cells. We evaluated how CD16 polymorphisms, and mAb with modified affinity for CD16, impact on NK cell phenotype and cytokine production. MAb consisted of R, anti-CD20 with enhanced affinity for CD20 (AME-133), and AME-133 with Fc engineered to have enhanced affinity for both CD20 and CD16 (Fc-eng-133). Peripheral blood mononuclear cells were obtained from normal subjects that were homozygous for high affinity CD16 (VV at position 158), homozygous low affinity (FF at 158), and heterozygotes and were mixed with mAb and Raji lymphoma cells at an effector:target ratio of 1:1. Higher concentrations of mAb were needed to induce CD16 modulation, CD54 upregulation, and IFNg production on NK cells from subjects with the lower affinity CD16 polymorphism. The dose of mAb needed to induce NK activation was lower with Fc-eng-133 irrespective of CD16 polymorphism.
Measure of activation - % of NK cells CD54 bright: EC50 ng/ml (mean moAb concentration +/− SEM): N=4 per polymorphism
R AME-133 Fc-eng-133 p&lt;0.01 - R vs AME-133; R vs Fc-eng-133; AME-133 vs Fc-eng-133 for each polymorphism VV 6.2 +/− 1.0 3.9 +/− 0.5 1.0 +/− 0.3 VF 11.6 +/− 1.7 5.7 +/− 0.6 1.4 +/− 0.4 FF 29.9 +/− 10.6 11.5 +/− 2.5 2.8 +/− 0.3 p value VV vs FF &lt;0.05 &lt;0.05 &lt;0.05
At saturating mAb concentrations, peak NK activation was greater for Fc engineered AME-133 when compared to R mAb for all samples studied
Ratio of Fc-eng-133 to R % NK cells CD54 bright 1.35 +/− 0.07 IFNg production 2.64 +/− 0.43
These data indicate tumor cells coated with mAb with enhanced affinity for CD16 are more effective at activating NK cells at both low and saturating mAb concentrations irrespective of CD16 polymorphism, and provide further evidence for the clinical development of such mAb with the goal of improving clinical response to mAb.
Correlation of Suppression of FDG PET Uptake with Serum Free Light Chain Levels - Both FDG PET-CT and Serum Clonal Free Light Chain Response Precede and Predict the Likelihood of Subsequent Complete Remission in Newly Diagnosed Multiple Myeloma.
Authors: Walker Ronald , Rasmussen Erik , Cavallo Federica , Jones-Jackson Laurie , Anaissie Elias , Alpe Terri , Epstein Joshua , Van Rhee Frits , Zangari Maurizio , Tricot Guido , Shaughnessy John , Barlogie Bart 
Published: April 2020

Abstract
18F-labeled Fluorodeoxyglucose Positron Emission Tomography - computed tomography (PET-CT) imaging results from newly diagnosed patients with multiple myeloma (MM) were correlated with serum free light chain (FLC) results from Total Therapy 3 (TT3). Patients had baseline PET-CT scans and serum kappa and lamda FLC analysis at diagnosis and at 2, 14 and 30 days post first cycle of VDTPACE and at pre-transplant 1 of two planned autologous stem cell transplants (TX1), correlating PET-CT response, clonal and nonclonal FCL response, and likelihood of subsequent complete clinical response (CR). Patients were selected with serum clonal FLC levels above the normal range that had been enrolled for sufficient time for a complete response (CR) to be recorded. Correlations between the serum clonal and nonclonal FLC levels and PET-CT response defined by both reduction in number of PET-defined focal lesions (PET-FL) (5 mm or larger areas of circumscribed uptake visually definable above background activity) and reduction in the maximum standardized uptake value normalized to the patients’ lean body mass (max SUV) were performed.
Results: Reduction in FLC and PET-FL and max SUV are correlated, as was normalization of clonal FLC levels with likelihood of subsequent CR (Fig 1). The Spearman’s correlation coefficient between percent reduction in clonal FLC and percent reduction in max SUV was 0.04 (n=49, p=0.79) at 2 days post-VDTPACE, 0.32 (n=48, p=0.028) at 30 days post-VDTPACE, and 0.52 (n=33, p=0.002) prior to TX1. The greatest reduction in max SUV occurred by 2 days post-VDTPACE while the maximum reduction in clonal FLC occurred at 30 days post-VDTPACE, indicating that the PET response lead and predicted the FLC response in near “real time.” While normalization of the clonal FLC levels at 30 days was strongly predictive of subsequent CR (p&lt;0.001, n=125), normalization of the non-clonal FLC levels was not (p=0.41, n=124).
Conclusion: We conclude that normalization of serum clonal FLC correlates both with improvement in FDG PET-CT scanning in both max SUV and number of PET-defined focal lesions and with likelihood of subsequent CR, though it “lags” behind FDG PET response by nearly a month, with FDG PET following the clinical course in near real time.
Figure 1: Kaplan-Meier Analysis Correlating Normalization of Clonal FLC Levels by 30 Days Post Initiation of Treatment with Likelihood of Subsequent Clinical Remission (p=0.0001, n=125). Figure 1:. Kaplan-Meier Analysis Correlating Normalization of Clonal FLC Levels by 30 Days Post Initiation of Treatment with Likelihood of Subsequent Clinical Remission (p=0.0001, n=125).