bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lenalidomide in High-Risk Myelodysplastic Syndrome and Acute Myelogenous Leukemia Associated with Chromosome 5 Abnormalities.
Authors: Borthakur Gautam , Garcia-Manero Guillermo , Faderl Stefan , Estrov Zeev , Verstovsek Srdan , Hood Mary , Kantarjian Hagop 
Published: April 2020

Abstract
Background: Treatment with lenalidomide produces remarkable responses in patients with myelodysplastic syndrome and deletion 5q 31 (5q-) cytogenetic abnormality; transfusion independence in 67% of patients, major cytogenetic responses in 44% and a median duration of response of 115.9 weeks (List et al. Abstract #251; Blood: 108). Responses have been reported in few patients with MDS and complex karyotype as well. Since chromosome 5 abnormality is considered to be an early genetic event, we designed a phase 2 study of lenalidomide in patients with high-risk MDS and relapsed/refractory acute myelogenous leukemia (AML) with any chromosome 5 abnormality.
Method: The dose of lenalidomide was 25 mg/day, orally for 21 days with 7 days rest (28 day cycle). For patient with platelet count &lt; 50,000/μl, the starting dose is 15 mg/day orally for 21 days with 7 days rest (28 day cycle). Dose interruptions/reductions are planned for ≥ grade3 neutrpenia with fever or ≥ grade 4 neutropenia (for patients starting with absolute neutrophil count [ANC] &gt;1000/μl) and for grade 3 thrombocytopenia (for patients starting with platelet count ≥ 50,000/μl).
Results: Eleven patients out of this 30 patient study have been treated so far; 6 with AML and 5 with MDS. All patients with AML and 2 patients with MDS had previous therapy. Median age is 63 years (range, 44 to 80 years). Nine patients had complex karyotypic abnormality and 2 patients had single additional chromosomal abnormality (+8 in one and 11q abnormality in another) along with 5q deletion. One patient with AML (previously treated with decitabine) and with karyotypic abnormality of trisomy 8 in addition to 5q deletion, achieved complete remission after 3 cycles of therapy. Another patient had reduction in bone marrow blasts from 21 to 5%, but could not continue lenalidomide due to profound thrombocytopenia complicated by gastrointestinal bleeding and eventually had disease progression. Four (4/6) patients with a platelet count of ≥ 30,000/μl and 5 (5/6) patients with ANC of ≥500/μl at study entry experienced worsening cytopenias. No grade 3/4 non-hematological toxicities were seen. One patient in CR currently remains on study. Reasons off study include disease progression (8 patients), stem cell transplant (1 patient) and one patient died while on study from sustained pancytopenia, infection and multiorgan failure.
Conclusion: The role of lenalidomide in the treatment of high-risk MDS and AML with chromosome 5 and additional karyotypic abnormalities needs to be defined in the context of clinical trials. Future studies are needed to examine lower doses with or without growth factor support.
Outcome of Allogeneic Stem Cell Transplantation after Hypomethylating Therapy with 2′-Deoxy-5 Azacytidine for Patients with Myelodysplastic Syndrome.
Authors: De Padua Silva Leandro , de Lima Marcos , Kantarjian Hagop , Champlin Richard , Faderl Stefan , Giralt Sergio , Kebriaei Partow , Davisson Jan , Estey Eli , Garcia-Manero Guillermo , Issa Jean-Pierre , Ravandi Farhad 
Published: April 2020

Abstract
Decitabine is approved for the treatment of patients with intermediate- and high-risk myelodysplastic syndrome (MDS). In vitro studies have demonstrated an increased expression of MHC class I molecules, HLA-DR and beta-2-microglobulin on the surface of MDS cells after decitabine therapy, potentially increasing their susceptibility to immune surveillance mechanisms and a graft-versus-leukemia effect. We analyzed the outcome of 12 patients with MDS with a median age of 58.5 years (range, 37 – 66) who underwent an allogeneic stem cell transplant (5 sibling, 5 unrelated, 2 cord blood) after prior therapy with decitabine. At diagnosis, 2 patients had intermediate-1, 7 intermediate-2 and 3 high risk MDS by the international prognostic scoring system. Nine had a non-myeloablative and 3 an ablative regimen. The source of stem cells was marrow in 2, peripheral blood in 8 and cord blood in 2. The patients had received decitabine for a median of 5.5 cycles (range, 1 – 20) and a median duration of treatment of 7.1 months (range, 1 – 27). Decitabine was well tolerated with reversible gastrointestinal toxicity and neutropenic infections as the main toxicity. Best response to decitabine was CR in 4 patients, PR in 6, and hematological improvement in 2. Eight had disease progression on decitabine. Five patients received additional chemotherapy and achieved a CR before transplant. The median time between completion of decitabine and transplant was 3.4 months (range, 0.2 – 11.7). At the time of transplant, 8 patients were in CR and 4 in PR. The median CD34+ cell number was 4.17 x 106 cells/kg (range, 0.58 – 10.1 x 106 cells/kg). Eleven patients engrafted and one had secondary engraftment failure. Median times to neutrophil and platelet engraftment were 13.7 days (range, 7 – 27) and 17.3 days (range, 7 – 28), respectively. No unusual toxicity was encountered. Nine patients (75%) developed acute GVHD, and 6 chronic GVHD (55% of those alive beyond 100 days). Ten patients were in CR and 1 in PR at day 100 post transplant. With a median follow 11.5 months (range, 3 – 26), 9 patients are alive (8 in CR and 1 with progressive disease) and 3 have died (2 after relapse and 1 from GVHD and sepsis). We conclude that prior therapy with hypomethylating agents may potentially improve the outcome of allogeneic transplant in MDS through enhancement of graft versus leukemia effect and should be examined prospectively.
A Clinical Test for the Identification of Amyloid Proteins in Biopsy Specimens by a Novel Method Based on Laser Microdissection and Mass Spectrometry.
Authors: Vrana Julie A. , Gamez Jeffrey D. , Theis Jason D. , Plummer Timothy B. , Bergen Robert H. , Zeldenrust Steven R. , Kurtin Paul J. , Grogg Karen L. , Dogan Ahmet 
Published: April 2020

Abstract
The management of systemic amyloidosis relies on the treatment of the underlying etiology and differs radically for different amyloid types. Therefore, given that at least 25 different proteins have been associated with amyloidosis, accurate identification of proteins deposited as amyloid fibrils is an important clinical problem. In this study, we describe a novel method that can characterize amyloid subtypes using laser microdissection (LMD) and mass spectrometry (MS) on routinely processed paraffin-embedded tissues. The study used 60 cases consisting of 16 transthyretin, 9 serum amyloid-associated protein, 20 immunoglobulin light chain lambda, 5 immunoglobulin light chain kappa, and 10 amyloid negative control samples. The biopsy specimens studied included heart, kidney, gastrointestinal tract, lung and decalcified bone marrow specimens. The amyloid type in all cases was previously characterized based on clinical findings, immunohistochemistry and, where indicated, by molecular testing for transthyretin mutations. Amyloid plaques were captured from an 10 micron paraffin section exhibiting positive Congo Red staining using LMD. Proteins were extracted, digested with trypsin and identified following MS/MS using the Mascot search algorithm analysis. MS correctly identified each of the 4 types of amyloidosis analyzed. Serum Amyloid P component and Apolipoprotein E were also identified as constituents of the amyloid deposition. The analysis was successful on all tissue types including decalcified bone marrow specimens and small biopsy specimens such as endomycardial biopsies and renal biopsies. The use of LMD from paraffin embedded biopsies and subsequent analysis by MS allows identification of the type of amyloid protein deposited with high specificity and sensitivity. This method promises to be a clinical test for accurate identification of amyloid proteins in routinely processed biopsy specimens and overcomes many of the specificity and sensitivity issues associated with current methods such as immunohistochemistry.
Immunoglobulin Free Light Chain Ratio Is an Independent Risk Factor for Progression of Smoldering Multiple Myeloma.
Authors: Dispenzieri Angela , Kyle Robert A. , Katzmann Jerry A. , Larson Dirk , Benson Joanne , Clark Raynell , Melton L. Joseph , Gertz Morie A. , Kumar Shaji K. , Fonseca Rafael , Jelinek Diane F. , Rajkumar S. Vincent 
Published: April 2020

Abstract
Background: Smoldering multiple myeloma (SMM) is an asymptomatic plasma cell proliferative disorder with a high risk of progression to symptomatic multiple myeloma. Identification of risk factors that predict progression of SMM to symptomatic MM could identify higher risk patients who might benefit from chemoprevention or more intensive surveillance. We hypothesized that increased monoclonal free kappa or lambda immunoglobulin light chains in smoldering myeloma (SMM), as detected by the serum free light chain (FLC) assay, indicates an increased the risk of progression to active myeloma.
Methods: Of 276 pathologically confirmed SMM patients seen at the Mayo Clinic from 1970 to 1995, baseline serum samples obtained within 30 days of diagnosis were available in 273.
Results: At a median follow-up of surviving patients of 12.4 years, transformation to active disease has occurred in 161 (59%) patients. An abnormal FLC ratio was present at baseline in 90% of patients. The best break-point for predicting risk of progression was a FLC ratio less than or equal to 0.125 or greater than or equal to 8 (hazard ratio, 2.3; 95% CI, 1.6–3.2) [Figure 1]. The extent of abnormality of FLC ratio was independent of SMM risk categories defined by number of plasma cells in the bone marrow and size of serum M-proteins (bone marrow plasma cells ≥ 10% and serum M protein ≥ 3 g/dL; bone marrow plasma cells ≥ 10% but serum M protein &lt; 3 g/dL; and serum M protein ≥ 3 g/dL but bone marrow plasma cells &lt; 10%). Incorporating the FLC ratio into the risk model, the division of patients into high-, intermediate-, and low-risk groups is 28, 42, and 30% with 5 year progression rates of 76, 51, and 25%, respectively [Figure 2].
Conclusions: The serum immunoglobulin FLC ratio is an important additional determinant of clinical outcome in patients with SMM.
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Polymorphisms of the Transforming Growth Factor Beta 1 (TGFB1) Gene Define a Subgroup of Patients with Late Onset of Disease and Poor Outcome in Multiple Myeloma.
Authors: Neben Kai , Mytilineos Joannis , Heiss Christiane , Ho Anthony D. , Benner Axel , Opelz Gerhard , Goldschmidt Hartmut 
Published: April 2020

Abstract
Common genetic variants in immune and inflammatory genes can affect the risk of developing multiple myeloma. The aim of this study was to determine the effect of polymorphic variants in the interleukin 10 (IL10), transforming growth factor beta (TGFB1) and tumor necrosis factor alpha (TNFa) genes on the predisposition to multiple myeloma and the effect of these polymorphisms on survival after treatment with chemotherapy. Genotype data of 7 single-nucleotide polymorphisms and clinical follow-up information were available for 239 consecutive patients with multiple myeloma who presented at our outpatient department. All patients were treated with chemotherapy, including high-dose chemotherapy and peripheral blood stem cell transplantation in 191 patients. In particular, there were 155 males and 84 females with a median age of 55 years (range, 30 – 84). The median follow-up period for the entire cohort was 75 months (range, 3 – 190 months). The TGFB1 low producer genotype 10Leu/Leu (n=78) was associated with a 3.5 years older age at start of treatment (57.5 versus 54 years, p=0.005), a higher level of beta2-microglobulin (3.4 versus 2.5 mg/l, p=0.01), a higher frequency of ISS-scores II and III versus I (OR=0.50, p=0.01) and an inferior median survival rate of 71.8 versus 91.6 months (p=0.02) as compared to other genotypes, respectively. In a multivariate analysis, the TGFB1 low producer genotype 10Leu/Leu was identified as an independent factor for survival (p=0.03), indicating that the poor prognosis of these patients is not due to the older age at the start of treatment. All other gene polymorphisms analyzed (IL10–1082A&gt;G, IL10–819C&gt;T, IL10–592C&gt;A, TGFB1 Arg25Pro, TNFa -308G&gt;A, TNFa -238G&gt;A) showed no statistically significant effect on overall survival. Our findings suggest that patients carrying the TGFB1 low producer polymorphism show a later onset of disease and need for chemotherapy treatment as compared to other genotypes. In addition, our findings indicate that TGFB1 genetic variants influence prognosis in patients with multiple myeloma who received chemotherapy and suggest that genetically determined cytokine production affects the clinical course of the disease possibly through regulation of immune surveillance.
Combination of Bortezomib with Tipifarnib (R115777) Induces Synergistic Myeloma Cell Apoptosis Via Down-Regulation of HDAC6 and Inhibition of Aggresome Formation.
Authors: David Ebenezer , Schafer-Hales Katherine , Marcus Adam I. , Kaufman Jonathan L. , Lonial Sagar 
Published: April 2020

Abstract
Background: We have previously demonstrated that the combination of bortezomib and a farnesyl transferase inhibitor (FTI) induce synergistic myeloma cell apoptosis, and that maximal cell death is sequence dependent (AACR meeting-2004, ASH meeting -2005, David, E. et.al., Blood, (2005)106: 4322–4329,). The mechanism responsible for this synergy has been poorly understood. As FTI’s are known to downregulate HDAC6 expression, we further evaluated alternative mechanisms responsible for the observed synergy between bortezomib and tipifarnib.
Methods: Myeloma cell lines and primary human myeloma cells were used for the analysis. Confocal microscopy was used to assess ubiquitination in RPMI8226 cells exposed to each drug combination. Western blot analysis and flow cytometry were used to assess phospho-protein expression and apoptosis in MM.1S cell line.
Results: The use of tipifarnib and bortezomib resulted in a time dependent reduction in HDAC6 expression as measured by western blot. Confocal images revealed that bortezomib treatment, both alone and in combination with tipifarnib resulted in an accumulation of ubiquitinated proteins, suggesting a disruption of the aggresome complex. Moreover, a potential sequence-dependent antagonistic affect was observed when tipifarnib was given prior to bortezomib resulting in the reduction of ubiquitination. Additional experiments also demonstrated that apoptosis induced by the combination was only partially inhibited by the pan-caspase inhibitor ZVAD-FMK. This led us to explore caspase independent causes of apoptosis. Inhibition of cathepsins demonstrated overall apoptosis was significantly inhibited, suggesting that both caspase independent and dependent pathways are important in myeloma cell apoptosis induced by this potent combination. Although FTI alone inhibited GO/G1/S phases to some level, only the combination of bortezomib and tipifarnib resulted in the enhanced level of apoptosis which was confirmed by different biochemical methods.
Conclusion: The use of a FTI represents an important and alternative mechanism by which to inhibit HDAC6 and thus the aggresome pathway of protein degradation. Given that combinations of bortezomib and aggresome inhibition have been shown to be very active in vitro and in animal models, this approach represents an attractive method by which to target both critical pathways of protein degradation within malignant cells.
Polymorphisms of NOS3 and FCGR2A Genes Contribute to Thrombotic Risk in Essential Thrombocythemia
Authors: Olcaydu Damla , Gisslinger Heinz , Esterbauer Harald , Wiesbauer Franz , Kralovics Robert 
Published: April 2020

Abstract
Thrombosis is the most frequent complication in essential thrombocythemia (ET) that impairs quality of life and contributes to mortality and morbidity of patients. The molecular basis of thrombosis in ET remains unclear. Polymorphisms (SNPs) in coagulation factors (FII-G20210A, FV-Leiden) and the V617F mutation of JAK2 gene (JAK2-V617F) have previously been shown to associate with thrombosis in ET. Since thrombosis occurs frequently in JAK2-V617F negative ET and FII-G20210A and FV-Leiden polymorphisms occur only in up to 5% of patients, we examined the hypothesis that other genetic factors contribute to the thrombotic risk in ET. In addition to FII-G20210A and FV-Leiden we studied thirteen candidate SNPs of genes involved in coagulation (F7, F13A1, VWF, THSD1, ATIII), platelet aggregation (P2Y12, GPIIIA, GPIIB, FCGR2A) and endothelial function (NOS3, ADRA2B) previously implicated in thrombosis in non-MPD patients. We studied 94 patients with the diagnosis of ET retrospectively observed for clinical variables such as venous and arterial thrombosis, stroke, myocardial infarction, deep vein thrombosis (DVT), bleeding, angina pectoris, superficial venous thrombosis (SVT), and pulmonary embolism. SNP genotyping was performed using allele-specific PCR and sequencing. We examined the association of individual SNPs with the presence or absence of clinical variables (adjusted for age, sex, and BMI). In our ET cohort, JAK2-V617F showed significant correlation with the presence of DVT (OR 10.08; 95%CI 1.15–88.49; P=0.01). Both FII-G20210A and FV-Leiden were rare in this cohort (2% and 4%, respectively) and thus did not show any significant associations. Homozygosity for the 298Asp variant of NOS3 gene (encoding the endothelial nitric oxide synthase) showed significant correlation with stroke (OR 46.75, 95%CI 1.38–147, P=0.014). Similarly, homozygosity for the 131Arg variant of Fc receptor gamma 2A (FCGR2A) involved in platelet GPVI signaling correlated with the presence of SVT (OR 9.46, 95%CI 1.48–60.82, P=0.014). Our results indicate that genetic factors other than JAK2-V617F, FII-G20210A and FV-Leiden contribute to thrombotic risk in ET. Furthermore, the nitric oxide and the platelet GPVI signaling pathways are likely to play an important role in the pathogenesis of thrombosis in ET.
Microparticles (platelet, Endothelial, and Leucocyte) Are Elevated in Patients with Myeloproliferative Disorders.
Authors: Lynch Susan , Stirling David , Ludlam Christopher A. 
Published: April 2020

Abstract
Patients with myeloproliferative disorders (MPD) have an excess of thrombotic complications which is not fully corrected by normalisation of their elevated haematocrit or thrombocytosis. The mechanisms responsible for this are not completely understood but platelets, leucocytes and endothelial cells have all been implicated. Plasma microparticles are observed to be elevated in other diseases associated with thrombotic complications. We measured platelet (PMP), endothelial (EMP), red cell (RMP) and leucocyte (LMP) microparticles in patients with MPD compared to healthy controls. Microparticles, of less than 1μm were measured by flow cytometry on platelet poor plasma obtained from citrated peripheral blood samples. Subtypes were identified using fluorescent conjugated monoclonal antibodies; EMP CD31+CD42-, PMP CD31+42+, RMP Glycophorin A + and LMP CD45+ events. Absolute concentrations of microparticles per μl plasma were calculated using Trucount beads. Results are currently available for 27 MPD patients, 9 polycythaemia vera (PV), and 18 primary thrombocythaemia and 14 healthy controls. All patients except 1 PV were on current treatment with hydroxycarbamide and all but 3 were receiving aspirin. PMP and LMP were elevated in MPD patients compared to healthy controls, PMP 12432 (1653) v 2371 (278) per μl, mean (SEM) p&lt;0.001, LMP 98.7 (14) v 62.6 (7.7) per μl, p=0.03. EMP were elevated in MPD patients at 202 (34) v 34 (14) per μl, p&lt;0.001. There was no significant difference in RMP levels 646 (131) v 589 (241) per μl, p=0.84. There were no significant differences in MP numbers between the PT and PV patients or by Jak2 V617F mutation status. Platelet count (x109/l) was higher in the MPD patients, PT 503 (39) v PV 372 (43) v control 236 (12), all comparisons p&lt;0.04. PMP/platelet count was calculated to adjust for this and the ratio remained elevated in the MPD patients, PV 28.5 (4.5) and PT 26.1 (3.7) v controls 9.9 (1.2), both p=&lt;0.03. In conclusion, these results demonstrate that PMP, EMP and LMP are elevated in MPD despite standard treatment, with the PMP elevated disproportionately to the platelet count. This is further evidence for platelet activation, endothelial dysfunction and a role for leucocytes in the thrombotic risk of MPD and that the microparticles also play an active role in promoting the prothrombotic state.
Activating JAK3 Mutations in Transient Myeloproliferative Disorder and Acute Megakaryoblastic Leukemia Accompanying Down Syndrome.
Authors: Sato Tomohiko , Toki Tsutomu , Kanezaki Rika , Xu Gang , Terui Kiminori , Kamio Takuya , Kojima Seiji , Kiyoi Hitoshi , Mano Hiroyuki , Ito Etsuro 
Published: April 2020

Abstract
Children with Down syndrome (DS) have an approximately 20-fold higher incidence of leukemia than the general population. The majority of leukemia cases associated with DS are acute megakaryoblastic leukemia (AMKL). Although GATA1 mutations have been found in almost all cases of transient myeloproliferative disorder (TMD) “a preleukemia” that may be present in as many as 10% of newborn infants with DS and AMKL accompanying DS (DS-AMKL), GATA1 mutation alone may not be sufficient for development of leukemia. Following identification of acquired activating JAK3 mutations in DS-AMKL, JAK3 mutations have been reported also in TMD patients. However, the frequency and functional consequence of JAK3 mutations in TMD remain unknown. To further understand how JAK3 mutations are involved in the development and/or progression of leukemia in DS, we screened TMD patients and two DS-AMKL cell lines for JAK3 mutation, and examined the functional consequences of these JAK3 mutations. In one out of the two DS-AMKL cell lines, MGS, we identified novel JAK3 mutations (JAK3Q501H mutation in the SH2 domain and JAK3R657Q mutation in the psuedokinase domain in the same allele). JAK3Q501H and JAK3R657Q each constitutively phosphorylates STAT5 and transformes Ba/F3 cells to factor-independent growth, whereas the double mutant (JAK3Q501H and JAK3R657Q) has more potent transforming activity than each mutant. Biochemichal analysis in Ba/F3 cells revealed that the degrees of phosphorylation of STAT5 in the cells transduced with each JAK3 mutant were correlated with its transforming activity. Although we previously identified a JAK3I87T mutation in one of two TMD patients, no JAK3 mutations were detected in another 9 TMD patients. Together with the previous results, we found JAK3 mutations in each of 11 TMD and 11 DS-AMKL patients. Although the number of the patients analyzed was small, these results indicate that there are no significant differences in the frequency of JAK3 mutations between TMD and DS-AMKL. In this study, we showed for the first time that the TMD patient-derived JAK3 mutation was also an activating one. JAK3I87T transformed Ba/F3 cells to factor-independent growth. Treatment with JAK3 inhibitors (WHI-P131 and WHI-P154) resulted in a significant decrease in the growth and viability of Ba/F3 cells expressing each activating JAK3 mutant. These results suggest that the JAK3 activating mutation is an early event during the development of AMKL in DS. Furthermore these results provide a proof-of-principle that JAK3 inhibitor should have therapeutic effects on the AMKL and TMD patients carrying the activating JAK3 mutations.