bluuebunny/crossref_metadata_2025_split huggingface dataset:
Depletion of the Shwachman-Diamond Syndrome Gene Product, SBDS, Leads to Growth Inhibition and Increased Expression of OPG and VEGFA.
Authors: Nihrane Abdallah , Sezgin Gulay , Dsilva Sehba , Ellis Steven , Liu Johnson M. 
Published: April 2020

Abstract
Shwachman-Diamond syndrome (SDS) is an autosomal recessive disorder characterized by bone marrow failure and leukemia predisposition, pancreatic exocrine dysfunction, and skeletal abnormalities, manifesting as skeletal dysplasia and osteoporosis. Mutations in SBDS have been shown to cause SDS, but the function of the SBDS gene product is unclear. Accelerated angiogenesis has recently been described in bone marrow cells from SDS patients. To clarify the unknown function of SBDS, we performed experiments analyzing the cellular effects of depleting SBDS by constitutive or inducible RNA interference. The growth of HeLa and NIH3T3 cells constitutively depleted of SBDS was markedly hindered when compared to cells stably transfected with siRNA against an irrelevant control gene. However, the growth abnormality was related to a significantly increased propensity to apoptosis in NIH3T3 more than in HeLa cells. We believe these differences are related to inactivation of p53 in HPV-18 positive HeLa cells, suggesting that depletion of SBDS may partially sensitize cells to p53-mediated apoptosis. As in constitutive knockdown, HeLa cells induced by doxycycline to express siRNA against SBDS exhibited growth inhibition, which was associated with modestly increased levels of apoptosis, suggesting a partial contribution of this process in inducible knockdown cells. By microarray analysis of knockdown cells, we found marked differences in expression of genes in multiple pathways, and we chose to examine a selected subset more closely using quantitative PCR arrays. In constitutive and inducible SBDS-depleted HeLa cell clones, we found 3- to 6-fold elevated mRNA levels of osteoprotegerin (OPG or TNFRSF11B) and vascular endothelial growth factor-A (VEGF-A). We confirmed significant overexpression of both secreted proteins by ELISA from supernatants of SBDS-depleted HeLa cells. Osteoprotegerin and VEGF-A are known to have diverse effects on osteoclast differentiation, angiogenesis, and monocyte/macrophage migration, all processes that may be aberrant in SDS, and we propose that overexpression of these factors may contribute to its pathology.
Inhibition of BCR Signaling Using the SYK Inhibitor R406 Antagonizes Migratory and Pro-Survival Responses in CLL Cells.
Authors: Quiroga Maite P. , Kurtova Antonina , Wierda William , Keating Michael , Burger Jan A. 
Published: April 2020

Abstract
Disrupting B cell receptor- (BCR-) induced signaling by inhibiting the spleen tyrosine kinase (SYK) represents a novel therapeutic approach in CLL. R406 is an ATP-competitive SYK inhibitor, and its prodrug, R788 (Fostamatinib disodium), displays clinical activity in patients with B cell lymphoma and CLL (Friedberg J. et al. Ann Oncol. 2008 Jun; 19 Suppl 4). In normal lymphocytes BCR engagement and SYK activation modulate cell adhesion and migration, suggesting that this pathway affects chemokine receptor and adhesion molecule function/signaling. The effects of SYK on CLL cell migration, activation and survival are largely unknown. Therefore, we examined the effects of BCR stimulation and its inhibition with R406 on CLL cell chemotaxis, expression of adhesion- and costimulatory- molecules, and survival. Activation of BCR with polyclonal goat F(ab’)2 fragments to human IgM significantly increased chemotaxis towards CXCL12 to 135.4% ±5.2% of controls and towards CXCL13 to 170.5% ± 31.5% of controls (Mean ± SEM, n=15). Engagement of BCR also significantly increased expression of CD40, CD44, CD54, and CD62L and decreased the expression of CXCR4. Pre-treatment with R406, which was kindly provided by Rigel Pharmaceuticals, Inc., abrogated the increased chemotaxis, adhesion- and co-stimulatory molecule expression, and pro-survival signals seen in response to BCR engagement. As displayed in Fig. 1, CLL cell viability at 24 hours was significantly increased to 60±5.2% with anti-IgM, compared to controls (41.8±6.7%) or CLL cells treated with anti-IgM plus R406 5 μM (39±6.6%, mean± SEM, n=9). Moreover, we found that R406 induced apoptosis in CLL cells in suspension culture and co-cultures with nurselike cells (NLC) and marrow stromal cells (MSC). When compared to CLL cells cultured with NLC (100%), the mean relative viability of CLL cells at 48h was reduced to 71.2±6.4% with 5 μM R406 and to 75.4±5.6% with 2.5 μM R406, whereas the same CLL samples cultured without NLC displayed a mean relative viability of 87±2.9% in controls, 58.9±6.8% with 5 μM R406, and 59.9±7.3%% with 2.5 μM R406 (mean±SEM, n=10). Similar levels of apoptosis were observed when CLL cells were incubated with R406 in MSC co-cultures.
Collectively, this study demonstrates that BCR engagement increases CLL cell migration, adhesion- and co-stimulatory-molecule expression, and survival. R406 effectively antagonizes these responses and induces CLL cells apoptosis, providing an explanation for the clinical activity of R406 in CLL patients.
Figure 1 Figure 1.
Dual Action of CD160 on CLL Cells: Giving Life to Prepare for Death!.
Authors: Agrawal Sami r G , Liu Feng-Ting , Giustiniani Jerome , Jia Li , Gribben John G. , Bensussan Armand 
Published: April 2020

Abstract
CD160 is a cell surface antigen expressed on normal human NK cells, as well as a subset of T cells. We have previously reported that almost all cases of chronic lymphocytic leukaemic (CLL) express CD160 and the activation of CD160 by monoclonal anti-CD160 antibodies (mAb) induces cellular activation (proliferation and cytokine production). In this study, using fresh, peripheral blood mononuclear cells from patients with lymphocyte counts of &gt; 40 x 109/L, we found that the activation of CD160 protected CLL cells from spontaneous cell death (n = 18). The rates of spontaneous cell death of CLL cells ranged from 5 to 64% (average 33 ± 18%) after culture for 18 hours. However, the spontaneous death rate was reduced to 20 ± 11% (p=0.012) when the CLL cells were incubated with anti-CD160 MAb. The protective effect on cell survival was more marked in the cases showing the greatest spontaneous cell death: in those cases with &gt; 30% cell death (ie, greater than the average for the whole population), cell death was reduced from 48±9.9% to 28±8.2% (p=0.0001). In the cases which were resistant to spontaneous cell death (&lt; 30%), CD160 reduced cell death from 16 ± 6.6% to 11 ± 5.9% (p = 0.15). The PI3- kinase/AKT pathway is a well known survival pathway and CD160 activity in NK cells has been shown to involve PI3-kinase. CLL cells were incubated with anti-CD160 mAb and found to show AKT phosphorylation, I-kB degradation, Bcl-2 upregulation, but also Bax upregulation. Interestingly, the PI3-kinase inhibitor, wortmannin, completely blocked AKT phosphorylation, but had no effect on CD160-mediated cell survival and did not prevent activation of NF-kB nor upregulation of Bcl-2 or Bax. In view of the upregulated Bax expression, we hypothesised that despite CD160-induced increases in Bcl-2 and the improved survival, CLL cells may have simultaneously become more sensitive to death signals. CLL cells (from 9 cases) were treated with chlorambucil (20mg/ml) with or without the anti-CD160 antibody overnight and apoptosis was determined with Annexin-V/propidium iodide by flow cytometry. Chlorambucil alone induced 63%±16 apoptosis with 34%±23 spontaneous cell death. Co-treatment with chlorambucil and anti-CD160 mAb increased apoptosis to 72±14% with spontaneous rate at 23%±13.
In summary, the activation of CD160 protected CLL cells from spontaneous cell death in vitro via a PI3-kinase/AKT independent pathway. However, this improved survival was also associated with increased Bax and sensitisation to chemotherapeutic agents. The mechanism by which the activation of CD160 plays dual roles in the destiny of CLL cells remains unclear, but this represents a potential novel therapeutic approach. Data will be presented showing the activity of a chimeric, humanised anti-CD160 mAb on CLL killing.
Stem Cell Transplant (SCT) for Patients (pts) with Chronic Myeloid Leukemia (CML) Resistant to Tyrosine Kinase Inhibitors (TKI) with BCR-ABL Kinase Domain (KD) Mutation T315I.
Authors: Velev Nikolai , Cortes Jorge , Champlin Richard , Kantarjian Hagop M. , Rondon Gabriela , Giralt Sergio , Borthakur Gautam , De Lima Marcos 
Published: April 2020

Abstract
Background: Resistance to TKI therapy is associated with development of KD mutations in approximately 50–60% of pts. Although many imatinib-resistant mutations respond well to second generation TKI, T315I is insensitive to all currently available TKI (imatinib, dasatinib, nilotinib) in vitro and in the clinic. SCT is frequently recommended for these pts but there is no available data about the efficacy of SCT in such pts.
Aims: To investigate the efficacy and safety of SCT for patients with TKI-resistant CML with a T315I mutation.
Methods: We reviewed the outcome of all pts with T315I that have received a SCT at MD Anderson Cancer Center.
Results: Seven pts received 8 transplants. Their median age was 44 years (yrs) (range, 26 to 64 yrs). The median time from diagnosis to SCT was 42 months (mo) (range, 9–160 mo). All pts had become resistant to with imatinib; 5 received dasatinib and 1 nilotinib after imatinib failure, and 6 pts received other additional therapy prior to SCT. At the time of SCT 2 pts were in chronic phase (CP), both in partial cytogenetic response; 2 in accelerated (AP) with active disease; and 3 in second or greater CP from lymphoid blast phase (BP) (1 in minor cytogenetic response, 2 major molecular response, 1 complete molecular response –CMR-). Six transplants were from matched unrelated donors and 2 from cord blood. Best response after SCT was CCyR in 3 (2 AP, 1 BP), CMR in 4 (2 CP, 2 BP), and 1 unknown (died early). After a median follow-up of 11 months from SCT, 4 pts are alive; the 2 transplanted in CP are alive after 11 and 42 months after SCT and in CMR; 1 pt transplanted in AP has a sustained CCyR 20 mo after SCT with persistent T315I representing 94% of transcripts by pyrosequencing; and 1 in BP has a CMR sustained 6 mo after a second SCT (relapsed 5 months after first SCT) 3 pts have died: 1 AP and 2 BP, all with relapse.
Conclusion: SCT appears to be an effective strategy for pts with CML with T315I, although longer follow up is needed. Results are significantly better when pts are transplanted in CP. Thus, SCT should be considered in pts with resistance to TKI once T315I is identified, ideally in CP.
Inhibition of BCR/ABL-T315I by Dismantling the Hydrophobic Spine.
Authors: Azam Mohammad , Seeliger Markus , Powers John T , Gray Nathanael , Kuriyan John , Daley George Q. 
Published: April 2020

Abstract
Mutation in the ABL kinase domain is the principal mechanism of imatinib resistance (IMR) in patients with chronic myelogenous leukemia (CML). The second generation BCR/ABL inhibitors, Nilotinib and Dasatinib, are effective in inhibiting essentially all IMR variants, but not the gatekeeper mutant T315I. Substitution of a bulky hydrophobic residue for the gatekeeper threonine not only causes steric blockade to the inhibitor but also stabilizes the active kinase conformation through a network of hydrophobic connections dubbed the hydrophobic-spine. In this study we describe the molecular mechanisms employed by the gatekeeper mutation to stabilize the active conformation, and demonstrate that these structural components can be targeted by the small molecule inhibitor compound #14, which efficiently inhibits native and gatekeeper mutant forms of the BCR/ABL kinase. Structural modeling and mutagenesis of residues constituting the spine suggests that compound #14 inhibits the kinase by disrupting the hydrophobic spine. Screening for drug resistance in vitro selected for clones having compound mutations involving both the P-loop and gatekeeper residues. Our studies provide structural guidance for the design of inhibitors against the gatekeeper mutant, and suggest that combination therapy may be required to prevent the emergence of compound resistance mutations.
Pre-Transplant Bone Status Evaluation in Patients Undergoing Allogeneic Stem Cell Transplantation.
Authors: Yakoub-Agha Ibrahim , Wibaux Cecile , Magro Leonardo , Juilliard Audrey , Legroux-Gerot Isabelle , Coiteux Valerie , Jouet Jean-Pierre , Cortet Bernard 
Published: April 2020

Abstract
In view of the high observed frequency of bone events following allogeneic haematopoietic stem cell transplantation (allo-SCT), the aim of this prospective study was to evaluate pre-transplant bone status in allo-SCT patients. In the month before transplantation, bone-loss risk factors were documented for 27 patients. We measured the levels of bone remodelling markers (BRMs: plasma osteocalcin and bone & total alkaline phosphatases for bone formation; CTX and telopeptides for bone resorption), together with plasma creatinine, intact PTH, TSH, testosterone, LH, FSH, 25 OH vitamin D, serum calcium & phosphorus and calciuria. In addition, bone mineral density (BMD) at the lumbar spine, femoral neck and hips was measured using double-energy X-ray absorptiometry (DEXA). Spine X-ray measurements were also made. Bisphosphonate, oral calcium and vitamin D or testosterone were administered according to the results of the evaluation.Between June 2006 and March 2007, 13 males and 14 females underwent allo-CST for haematological malignancies. The median age at transplantation was 44 years (range: 22–60). Eighteen had received prior corticosteroid therapy, 10 were smokers and 2 had a history of alcohol abuse. All had received prior chemotherapy, including 2 patients having already undergone autologous SCT. Ten of the 14 women were post-menopausal but none was on hormone replacement therapy. The median BMI was 24 kg/m2 (19–35). The daily calcium intake was low, with a median value of 950 mg/day (467–1852). While serum calcium, phosphorus and creatinine levels were within the corresponding normal ranges for all patients, 15 individuals displayed vitamin D deficiency and 6 had calciuria &lt; 100 mg/day. Two patients suffered from hyperparathyroidism. In terms of BRMs, the patients respectively displayed a normal profile (n=9), high bone resorption activity (n=12) or high bone formation/resorption activity (n=5). One patient had hyperthyroidism and another presented hypotestosteronaemia. Bone density results were normal in 16 patients and abnormal in 11 (41%), including 8 with osteopaenia and 3 with osteoporosis. Vertebral fractures were observed in 4 patients. Overall, 18 patients (67%) were considered as having a pathological bone status and required treatment with bisphosphonate alone (n=5), vitamin D supplementation alone (n=11) or both (n=2).This study revealed that a large proportion of allo-CST patients have pre-existing abnormal bone status and thus demonstrates the importance of pre-transplant bone status evaluation in allo-CST candidates. The implementation of appropriate bone-related treatments may reduce the frequency of post-transplant bone events.
TNF-a Polymorphism Modulates the Outcome of Multiple Myeloma Patients Treated with Bortezomib
Authors: Buda Gabriele , Martino Alessandro , Maggini Valentina , Orciuolo Enrico , Galimberti Sara , Gentile Massimo , Brunori Marino , Morabito Fortunato , Rossi Anna Maria , Petrini Mario 
Published: April 2020

Abstract
TNF-a is a potent mediator of inflammation and bone resorption that seems to be involved in the malignant transformation of plasma cells, since mononuclear cells, obtained from Multiple Myeloma (MM) patients and exposed in vitro to TNF-a and interleukin-4, produced monoclonal plasma cells. In addition, TNF-a can stimulate plasma cell proliferation, by triggering interleukin-6 secretion, and shows proangiogenic properties in vitro (1). In addition, TNF-a determination was reported to be a good parameter for estimating tumour mass and for monitoring therapy outcome during treatment with different protocols (2). In particular, TNF-a is able to activate NF-kB, which is the main target of bortezomib (3). Variation in TNF-a levels can be related to gene expression, which is regulated at transcriptional level, as well as to genetic polymorphism. Single nucleotide polymorphisms (SNPs) have been identified at position -308 and -238 in the gene promoter. In particular, a G to A substitution at position -308 is associated with higher levels of TNF-a (1). The aim of this study was to investigate the effect of this TNF-a polymorphism on the outcome of refractory and relapsed MM patients (pts), receiving Bortezomib containing regimen as second line therapy. From September 2005 to April 2008 we selected 90 MM pts, who received at least one cycle of chemotherapy before treatment with Bortezomib and at least one cycle of high dose chemotherapy with peripheral blood stem cell transplantation in 25 pts. TNF a polymorphism at position -308 was determined on genomic DNA extracted from blood samples. The genotype frequencies obtained (71/90 G/G; 19/90 G/A) were in agreement with Hardy–Weinberg equilibrium. Patients were categorized as responders (Complete + Partial response =R) or non responders (Stable + Progression disease=NR) to treatment. The overall response in our patients was about 74% (67/90). In MM pts carrying the rarest A allele the overall response was reduced to 58% (11/19 pts), whilst GG carriers showed a better response rate of 79% (56/71 pts) (OR = 0.37). Our data indicate that the SNP at position -308 of the TNF-a gene promoter may be associated with different outcome in patients affected by MM and treated with bortezomib containing regimen. Similar conclusion was reached in a previous study where TNF-a gene promoter could predict the outcome after thalidomide therapy (1). To investigate whether TNF-a SNP is an independent predictor for response to MM therapy, larger studies are necessary, however if these preliminary results will be confirmed, they may represent a rationale for targeting TNF-a in novel therapeutic approaches to MM (4).
15 Year Follow-up of ABMT for NHL: Surprisingly Favorable Outcome for Younger Adults.
Authors: Duong Hien , Rybicki Lisa , Andresen Steven , Copelan Edward , Dean Robert , Kalaycio Matt , Pohlman Brad , Sobecks Ronald , Sweetenham John , Chan Josephine , Sungren Shawnda , Bolwell Brian 
Published: April 2020

Abstract
Despite the fact that autologous bone marrow/stem cell transplantation (ABMT) has been successfully used to treat patients with relapsed Non Hodgkin’s Lymphoma (NHL) for over 25 years, few series report follow-up beyond 10 years. We have followed a cohort of 110 NHL patients who underwent ABMT from 9/1988 through 12/1993, all treated with a uniform preparative regimen of CBV (Cyclophosphamide, BCNU, Etoposide (VP-16)). At the present time, 38 (35%) are alive and 72 patients have expired. Among the 38 living patients, the median follow-up is 15 years. NHL histology for all transplanted patients was based on the Working Formulation, and 21% had low grade, 54% intermediate grade, 25% high grade (the majority of the high grade group had diffuse immunoblastic large cell lymphoma). Median age at transplant was 47 years; 85% were sensitive to salvage chemotherapy; 31% had elevated LDH at the time of diagnosis; age adjusted IPI at ABMT was 0 (7%), 1 (56%), 2 (29%), 3 (7%). Multivariable analysis analyzing prognostic factors for survival predictably revealed that sensitivity to chemotherapy, and LDH status at the time of transplant, had significant impact on survival. Additionally, age at transplant was also highly significant (p=0.002). This is shown graphically below:
Figure Figure
The incidence of low grade histologies was similar for all three groups. The younger group had a lower incidence of relapse mortality, as shown below:
Figure Figure
It is commonly believed that age is of limited importance with respect to survival after ABMT. However, this data suggests that age may play a very important prognostic role. The causes are not well defined, but the continued relapse and mortality risk may warrant surveillance indefinitely for patients undergoing ABMT for NHL.
The Promising Strategy of Mycophenolate Mofetil Dosing to Prevent Moderate- to Severe-Acute Graft-Versus-Host Disease.
Authors: Nishikawa Shinichiro , Okamura Atsuo , Yamamori Motohiro , Katayama Yoshio , Shimoyama Manabu , Matsui Toshimitsu 
Published: April 2020

Abstract
To prevent acute graft-versus-host disease (GVHD) and graft failure, mycophenolate mofetil (MMF) has been employed as a substitute of methotrexate in allogeneic hematopoietic stem cell transplantation (all-SCT). However, the dosing strategy of MMF after all-SCT remains to be established.
In this study, we investigated the optimal MMF dosing until day 30 based on pharmacokinetic studies in 28 Japanese allo-SCT patients. For better clinical outcomes of MMF, higher mycophenolic acid (MPA) plasma levels are proposed to be desirable. Therefore, the first 11 patients (group A) received MMF orally every 12 hours at an escalated dose from 15 mg/kg to 25 mg/kg (maximum total daily dose 3000 mg), according to real-time pharmacokinetic monitoring of the total MPA area under the curve (AUC). However, the dose escalation in each individual did not always increase the AUC. Then, MMF was given orally at a fixed dose of 1000 mg every 8 hours in the subsequent 17 patients (group B). The pharmacokinetic data revealed that the increase of dosing frequency could statistically keep higher MPA plasma levels, as reflected in concentration at steady state (Css: group A vs. group B; 1.12 vs. 2.18 μg/ml on day 16, P = 0.003) or trough value (Ctrough: 0.18 vs. 0.56 μg/ml on day 16, P = 0.019). These results indicate that MMF administration of every 8 hours would be better than that of every 12 hours even in the same total daily dose until day 30.
We further assessed the safety and efficacy of extended MMF administration beyond day 30 retrospectively. Twenty-five patients ceased MMF at day 30, whereas 16 patients were subjected to extended regimen depending on the individual risk factors for GVHD (median dosing period 64.5 days, 50–94 days). No severe adverse events were observed in both groups. While the cumulative incidence (CI) of more than grade I acute GVHD at day 100 was comparable between the two groups (72.1 % vs. 62.5 %, P = 0.63, Figure 1A), the CI of grade II to IV acute GVHD was lesser in the latter group (42.3 % vs. 12.5 %, P = 0.045, Figure 1B).
These results suggest that MMF should be administered every 8 hours until day 30 after allo-SCT, followed by extending MMF administration at least until day 60 as a preemptive therapy for moderate- to severe-acute GVHD. This strategy might be very useful for double UCB transplantation under non-myeloablative conditioning especially, because the transplantation is reported to develop severe acute GVHD frequently.
Figure Figure