bluuebunny/crossref_metadata_2025_split huggingface dataset:
Familial Chronic Lymphocytic Leukemia(CLL): The Mayo Clinic Experience.
Authors: Haddad Tufia C. , Shanafelt Tait D. , Slager Susan L. , Phyliky Robert L. , Zent Clive S. , Dewald Gordon W. , Kay Neil E. , Call Timothy G. 
Published: October 2019

Abstract
Background: Over the last decade, increasing numbers of families with at least 2 cases of CLL or other B-cell lymphoproliferative disorders have been described.
Methods: Families seen at Mayo Clinic with at least one individual diagnosed with CLL and one or more first or second degree relative diagnosed with CLL or a B-cell lymphoproliferative disorder(LPD) were identified. Clinical characteristics and risk stratification test results (cytogenetic abnormalities by FISH, level of CD38 expression, and IgVH gene mutation status) were extracted and reviewed. Serum, intracellular and CLL B cell secreted levels of pro- and anti-angiogenic cytokines (VEGF, BFGF, and TSP) were also compared between cases of familial and sporadic CLL.
Results: Seventy-one families seen at Mayo Clinic were identified meeting criteria for familial CLL. Of the 71 index CLL families, 78.9% (n=56) had at least one other first or second-degree relative with CLL or a B cell LPD, 15.5% (n=11) had 2 other affected members, and 5.6% (n=4) had 3 or more affected members. Of affected family members with B-cell LPDs other than CLL, 64% (n=25) had non-Hodgkin lymphoma, 21% (n=8) had Hodgkin disease, 10% (n=4) had multiple myeloma or Waldenstrom macroglobulinemia, and 5% (n=2) had another lymphoid leukemia (1 ALL, 1 hairy cell leukemia). The median age was 62, with 63% males and 37% females. The majority of individuals had an early Rai stage at diagnosis (stage 0–66%, stage I-19%). Results of cytogenetic testing by FISH, CD38 status, IgVH gene mutation status appear similar in familial and sporadic CLL cases seen at Mayo clinic during the same time interval. No clear pattern of IgVH gene mutation status, IgVH gene family usage, level of CD38 expression, or cytogenetic abnormalities by FISH was seen among multiple affected members from the same family. There were also no significant differences noted in serum, intracellular, and CLL B cell secreted levels of VEGF, BFGF or TSP in the familial samples when compared to a cohort of non-familial samples.
Conclusions: Expression of cytogenetics by FISH, immunophenotype, IgVH mutation status and levels of serum, intracellular and cytoplasmic pro- and anti-angiogenic factors(VEGF, BFGF, and TSP) were similar in individuals with familial and sporadic CLL.
Molecular Mechanisms Involved in Homing and Migration of B-Chronic Lymphocytic Leukemia (CLL) in Response to CXCR4 Stimulation and Downstream Activation of the PI3K Pathway.
Authors: Ghobrial Irene M. , Ghobrial Joanna M. , Bramati Patricia , Timm Michael M. , Kumar Ashock , Witzig Thomas E. , Kay Neil , Hedin Karen E. 
Published: October 2019

Abstract
Malignant B-cells characteristically home to the bone marrow and lymph nodes. However, the mechanisms by which cells are recruited into and mobilized from the bone marrow/lymph nodes into the peripheral blood are not well understood. Chemokines such as SDF-1 and it receptor CXCR4 play a central role for lymphocyte trafficking and homing. Downstream activation of the PI3K pathway by chemokines has been implicated in the migration of many cell types. We recently demonstrated that CXCR4 expression correlates significantly with disease progression in CLL. Patients with Rai stage 4 had a higher expression of CXCR4 when compared to patients with Rai stage 0. The expression of CXCR4 was not found on CLL B cells resident within the lymph nodes implying that there may have been downregulation of CXCR4. In this study, we explore the molecular mechanisms involved in homing and migration of B-CLL cells in response to CXCR4 and investigated the role of the PI3K pathway in migration of CLL cells in response to SDF-1. Boyden chamber in vitro migration assays were used with primary CLL peripheral blood cells and the MEK1 CLL cell line. In addition, we expressed fluorescent YFP-CXCR4 in the cells and used confocal microscopy to visualize changes in the subcellular location of the fluorescent CXCR4 before and after SDF-1 stimulation. Inhibitors of the PI3K pathway such as the PI3K inhibitor LY294002 (10uM, one hour pretreatment) and the mTOR inhibitor, rapamycin (200nM overnight) were used. SDF-1 induced a dose dependent migration of primary CLL cells and MEK1 cells, indicating functional CXCR4 receptors. MEK 1 cells transfected with YFP-CXCR4 demonstrated surface localization on the cells. 3-Dimensional and continuous live imaging after SDF-1 stimulation for 30 minutes demonstrated alterations in the CXCR4-YFP leading to its capping and internalization subcellularly. Pretreatment of the cells with PI3K pathway inhibitors led to inhibition of migration with LY294002 but not with rapamycin. Confocal microscopy demonstrated abrogation of subcellular localization in the presence of LY294002 but not with rapamycin with the addition of SDF-1. These results demonstrate that CLL cells express functional CXCR4 receptora and that the PI3K pathway is essential for SDF-1 dependent CLL migration. In addition, we demonstrate that the CXCR4 receptor is internalized in response to SDF-1 stimulation indicating that CLL cells downregulate the CXCR4 receptor once they home to the bone marrow or lymph nodes where there is abundant SDF-1 in the microenvironment. This leads to the confinement of the cells in the microenvironment preventing further migration of the cells outside of the bone marrow/lymph nodes. Supported in part by CA97274
Inhibition of Tumor Growth by RGD Peptide Directed Delivery of Truncated Tissue Factor to the Tumor Vasculature.
Authors: Kessler Torsten , Bieker Ralf , Padro Teresa , Herrera Federico , Ruiz Sandra , Serve Hubert , Kreuter Micheal , Berdel Wolfgang E. , Mesters Rolf M. 
Published: October 2019

Abstract
Selective activation of blood coagulation in tumor vessels with subsequent tumor infarction is a promising anticancer strategy. To this end, a fusion protein consisting of the extracellular domain of tissue factor (truncated tissue factor, tTF) was fused to the peptide GRGDSP selectively targeting avb3 and avb5 integrins on tumor endothelial cells. The fusion protein tTF-RGD retained its thrombogenic and integrin binding activity as demonstrated by coagulation assays and binding assays with purified avb3 and endothelial cells. In vivo studies in mice bearing established human adenocarcinomas (CCL185), human melanoma (M21) and human fibrosarcoma (HT1080) revealed that i.v. administration of tTF-RGD induced partial or complete thrombotic occlusion of tumor vessels as indicated by histological analysis. Furthermore, treatment studies showed that tTF-RGD but not untargeted tTF induced significant tumor growth retardation or regression in all three types of solid tumors in mice without apparent side effects such as thrombosis in liver, kidney, heart or lung.
Thus, selective thrombosis in the tumor vasculature induced by tTF-RGD may be a promising strategy for the treatment of cancer.
Functional Pan-Universality of the Age-Related Regulatory Mechanisms of Gene Expression.
Authors: Kurachi Kotoku , Fontaine Jean-Marc , Abe Takahiro , Kurachi Sumiko 
Published: October 2019

Abstract
Through analyses of human factor IX (hFIX) and protein C (hPC) gene expressions in transgenic mice, we recently discovered the first molecular mechanism of age-dimension gene regulation, involving two critical genetic elements ASE and AIE required for age-related stable and increase patterns of gene expression, respectively. The next most critical issue to be tested was whether or not these elements, particularly ASE due to its possible utility, similarly functions with grossly different genes. We constructed hFIX expression vectors carrying a CMV promoter with or without ASE, and tested them in transgenic mice. Vectors with no ASE showed an age-dependent gradual decrease in hFIX expression, reaching the background levels in 3–9 months. Vectors with ASE, however, showed age-stable hFIX expression over the same time period. These findings supported the pan-universality of ASE function. As we previously reported, ASE regulates gene expression not only temporally but also spacially. With ASE, CMV-driven expression showed the highest in the heart muscle, whereas without ASE, the highest in the skeletal muscle. These results led to development of the Age Dimension Technology, a new field for exploring the unique applications of the age-related knowledge.
Intermediate Dose Methotrexate in Low-Risk Acute Lymphoblastic Leukemia. Results of the FRALLE 93A Protocol.
Authors: Perel Yves , Chambost Hervé , Gandemer Virginie , Leblanc Thierry , Pascale Schneider , Claudine Schmitt , Debré Mariane , Vermylen Christiane , Couillault Gérard , Lamagnere Jean-Paul , Auclerd Marie-Françoise , Leverger Guy , Baruchel André ,   the FRALLE group
Published: October 2019

Abstract
The FRALLE 93A protocol aimed to identify a very low-risk ALL subgroup, to limit treatment morbidity and to investigate the value of intermediate dose methotrexate. Inclusion criteria were : B-cell precursor ALL, age 2-7 years, leukocyte count <10G/l ; patients with CNS or testicular leukemia, bulky disease, t(9;22), t(4;11), t(1;19) or hypodiploïdy (<44 chromosomes) were not included.
181 children were included (13.5% of all the ALL included in the FRALLE 93 protocol) between June 1993 and Dec 1999. After a predisone prophase and induction phase (4-week daily prednisone, 4 weekly vincristine, thrice weekly 10,000 units of E coli asparaginase for 6 doses), during the 12-week antimetabolite consolidation therapy, patients were randomized to receive either low-dose methotrexate (25 mg/m2 p.o.) (lmtx) or 6 doses of intermediate dose methotrexate (IMTX) at 1500 mg/m2 by 24 hours infusion. Further therapy included a CCG type delayed intensification (dexamethasone, weekly vindesine, 3 doses of weekly doxorubicin and asparaginase, cytarabine) but without cyclophosphamide and a two years maintenance treatment. All pts received 16 triple intrathecal injections. The complete remission rate was 99.5%. The overall survival and event-free survival at 5 years were 93 % ± 4 % and 82 % ± 6 % (point date : 01.01.04). Only 1pt was considered prednisone-resistant (0.5%) D21 M1 marrow response is associated to a better 5 y EFS (86 + 5% vs 44 + 21% for M2/M3 pts (18 pts); p=0.001). Among 158 randomized patients, 9 testicular/meningeal relapses (6 isolated, 3 combined) occurred in the lmtx arm versus 1 in the IMTX arm (p=0.03) ; six BM relapses occurred in the lmtx group vs 9 in the IMTX group (p= NS). The 5-year DFS is not different 83% ± 8% and 85% ± 8% (p=0.47).
Conclusion: a high cure rate could be achieved in low-risk ALL without cranial radiation, alkylating agents or epipodophyllotoxin and with a low cumulative dose of anthracyclins (75 mg/m2). Prednisone response is of no help in this subgroup of BCP-ALL. D21 marrow response allows to identify a very low-risk subgroup in addition to initial criteria. IMTX proved superior to lmtx in preventing testicular or meningeal relapse in low-risk ALL.
Somatic Mutations and Polymorphisms of the Glucocorticoid Receptor Gene Are Not Related to Glucocorticoid Resistance in Childhood ALL.
Authors: Tissing Wim , Meijerink Jules P.P. , Den Boer Monique L. , Brinkhof Bas , van Rossum Liesbeth , van Wering Elisabeth R. , Koper Jan Willem , Sonneveld Pieter , Pieters Rob 
Published: October 2019

Abstract
In vivo and in vitro response to glucocorticoids (GC) are important prognostic factors in childhood ALL. To induce apoptosis, GC bind to the intracellular GC receptor (GR), which subsequently triggers transactivation and repression of downstream genes. As somatic mutations and polymorphisms have been linked to both increased and decreased GC sensitivity in healthy individuals, we studied the relationship between these gene alterations and GC sensitivity in childhood ALL.
Methods: We analysed leukemic cells taken at diagnosis and normal lymphocytes at time of complete remission (CR) from 57 children. DNA mutations were screened for using a PCR - SSCP technique. PCR products with abnormal SSCP patterns were sequenced. In vivo response to prednisone was determined according to BFM criteria as prednisone good response (PGR) or prednisone poor response (PPR). The in vitro prednisolone sensitivity was determined by MTT assay.
Results: The polymorphism ER22/23EK was found in 4% of cases, N363S in 12%, Bcl1 in 55%, intron mutation 16 basepairs upstream of exon 5 in 40%, H588H in 2% and N766N in 21%. They were proven to be polymorphisms as they were found in normal lymphocytes of the same patients at CR as well. The incidence of the polymorphisms was comparable to reports in the literature in the healthy population, except for N363S, which was found at a 2–4 fold higher incidence in ALL patients. Using the Chi-square test and the Mann-Whitney U test, no relation could be found between the occurrence of polymorphisms and in vivo prednisone response or in vitro sensitivity to prednisolone.
Conclusion: GC resistance in childhood ALL can not be attributed to polymorphisms of the hGR. The association between a higher incidence of the N363S polymorphism in childhood ALL as compared to the normal population needs further investigation.
Chromosomal Rearrangements Induced by DNA Double-Strand Breaks.
Authors: Varga Tamas , Aplan Peter D. 
Published: October 2019

Abstract
It has been suggested that introduction of double-strand DNA breaks into human chromosomes can lead to gross chromosomal rearrangements (GCRs) through improper repair mechanisms, and that these GCRs may be leukemogenic through the loss of tumor suppressor genes or the creation of fusion oncogenes. Using the human histiocytic lymphoma cell line U937, we developed a model system in which we were able to introduce a double-strand DNA break (DSB) in vivo at a predetermined location, and subsequently examine the ensuing chromosomal changes flanking the breakage site. In this model, U937 cells are stably transfected with a construct that contains the 18 bp recognition site for a rare cutting endonuclease, I-SceI, whose recognition sequence is not normally present in the human genome. The I-SceI site is placed between a strong constitutive promoter (EF1alpha) and the Herpes simplex virus thymidine kinase (HSV-tk) gene, which serves as a negative selectable marker by conferring ganciclovir (GCV) sensitivity to the cells. Following transfection of an I-SceI expression vector, double-strand DNA breakage occurs and abrogatesHSV-tk expression, leading to a dramatic increase in GCV resistant cells. We showed that, even in the abscence of GCV selection, the IsceI enzyme was able to cleave its recognition sequence in at least half of the transfected cells. The predominant type of rearrangement detected following the appearance of a double-strand break was an interstitial deletion. These deletions typically displayed uneven recession of free DNA ends flanking the cleavage site, generally resulting in either very short (<10 bp) or very long (>1000) bp deletions extending into the EF1alpha promoter and homogeneous deletions of 100–1700 bp into the HSV-tk gene. Moreover, in many clones, nucleotide microhomologies of 1–6 basepairs, palindromes, or both were detected at the breakpoint junctions, suggesting repair via a non-homologous end joining (NHEJ) mechanism.. We also showed that in 25 % of the recovered clones the DSB was repaired by formation of direct or inverted repeats derived from the breakpoint junction. Several clones showed similarly complex rearrangements, including short nucleotide insertions, inversions and duplications. Finally, approximately 4.5% (7/150) of the clones demonstrated insertion events that involved integration of nucleotide sequences derived from distant genomic regions, including LINE elements, coding regions of endogenous genes and sequences exhibiting known genomic instability. These insertions may have been derived via reverse transcription of mRNA sequences; alternatively the distant genomic regions may have served as templates for the inserted sequences, or genomic DNA fragments may have been directly incorporated into the I-SceI-induced break. A step-by-step modification of this loss of function selection scheme will provide a platform for studying DNA rearrangements induced by faulty dsDNA repair.
STI571 Suppresses Proliferation by Restoring Nuclear Cyclin Dependent Kinase Inhibitors (CDKIs) while STI571+TRAIL Promotes Cell Death by Decreasing Cytoplasmic CDKIs.
Authors: Dao Mo A. , Verfaillie Catherine M. 
Published: October 2019

Abstract
Cyclin dependent kinase inhibitors (CDKIs), p27Kip1 and p21Cip1, function as cell cycle inhibitors when located in the nucleus. When localized to the cytoplasm, these CDKIs can function as anti-apoptotic molecules by sequestering/preventing the activation of pro-apoptotic proteins such as ASK1 and procaspase-3. Our lab has reported elevated cytoplasmic CDKIs and decreased nuclear CDKIs in hematopoietic cells expressing BCR/ABL, the oncogene found in more than 90% of cases of chronic myeloid leukemia. Within the past decade, STI571 has been shown highly promising for CML treatment. However, there is increasing evidence suggesting that the drug might function more as a suppressor of proliferation and less as a promoter of cell death. In the current studies, we differentiate the effect of STI571 on proliferation vs. survival by tracking the subcellular increase/decrease in CDKIs using MO7e cells engineered to express BCR/ABL. To determine if a correlation exists between STI571 resistance and levels of cytoplasmic anti-apoptotic CDKIs, we also investigated changes in levels of nuclear vs. cytoplasmic CDKIs in LAMA84 -S (sensitive to STI571) vs. LAMA84-R (resistant to STI571). And lastly, we tested whether activation of TRAIL would enhance cell death in STI571-resistant cells. STI571 treatment increases nuclear CDKIs, correlating directly with a decrease in proliferation of MO7e/p210 cells. However, the high levels of cytoplasmic CDKIs in MO7e/p210bcr/abl was not modulated following STI571 treatment and cell death was not prominent, unless growth factors were removed. Moreover, cytoplasmic p21Cip co-immunoprecipitated with ASK1 and procaspase 3. When compared with LAMA-S cells, LAMA-R cells expressed even higher levels of cytoplasmic CDKIs. Treatment with STI571 decreased cytoplasmic CDKIs in LAMA84-S cells and resulted in cell death. As hypothesized, LAMA84-R cells did not show reduction in cytoplasmic CDKIs and did not enter apoptosis. However, when treated with STI571 and TRAIL, LAMA84-R cells showed a decrease in cytoplasmic CDKIs, and increase in apoptosis. Based on these observations, we conclude that: 1. BCR/ABL expression reduces nuclear CDKIs but increases cytoplasmic CDKIs. 2. STI571 treatment restores nuclear CDKIs and reduces cell proliferation of BCR/ABL expressing cells under physiological conditions. 3. Treatment of STI571+TRAIL reduces cytoplasmic CDKIs and increases cell death of BCR/ABL expressing cells, most notably, the STI571-resistant cells. In conclusion, we show that the imbalance between nuclear (cell cycle inhibitor) and cytoplasmic (cell survival enhancer) CDKIs exist in BCR/ABL-hematopoietic cells.
Enhanced Killing of Chronic Myelogenous Leukemia Cells by Rapamycin and Imatinib Is Associated with Differential Inhibition of 4E-BP1 and eIF4E Phosphorylation and Decreased Protein Expression by Non-Overlapping Mechanisms.
Authors: Moore James C. , Ly Chi , Luke Halbur , Ong S. Tiong 
Published: October 2019

Abstract
The Bcr-abl tyrosine kinase is known to promote transformation by dysregulating gene transcription, but its role in dysregulating translation is less well documented. Our recent work has implicated the mammalian target of rapamycin (mTOR) signaling as a downstream target of Bcr-Abl, since we find that the mTOR effectors, 4E-BP1 and S6, are phosphorylated in a Bcr-Abl kinase-dependent manner (Ly et al., Cancer Research, 2003). Because mTOR is a central regulator of eukaryotic translation, and inhibitors of mTOR act synergistically with imatinib mesylate (imatinib) to kill CML cells, these results suggest that, like transcription, translation may be a general cellular process dysregulated by Bcr-Abl activity. If this were so, then components of the cellular apparatus co-opted by Bcr-Abl to increase translation would constitute rational therapeutic targets. These would include signaling pathways mediating increased translation, components of the cap-binding complex (eIF4E, eIF4GI, and eIF4A) that regulate cap-dependent mRNA translation, as well as proteins whose translation is increased by Bcr-Abl kinase activity. Here we identify eIF4E as well as cyclin D3 as potential therapeutic targets in CML. Since eIF4E is essential for cap-dependent translation, and increased translation parallels eIF4E phosphorylation at Ser209, we determined the status of eIF4E phosphorylation in murine hematopoietic Ba/F3 cells expressing Bcr-Abl (Ba/F3-Bcr-Abl), and its dependence on Bcr-Abl kinase activity. Using phosphospecific antibodies to eIF4E, we found that Bcr-Abl kinase activity was essential for phosphorylation of eIF4E at Ser209, but had no effects on total levels of the protein. In contrast, rapamycin had no effect on the degree of eIF4E phosphorylation, although it was able to inhibit phosphorylation of 4E-BP1 completely (unlike imatinib). By examining total mRNA and protein levels of known targets of Bcr-Abl, we determined that cyclin D3, but not cyclin D2, was post-transcriptionally regulated by Bcr-Abl. Metabolic labeling studies were also conducted in Ba/F3-Bcr-Abl cells treated with media alone, imatinib, rapamycin, or both. Our results demonstrated that translation of cyclin D3 protein is regulated by the mTOR kinase in Bcr-Abl-expressing cells, and that combined inhibition of mTOR and Bcr-Abl resulted in an additional decrease in protein levels. Together, these results demonstrate that Bcr-Abl promotes protein translation of specific genes via mTOR, and that the activity of both Bcr-Abl and mTOR kinases contribute to dysregulated protein expression via non-overlapping mechanisms in CML cells. Ongoing studies are being conducted to determine the role of both eIF4E and cyclin D3 in the pathogenesis of CML.