bluuebunny/crossref_metadata_2025_split huggingface dataset:
Non-Total Body Irradiation (TBI) Containing Preparative Regimen in Alternative Donor Bone Marrow Transplantation (BMT) for Severe Aplastic Anemia (SAA).
Authors: Lee Je-Hwan , Choi Seong-Jun , Lee Jung-Hee , Lee Young-Shin , Seol Miee , Lee Jung-Shin , Kim M. Wookun , Lee Kyoo Hyung 
Published: April 2020

Abstract
Most preparative regimens in alternative donor BMT for SAA have included TBI to overcome high incidence of graft rejection, which is the major problem in this setting. With routine use of irradiated leukocyte-poor blood products, however, the incidence of graft rejection due to pre-transplant allo-sensitization has decreased. Recently, efforts to reduce dose intensity of the preparative regimen for alternative donor transplants have been made with promising results. Using non-TBI containing preparative regimens, 13 patients with SAA were transplanted from alternative donors in our center and we report on the results of BMT in these patients. Median age of the patients, 6 males and 7 females, was 22 years (range, 15-34). Twelve donors were unrelated volunteers and one was an HLA one-locus mismatched sibling. Median time from diagnosis of SAA to BMT was 304 days (range, 49–5,403). Nine patients had received one cycle of immunosuppressive treatment with ATG (± cyclosporine) before BMT, while 4 had not. All patients except one had been transfused before BMT. Preparative regimens consisted of cyclophosphamide (50 mg/kg x 4) plus ATG (Atgam® 30 mg/kg x 3) in 9 patients, cyclophosphamide (50 mg/kg x 4) plus fludarabine (30 mg/m2 x 3) in 2 patients, and cyclophosphamide (50 mg/kg x 2) plus fludarabine (30 mg/m2 x 5) plus ATG (Atgam® 30 mg/kg x 3 or Thymoglobuline® 3 mg/kg x 3) in 2 patients. All patients received non-T-cell depleted bone marrow graft form the donor. Cyclosporine plus methotrexate were given for GVHD prophylaxis. All patients engrafted on a median of day 21 (range, 15–27) with complete donor chimerism. Grade III-IV acute GVHD developed in 3 (23%) of 13 patients and extensive chronic GVHD in 4 (31%) of 12 evaluable patients. With a median follow-up duration of 1,138 days (range, 118–1,553), 10 patients are alive with durable engraftment showing 75% of survival rate. Causes of death were chronic GVHD in 2 and CNS bleeding in 1. In conclusion, non-TBI containing preparative regimen could ensure durable engraftment in alternative donor BMT for SAA and showed promising results.
Bypassing Cellular Senescence by Ezh2.
Authors: Kamminga Leonie M. , Bystrykh Leonid , Houwer Sita , Douma Jose , Weersing Ellen , Dontje Bert , de Haan Gerald 
Published: April 2020

Abstract
In this study mouse embryonic fibroblasts (MEFs) and hematopoietic stem cells (HSCs) were used to identify genes involved in the process of cellular aging. MEFs are able to undergo a distinct number of population doublings, after which they enter a state of irreversible growth arrest termed replicative senescence. Previous studies of others and us have shown that aging is regulated by an intrinsic genetic program. In order to identify candidate genes involved in senescence we assessed gene expression profiles of proliferating and senescent MEFs. Interestingly, a significant number of genes that was higher expressed in proliferating vs. senescent MEFs, are involved in epigenetic regulation of gene transcription by modulating histone methylation and deacetylation. The top candidate gene was Enhancer of zeste homolog 2 (Ezh2), a Polycomb group protein (PcG), involved in histone methylation and deacetylation and known to function as a negative regulator of gene transcription. It has been reported that Ezh2 is essential during development, since Ezh2-deficient mice die early during embryonic development. Ezh2 expression was rapidly downregulated during senescence in primary MEFs, but was readily detectable in spontaneously transformed cells. Retroviral overexpression of Ezh2 in primary MEFs resulted in bypassing the senescence program. In addition, overexpression of Ezh2 in spontaneously transformed MEFs increased proliferation rates. We found that Ezh2 is highly expressed in purified Lin−Sca-1+c-kit+ (LSK) HSCs. In agreement with MEF studies, terminal differentiation of stem cells in vitro resulted in rapid downregulation of Ezh2 expression. Furthermore, overexpression of Ezh2 in purified HSCs results in increased proliferation rate in vitro. Moreover, HSCs transduced with Ezh2 were enriched for CFU-GM activity. Ongoing studies will address whether overexpression of Ezh2 in stem cells will affect their self-renewal potential in vivo. In an extensive genetic screen we identified genes that correlate with Ezh2 expression in stem cells, potential targets or partners of Ezh2, among others Top2a, MKi67, Pcna, and Eed. These data were deposited in an on-line resource (www.WebQTL.org), to uncover genetic networks that are involved in regulating the process of aging.
Methylation of the Tumor Suppressor Gene PTPRO (Receptor-Type Protein Tyrosine Phosphatase) Is Associated with Expression of Important Apoptosis-Related Proteins in Chronic Lymphocytic Leukemia (CLL).
Authors: Motiwala T. , Ghoshal K. , Majumder S. , Kutay H. , Neuberg D. S. , Kitada S. , Reed J. C. , Dewald G. W. , Flinn I. W. , Tallman M. A. , Liu T. , Plass C. , Lucas D. M. , Byrd J. C. , Jacob S. T. , Grever M. R. 
Published: April 2020

Abstract
We previously demonstrated that apoptosis-inhibiting proteins (e.g., Bcl-2, Mcl-1, and XIAP) are important in predicting response to chemotherapy and antibody therapy for CLL. Additionally, genetic abnormalities and epigenetic control of gene expression have become increasingly important in understanding the prognosis and potential therapeutic opportunities in this disease. Greater than 80% of patients with CLL have genetic abnormalities detectable by FISH. Important subsets include those with a less favorable prognosis (deletions of 17p, 11q), an intermediate prognosis (trisomy 12), or a favorable outcome (deletion of 13q). PTPRO, encoding a receptor-type tyrosine phosphatase, is a specific tumor suppressor gene located in chromosomal region 12p12.3. PTPRO has been highly characterized as inhibiting anchorage-independent tumor growth in human lung carcinomas. This gene has been found to be silenced by promoter methylation, and can be re-expressed by hypomethylating agents in human lung tumor models. Thus, we examined the methylation profile of the promoter region of PTPRO in primary leukemic cells from patients with CLL, and in B cells obtained from normal volunteers. Analysis of the PTPRO CpG island by methylation-specific PCR and combined bisulfite restriction analysis (COBRA) demonstrated that this region was methylated in 91% of samples from 46 patients, whereas it was unmethylated in control B cells from normal volunteers. Specific quantitative determinations of important proteins related to apoptosis in the leukemic cells from 20 of these patients showed that, in the group with methylation of the PTPRO promoter, there was a significant elevation of Bcl-2 (p=0.079), Mcl-1 (p=0.004), XIAP (p=0.048), Bcl-2/BAX ratio (p=0.0024), and Mcl-1/BAX ratio (p=0.0017). There was no apparent relationship with FISH category or immunoglobulin VH mutational status. Thus, PTPRO promoter methylation analysis may identify a subset of CLL patients less likely to respond to therapy, and provide a potential reversible target for new therapeutic approaches to this disease. Finally, analysis of methylation of CLL samples by restriction landmark genome scanning also revealed methylation of genes for three additional protein tyrosine phosphatases: PTPRN2, PTPRZ2, and PTPN11. Increases in methylation of two of these (PTPRN2 and PTPRZ2) in CLL samples relative to control B cells was confirmed by COBRA. These results provide additional potential molecular targets for CLL therapy.
Thrombocytosis Increases the Risk of Chemotherapy-Associated Thrombosis.
Authors: Khorana Alok A. , Francis Charles W. , Culakova Eva , Lyman Gary H. 
Published: April 2020

Abstract
Background: Venous thromboembolism (VTE) is a frequent and significant complication of cancer and of chemotherapy, with an estimated risk of 0.04%/month (0.5%/year). Patients with thrombocytopenia are widely perceived to be at low-risk for VTE. Conversely, thrombocytosis may contribute to risk of VTE, but this has not been well-studied. We conducted a prospective analysis to determine the association of platelet counts with the incidence of symptomatic VTE in cancer patients receiving chemotherapy.
Methods: The Awareness of Neutropenia in Cancer (ANC) Study Group is currently enrolling patients with breast, lung, colon, ovary and other cancers, at 137 sites nationwide, with an accrual goal of 4,500 patients. Patients are registered at the time of initiation of a new chemotherapy regimen, and followed for 4 cycles. The association of VTE with platelet counts and other clinical variables was studied in univariate analysis and in a multiple logistic regression model.
Results: A total of 2,151 patients treated with at least one cycle of chemotherapy were available for analysis. Of these, 18.1% had platelet counts &lt;200,000 and 21.5% had counts ≥350,000 prior to initiating chemotherapy. Symptomatic VTE occurred in 46 patients, for an incidence of 2.14% over a median follow-up of 2.5 months (0.85%/month). The incidence of VTE by quartiles of pre-chemotherapy platelet counts is shown in the figure. VTE occurred in 4.34% (1.73%/month) of patients with platelet counts of ≥ 350,000 prior to initiation of chemotherapy. This was significantly greater than 1.8% (0.72%/month) incidence in those with baseline counts of &lt;200,000 (p=0.035). Similarly, the mean platelet count prior to chemotherapy was higher for patients who developed VTE than for those who did not (351,000 versus 288,000, p=0.007). Platelet counts during chemotherapy were also associated with risk for VTE. The mean platelet count at nadir during the cycle VTE occurred was 202,000. This was significantly greater than the mean nadir count of 157,000 for corresponding cycles in patients who did not develop VTE (p=0.007). Other clinical variables associated with development of VTE included primary site of cancer, ECOG performance status ≥ 2, and baseline use of erythropoietin or colony-stimulating factors. A baseline platelet count of ≥ 350,000 continued to be significantly associated with VTE after adjusting for other risk factors in a multivariate analysis (odds ratio 3.00, 95% CI 1.61–5.55, p=0.0005).
Conclusion: This is the first report describing an independent association of platelet counts with symptomatic VTE in cancer patients receiving chemotherapy. Patients with platelet counts of ≥ 350,000 prior to initiation of chemotherapy are at a three-fold greater risk for developing VTE. The role of platelets in the pathogenesis of chemotherapy-associated thrombosis warrants further study. Platelet counts should be taken into account when defining high-risk patients for thromboprophylaxis.
Figure Figure
Investigation of TCR-Vβ CDR3 Spectrotypes in CD4 and CD8 Lymphocytes from Paroxysmal Nocturnal Hemoglobinuria (PNH) Patients.
Authors: Nakamura Akiko , Shichishima Tsutomu , Noji Hideyoshi , Ikeda Kazuhiko , Maruyama Yukio 
Published: April 2020

Abstract
PNH is one disorder of bone marrow failure syndromes, including aplastic anemia and myelodysplastic syndrome. It is considered that immunologic mechanisms by cytotoxic T lymphocytes (CTLs) and interferon-γ (IFN-γ) contribute to hypoplastic bone marrow of these disorders. In addition, PNH is an acquired clonal disorder of the hematopoietic stem cell. Recently, it has been reported that analysis of T cell-antigen receptor (TCR)-Vβ repertoires, especially TCR-Vβ CDR3 (complementarity- determining region 3) spectrotypes, is an effective tool to study immunologic mechanisms by CTLs in pathophysiology of PNH (Karadimitris et al, Blood, 2000; Kook et al, Blood, 2002; Risitano et al, Blood, 2002). In the present study, we investigated 21 kinds of TCR-Vβ repertoires by flow cytometry in CD4 and CD8 lymphocytes from 5 PNH patients and a healthy volunteer and the TCR-Vβ CDR3 spectrotypes using polymerase chain reaction assay in CD4 and CD8 lymphocytes from 3 of 5 PNH patients and the control. We also quantitated intracellular IFN-γ in CD4 and CD8 lymphocytes from 5 PNH patients and the control according to the method by Sloand et al (Blood, 2002). We found no specific TCR-Vβ repertoires in CD4 and CD8 lymphocytes from PNH patients compared with the control. The TCR-Vβ repertoires with relative increase of CD4 or CD8 lymphocytes (over 10 of ratio of the proportion of each TCR-Vβ repertoire in a PNH patient/the proportion of the same TCR-Vβ repertoire in a healthy volunteer) were 13.6 or 4 and 22 in Case 1, 3 and 11 or 1 in Case 2, 3 and 13.6 or 3 in Case 3, 5.3 and 7.2 or 2, 3, 7, and 18 in Case 4, and 4, 5.2, 13.6, 16, and 23 or 1 and 14 in Case 5, respectively. TCR-Vβ CDR3 spectrotyping showed that in CD4 lymphocytes most CDR3 patterns were chiefly polyclonal, except for one oligoclonal (Case 1) and one monoclonal (Case 3) patterns of TCR-Vβ25; in CD8 lymphocytes most CDR3 consisted of polyclonal, oligoclonal, and/or monoclonal patterns, suggesting the possibility that CD8 lymphocytes recognize much more antigens of abnormal cells, probably including PNH clones, than CD4 lymphocytes. Unfortunately, we found the same patterns as described above in CD8 lymphocytes from the control, although CD4 lymphocytes from the control presented only polyclonal pattern of CDR3. Quantitative analyses of IFN-γ showed that index values of IFN-γ in CD4 and CD8 lymphocytes from PNH patients were higher than those from the control. However, we did not find any significant correlations between the spectrotypes of TCR-Vβ CDR3 and the index values of IFN-γ in PNH patients, suggesting that TCR-Vβ repertoires with monoclonal and oligoclonal CDR3 patterns do not necessarily produce much IFN-γ. In conclusion, our findings suggest that TCR-Vβ CDR3 spectrotyping is more effective tool to resolve some immune mechanisms of pathophysiology in PNH, especially by auto-reactive CTLs.
Sustained Polyclonal Hematopoietic Repopulation after Successful SCID-X1 Gene Therapy by Means of a Non Random Integrating Pseudotyped Gammaretrovector.
Authors: Schwarzwaelder Kerstin , Schmidt Manfred , Howe Steven , Prinz Claudia , Wissler Manuela , Schmidt Sonja , Deichmann Annette , Glimm Hanno , Gaspar Bobby , Trasher Adrian , von Kalle Christof 
Published: April 2020

Abstract
The integration of a retroviral vector as used in hematopoietic gene therapy trials produces a transition sequence from the vector DNA into the genomic DNA and may thus serve as a stable molecular marker unique for each cell clone. High sensitive linear amplification mediated PCR (LAM-PCR) allows the detection of specific retroviral integration sites. Thus it is possible to determine the clonal composition of the hematopoietic system in vivo (1). We could show that the hematopoietic repopulation in human SCID-X1 patients was derived from various, long-lived progenitor cell clones indicating retroviral transduction into pluripotent cells (manuscript submitted). In two cases of lymphoproliferative disorder after successful SCID-X1 gene therapy integration of the retroviral vector into the LMO-2 oncogene was probable the main reason for malignancy (2). The distribution of integration sites over the whole genome, the potential preference for integration at certain loci and which cells receive genetic correction and engraftment are therefore of considerable interest. Recently, another gene therapy trial has successfully corrected 4 infants suffering from SCID-X1. A GALV-pseudotyped MLV-based vector carrying the therapeutic common gamma chain gene was used for transduction of autologous CD34+ cells. The patients did not receive any conditioning therapy before transplantation. We analyzed lymphoid and myeloid DNA from all patients. The transduction efficiency of T lymphocytes and myeloid cells was up to 100 and 1 %, respectively. In vivo clonality analysis of CD3+ cells showed a polyclonal composition 1 to 2 years after transplantation. The myeloid repopulation also consisted of various different clones. These data may indicate transduction of pluripotent and long term active stem or progenitor cells. We here report on more than 300 sequenced integration sites of the patients, whereas 250 sequences could be assigned unequivocally to a unique locus. So far, no vector integration in the LMO-2 oncogene could be detected in this trial, and the patients do not reveal any other evidence for malignancy or clonal deformation of their stem cell compartment. We could show that integration of the mammalian gammaretroviral vector in this gene therapy trial is not random. Integration of the vector happens generally within or close to specific regions of genes. We found common integration sites (CIS) in RefSeq gene regions. The targeted RefSeq genes were classified according to the Gene Ontology database. Our data strongly support the presumption that curative gene therapeutic treatment requires a sustained polyclonal contribution of ex vivo manipulated stem and progenitor cells and provide an important insight into the integration manner of GALV-pseudotyped MLV-based vectors.
Imatinib Mesylate Disrupts Cell Cycle Progression, Modifies the Nucleoskeleton and Suppresses Activation-Induced Transcription in Human T Cells.
Authors: Dietz Allan , Johnson William B. , Knutson Gaylord J. , Bulur Peggy A. , Schulenberg Bertie , Beechem Joe M. , Prendergast Franklyn G. , Vuk-Pavlovic Stanimir 
Published: April 2020

Abstract
Imatinib mesylate (imatinib, Gleevec®, Novartis, Basel, Switzerland) inhibits T cells in vitro and in vivo (Dietz et al., Blood104: 1094–1099, 2004; Cwynarski et al., Leukemia18: 1332–1339, 2004). The drug blocks T cell cycle progression rather uniquely as it neither inhibits expression of CD69, an early marker of T cell activation, nor induces apoptosis. To characterize the molecular effects of imatinib leading to this mode of T-cell inhibition, we measured the changes in transcriptome (by Affymetrix U133 chips), proteome and phosphoproteome (by Western blotting, differential phosphoprotein expression and mass spectrometry). We found that phytohemagglutinin activated T cells pre-treated with imatinib had reduced expression of 983 transcripts and increased expression of 271 transcripts when compared to untreated PHA activated T cells by the factor of 1.5 or more (p&lt;0.05). Among the prominently down-regulated transcripts were granzyme B, CTLA-4 and IL-2-receptor α-chain (CD25), all characteristic of activated T cells, as well as cyclins D2 and D3 and cyclin-dependent kinases 3, 4 and 7, the molecules regulating cell cycle progression. Among the up-regulated transcripts were Kruppel-like transcription factors 2 and 7, and p27, a finding compatible with the observed cell cycle inhibition. Furthermore, we selected and identified 30 proteins from 2-D gels that were up-regulated and/or hyperphosphorylated in imatinib treated activated T cells. Among these were four heterogeneous ribonucleoproteins, three lamins and γ-actin, all components of the nucleoskeleton at the interface of chromatin and inner nuclear membrane and involved in replication and transcription (Herrmann and Foisner, Cell. Mol. Life Sci.60: 1607–1612, 2003; Shumaker et al. Curr. Opinion Cell Biol.15: 358–366, 2003). Thus, imatinib-borne interference with T cell signal transduction affects the nuclear structure indicating for the first time that nucleoskeleton structural changes are associated with T cell activation status.
Sudden Blastic Transformation (SBT) in Patients (pts) with Chronic Myeloid Leukemia (CML) Treated with Imatinib Mesylate.
Authors: Vidal Marieberta , Kantarjian Hagop , O’Brien Susan , Rios Mary Beth , Verstovsek Srdan , Cortes Jorge 
Published: April 2020

Abstract
Imatinib has become standard therapy for pts with CML in chronic phase (CP). SBT has been reported in pts receiving IFN-α, at a rate of 0.5% to 2.5% during the first 3 years of therapy. There is little information on the occurrence of SBT among pts treated with imatinib. Here we report 3 pts who developed SBT, defined as occurring after being found in complete cytogenetic remission (CG CR). These 3 pts represent 0.5% of 557 pts who received imatinib at MD Anderson for chronic phase CML. The total population has been followed for a median of 33 months (range, 1 to 56). Pt #1 was a 54 yr old female with a variant Ph translocation, t(9;22;19;10) (q34;q11.2;p13.1;q22). Sokal risk was low. She started imatinib 400mg/day within 4 mos from diagnosis, achieved a CG CR within 3 mos and was still in CG CR after 9 mos with BCR-ABL/ABL 0.66%. On routine follow-up at 12 mos she was found in myeloid blast phase with 48,XX,+8,t(8;21)(q22;q22),t(9;22;19;10) (q34;q11.2;p13.1;q22), +der(22)t(9;22;19;10) in all 20 metaphases. She received standard induction chemotherapy followed by bone marrow transplantation and is alive and in CR after 24+ months. Pt #2 was a 51 yr old female initially treated with IFN-α. She achieved a CG CR but discontinued therapy because of toxicity after 18 mos. Fourteen mos after stopping IFN-α she had a CG recurrence (35% Ph-positive) and was started on imatinib 400mg/day. Three mos later she was in CG CR and BCR-ABL was undetectable by nested PCR; a similar result was found after 9 mos of therapy. 9 mos later (18 mos after start of imatinib) she had a lymphoid SBT (precursor B-cell). CG showed 45,XX,-8, t(9;22) (q34;q11.2), −20,+mar in one metaphase, and 11 metaphases were diploid. No ABL mutations were identified. She achieved CR with HCVAD + imatinib and is currently receiving a BMT. Patient #3 was a 27 yr old pt who received imatinib 800 mg/d as first line of therapy for CML in CP with intermediate Sokal risk score. Three mos after start of therapy he was in CG CR and repeated analyses showed continued CG CR up to mo 15, with the lowest BCR-ABL/ABL 0.12 at mo 6. On mo 18 he had a lymphoid SBT (precursor B phenotype) with 43–46,XY, −7, −8,i(9)(q10),t(9;22)(q34;q11.2), −13, −17, −21,+der(22)t(9;22),+mar in 13 metaphases and one diploid metaphase. No ABL mutations were identified. He was treated with HCVAD + imatinib and achieve a CR, becoming undetectable for BCR-ABL by nested PCR. He is undergoing a BMT. In pts 2 and 3, transformation was associated with a sudden drop in platelet counts. These isolated events should alert the physicians to continued monitoring of pts on imatinib and consider marrow studies in the event of late, unexpected peripheral blood count changes. Still, SBT is rare, probably less common that seen with IFN-α therapy.