bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of Activation Marker, CMRF-44, on Blood CD11c+ Dendritic Cells Is Closely Associated with the Severity of Acute GVHD after Allogeneic Haematopoietic Stem Cell Transplant.
Authors: Lau Jenny , Sartor Mary , Vuckovic Slavica , Munster David , Hart Derek N. , Bradstock Kenneth F. 
Published: April 2020

Abstract
AIM: Dendritic cells (DC) are considered central to the development of graft versus host disease (GVHD) following allogeneic haematopoietic stem cell transplantation (alloHSCT). Both donor and host DC are thought to initiate allogeneic immune responses by presenting host antigens to donor T lymphocytes. We studied the potential of CMRF-44, a novel monoclonal antibody identifying activated circulating blood DC, as a predictive marker of acute GVHD.
METHODS: In a prospective study, peripheral blood was taken from 40 patients twice weekly up to 100 days post-alloHSCT. Circulating myeloid (CD11chi) and plasmacytoid (CD123hi) DCs were enumerated and the expression of CMRF-44 was assessed on CD11chi DC by four colour flow cytometry. Multivariate analyses were performed using a non-parametric Mann-Whitney U-test and Receiver Operating Characteristic (ROC) curves.
RESULTS: Following alloHSCT, the severity of acute GVHD correlated with the number of total DC in the blood (p=0.035). Furthermore, low myeloid and plasmacytoid DC numbers were significantly associated with grade 2-4 acute GVHD (p=0.046 and 0.017 respectively). In 40 alloHSCT patients, 27 developed acute GVHD. CMRF-44 was expressed on CD11c+ DC in all cases prior to the onset of acute GVHD. Of the 13 patients without GVHD, 8 had no circulating CMRF-44+ CD11c+ DC. CMRF-44 expression was independent of the reconstitution of myeloid DC (p=0.73). Patients who had CMRF-44+ CD11c+ DC in more than 20% of their post-transplant monitoring samples were more likely to develop acute GVHD (p=0.001, OR=37.1). In addition, patients with more severe grade 2–4 GVHD had significantly higher percentages of CMRF-44+ CD11c+ DCs (p=0.001). CMRF-44 expression at greater than or equal to 12% of CD11chi DCs had a sensitivity of 87.5 for prediction of grade 2–4 acute GVHD, and a specificity of 91.7.
CONCLUSION: CMRF-44 expression on blood CD11c+ DC is highly associated with the onset of acute GVHD. These results suggest that CMFR-44 may be used as a predictive tool to identify patients at risk of severe GVHD and to direct therapy. Furthermore, it reinforces the potential application of suitably engineered CMRF-44 antibodies for the prevention or treatment of graft versus host disease.
Risk and Clinical Implications of Transformation of Follicular Lymphoma (FL) to Diffuse Large Cell B-Cell Lymphoma (tFL).
Authors: Montoto Silvia , Davies Andrew J. , Matthews Janet , Calaminici Maria , Norton Andrew J. , Amess John , Vinnicombe Sarah , Rohatiner Ama Z.S. , Lister Andrew 
Published: April 2020

Abstract
Between 1971 and 2000, 330 consecutive adults (176M/154F) with newly diagnosed FL were referred to St Bartholomew’s Hospital. The median age was 55 (range 23–90) years; 50 Stage I, 47 Stage II, 56 Stage III, 177 Stage IV. The FLIPI score was ‘low’: 38%, ‘intermediate’: 27%, ‘high’: 35%, with 111 missing observations (LDH or number of nodal sites). 100 patients are still alive, 230 having died between 6 months and 30 years, the causes of death being: disease 106, treatment-related complications 37, other malignancy 30, cardiac disease 20, ‘other’ 18, unknown 19. With expectant management if appropriate and intervention with irradiation, chemotherapy or immunotherapy at times of progression, the response rate was 84% at 1st treatment, declining to 43% at fifth, with duration of remission declining correspondingly. With a strict policy of documenting histology at each progression, a repeat biopsy rate of 70% was achieved. tFL has occurred in 89/330 cases between diagnosis and 16 years, in 25/330 (8%) whilst being managed expectantly or during first therapy and at a relatively constant rate at 2nd, 3rd, 4th and 5th progression, (17%, 16%, 15%, 14% respectively). To date, no patients have transformed after 16 years, with 41 patients ‘at risk’ between 16 and 35 years (Figure 1), 18 in continuous first remission and 23 having had a recurrence of FL. On univariate analysis, risk of transformation is higher with high FLIPI score (p= .009) and advanced stage (p= .02) at presentation. Survival of patients is shorter in whom transformation occurs (p= .0003). Median survival from transformation was 1.3 years, and correlated with ‘stage’ (p= .008) and LDH (p= .03) at the time of transformation, but not with time from diagnosis to transformation. These observations highlight the significance of transformation of FL to t-FL, demonstrating its major clinical importance. The actuarial prediction is that transformation will occur in at least 50% of patients overall with a very poor prognosis for the majority. The ‘time to transformation curve’ supports the hypothesis that there may be two distinct populations of patients who develop FL, one in which transformation is programmed to occur and one in which it is not.
Figure 1. Time to trasnformation. Figure 1. Time to trasnformation.
Signatures of Response to the Proteasome Inhibitor Bortezomib in Diffuse Large B-Cell Lymphoma.
Authors: Feuerhake Friedrich , Monti Stefano , Blank Jonathan , Wu Erxi , Chen Wen , Golub Todd R. , Shipp Margaret A. 
Published: April 2020

Abstract
The proteasome inhibitor, bortezomib (VELCADE®, formerly PS341), has significant anti-tumor activity in several lymphoid malignancies. Reported targets of this broad-based inhibitor include the NF K B pathway (I K B A). Recently defined subtypes of large B-cell lymphoma (LBCL) exhibit constitutive activation of NF K B, prompting us to analyze the efficacy of bortezomib in a panel of 10 DLBCL cell lines. Six of the diffuse LBCL cell lines were sensitive to bortezomib treatment at doses below 10 nM (range IC50 = 2.9 to 6.9 nM) whereas 4 cell lines were resistant at 10 nM (IC50 = 14.8 to 70.2 nM). Baseline proteasomal function, as defined by cleavage of the 20S proteasome-specific fluorogenic peptide LLVY-AMC, was similar in sensitive and resistant DLBCLs; however, the IC50 for bortezomib proteasomal inhibition was somewhat lower in sensitive vs. resistant lines (sens. vs res., p = .04, one-sided t test). Baseline NF K B activity varied widely in the DLBCL cell lines and did not differ in cell lines that were sensitive vs. resistant to bortezomib. Ten nM bortezomib did not inhibit NF K B activity in resistant DLBCL cell lines whereas the same dose reduced NF K B activity in sensitive DLBCL cell lines (sens. vs. res., p &lt; .005, rank test [Mann-Whitney]). However, 5 of 6 sensitive DLBCL cell lines had very low baseline NF K B levels (&lt; 0.5 relative absorbance units) suggesting that NF K B inhibition was not a major factor in bortezomib response and prompting further analysis of additional bortezomib targets. Three sensitive and 1 resistant DLBCL cell line were selected for detailed analyses of transcriptional profiles following bortezomib treatment. We developed an algorithm for identifying genes that were significantly up- or down-regulated in the bortezomib-sensitive cell lines but unchanged in the resistant line. In addition, we utilized gene set enrichment analysis (GSEA) and gene ontogeny (GO) termed enrichment to interpret the molecular signatures of response. Genes down-regulated in response to bortezomib included critical B-cell transcription factors, components of the B-cell receptor signaling cascade and genes regulating mitosis and cell cycle control; up-regulated genes included heat shock proteins (HSP) and multiple proteasomal components. Consistent with the functional data, down-regulation of NF K B target genes was not a common feature in all bortezomib-sensitive cell lines. In contrast, target genes of the c-MYC transcription factor were significantly down-regulated and c-MYC activity was decreased in sensitive (but not resistant) DLBCL cell lines following bortezomib treatment (sens. vs. res., p &lt; .005, rank test). Taken together, the results provide insights into likely mechanisms of action of bortezomib in DLBCL, highlighting c-MYC as a potentially important target and identifying HSP as a complementary target to overcome bortezomib resistance.
Targeting β2-Microglobulin for Induction of Tumor Apoptosis in Human Hematological Malignancies.
Authors: Yi Qing , Yang Jing , Qian Jianfei , Wang Siqing , Wezeman Michele , Kwak Larry W. 
Published: April 2020

Abstract
We discovered that monoclonal antibodies (mAbs) specific to human β2-microglobulin (β2M) induce programmed death of myeloma and other hematological tumor cells. The mAbs exhibited potent in vitro tumoricidal activity on all 14 β2M/HLA-ABC-bearing myeloma, lymphoma, and leukemia cell lines and primary myeloma cells isolated from eight patients. Cell death occurred rapidly, without the need for exogenous immunological effector mechanisms. Although the expression of β2M on normal hematopoietic cells is a potential safety concern, the mAbs seem to be selective to tumor-transformed cells and did not induce apoptosis of normal cells, including T and B lymphocytes and CD34+ bone marrow stem cells. Furthermore, the mAbs were able to selectively kill myeloma cells without damaging normal stromal cells in their cocultures. After binding to cell surface, the mAbs mediated internalization and down-modulation of surface β2M and HLA-ABC molecules. The mAbs induced cell death via inhibiting PI3K/Akt and ERK, activating JNK, upregulating Bad and Bax protein expression, inducing phosphorylation of Bcl-2, and decreasing phosphorylation of Bad, all of which compromised mitochondrial integrity, leading to cytochrome c release into cytosol and activation of the caspase-9-dependent cascade. Inhibitors to pan-caspases or caspase-9, but not to caspase-8, or to JNK, and knockdown of surface β2M and HLA-ABC expression by β2M-specific siRNA, but not control siRNA, abrogated apoptosis of tumor cells induced by the mAbs. Furthermore, anti-β2M mAbs were also active and therapeutic in vivo; After administration subcutaneous or intraperitoneal, the mAbs substantially reduced tumor burdens and retarded tumor growth in SCID mice subcutaneously implanted with human myeloma cell lines ARP-1 or MM.1S. Therefore, such mAbs offer the potential for a novel therapeutic approach to hematological malignancies and possibly to other cancers that express surface β2M. This study also suggests that differential expression of β2M/MHC class I molecules by normal and cancer cells might be a crucial point in the sensitivity and selectivity of anti-β2M mAb-induced apoptosis.
Targeting p62ZIP in Marrow Stromal Cells Is Highly Effective at Inhibiting Myeloma Cell Growth and Osteoclast Formation.
Authors: Kurihara Noriyoshi , Honjo Tadashi , Windle Jolene J. , Shin J. , Roodman G. D. 
Published: April 2020

Abstract
Marrow stromal cells play a critical role in osteolytic bone destruction in multiple myeloma, and promote tumor growth. In particular, adhesive interactions between myeloma cells and marrow stromal cells increase RANK ligand (RANKL), a potent inducer of osteoclast formation, IL-6 and TNF-α production by marrow stromal cells. IL-6 enhances the growth and prevents apoptosis of myeloma cells, and TNF increases production of RANKL and IL-6. Recently, a new member of NF-κB signaling pathway, p62ZIP, has been identified. p62ZIP plays a critical role in NF-κB activation induced by TNF-α and RANKL and is involved in multiple signaling pathways that result in enhanced IL-6 production, tumor cell survival and bone destruction. It is our hypothesis that inhibiting p62ZIP expression will profoundly diminish myeloma growth by blocking the effects of IL-6 produced in the tumor-bone microenvironment in response to TNF-α. Therefore, we used p62ZIP−/− mice to determine the effects of deleting p62ZIP in stromal cells on the growth of myeloma cells. Marrow cells from p62ZIP −/− or wild type mice were used to establish long-term Dexter-type marrow cultures to isolate marrow stromal cells. Marrow stromal cells from p62ZIP −/− or wild type mice were cocultured for 48 h with a GFP-labeled human MM.1S myeloma cell line (MM.1S) and murine and human RANKL, IL-6 and TNF-α levels were determined in conditioned media from these cocultures using commercial ELISA assays. Cocultures of MM.1S with wild type marrow stromal cells resulted in much greater upregulation of murine IL-6 than p62−/− stromal cell coculture (IL-6 in p62−/− stromal cell cultures; 114+70 vs. WT 1900+9 pg/ml). In addition, deleting p62ZIP in stromal cells markedly decreased the growth of tumor cells. Coculture with wild type stromal cells induce 1.4-fold greater increase in MM.1S cell growth at 72 h compared to p62−/− stromal cells. Further, addition of neutralizing antibodies to TNF-α and IL-6 to the cocultures of MM.1S cells with WT stromal cells similarly affected the growth of the MM.1S. Since TNF-alpha can increase the expression of adhesion molecules on stromal cells and tumor cells, we measured expression levels of ICAM-1 and VCAM-1 by Western blot. VCAM-1 and ICAM-1 levels on p62ZIP−/− bone marrow stromal cells were not changed compared to WT stromal cells. We then determined the capacity of p62 −/− cells to support OCL formation by normal spleen CFU-blast. OCL formation was decreased about 50 % in cocultures containing p62−/− stromal cells treated with PTH-rp, IL-6 and TNF-α compare with WT stromal cell culture. These results show that p62 plays an important role in myeloma cell growth and OCL formation induced by cytokines that are upregulated in the marrow microenvironment in patients with myeloma.
GRN163L, a Novel and Potent Telomerase Inhibitor, Inhibits Myeloma Cell Growth In Vitro and In Vivo.
Authors: Shammas Masood A. , Koley Hemanta , Tassone Pierfrancesco , Neri Paola , Protopopov Alexei , Gryaznov Sergei , Chin Allison , Bertheau Robert C. , Mitsiades Constantine , Anderson Kenneth C. , Munshi Nikhil C. 
Published: April 2020

Abstract
Telomerase activity is either low or completely absent in most normal somatic cells; while it is elevated in most cancer cells providing unlimited proliferative potential by preventing telomere shortening. The inhibitors of telomerase, therefore, induce telomere shortening leading to apoptotic cell death in tumor cells while having little or no effect on normal diploid cells. We have evaluated the in vitro and in vivo efficacy of thio-phosphoramidate oligonucleotide specifically targeting the RNA component of telomerase (GRN163L) with demonstrated nuclear uptake by &gt;99% cells without the transfection enhancer. Delivery of GRN163L (1 μM) to MM cells (INA6 and ARP) was specifically associated with complete loss of telomerase activity as early as 6 hrs following exposure and was accompanied by a reduction in myeloma cell growth and survival. Treatment of INA6 cells with GRN163L for three weeks induced 96±4% and 100% cell death at 0.5 and 1 μM concentrations, respectively. ARP cells, which express higher levels of telomerase activity and have longer telomeres, showed 67±4% cell death at 5 weeks with 0.5 μM inhibitor and 82±3% and 100% cell death at 4 and 5 weeks, respectively, with 2 μM GRN163L. The apoptotic cell death was confirmed in 51% INA6 cells at two weeks and in &gt;80% ARP cells at four weeks. Apoptosis was associated with reduction in mean Telomere Fluorescence Intensity (TFI) on interphase chromosomes from 87.1±6.2 in control oligo treated INA6 cells to 36.2±2 (2.4 fold) in GRN163L treated cells. Moreover, GRN163L treatment was also associated with a similar reduction in number of chromosomes with detectable telomeres, indicating development of telomere-free ends. We have confirmed in vivo efficacy of GRN163L in a SCID-hu murine model of multiple myeloma. Following growth of GFP-transduced myeloma cells in the fetal bone chip introduced into the mice, GRN163L was injected on alternate days. In two independent experiments significant reduction in tumor cell growth, as measured by reduction in human myeloma related protein, and better survival than mice injected with control oligo was observed. We have now evaluated efficacy of combination of GRN163L with other novel agents. We have observed synergistic activity with Hsp90 inhibitor 17AAG on myeloma cell death. Addition of 17AAG (0.05 μM) to myeloma cells pre-treated with GRN163L (1 μM) for one week led to complete growth arrest within four days compared to continued growth of cells not pre-treated with GRN-163. These data provide the preclinical rationale for clinical evaluation of GRN163L in myeloma and in combination with Hsp90 inhibitor.
Protein Palmitoylation in Platelet Function: Role in G Protein-Mediated Platelet Activation and Platelet Recruitment into Thrombi.
Authors: Flaumenhaft Robert , Dilks James R. , Sim Derek S. 
Published: April 2020

Abstract
Protein palmitoylation represents the covalent linkage of a 16-carbon saturated fatty acid to a protein. This reversible post-translational modification directs protein-protein interactions as well as protein association with membranes and lipid rafts. Protein palmitoylation participates in ligand-induced signal transduction in several nucleated cells. Its role in platelet activation, however, has not previously been evaluated. We have found that platelets contain the palmitoyl transfer proteins GODZ and HIP14 as well as the palmitoyltransferase, acyl-protein thioesterase 1 (APT1). Thus, platelets possess the basic machinery for regulated palmitoylation. Studies using [3H]-labeled platelets identified several platelet proteins that were palmitoylated following exposure to the protease-activated receptor 1 (PAR-1) ligand, SFLLRN. To determine whether protein palmitoylation functions in activation-induced platelet functions, we infused recombinant APT1 into permeabilized platelets prior to activation with SFLLRN. Infusion of APT1 inhibited platelet protein palmitoylation and completely blocked platelet α-granule secretion induced by SFLLRN. Similarly, the protein palmitoylation inhibitor cerulenin blocked SFLLRN-induced platelet protein palmitoylation, α-granule secretion, and platelet aggregation in intact platelets. To assess the mechanism by which protein palmitoylation affects platelet function, we evaluated the effect of inhibitors of protein palmitoylation on G protein activity. Gαq is essential to PAR-1-mediated platelet activation and is palmitoylated in an activation-dependent manner in nucleated cells. Immunoprecipitation of Gαq from [3H]-labeled platelets showed that it is palmitoylated following activation of platelets with SFLLRN. Both APT1 and cerulenin inhibited SFLLRN-induced palmitoylation of Gαq. In addition, APT1 and cerulenin inhibited SFLLRN-induced GTPase activity as detected using [γ-32P]GTP-labeled platelet lysates. These results show that palmitoylation of Gαq participates in PAR-1-mediated signal transduction. We next used intravital microscopy to determine if protein palmitoylation functions in thrombus formation in vivo. For these experiments, platelets from a donor mouse were incubated with cerulenin and labeled with calcein-AM (green) or incubated with vehicle alone and labeled with calcein-AM red-orange (red). Equal numbers of green and red labeled platelets were then infused into a recipient mouse. The accumulation of cerulenin- and vehicle-treated platelets into thrombi following laser-induced injury of the mouse cremaster muscle was quantified using high speed, digital videomicroscopy. Incubation of platelets with cerulenin resulted in an approximately 50% reduction in their ability to accumulate into thrombi. These studies show that platelet protein palmitoylation is required for thrombus formation as well as for normal platelet function.
Outcomes of Older Patients with Myeloid Leukemias Treated with Myeloablative Intravenous Busulfan-Based Conditioning Regimens and Allogeneic Blood or Marrow Transplantation.
Authors: de Lima Marcos , Couriel Daniel , Ghosh Shubhra , Jones Roy B. , Shpall Elizabeth J. , Giralt Sergio , Hosing Chitra , Kebriaei Partow , Champlin Richard , Andersson Borje S. 
Published: April 2020

Abstract
Older age (e.g. &gt; 55–60 yrs) is a commonly cited indication for reduced-intensity conditioning regimens (RIC) in the treatment of myeloid leukemias. These diseases, however, may benefit from the highest possible dose intensity that can be safely delivered. Here we analyze our experience treating patients (pts) older than 55 yrs. with intravenous (IV) busulfan (Bu)-based preparative regimens.
Methods: Fifty-six pts older than age 55 yrs. with MDS (n=13), AML (n=33), or CML/myeloproliferative disorder (n=10) were transplanted from 6/1997 to 12/2004 on two consecutive protocols. Conditioning were IV fludarabine (Flu) 40 mg/m2 and IV Bu 130 mg/m2 over 3 hr (days -6 till -3) given once daily (BuFlu; n=36), and IV Bu 0.8 mg/kg over 2 hr every 6 hr for 16 doses (days -7 till -4), followed by Cyclophosphamide (Cy) 60 mg/kg on days -3 and -2 (IV BuCy2; n=20). Pts with an unrelated (UD) or mismatched donor received ATG. Graft-versus-host disease (GVHD) prophylaxis was based on tacrolimus and mini-methotrexate.
Results: Median age was 58.4 yrs (55–66.8); 14 pts were older than 60 yrs. Most pts were not in complete remission (CR) at BMT (n=38, 68%, with similar distribution in pts older and younger than age 60). Donors were HLA-compatible siblings (SIBS) in 59% (n=33) and UD in 41% (n=23). UD were used in 5 pts &gt;59 yrs (36%) and in 18 &lt; 60 yrs (43%). Stem cell source was bone marrow (n=23) or peripheral blood (n=33). All but one pt had documentation of donor cell engraftment. Median follow-up of surviving pts (June 2005) was 15 mos (range, 4–49; n=23). Grade II-IV acute (a) and chronic (c) GVHD rates were as follows (pts &lt;60 vs &gt;59 yrs): 38% vs 64% and 45% vs 54%, respectively. 100-day treatment-related mortality (TRM) was 11% (due to graft rejection (n=1), toxicity (n=2); pulmonary hemorrhage with fungal pneumonia (n=1) and aGVHD (n=2). All early 100-day TRM pts were &lt; 60 yrs. Overall, 17 pts relapsed and 33 pts have died. One-year TRM: 29%, n=16 (due to GVHD, n=7, infections, n=4; rejection, n=1; unknown causes, n=1; toxicity, n=2, bleeding, n=1). Only 2 pts &gt; 59 yrs are alive, while the others died of cGVHD (n=5) and disease relapse (n=7).
Conclusion: In this older population of predominantly relapsed pts with myeloid leukemias, major cause of treatment failure was not the preparative regimen employed, but GVHD and disease relapse. An association between intensity of the regimen, age and GVHD cannot be ruled out in this analysis. These results would suggest that age per se should not be used as the main reason for selecting a reduced intensity regimen in patients up to at least age 60 with myeloid leukemias.
Figure Figure