bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Non-Hematopoietic Stem Cells in Mouse Bone Marrow Osteoblastic Zone.
Authors: Sato Hidetaka , Arai Fumio , Shibata Mami , Hirao Atsushi , Suda Toshio , Sakurada Kazuhiro 
Published: October 2019

Abstract
Bone marrow contains hematopoietic and non-hematopoietic stem cells. Although the character of hematopoietic stem cells (HSCs) in vivo has been extensively studied, our understanding of non-hematopoietic stem cells is derived only from in vitro expanded culture such as mesenchymal stem cells (MSCs). Since the number of non-hematopoietic stem cells in bone marrow is small, their characteristics, anatomical location and physiological role in vivo remain unknown. We attempted to overcome this problem by introducing a novel method. We extracted bone marrow cells from osteoblastic zone by enzymatic digestion. Bone marrow extracted cells (BMECs) contained non-hematopoietic cells (CD45−/Ter-119− cells) more than flushed out bone marrow cells (BMCs). Non-hematopoietic cells in BMECs were divided into two types, CD34+ (BME 34+) cells and CD34− (BME 34−) cells.
When BME 34− cells were cultured to expand to a confluent layer in MSC culture condition, they formed fibroblastic cells, which have the ability to differentiate into osteocytes, chondrocytes and adipocytes, indicating that BME 34− cells are the in vivo origin of MSCs.
On the other hand, BME CD34+ cells contained mostly non-hematopoietic SP fraction and they were Sca-1+ cells. Immunohistochemical analyses show that CD31-BME 34+ cells localized on osteoblasts and endothelial cells in bone marrow. When BME 34+ cells were co-cultured with stromal (OP9) cells, they formed a colony of myogenic cells, endothelial cells or adipogenic cells. However, different from BME 34− cells, fibroblastic cells expanded from BME 34+ cells did not differentiate to osteocytes, chondrocytes or adipocytes in MSC culture condition. When 4,000-6,000 BME 34+ cells were transplanted into irradiated mice, these cells engrafted as non-hematopoietic Vimentin+ fibroblastic-like cells in various organs such as liver, spleen, pancreas, colon, intestine, kidney, skin, brain, lung, heart, skeletal muscle, and bone marrow. In spleen and liver, transplanted cells formed clusters of Vimentin+ fibroblastic cells indicating that BME 34+ cells have ability to expand in vivo to some extent. When BME 34+ cells were directly injected into tibial muscles which were damaged by cardiotoxin, these cells gave rise to mature skeletal muscle cells and endothelial cells.
In summary, these studies show distinguishable two types of non-hematopoietic stem cells in bone marrow. It is possible that mobilization and recruitment of these bone marrow-derived non-hematopoietic stem cells play a major role in the setting of wound healing, organ regeneration and tumor growth. In addition, non-hematopoietic two types of BMECs may contribute to clarify the debate on the stem cell plasticity of bone marrow cells.
Outcome for Adolescent and Young Adults 16–21 years of age (AYA) with Acute Lymphoblastic Leukemia (ALL) Treated on the Children’ s Cancer Group (CCG) 1961 Study.
Authors: Nachman J. , Siebel N. , Sather H. , Steinherz P. , DeLaat C. , Fryer D. , Mattano L.A. , Hastings C. , Rubin C. , Bertolone K. , Franklin J. , Mitchell T. , Ettinger L. , Pyesmany A. , Gaynon P.S. ,   for the Children’s Oncology Group (COG), Arcadia, CA
Published: October 2019

Abstract
AYA Patients 16–21 years of age with ALL are treated on both pediatric and adult ALL protocols. Recent publications have suggested that AYA patients treated on pediatric protocols have a significantly better outcome than those treated on adult protocols. From November 1996 to May 2002, 262 AYA ALL patients were entered on the CCG 1961 protocol. Treatment was based on the following observations from the previous CCG 1882 Protocol (1989–1995). 1) For patients with rapid early response (< = 25% blasts in a day seven bone marrow aspirate-RER), in the context of CCG modified BFM therapy (S-BFM), pre-symptomatic central nervous system therapy with intensive intrathecal (IT) methotrexate (MTX) alone provided equivalent results to standard IT MTX and cranial radiation (CRT). 2) For patients with slow early response (> than 25 blasts on day 7-SER), the augmented BFM (A-BFM) regimen produced a significant improvement in outcome compared to S-BFM. A-BFM featured a significant increase in dose intensity for vincristine (VCR), L-asparaginase (L-ASP), and steroid, the use of intravenous MTX without rescue in conjunction with VCR and L-ASP in interim maintenance (IM), and a second IM and delayed intensification (DI) phase. RER patients on 1961 were randomized to one of four arms; S-BFM, S-BFM with a second standard IM and DI phase, A-BFM with only 1 IM/DI phase, and full A-BFM. SER patients were randomized to A-BFM or A-BFM with Idarubicin/Cytoxan pulses added to the 2 DI phases. One hundred ninety patients had < 50K WBC (73.4%) and 21% had T cell immunophenotype. Seven patients had a t (9/22) and three patients had a t (4/11). Five year EFS for AYA patients treated on CCG 1961 was 68% ± 3.7% and the five-year survival was 77% ± 3.2%. The majority of events were isolated bone marrow relapses. For RER AYA patients, five year EFS for patients treated on the augmented intensity arms (N=87) was 83.2% compared to 60.8% for patients treated on the standard intensity arms (N=76) (P = .10). Within the augmented intensity arms, there was no difference in EFS for patients randomized to one or two DI phases. There was no difference in EFS for the 2 SER treatment arms. 5 year EFS was 78% for the Idarubicin/Cytoxan arm and 74% for the ABFM arm. Five year EFS for AYA patients with < 50K WBC was 73.2% (N=190) versus 54% for AYA patients with > 50K WBC. Unlike younger patients, there was no difference in outcome for males and females. Conclusions: 1) AYA patients with ALL have a 68% 5 year EFS when treated with BFM based chemotherapy. 2) WBC < 50K predicts for a favorable outcome. 3) Early augmented intensity therapy appears to improve outcome for AYA ALL patients showing a rapid response to induction therapy.
Phase II Trials of Clofarabine in Relapsed or Refractory Pediatric Leukemia.
Authors: Jeha Sima , Razzouk Bassem I. , Rytting Michael E. , Gaynon Paul S. , Kadota Richard , Rheingold Susan , Luchtman-Jones Lori , Shen Violet , Arceci Robert J. , Fernandez Manuel , Weitman Steven , Steinherz Peter G. 
Published: October 2019

Abstract
Background: Clofarabine, a next generation nucleoside analogue, was well tolerated and demonstrated activity in adult and pediatric Phase I trials conducted in heavily pretreated leukemia patients. Multicenter Phase II studies in pediatric leukemia have completed accrual in the US and are reported here.
Methods: Two Phase 2, multicenter, open-label studies were conducted with clofarabine in children with refractory or relapsed ALL or AML. Clofarabine was administered intravenously over 2 hours at 52 mg/m2/day for 5 consecutive days. Cycles were repeated every 2 to 6 weeks based on response and toxicity.
Results: The studies enrolled 100 patients (60 ALL and 40 AML). Currently, data are available for 84 patients (49 ALL, 35 AML). Median age is 12 years (range 1 to 22 years) and median number of prior regimens is 3 (range 1 to 6). Thirty-nine percent had received prior bone marrow transplant (BMT). As determined by independent review, preliminary data indicate overall response rates of 31% in ALL (6 CR, 4 CRp, and 5 PR) and 26% in AML (1 CRp and 8 PR). Median duration of remission for ALL is 9.7 weeks (range 1.0 to 28.6) and for AML is 16.2 weeks (range 1.7 to 56.6+). Thirteen of 24 responding patients (54%) proceeded to BMT. Median survival was 42 weeks (range 7.0 to 63.1+) for responding ALL patients (CR+CRp+PR) and 39 weeks (range 7.7 to 93.6+) for responding AML patients (CRp+PR). Patients who failed treatment or were non-evaluable had shorter median survival; 7.4 weeks (range 0.9 to 40.1+) and 12.4 weeks (range 1.6 to 84.9+) for ALL and AML, respectively. Among the patients who were refractory to the last prior chemotherapy, 7/30 (23%) with ALL and 4/22 (18%) with AML achieved a response with clofarabine. Median duration of remission in these patients is 4.6 weeks (range 2.3 to 24.4+) for ALL and 20 weeks (range 1.7 to 56.6+) for AML. Most drug-related adverse events were transient including febrile neutropenia, diarrhea, nausea/vomiting, fever, skin rash, headache, elevation in liver enzymes and bilirubin, and infusion-related flushing and anxiety.
Conclusions: Clofarabine is active as a single agent in pediatric ALL and AML that are refractory to intensive salvage regimens. The overall safety profile is similar to that reported in other pediatric salvage studies. Clofarabine in combination with standard chemotherapy is currently under investigation in children.
In Vivo Evidence of Modulation of the Hemophilia Phenotype by the Factor V Leiden.
Authors: Schlachterman Alexander , Liu Jianhua , Liu Yi-Lin , High Katherine , Arruda Valder 
Published: October 2019

Abstract
The amelioration of hemophilia phenotype or delayed onset of the bleeding episodes in subjects with severe hemophilia A (FVIII deficiency) has been associated with inherited resistance to activated protein C due to factor V Leiden (R506Q), FVL. These observations were confirmed by in vitro systems in which homozygous phenotype of FVL increased thrombin formation in presence of < 1% FVIII by nearly 5-fold (Blood 90:3067). Here we provide in vivo evidence of the beneficial interaction of FVL on the hemophilia phenotype in mice. Animals with severe deficiency of factor VIII due to deletion of intron 16 of factor VIII gene (HA) or large gene deletion of factor IX (HB) were crossed with FVL homozygous mice [+/+] on C57Bl6 strain [Cui et al. (Blood 96:4222)] We used a modified activated partial thromboplastin time (aPTT) assay to compared clotting times among male HA (n=30), HA/FVL [+/+] (n=7), FVL [+/+] (n=5), and litermate WT mice (n=6) with age ranging from 6–12 weeks. Blood samples were collected by tail vein transection into 3.8% sodium citrate. Values for the aPTT in male HA were 67.3 ± 4 sec, and among WT or FVL [+/+] values were 37 ± 4 sec or 31± 2 sec, respectively. Whereas intermediate aPTT values of 53 ± 3.7 sec were determined in HA/FVL [+/+], which differs from HA mice (p<0.0001) but also from WT or FVL (p<0.0001). A similar shortening of aPTT was also determined among HB/FVL[+/+] which were compared to HB, 55 ± 7 sec vs. 64 ± 0.9. Hemostatic challenge by tail clipping assay failed to revealed differences in bleeding times/blood loss among hemophilia animals with or without FVL mutation. To test whether a more sensitive technique would provide further evidence of the improved hemostasis in HA/FVL mice, we assessed real-time in vivo thrombus formation by confocal and widefield microscopy. Mice were anesthetized and the cremaster muscle was exposed for intravital microscopy. Infusion of fluorescently labeled antibody to murine platelet glycoprotein IIb/IIIa complex via the jugular vein allowed monitoring of platelet deposition upon laser-mediated endothelial injury at several sites of the arterial vessel wall. No thrombus formation was observed in severe HA mice following successive vascular injuries, a finding also common in severe HB mice. However, infusion of factor VIII concentrated clearly induced the thrombi formation upon vascular injury. HA/FVL mice tested presented thrombus formation in a comparable fashion of HA-FVIII transfused mice. These in vivo data provide support to the hypothesis that the FVL mutation has the potential to improve the phenotype of severe hemophilia and may offer a novel therapeutic target for hemophilia.
Intermediate Dose Intravenous Melphalan and Dexamethasone Treatment in 144 Patients with Systemic AL Amyloidosis.
Authors: Goodman Hugh J.B. , Lachmann Helen J. , Bradwell Arthur R. , Hawkins Philip N. 
Published: October 2019

Abstract
The optimal intensity of chemotherapy in AL amyloidosis remains contentious. Traditional low dose oral melphalan and prednisolone therapy is ineffective in most patients, whereas high dose therapy with autologous stem cell rescue carries substantial procedure related mortality. We report here the outcome of intermediate dose intravenous melphalan (IDM) (25mg/m2 d1 + dexamethasone 20mg po d1-4, repeated every 21-28d) among 144 patients with amyloidosis (83M:61F, median age 62yrs, range 30–79). The median number of organs involved by amyloid was 2, including kidneys in 74% of patients, heart in 56%, neuropathy in 25% and liver in 20%. Median follow-up was 13mo (0.5–53). Patients received a median of 3 cycles of treatment (range 1–8). Dexamethasone was omitted in 51 cases, principally due to concern about fluid retention associated with nephrotic syndrome or cardiac amyloidosis. There were 52 deaths of which 3 (2%) were considered treatment-related (1 neutropenic sepsis, 2 dexamethasone-induced pulmonary oedema). Serum free light chain (sFLC) measurements were available before and after at least one cycle of IDM in 140 patients; 23% achieved complete response (CR, sustained normalization of sFLC ratio), 31% a partial response (PR, sustained >50% reduction in pre-treatment clonal isotype) and 46% did not respond (NR). Rates of sFLC CR+PR were higher among patients who also received dexamethasone at 64% vs 37% (p<0.0004). Paraproteins were detectable by electrophoresis and immunofixation of serum and/or urine in 115pts (80%); 85 of these pts had post-treatment data, showing CR (complete resolution of serum and urine paraprotein) in 17 (20%), PR (50% reduction) in 24 (28%) and NR in 44 (52%). Cycle by cycle assays of sFLC allowed rapid assessment of early clonal response, which was evident within 2 cycles in almost all responders. Maximal response was usually achieved by the fourth cycle. Median survival was 44 months among patients with CR or PR, and 18 months among non-responders (p<0.003). Amyloidotic organ dysfunction improved in 14%, remained stable in 51%, and deteriorated in 19% of evaluable cases. The findings support the use of IDM coupled with cycle by cycle sFLC measurements as a valid treatment strategy in patients with AL amyloidosis. Although clonal response rates in this series may be lower than selected patients treated with high dose therapy and autologous stem cell rescue, IDM can be delivered to a wider range of patients, in whom the rapidity of its action, relative simplicity of the regimen, and much lower treatment related mortality favor its continued use and further investigation.
Ectopic Fibroblast Growth Factor Receptor 3 (FGFR3) Expression Mediated by Isotype Switch Recombination.
Authors: Affer Maurizio , Robbiani Davide , Chesi Marta , Bergsagel Peter L. 
Published: October 2019

Abstract
FGFR3 is disregulated in approximately 15% of cases of multiple myeloma (MM) occurring by chromosomal translocation in the Igh switch region which results in the juxtaposition of FGFR3 next to the regulatory elements of the IgH locus, causing its ectopic expression in plasma cells. The acquisition of FGFR3 activating mutations in some tumors indicate a role for FGFR3 in tumor progression. In order to study its role in MM progression we developed a strain of transgenic mice in which the expression of the activated form of FGFR3 is mediated by isotype switch recombination by replacing the IgH gamma1 costant region with an FGFR3-IRES-EGFP cassette in a BAC that covers the entire murine IgH locus. We predict that in the transgenic mice B cells that undergo switch recombination to g1 on the productive allele will also undergo switch recombination to g1 at the transgenic locus and express FGFR3 and EGFP. To evaluate FGFR3 expression in vitro, we collected splenocytes from our transgenic mice and induced their proliferation and differentiation to plasma cells (mainly IgG1) using LPS and IL4. After four days of stimulation, the average number of EGFP positive cells went from 0.2% to 80%. We then analyzed FGFR3 expression by RT PCR, Northern and Western blot and detected LPS/IL4 inducible FGFR3 expression in plasma cells mediated mainly by the Igamma1 promoter and in a subset of them by the Vh promoter. We then tested if forced FGFR3 expression in B cells would affect their proliferation rate and ability to differentiate in vitro by MTT assay, cell cycle analysis and PI staining on LPS/IL4 stimulated splenocytes at different time points. With this system, we did not observe any FGFR3 mediated alteration of splenocytes growth parameters when compared to wild type controls. Similarly, in vivo no monoclonal gammopathy has been observed by serum protein electrophoresis and all the transgenic mice remaine tumor free at 1.5 years. Although we have not been able to demonstrate a tumorigenic role for FGFR3 in switched plasmacells we were able to obtaine regulated FGFR3 expression in them. It is possible that FGFR3 expression alone is not sufficient to induce plasma cells transformation and that FGFR3 has a dispensable role in tumor initiation, but that it could still play a role in tumor progression. Consistently, FGFR3 expression is lost in about 25% of multiple myeloma wih a t(4;14). To address this hypotesis we are currently crossing these mice with MMSET (the other gene disregulated in the t(4;14)) transgenic mice to investigate a possible collaborative role between these two genes in the development of MM.
Sequencing the Multiple Myeloma Kinome: Absence of Mutation in Known Malignancy-Associated Kinases.
Authors: Claudio Jaime O. , Khaja Razi , Zhuang Lihua , Bali Meenakshi , Sivananthan Kamalanayani , Christopoulos Constantine C. , Fonseca Rafael , Bergsagel P. Leif , Scherer Stephen W. , Stewart A. Keith 
Published: October 2019

Abstract
In approximately 50% of Multiple Myeloma (MM), translocation of non random genes into the IgH locus is believed to be the seminal event in the pathogenesis of the disease. Another 50% of cases are hyperdiploid and trisomic to certain autosomes, but do not harbor any translocation and thus are believed to have genetic alterations in unidentified loci. These observations, together with the finding of somatic mutations in FGFR3, N- and K-RAS, MYC, TP53 and CDKN2C/p18INK4C during the later stages of MM indicate that defective signaling pathways likely play a role in the progression of this malignancy. Of relevance then, in recent years recurrent mutations in kinases have frequently been implicated in malignancies including notably colon cancer and melanoma. We have therefore begun a comprehensive effort to sequence the tyrosine kinome for mutations and genetic polymorphisms in MM. Of particular interest are 90 receptor tyrosine kinases, 43 receptor tyrosine kinase-like, 5 receptor guanylate cyclase, and a lipid kinase. We report here results from our pilot high throughput exon scanning in 32 human MM cell lines which initially focused on 13 kinases known to be somatically mutated in human cancers. To date we have expanded this effort to assess 30 genes with sequence obtained which currently spans 80% coverage of the kinase domains of these genes. A total of 1.9 million bp have been sequenced across 235 exons. No recurrent mutations have been identified in the kinase domains of the cancer-associated genes: SRC, ILK1, KIT, GUCY2F, PDGFRA; in the genetic disorder-associated kinases: BTK, EPHA4, LAMA2, EPHB6, ACVR2; and in the mutation hot spots of frequently mutated cancer gene PIK3CA. A novel missense mutation is however identified upstream of the kinase domain of FGFR3 changing a Ser residue to Arg at codon 433. This residue, which is conserved across species and in FGFR1 and FGFR2, has not been reported in myeloma and in thanatophoric dysplasia, but the biological significance of this mutation is unknown. Several single nucleotide polymorphisms were identified in the coding regions of some of these kinases. Notably, synonymous polymorphisms in the kinase domains of EPHA4, PDGFRA3, KIT, MLK1, ILK1, NTRK3, FLT3, ABL1, FES, MLK4, and EGFR1 were identified that changed a codon but not the amino acid. More importantly, we identified non-synonymous amino acid variations in the kinase domains of EPHA4, GUCY2F, PTK2, and PIK3CA genes that are more likely to effect variability in the activity of these kinases. In summary, no recurrent kinase mutations of significance in Myeloma development or progression have yet been identified. Sequencing of the known cancer associated kinases in MGUS and hyperdiploid MM patients is now underway and our data set is being expanded to include all 139 kinases.
Comprehensive Genome-Wide Profile of Regional Gains and Losses in Multiple Myeloma Using Array-CGH: The 1q21 Amplification and Potential Role of the BCL-9 Gene in Multiple Myeloma Pathogenesis.
Authors: Carrasco Ruben , Tonon Giovanni , Brennan Cameron , Protopopov Alexei , Sinha Raktim , Feng Bin , Leo Christopher , Zhang Yunyu , Zhang Jean , Zhan Fenghuang , Protopopova Marina , Mitsiades Constantine , Munshi Nikhil , Chin Lynda , Bergsagel Peter , Kuehl Michael , Shaughnessy John , Anderson Kenneth , DePinho Ronald 
Published: October 2019

Abstract
Multiple Myeloma (MM) is characterized by a clonal proliferation of abnormal plasma cells in the bone marrow and is among the most frequent and lethal hematological diseases. In spite of significant effort towards the identification of the molecular events leading to this malignancy, the genetic alterations responsible for the pathogenesis of this disease remain poorly understood. Regional copy number alterations (CNAs) in cancer genomes have been among the most informative structural changes in cancer and have led to the discovery of many oncogenes and tumor supressor genes. Using array comparative genomic hybridization (array-CGH) and expression microarray technologies we have analyzed a large collection of cell lines and clinically annotated primary tumors. This high-resolution genomic analysis has identified all previously reported regional gains and losses as well as many novel highly recurrent genetic loci with potential biological and clinical relevance. In particular, we have identified an amplification at chromosome 1q21 as one of the most recurrent genetic changes in cell lines and in a subgroup of primary tumors. This chromosomal change has been previously implicated with disease progression. Analysis across several cell lines has allowed the identification of a Minimal Common Region (MCRs) of amplification at 1q21. Correlation between DNA copy number changes and expression profiling data has identified a limited set of candidate genes within this MCR that are amplified and overexpressed. Using shRNAi technology we have identified BCL-9 as a candidate gene residing at the 1q21 MCR. In vitro and in vivo functional data about the role of BL-9 will be presented. These data will provide critical understanding on the diverse pathways leading to Multiple Myeloma progression.