bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Cytomegalovirus Can Induce Alteration of β-Actin mRNA and Microfilaments as Well as Induce Apoptosis in Mesenchymal Stem Cells.
Authors: Wei Guoqing , Lin Maofang , Cai Zhen , Huang He
Published: October 2019

Mesenchymal stem cells (MSCs) is a very important cell system in bone marrow stromal and human cytomegalovirus (CMV) infection is one of the most common complications following stem cells transplantation. Our studies were to investigate the cytoskeleton change as well as the apoptosis induction effects of CMV infection on MSCs. In our study, MSCs were infected by 100,10,1 TCID50 of CMV. After 48h of culture with DMEM supplemented with 10% (v/v) fetal calf serum, MSCs shapes were observed with microscope. CMV particles and ultra structure in the cells were detected with transmission electron microscope. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene,β-actin gene and GAPDH genes. Flow Cytometry was used to detect the apoptotic cells. After the infection of CMV, shape of MSCs changed, turning from thin shuttle shape to round and thick ball shape, even escaping from wall. Cell shape changed more dominantly as the CMV titer increased. With transmission electron microscope, CMV particles and typical cellular apoptotic character as well as the ruptured- Microfilaments could be seen in the cells infected by CMV. MSCs infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time- and titer- dependent manner compared with the uninfected cells whose expression of GAPDH mRNA did not change much. With Flow Cytometry, it was found there were more apoptotic and necrosis cells in the cells infected by CMV. These results indicate that CMV can not only infect MSCs and destroy the cytoskeleton in the cells but also can induce apoptosis of the cells, much more attention of CMV infection on MSCs should be paid after stem cells transplantation.
Positive Expression of MUC1 Gene Is a Poor Prognostic Factor in Acute Myelogenous Leukemia.
Authors: Huang Ren-wei , Li Guo-wei , Wang Dong-ning , Li Xu-dong , Lin Gui-zhen , Lin Dong-jun
Published: October 2019

Objective: To study the expression of MUC1 in acute leukemia and its clinical significance.

Methods: Expression of MUC1 mRNA was detected in 73 newly diagnosed and relapsed patients of acute leukemia by reverse transcriptase polymerase reaction(RT-PCR). The MUC1 positive PCR products were analyzed by digestion with pst I. With clinical observation, the relationship of expression of MUC1 gene and treatment results were done.

Results: The expression of MUC1 was positive in 36 of 73 AL patients (49.3%). The MUC1 expression rate was 39.1% in ALL and 52.2% in AML and they show no significance difference. MUC1 gene was undetectable in 23 healthy subjects. Nineteen of 21 (90.5%) MUC1-negative non-M3 acute leukemia patients got CR while 14 of 22 (63.6%) MUC1-positive non-M3 acute leukemia patients got CR which showed significance difference (p<0.05); Only 9 of 16 cases of MUC1-positive non-M3 AML patients got CR(56.3%), while 10 of 11 patients of MUC1-negative non-M3 AML got CR(90.9%). The expression of MUC1 could turn negative after CR. There was no mutation of MUC1 gene correlating with digested site of pst I.

Conclusion: MUC1 expression is observed in some acute leukemia patients. The positive expression of MUC1 is not different between AML and ALL. The AML patients whose MUC1 gene showed positive got lower CR rate. It indicates that the expression of MUC1 can be used as a poor prognostic factor in AML and a marker for detection of minimal residual disease in AL.
Upregulation of FZ5 Results in Abberant Expression of Beta-Catenin and LEF/TCF Complex in Acute Myeloid Leukemia.
Authors: Atalar Fatmahan , Hatirnaz Ozden , Erbilgin Yucel , Ozbek Ugur , Sayitoglu Muge
Published: October 2019

Wnt proteins form a family of secreted signalling proteins that play a key role in various developmental events such as cell differentiation, cell migration, cell polarity and cell proliferation. Activation of the Wnt/beta-catenin through canonical and non-canonical pathways has recently been shown to be crucial to the self-renewal of hematopoietic progenitors and stem cells. WNT proteins initiate the canonical (beta-catenin-regulated) signaling cascade by binding to seven-transmembrane spanning receptors of the Frizzled (FZ) family. WNT stimulation leads to beta-catenin accumulation, nuclear translocation and interaction with T-cell factor/lymphoid enhancer factor (TCF/LEF) to regulate genes important for embryonic development and proliferation. Also beta-catenin is thought to have a role in the self renewal potential of leukemic cells in AML. To determine the role of the WNT pathways in the development of acute myeloid leukaemia (AML), we studied seven WNT pathway genes (WNT10b, WNT5a, FZ5, Beta-catenin, APC, TCF1 and LEF1) in 34 AML patients by quantitative real-time PCR. Results were compared by peripheral blood samples from healthy donors by using Ct values. FZ5 gene expression showed 10 fold and LEF1gene expression showed 3 fold increase when compared to controls (p>0.01 and p=0.02 respectively). Beta-catenin was 5 fold higher in patients than normal peripheral blood samples. The WNT10b expression was slightly lower (1.5 fold) comparing to other genes. TCF1 expression level also seemed to be higher in AML patients (OR=4.2 95%CI=0.04–2.82, p=0.06). These results may represent that canonical WNT signaling takes place in AML development and provide the evidence for the involvement of FZ5 upregulation in the pathogenesis of AML through activation of the TCF/LEF mediated signaling.
LMP1, LMP2A and CD40 Are All Essential for EBV-Associated Lymphoma Cell Survival.
Authors: Guasparri Ilaria , Cesarman Ethel
Published: October 2019

LMP1, LMP2A and CD40 induce signals that lead to activation of the transcription factor NF-kB. Since inhibition of NF-kB leads to the apoptosis of EBV-infected lymphoblastoid

and lymphoma cell lines, we sought to determine whether elimination of one or more of these signals is sufficient to induce apoptosis of lymphoma cells expressing this proteins.

We developed and tested siRNA targeting LMP1 and LMP2A, which were transfected into two EBV positive (Type III)-lymphoma cell lines (IBL-1 and BCKN1) and two lymphoblastoid

cell lines (RPMI-8402 and LCL-9001). LMP1 or LMP2A suppression with siRNA resulted in inhibition of basal NF-kB by approximately 75% in the Type III cell

lines tested, indicating that a significant proportion of the constitutive NF-kB activity is being induced by both LMP1 and LMP2A. Simultaneous elimination of both LMP1 and

LMP2A showed no significant additive or synergistic effect. Elimination of LMP1 and or LMP2A was sufficient to induce apoptosis of EBV infected cells, as evaluated by flow

cytometry for Annexin V after suppression of these proteins. No effect was seen in LMP1 negative controls. Simultaneous suppression of both proteins resulted in similar levels of apoptosis, with no additive or synergistic effect. As EBV+ lymphoma cell lines express CD40, as well as CD154 (CD40 ligand) causing an autocrine loop, we hypothesized that inhibition of CD40 signaling may enhance the apoptotic effect induced by suppression of LMP1 and LMP2A. Treatment with blocking antibody to CD154 (CD40 ligand) in conjunction with LMP1 or LMP2A suppression increased apoptosis by approximately 20%. These data indicate that CD40, LMP-1 and LMP2A are critical for survival of EBV-infected lymphoma cells expressing these proteins. Thus CD40 represents a potential cellular target, and LMP1 and LMP2A may be useful viral targets for the treatment of EBV-associated lymphomas.
Decreased Expression of Telomere Length Regulating Factors TRF1, and Tankyrase 1 in Patients with Adult Acute Leukemia.
Authors: Qian Wenbin , Yang Xiudi , Xu Weilai
Published: October 2019

Telomeric repeat binding factor 1 (TRF1) and Tankyrase 1 (a human telomeric poly(ADP-ribose) polymerase) may play key roles in the maintenance of telomere function. TRF1 negatively regulates telomere elongation, while Tankyrase 1 acts as positive regulator of telomere length. In this study, we examined the expressions of TRF1 and Tankyrase 1 in 46 samples from patients with acute leukemia at diagnosis by quantitative reverse transcription real-time polymerase chain reaction and then analyzed the relation with the expression of protooncogene c-myc. TRF1 mRNAs were significantly down-regulated in patients with acute leukemia compared to controls. On the other hand, the mononuclear cells (MNCs) of patient with acute myelogenous leukemia (AML) expressed TRF1 at higher levels than that of patient with acute lymphoblastic leukemia (ALL) (p<0.01) or myelodysplastic syndrome (MDS). Compared to the patients with CD 34 negative AML, patients with CD34 positive AML expressed lower levels of TFR1 (p<0.01). The expression levels of Tankyrase 1 mRNA progressively decreased in patients with MDS, AML and ALL, although it was not significant. Moreover, there was a significantly correlation between the expression of TRF1 mRNA and Tankyrase 1 mRNA in AML cells (r = 0.394, p<0.01). Expression of c-myc mRNA was found gradually reduced in patients with AML, ALL and MDS. Otherwise, the expression level of TRF1 significantly correlated with those of c-myc in patients with AML(r=0.381, p=0.02) and CD34 positive AML (r=0.814, p<0.01). These results suggest that TRF1 gene expression in acute leukemia is down-regulation, and higher expression of TRF1 mRNA may be a reason for shorter telomere in some AML cells. There is dysfunction of TRF1 expression in the CD34 positive subset of AML. The patients with lower expression of TRF1 may have a poor prognostic.
Dynamic Regulation of Mis-Folded N-CoR by Aberrant Protease and Its Implication in UPR-Induced Apoptosis of APL Cells.
Authors: Khan Matiullah
Published: October 2019

We have recently reported that accumulation of mis-folded N-CoR as insoluble protein aggregates in APL cells induces Endoplasmic Reticulum (ER) stress and activates unfolded protein response (UPR). Although, accumulation of mis-folded proteins is known to trigger UPR-induced cytotoxic cell death in several neurodegenerative disorders, APL cells are notably resistant to UPR-induced apoptosis. The molecular basis for the paradoxical response of APL cells to UPR is not known. Here we report that a glyco-protease, selectively expressed in APL cells, regulates APL cells’ response to UPR-induced apoptosis through processing of mis-folded N-CoR protein. Results show that mis-folded N-CoR is cleaved selectively in APL cells, and cellular extracts of APL cells and human primary APL cells contain activity that cleaves N-CoR protein. Purification and spectrometric analysis of N-CoR cleaving activity from an APL cell line reveals that it is a glycoprotein endopeptidase known as OSGEP. Furthermore, the cleavage of N-CoR in APL cells could be blocked by the broad-spectrum protease inhibitor, AEBSF, and by RNAi-mediated down-regulation of OSGEP expression. AEBSF selectively inhibits growth and promotes apoptosis of APL cells, possibly through a mechanism involving AEBSF-induced accumulation of insoluble N-CoR protein and by triggering ER stress. Taken together, these finding suggest that selective induction of protease activity in APL cells may represent a novel cytoprotective component of UPR, which could be exploited by tumor cells to survive the toxic insult of mis-folded protein(s).
Predictive Factors of All Trans-Retinoic Acid (ATRA) Related Complications during Induction Course of Acute Promyelocytic Leukemia (APL).
Authors: Ramzi Jeddi , Karima Kacem , Hend Benneji , Samia Mnif , Raihane Belakhal , Lamia Aissaoui , Hela Benabid , Zaher Belhadjali , Balkis Meddeb
Published: October 2019

Background: The combination of ATRA and chemotherapy has made APL a highly curable leukemia. However several complications are reported with this treatement the most serious and life threatening being ATRA syndrome. We aimed at identifying factors that could predict complications caused by ATRA during induction treatement of APL.

Patients: 42 patients with comfirmed APL(by t(15,17) and/or PML/RARA) treated at our institution (University hospital of Tunis) between January 1998 and June 2006 using two consecutive protocols: European APL93 trial (24 patients) until February 2004 and Spanish PETHEMA LPA99 trial (18 patients) more recently. Induction regimen consisted of ATRA 45 mg/m2/d until CR combined to DNR 60 mg/m2/d×3 + Cytarabine 200 mg/m2/d×7 (APL93) and Idarubicin 12 mg/m2 d2,4,6,8 (LPA99). Prednisone (0,5 mg/kg d1–d15) was added if WBC > 10 000/mm3 to prevent ATRA syndrome in LPA 99.

Results: Median age was 36 yr (7–64yr), M/F=16/26(0,61), median WBC was 2400/mm3(range 600–100 000/mm3). WBC>10 000/mm3 was noted in 14 patients (33%). Additional cytogenetic abnormalities were seen in 12/42(28%). Median Body Mass Index(BMI= weight/length2:N 20–25) was 24kg/m2(range 16–40kg/m2), BMI>30 was noted in 9 patients(8F and 1M).33 patients achieved CR (78,57%):18/24 (75%) in APL93 vs 15/18(83%) in LPA99.9 patients(21,42%) had early death. Causes of early death were:ATRA syndrome(6) and CNS haemorrhage (3). Complications due to ATRA were:ATRA syndrome(10), Scrotal ulcerations(3), Sweet syndrome(2), Perineal ulcerations(1), Pseudotumor cerebri (1). Prognostic factors for complications of ATRA(Fisher exact test) were:BMI>35 (p=0,055), induction treatment without cytarabine(LPA99 trial) (p=0,047), whereas age(p=0,74), gender (p=0,51), initial WBC (p=0,47), and additional cytogenetic abnormalities were not predictive. ATRA syndrome was more reported in patients with initial WBC>10 000 (p=0,08).

Conclusion: We found high BMI (>35) in female and treatement without Cytarabine to increase the risk of developping complications with ATRA.
Pegilated Liposomal Doxorubicin in Combination with Cyclophosphamide, Vincristine, Prednisone and Rituximab in Patients with Newly Diagnosed Diffuse Large B Cell NHL: Clinical Efficacy and Related Cardiac Safety.
Authors: Geraci Leonardo , Colangelo Mauro , Ranalli Roberto , Puglielli Serina , Nenna Caterina , Celeste Franco
Published: October 2019

Pegylated liposomal doxorubicin, (CAELYX; Schering Plough) was introduced because of its low or totally absent cardiotoxicity and greater penetration in tumor tissue. Based on differences in clinical pharmacokinetics and clinical studies, the dose for PLD is lower than the standard dose for doxorubicin and epidoxorubicin. This feature makes it suitable for use in older patients and/or those with previous heart disease suffering from diffuse large B-cell NHL, by including Caelyx in 30 mg/mq doses in the classic CHOP regimen, in place of conventional doxorubicin. In this single-centre study, the endpoint were the therapeutic response and the incidence of cardiotoxicity, assessed by means of echocardiogram (LVEF),before and after chemotherapy and at a later stage following the last chemotherapy cycle. Pegylated liposomal doxorubicin was administered in a dose of 30 mg/mq in association with standard doses of Cyclophosphamide 750 mg/mq, Vincristine 1,4 mg/mq, Prednisone 100 mg/mq p.o. day 1–5, and Rituximab 375 mg/mq at 21 day intervals.The trial enrolled 24 patients, aged between 65 and 83 (average age 75 years), in stage II-IV (15 patients in stage IV, 6 in stage III and 3 in stage II); in 65% of the patients, the International Prognostic Index was high-intermediate. An average of six chemotherapy cycles were administered per patients. No dose-limiting cumulative toxicity was observed. The objective responses observed were 85%, with 70% complete responses and 15% partial responses. The average duration of the response was 14 months (1.7–41.5 months). There was no decrease in LVEF associated with administration of pegylated liposomal doxorubicin. Pegylated liposomal doxorubicin in combination with Cyclophosphamide, Vincristine and Prednisone in addition to Rituximab, was found to be active and safe in this series of patients with diffuse large B-cell lymphoma with complete remission in 70% of the cases, without this affecting cardiac safety.
Placenta Derived Adherent Cells (PDACs) Suppress Tumor Cells of Diverse Origin.
Authors: Harbacheuski Ryhor , Paludan Casper , Murray Rose Ann , Mendillo Megan , Soler Jorge , Wang Jia-Lun , Ye Qian , Hofgartner Wolfgang , Edinger James , Kaplan Gilla , Hariri Robert
Published: October 2019

The placenta is a readily available and ethically non-controversial source of large amounts of therapeutic stem cells. We isolated adherent cells from the Umbilical Cord (UC) and the Amnion Chorion (AC) of term placentas. These Placenta Derived Adherent Cells (PDACs) displayed a cell surface phenotype of CD29+, CD44+, CD73+, CD90+, CD105+, CD200+, CD34−, and constitutively secreted IL-6, IL-8 and Monocyte Chemoattractant protein-1 (MCP-1). PDACs demonstrated in vitro pluripotency and suppressed T cell proliferation in both the allogeneic mixed leukocyte reaction, and the autologous EBV regression assay. Because progenitor cells have been described to inhibit tumor growth in some systems, the role of PDACs in tumor suppression was investigated.

EBV transformed tumor cells were cultured either alone, or with AC or UC PDACs. After 6 days, live (7-AAD− Annexin-V−) tumor cells were counted on a flow cytometer. Free growing tumor cells numbered 40,000. When tumor cells were co-cultured with AC or UC PDACs, the growth was suppressed to 10,000 cells, a suppression of 75%. We designed and tested a panel of tumor cell lines that included retinoblastoma (RB), histiocytic lymphoma (HL), chronic myelogenous leukemia (CML), and colon adenocarcinoma (CAC). The tissue origin of the selected tumor cell lines included neuronal tissue (retinoblastoma), epithelium (carcinomas) and the hematopoietic lineage (lymphomas and CML). Co-culture experiments showed that the growth of all tumor lines was suppressed more than 40%. When a transwell was introduced separating PDACs and tumor cells, contact dependency of suppression was 12% for HL, 22% for CML, 42% for CAC, and 51% for EBV transformed tumor cells.

These results indicate that PDACs may have therapeutic value with respect to certain cancers.
Differentiation and Hematopoietic-Support of Clonal Cells in Myelodysplastic Syndromes.
Authors: Li Xiao , Wu Lingyun , Ying Shaoxu , Chang Chunkang , He Qi , Pu Quan
Published: October 2019

Studies were performed to obtain evidences on differentiating and hematopoieting potential of marrow clone-original cells in patients with myelodysplastic syndromes (MDS). Firstly, results of correlation analysis between bone marrow clonal cells and blast percentages for total 51 MDS showed that almost all cases had higher clonal (mean 48.2%) than blast proportion (mean 6.7%) (P<0.001). Wheras in 11 AML patients, mean clone/blasts desparity was nearly zero. Secondary, amount of clone-original individual cells were defined in mature hematopoietic cells, mean 45.9% in segmented granulocytes, 46.0% in orthochromatic normoblasts and 38.0% in mature megakaryocytes. In addition, FISH examination presented approximative proportions of clonal cells in peripheral blood for all 16 tested cases (mean 37.3% in blood vs 48.6% in marrow). Moreover, the neutrophils in MDSs peripheral blood oxidazed dihydrorhodamine123 (DHR) nearly as the same as neutrophils in normal donor’s circulations did, while obviously poor function-exerting were observed in neutrophils from AML blood. Of note, researches on clonality appeared an unexpectedly outcome that most of the typical morphological dysplastic cells possessed normal karyotypes. Our discoveries proposed that the biological features of MDS clones is obviously distinguished to AML clone, which perhaps provide some new consideration for MDS pathogenesis and treatment tactics.
G-CSF Priming before HA Chemotherapy Increased Apoptosis and Chemotaxis in Myelodysplastic Syndromes Cell Line MUTZ-1 by Activating the MEK/ERK Dependent Signal Pathway.
Authors: Cai Zhen , He Dong Hua , Hu Jie , Gao Liang
Published: October 2019

Myelodysplastic syndrome (MDS) represents a heterogenous group of clonal stem cell disorders with qualitative and quantitative abnormalities of blood cells and a high probability of evolving to acute leukemia. Intensive induction chemotherapy in order to reduce the malignant clone and reconstruct normal hematopoiesis is a classic therapy for MDS, especially high risk MDS. Homoharringtonine (HHT), one of the alkaloids from a Chinese natural plant, Cephalotaxus, incombination with cytarabine (A-rac) (HA chemotherapy) has been used successfully in the treatment of MDS. It has been recently reported that the effect of HA chemotherapy could be enhanced by granulocyte colony stimulating factor (G-CSF) (HAG chemotherapy). But the mechanism underlying the enhanced effectiveness in HAG remains unknown. We investigated the effect and mechanism of HAG chemotherapy on the apoptosis and chemotaxis in MDS cell line MUTZ-1. Cells were divided into HA group (HHT + Ara-C) and HAG group (treated with G-CSF before HA). Growth inhibition was measured by MTT method and 50% inhibiting concentration (IC50) was calculated. Cell cycle and apoptosis were measured by flow cytometry. Mitochondria membrane electric potential was determined by confocal microscopy. The mRNA expression of the chemokine receptor CXCR4 and the cysteinyl leukotriene receptor Cys-LT1 were detected by RT-PCR. The protein expression of Cys-LT1 and signaling molecular in MUTZ-1 cells was detected by western blot. The chemotaxsis of MUTZ-1 cells was detected by transwell migration assay. We found that the percentage of apoptotic MUTZ-1 cells increased and the mitochondria membrane electric potential in the cells decreased significantly more in HAG group than in HA group. The percentage in S phase of the cell cycle increased after exposure to G-CSF for 4 hours, and at its peak after 6 hours. Cys-LT1 mRNA and protein expression and chemotactic migration increased more in HAG group than in HA group, while no significant change was observed in CXCR4 expression. G-CSF activated both mitogen activated protein kinase kinase (MEK1/2) and extracellular signal-regulated kinase (ERK1/2). Chemotactic migration was partly blocked by the MEK1/2 inhibitor U0126. These results indicated that G-CSF priming before HA chemotherapy increased apoptosis and chemotaxis in MDS cell line MUTZ-1, and MEK/ERK signaling pathway is involved in G-CSF-induced chemotaxis.
Screening of JAK2 V617F Mutation in 84 Patients with Thrombocythemia and Erythrocytosis. Is Necesary for Diagnosis of a Chronic Myeloproliferative Disorders?.
Authors: Sobas Marta A. , Martinez M. , Perez-Encinas M. , Gonzalez T. , Quinteiro C. , Barros F. , Bendana A. , Ansoar E. , Rabunal M. , Gonzalez S. , Alonso N. , Diaz J. , Bello J.
Published: October 2019

BACKGROUND: The diagnosis of BCR-ABL negative chronic myeloproliferative disorders (CMPD) according to WHO/PVSG criteria may be difficult. The V617F mutation has been reported in 65–97% of patients with polycythemia vera (PV), 23–57% with essential thrombocythemia (ET) and 35–57% with idiopathic myelofibrosis (IM). The value of JAK2 mutation in unclassifiable CMPD, idiopathic erythrocytosis (IE) and idiopathic thrombocythosis (IT) is unknown.

AIMS: We wished to evaluate the utility of the V617F screening in patients with known or suspected myeloproliferative diseases.

METHODS: We applied the screening assay for JAK2V617F described by Baxter (Lancet2005;365) in 84 patients from our Clinical Department with a previous or suspected diagnosis of CMPD (retrospective and prospective cohort study). The variables were obtained from clinical records and the diagnosis were made according to WHO or PVSG criteria when was possible. There was: 14 cases of PV, 34 of ET, 14 of unclassifiable CMPD, 3 of IM, 13 of IT and 6 of secondary disorders.

RESULTS: The V617F screening was positive in 11/14 (79 %) of PV, 14/34 (41 %) of ET, 10/14 (71%) of unclassifiable CMPD, 3/3 IM, 1/13 (8%) of IT and 0/6 secondary disorders. At diagnosis, patients with mutation had significantly higher leucocyte and neutrophil counts and hemoglobin, hematocrit and LDH level. No significant interaction was noted between V617F mutation and the clinical data (gender, rates of transformation, splenomegaly, thrombosis and hemorrhage). The unclassifiable CMPD group was compared with the ET and PV groups. The unclassifiable CMPD group compared with the ET group showed a lower platelet count, a higher HCT level, and a higher frequency of JAK2 mutation.

CONCLUSIONS:

In case of suspected CMPD the JAK2 mutation screening seems to be useful . Some patients with IT had the mutation and these patients may represent an early phase of ET. According to the literature the V617F mutation was associated with some laboratory parameters but we don’t find significant clinical associations. The JAK2 mutation is frequent in the unclassifiable CMPD and these patients had clinical features similar to the PV group. The WHO or PVSG criteria should be changed to incorporate the JAK2 screening to reach a higher sensibility.
CD31 Expression and Gene Polymorphisms in Patients with B-Cell Chronic Lymphocytic Leukemia (B-CLL).
Authors: Mainou-Fowler Tryfonia , Norden Jean , Porteous Alistair , Davies Sian , Middleton Peter , Nicolle Annette , Proctor Stephen , Dickinson Anne , Summerfield Geoffrey
Published: October 2019

The CD31 antigen is expressed by a variety of cells including B lymphocytes from patients with B-CLL. The CD31 gene is polymorphic and CD31 gene polymorphisms involving single nucleotide substitutions (SNPs) such as C+373G (Leu125Val) and G+1688A (Ser563Asn) have been implicated in the pathogenesis of a number of diseases. CD31 is the physiological ligand for the CD38, a powerfull and independent prognostic marker for patients with B-CLL. A recent published study reports that the percentage of CD31 expression does not predict for poor prognosis in B-CLL (Ibrahim S, et al. Cancer.2003;97:1914–1918). A previous study showed that quantification of CD38 expression by flow cytometry significantly improves its prognostic value (Mainou-Fowler T, et al. Br J Haematol.2002;118:755–761). To re-assess the predictive power of CD31 in B-CLL, we quantified its expression and correlated its intensity with clinical outcome in a cohort of 120 patients (70M, 50F, age 47–91). The role in B-CLL of CD31 SNPs was also examined.

We report that Binet stage A patients had a significantly higher density of CD31 compared with stage B/C patients. A low CD31 predicted for poor disease outcome and identified those Binet stage A patients, and patients with low CD38, who followed a more aggressive clinical course. Disease-specific survival of patients with both low CD31 and high CD38 was significantly shorter than all other groups. Low CD31 expression significantly related with the homozygous GG and AA genotypes of the Leu125Val and Ser563Asn polymorphisms, respectively. The alleles found to be in strong disequilibrium but this was not 100%. There was no significant difference between observed and expected allele frequencies and no significant association was observed between patient genotype and clinical outcome.

Statistical comparisons are as follows: All patients:survival:high CD31 vs low CD31: P=0.0087,high CD31, low CD38 vs low CD31, low CD38: P=0.0425,low CD31, low CD38 vs low CD31, high CD38: P=0.0013,time to treatment: high CD31 vs low CD31: P=0.0064.

In stage A patients:P=0.0353,P=0.0721,P=0.0214,P=0.0716.

Distribution of CD31 expression within the genotypes: Leu/Leu vs Leu/Val, P=0.1442; Leu/Leu vs Val/Val, P=0.0466; Leu/Val vs Val/Val, P=0.0466; Asn/Asn vs Asn/Ser, P=0.1850; Asn/Asn vs Ser/Ser, P=0.0251; Asn/Ser vs Ser/Ser, P=0.0439.

In conclusion, low level of CD31 expression is an adverse prognostic indicator in B-CLL. The role of CD31 SNPs in B-CLL requires further investigation.
Bone Marrow Mesenchymal Stem Cells Exhibit a Transdifferentiation and Therapeutic Effects in a Murine Model of Orthotopic Hepatocellular Carcinoma.
Authors: Ren Jun , Jiang Hanfang , Di Lijun , Song Guohong
Published: October 2019

Background and Aim: Bone marrow stem cells can differentiate into mature hepatocytes in vitro and in vivo. Moreover, recent study shown bone marrow mesenchymal stem cells (MSCs) are the most potent component in hepatic differentiation, suggesting that the transplantation of MSCs is a promising treatment for liver disease. However, little information is available about the therapeutic potential of MSCs transplantation in cases of hepatic cell carcinoma (HCC). Here, we transplanted bone marrow-derived MSCs to testify their effects in a murine model of orthotopic HCC.

Methods:MSCs were obtained from tow male strains of β-galactosidase (β-gal) transgenic mouse(Rosa 26) and BALB/c mouse. MSCs were injected into tumor in BALB/c femal murine models of orthotopic HCC. Tumor growths were assessed by MRI on 7 days and survival rates were observed. When mouse was dying, the liver was removed from each treated mouse and evaluated by x-gal staining, and immunohistochemisty as well.

Results: MSCs transplantation increased the survival of hepatocellular carcinoma-bearing mice(25.5±4.5days verus 21.3±1.7days, p=0.025) and decreased tumor diameter slightly (7.7±2.9mm versus 9.4±2.8mm, p=0.284). MSCs transplanted directly into the tumor and/ or normal hepatic parenchyma in the same liver lobe localized mainly at the border between the tumor cells and normal liver parenchyma, induced a large area of coagulative necrosis in the tumor bed. Some engrafted MSCs were positive for albumin. There are in the carcinoma bearing BALB/c mice with MSCs implanted, whether MSCs from BALB/c mice or from Rosa 26 transgenic mice.

Conclusion: Our results suggest that the therapeutical effects of MSCs might be mediated not only by their differentiation into hepatocyte, but also mainly by they possess intrinsic antineoplastic properties.
Outcome Comparisons between Two Cord Blood Banks on Different Continents Using Identical Procedures.
Authors: Lin Allen , Yang Steven , Wang Brian , Tseng Peggy , Wu Tina , Chan Wendy , Gindy Laura , Brown Dick , Tonai Richard , Olmstead Jay , Sotomayor Gerry , Gjertson David , Petz Lawrence , Jaing Tang-Her , Tan Patrick , Chow Robert
Published: October 2019

Theoretically, cord blood (CB) banked using identical procedures by different banks should yield similar outcome if standard operating procedures (SOP) and quality programs are followed rigorously. To compare and validate the quality of the CB products, we studied whether all CB with available engraftment and/or survival transplant data from two banks, one in the USA and one in Taiwan, banked using virtually identical procedures, yielded similar transplantation outcome. Unadjusted engraftment of ANC500 were 91±3% for US (n=144) and 79±6% for Taiwan (n=49), while 1-year overall survival (OS) were 56±4% for US (n=153) vs. 68±7% for Taiwan (n=50). A number of obvious differences in transplants for the two banks were identified, for example, CB Transplants from the Taiwan CB bank (CBB) had more patients with benign diseases versus US (54% vs. 22%), which also influenced the proportion of pediatric patients and cell dose. Moreover, the Taiwan CBB had most units transplanted outside of the USA, whereas the USA CBB shipped most of its products within the USA. Another difference was that a higher proportion of CB were washed post-thaw for the USA CBB because USA transplant centers were more likely to practice post-thaw washing. We also analyzed the benign and malignant groups separately. For malignant patients, unadjusted engraftment of ANC500 were 92±3% for USA (n=114) and 73±10% for Taiwan (n=22), while 1-year OS were 50±5% for USA (n=120) vs. 40±11% for Taiwan (n=23). For benign indications, unadjusted engraftment of ANC500 were 89±6% for USA (n=30) and 85±7% for Taiwan (n=27), while 1-year OS were 66±9% for USA (n=33) vs. 92±5% for Taiwan (n=27). To adjust for some of these differences, a multivariate analysis was performed adjusting for malignancy, TNC dose, CD34+ dose, #HLA matches, recipient age, recipient sex, and washed status of CBU. Collectively, these data suggest that most of the apparent differences in outcome between the two banks may be explained by factors such as % malignant indications, and use of post-thaw wash. Overall, after adjustment for the differences between transplants using CB from the two banks, outcomes were quite similar, suggesting that if SOPs and quality programs were followed rigorously, results can be reproducible for CBB at disparate locations.
Association between Polyoma BK Virus Infection and Hemorraghic Cystitis in Allogenic Hematopoietic Stem Cell Transplantation Recipients.
Authors: Bouabdallah Kamal , Gomez Aurélie , Lafon Marie-Edith , Tabrizi Reza , Milpied Noel-Jean
Published: October 2019

BK Virus (BKV) infection is a common and benign infection occuring in childhood and resulting in viral latency. Because of immunosuppression, acute graft-versus-host disease (a-GVHD), and co infection with other viruses BKV infection often occurs after allogenic hematopoietic stem cell transplantation (HSCT) resulting, in some cases, in hemorraghic cystitis (HC).

We reviewed 258 patients who underwent allogenic HSCT in our institution between january 2001 and december 2005. One hundred and seven patients had a myeloablative conditioning regimen based on cyclophosphomide in combination with either total-body-irradiation (TBI/Cy) or Busulfan (Bus/Cy) with prophylaxis by MESNA. One hundred and fifty-one patients had a Fludarabine based reduced intensity conditioning regimen. BKV was detected by real-time PCR on urinary samples in most cases.

Macroscopic HC occured in 17 patients and BKV was detected in 15 patients out of 17. Among patients with HC and BKV viruria 11 (out of 107) received a conventional regimen compared with only 4 patients out of 151) positive for BKV in the non-myeloablative regimen group.

Although the number of patients is small, these data suggest strongly the role of the myeloablative regimen in the occurence of BKV induced HC. The high immunosuppression level induced by the conditioning regimen and the bladder injuries could explain this high frequency compared with the relative low frequency in patients with the less intensive regimen.

A prospective work is ongoing to better characterize the incidence, risk factors, clinical outcome and management of BKV induced HC after allogenic stem cell transplantation.
Preliminary Results from a Phase II Study of Lenalidomide Monotherapy in Relapsed/Refractory Aggressive Non-Hodgkin’s Lymphoma.
Authors: Wiernik Peter H. , Lossos Izidore , Tuscano Joseph , Justice Glen , Vose Julie M. , Pietronigro Dennis , Takeshita Kenichi , Ervin-Haynes Annette , Zeldis Jerome B. , Habermann Thomas M.
Published: October 2019

Background: Lenalidomide (Revlimid®), an immunomodulatory drug (IMiD®), was recently approved in the US for treatment of relapsed/refractory multiple myeloma and myelodysplastic syndromes associated with a deletion 5q[31] cytogenetic abnormality. Lenalidomide also has demonstrated activity in chronic lymphocytic leukemia and cutaneous T-cell lymphoma while thalidomide, a less potent IMiD®, has activity in non-Hodgkin’s lymphoma as both monotherapy and in combination with rituximab.

Aim: To assess the safety and activity of lenalidomide monotherapy in subjects with relapsed/refractory aggressive non-Hodgkin’s lymphoma (NHL).

Methods: Subjects with relapsed/refractory aggressive NHL following at least 1 prior treatment regimen with measurable disease are eligible. Subjects receive 25 mg lenalidomide orally once daily on Days 1–21 every 28 days and continue therapy for 52 weeks as tolerated or until disease progression. Response and progression are evaluated using the IWLRC methodology.

Results: As of July 25, 2006, 32 subjects of a planned 40 have enrolled and 31 have received drug. Twenty-two subjects are currently evaluable for tumor response. The median age of the 22 response-evaluable subjects is 65 (46–83) and 13 are female. Histology is diffuse large B-cell lymphoma [DLBCL] (n=12), follicular center lymphoma grade 3 [FL] (n=3), mantle cell lymphoma [MCL] (n=5) and transformed [TSF] (n=2). Median time from diagnosis to lenalidomide monotherapy is 2.3 (0.7–7) years and median number of prior treatment regimens per subject is 2 (1–6). Seven subjects (32%) exhibited an objective response (2 complete responses unconfirmed (CRu) and 5 partial responses (PR)), 6 had stable disease (SD) for a tumor control rate (TCR) of 59% and 9 progressive disease (PD). Responses were produced in each of the aggressive histologic subtypes studied: DLBCL (3/12), FL (1/3), MCL (2/5) and TSF (1/2). Five of 11 subjects (45%) with 1–2 prior treatment regimens had an objective response, as did 2 of 3 subjects (67%) who had received a prior stem cell transplant. Median time-to-response was 2 months. Grade 3 or 4 adverse events occurred in 18 of 31 (58%) subjects receiving drug. These were predominantly Grade 3 hematological events (neutropenia, thrombocytopenia) with only 4 subjects (13%) experiencing a Grade 4 adverse reaction.

Conclusion: Preliminary results indicate that lenalidomide monotherapy is active with manageable side effects in relapsed/refractory aggressive NHL.
Outcome of Invasive Aspergillosis (IA) in 30 Patients with Hematological Cancer (Hem-Ca): Immunity Is (Almost) Everything.
Authors: Miceli Marisa H. , Grazziutti Monica L. , Woods Gail L. , Diaz Jose A. , Krishna Somashekar G. , Kiwan Elias N. , Syed Nayyar , Barlogie Bart , Anaissie Elias J.
Published: October 2019

Background: Among IA pts receiving antifungal therapy, a strong relationship exists between outcome of IA and resolution of immunosuppression. Determining the respective contributions of immune reconstitution and antifungal therapy to response has not been attempted. Yet, treatment strategies for IA have focused almost exclusively on antifungal agents to the detriment of immune therapies. Galactomannan (GM) is an Aspergillus-specific polysaccharide released during aspergillosis and detected by the serum GM test. This FDA-approved test is an accepted diagnostic marker for aspergillosis and preliminary data suggest a correlation between GM index (GMI) and outcome.

Purpose: using GMI, to describe the contribution of immune recovery to outcome of IA in pts with hematological cancer (Hem-Ca).

Patients and Methods: From 11/03–2/06, pts at risk for IA underwent GMI screening during periods at risk. The clinical and radiological findings of pts with ≥ 2 positive (+) GMI (optical density ≥ 0.5) were reviewed.

Results: 30 pts had GMI (+) aspergillosis of the respiratory tract [myeloma 92%; median age: 59 years (27–75); 15 males]. Aspergillosis developed following stem cell transplant (SCT) [autologous (11), allogeneic (1)], or conventional chemotherapy (18). 25 pts were neutropenic (<1000/ml). The critical role of immune reconstitution was evident in 15/30 pts (50%). 7 of 25 (28%) neutropenic pts developed transient clinical and radiological worsening coinciding with immune recovery and rapid decline in GMI. All 7 responded without treatment modifications, suggesting an immune reconstitution inflammatory syndrome (IRIS). 6 pts had spontaneous clinical response with GMI normalization before commencing antifungal therapy (4 after neutrophil recovery, and 2 non-neutropenic pts after tapering immunosuppressants). 2 pts with IA refractory to prolonged and maximal antifungal therapy rapidly responded to interferon γ (1) and leukocyte transfusions (1) with GMI normalization within 48 h and subsequent clinical response.

Overall, all 3 pts who remained neutropenic died compared to 2 of 22 pts whose myelosuppression resolved and 1/5 non-neutropenic pts (p=0.005).

Conclusion: In addition to the strong association between neutrophil recovery and IA response, the critical role of immunity in IA is supported by common development of IRIS, spontaneous resolution of IA and response to immunomodulatory therapies. Daily GMI testing should be performed to accurately assess aspergillosis response. Declining GMI index obviates the need for invasive procedures and / or therapy modification. Immune therapies should be explored in IA.
Factors Associated with the Determination of Predominant Unit in Two Units Cord Blood Transplantation for Pediatric Acute Leukemia.
Authors: Kang Hyoung Jin , Koh Sang Hyeok , Jang Mi Kyung , Park Jeong Ah , Shin Hee Young , Ahn Hyo Seop
Published: October 2019

Cord blood transplantation (CBT) is a promising alternative means of allogeneic stem cell transplantation. However, limited cell doses may compromise outcome. To enhance engraftment, CBT has been conducted using two units with promising results. Our preliminary results showed the dominance of one unit which occurred early after CBT (Bone Marrow Transplant. 38:197). However, little is known about the mechanism of engraftment and factors influencing the dominancy. To improve the outcomes, CBT was performed with 2 partially HLA-matched units in pediatric acute leukemia with the analysis of factors that affect the determination of dominancy. The median age and body weight of patients (M:11, F:4) were 13 years (3–17) and 45.0 kg (10.5–62.9), respectively. Patients received various conditioning regimens according to the disease status. The median number of the infused nucleated cells by the sum of 2 units after thawing was 5.28×107/kg (0.72–15.09×107/kg). Engraftment achieved in 14/15 Patients. The median number of days required for ANC of more than 0.5×109 or 1.0×109/l were 18 days (13–31 days) and 21 days (14–34 days), respectively. Spontaneous platelet recovery to more than 20×109/l required a median 50 days (19–93 days). The early engraftment kinetics reveals that dominancy of one of two units determined from the day of engraftment (ANC > 0.5×109/l). The numbers of total nucleated cells, CD34+ cells, CD3+ cells, CFU-GM, order of infusion, and donor-recipient HLA disparities of the UCB units were viewed as factors that potentially influence dominancy. Among them infused CFU-GM per patient body weight was associated with the dominancy (P = 0.017). Interestingly, CFU-GM could not be found from each cord blood units in patient failed engraftment. Multiple factors associated with outcomes of CBT could influence the determination of dominancy, such as those factors that affect the recipient environment (underlying disease, previous treatment, conditioning regimen, use of ATG and the method of GVHD prophylaxis), the characteristics of each UCB unit and HLA disparity. For this reason, the selection of the two units requires consideration of the above factors. However optimal selection guidelines that take into account influencing factors are unavailable. CFU-GM was associated with the dominancy in our preliminary result but this kind of data is hard to get before CBT practically. Introducing the analysis of new parameters such as CFU-GM before freezing could help the selection of optimal cord blood unit and improve the outcomes with the consideration of cost-benefit.
Unrelated Donor Umbilical Cord Blood Transplantation in Children with Acquired Aplastic Anemia. Korean Experience of 12 Cases.
Authors: Kook Hoon , Baek Hee-Jo , Hwang Tai-Ju , Koo Hong-Hoe , Yoo Keon-Hee , Sung Ki-Woong , Lee Young-Ho , Seo Jong-Jin , Im Ho-Jun , Cho Bin , Kim Hack-Ki , Kim Heung-Sik , Park Jun-Eun , Kang Hyoung Jin , Shin Hee-Young , Ahn Hyo-Seop ,   for Aplastic Anemia Working Party of Korean Society of Pediatric Hematology/Oncology Cord Blood Committee of the Korean Society of Hematoipoietic Stem Cell Transplantation
Published: October 2019

Matched unrelated donor transplants have been reserved for patients with severe aplastic anemia (SAA) who fail a round of immunosuppressive therapy (IST). Despite the general acceptance of cord blood as an alternative stem cell source, unrelated donor cord blood transplantation (CBT) has not yet been recommended for SAA because of the high risk of graft failure and infectious complications. Only a few cases of successful CBT in SAA have been reported in the literature. From July, 2003 to Dec., 2005, twelve cases of unrelated donor CBTs in Korean children with acquired SAA were performed, and enrolled in this retrospective study. Two of them received double unit CBT. One patient who rejected a CBT received the second transplant with double unit CBT. All patients had no matched family donors. The median age and weight at the time of transplant were 7.0 years (2.8–18.8 years) and 23.3 kg (12.1–49.4 kg), respectively. Seven patients received previous IST including ATG/ALG plus cyclosporine (CyA). The median interval from the diagnosis to transplant was 20.5 months. The HLA discrepancy between the single unit umbilical cord blood and the patient was 0/6 in 1, 1/6 in 8, and 3/6 in 1. The conditioning regimen was heterogeneous, but included radiation in 3, and fludarabine in 3. The median infused nucleated cell dose was 3.3 × 107/kg (0.3–9.4 × 107/kg), and median CD34+ cell dose was 1.7 × 105/kg (0.4–6.6 × 105/kg) of recipient weight. Graft-versus-host disease (GvHD) prophylaxis was CyA plus methylprednisolone in 9 cases. Primary engraft failure was encountered in 3. The absolute neutrophil count ≥500/μL was achieved at 17.0 days (15–34 days), and platelet count ≥20,000/μL at 57.0 days (32–122 days), respectively. Complete donor chimerism was promptly observed in 8, with transient mixed chimerism in another patient. Acute GvHD ≥ II was observed in 5 cases, and extensive chronic GvHD was found in 2 among 8 evaluable cases. The 3-year overall survival was 74.1%, and 3-year estimated failure-free survival was 58.3%. The causes of death were sepsis, cytomegloviral pneumonitis, and chronic GvHD with intracranial hemorrhage in each patient. Cytomegaloviral disease was found in 3 cases. These results are comparable with those from unrelated donor BMT in refractory SAA. Considering the better tolerability of HLA mismatching, CBT should be considered as an alternative source of transplants. A randomized, prospective study comparing the 2nd-line IST, unrelated bone marrow transplants, or CBT is warranted to answer the best option for those who fail to 1st line IST without matched siblings.