bluuebunny/crossref_metadata_2025 huggingface dataset:
Tax Ubiquitylation and Sumoylation Control the Two Distinct Steps of NF-κB Activation.
Authors: Nasr Rihab , Chiari Estelle , El-Sabban Marwan , Mahieux Renaud , Kfoury Youmna , Abdulhay Mohammad M. , Hermine Olivier , de Thé Hugues , Pique Claudine , Bazarbachi Ali
Published: April 2020

Activation of the NF-κB pathway by the HTLV-I oncoprotein Tax plays a mandatory role in the proliferation and transformation of infected T cells. We have previously demonstrated that Tax is ubiquitylated, mostly on lysine residues located in its carboxy-terminal half. In this study, we investigated the contribution of Tax ubiquitylation on its ability to transactivate the NF-κB pathway. We show that the integrity of C-terminal lysines, and Tax ubiquitylation, are critical for Tax binding to IKK, IKK activation and nuclear translocation of NF-κB. We also report that Tax is post-translationally modified by SUMO on lysine residues also targeted by ubiquitin and that Tax sumoylation occurs in the nucleus and is required for the terminal steps of NF-κB activation. Conversely, Tax ubiquitylation and sumoylation are not involved in CREB activation. Thus, the differential ubiquitylation or sumoylation of the same lysines in Tax regulates essential events controlling the NF-κB pathway, revealing how distinct cellular modifications enrich the functions of this versatile oncoprotein.
Promoter Hypermethylation of Tetraspanin Members Contributes to Their Silencing in Myeloma Cell Lines.
Authors: Drucker Liat , Tohami Tali , Tartakover-Matalon Shelly , Zismanov Victoria , Shapiro Hava , Radnay Judith , Lishner Michael
Published: April 2020

Multiple myeloma (MM) cell interactions with their microenvironment modulate acquired drug resistance and disease progression. Indeed, reported aberrant gene methylation underscores the possible role of epigenetic events in MM’s molecular profile. Membranal tetraspanins are often inversely correlated with cancer prognosis and metastasis, however mutations were unidentified hitherto. Their promoter characteristics and frequent down-regulation conform to transcriptional silencing by chromatin remodeling. We delineated the baseline expression of select tetraspanins in MM cell lines (RPMI 8226, U266, ARP1, ARK, CAG, and EBV transformed ARH77) and fresh bone marrow samples (n=9) for the first time and determined reduced expression of CD9, CD81, and absence of CD82. Thus, we aimed to assess their promoter methylation status. Indeed, we established CD9, CD81, and CD82 promoter methylation in MM cell lines employing: Methyl-specific-PCR of bisulfite modified G-DNA; PCR of G-DNA digested with methylation sensitive restriction enzyme (Hin6I). Re-transcription of assayed genes in the cell lines following demethylation (5-aza-dC) confirmed the mechanistic significance of methylation to their regulation. Combined de-methylation and de-acetylation (TsA) induced synergistic elevation of CD82 mRNA. We conclude that chromatin remodeling contributes to tetraspanin silencing in MM.
FDG-PET Predicts Response to Fractionated Radioimmunotherapy with 90Y-Epratuzumab Anti-CD22 MAB in Patients with NHL.
Authors: Bodet-Milin Caroline , Rousseau Caroline , Campion Loic , Ansquer Catherine , Dupas Benoit , Milpied Noel , Mahe Beatrice , Chatal Jean-Francois , Wegener William A. , Goldenberg David M. , Kraeber-Bodere Francois
Published: April 2020

Objective: To evaluate FDG-PET imaging for early prediction of response in patients with NHL treated with fractionated radioimmunotherapy (RIT).

Methods: Ten patients from a larger ongoing, multicenter, Phase I/II trial of fractionated RIT (2–3 injections 1-week apart of humanized anti-CD22 antibody, epratuzumab, labeled with 90Y) underwent FDG-PET imaging together with CT scans of the chest, abdomen and pelvis at baseline and 6 weeks post-RIT, and then every 3 months until progression. Tumor responses evaluated from CT images were classified using Cheson lymphoma criteria as complete response (CR), unconfirmed CR (CRu), partial response (PR), stable disease (SD) or progression of disease (PD). PET images were evaluated for abnormal focal uptake visually, using standard uptake value (SUV) quantitation, and were classified as CR when all tumor foci disappeared, incomplete response (IR) when FDG uptake decreased with persistent foci, or PD when FDG uptake increased or new foci developed.

Results: A total of 36 paired imaging studies were obtained post RIT (including 3 patients after retreatment) and evaluated as CR (n=7), CRu (n=14), SD (n=5) or PD (n=10) by CT and CR (n= 13), IR (n= 8) or PD (n=15) by PET. Of the 14 studies evaluated as CRu by CT, 7 were definitively evaluated by PET as CR, 3 as IR, and 4 as PD. Of 22 studies not evaluated as CRu by CT, PET identified PD in one case evaluated as CR by CT and was otherwise concordant with CT (10 PD/PD, 6 CR/CR, 5 SD/IR). Among PET images acquired at 6 weeks post-RIT, the mean time-to-progression (TTP) was 9.6 months for negative PET images (CR) compared to 4.1 months for positive PET findings (IR, PD) (P=0.16).

Conclusion: In our study, FDG-PET appeared superior to conventional CT in evaluating response to fractionated RIT. For CT scans frequently evaluated as CRu, PET resolved uncertainty regarding residual disease, and PET images acquired 6 weeks after RIT predicted later relapse.
Adoptive Immunotherapy with Donor Allodepleted T Cells.
Authors: Dal Cortivo L. , Mahlaoui N. , Picard C. , Neven B. , Andre-Schmutz I. , Luby J.M. , Martinache C. , Paquet T. , Ledeist F. , Fischer A. , Cavazzana-Calvo M.
Published: April 2020

Allogeneic hematopoietic stem cell transplantation (HSCT) is the treatment of choice for many hematological malignancies or inherited disorders. In the partially incompatible HLA setting, ex vivo T-cell depletion of the graft and post-transplantation immunosuppression effectively prevent development of graft-versus-host-disease (GVHD), but lead, in turn, to a delay in immune reconstitution and a concordant increase in the rate of incidence of opportunistic infections and disease relapse. In order to selectively deplete T cells responsible for GVHD, we have designed an ex vivo procedure to eliminate CD25 expressing alloactivated donor T cells. In a first phase I/II study based on the use of an anti-CD25 immunotoxin (IT) 15 pediatric patients with acquired or congenital hematopoietic disorders who had received HSCT were infused with 1 to 8x105 allodepleted T cells/kg. No cases of severe acute (>grade II) GVHD occurred. aGVHD of grade I or II, which developed in four patients, correlated with anti-host residual proliferation above 1% in a mixed lymphocyte reaction. Evidence for early T cell expansion was shown in 3 patients with ongoing viral infections. In an attempt to improve allo-depletion and infuse larger numbers of cells, we have developed a new method based on immuno-magnetical depletion of CD25 expressing cells. In pre-clinical tests, this method lead to a more specific and efficient allo-depletion than that obtained with IT. This new procedure is currently used in a phase I/II clinical trial that will include 25 pediatric patients with inherited disorders of the immune system. 3x105 to 5x106 allo-depleted donor T cells/kg will be infused, with three to five patients per dose. Allodepleted donor T cells are cryopreserved until the obtention of quality control results Inclusion, i.e. infusion of allodepleted T cells is performed only if:1) engraftment is proven, 2) absence of GVHD 3)residual antithymoglobulin <0.2microg/ml,4) allodepletion inhibition more than 99% of residual proliferation against host cells in a mixed lymphocyte reaction. Included patients will be stratified in two groups depending on the presence or absence of a viral infection with clinical symptoms. Toxicity is evaluated by occurrence of aGVHD above grade II. Efficacy is measured by CD4+CD3+ T cell counts in the blood two months post HSCT. A real time bayesian statistical approach will allow to test both criteria as primary objectives and to increase or decrease the dose according to the results obtained. Two patients have been so far included in the “infected” group and infused with 3x105 allodepleted T cells/kg. One patient developed grade II cutaneus GVHD. For both, efficacy was reached, since they presented more than 200 CD4+CD3+ T cells /mm3 as early as 60 days post HSCT. These preliminary results are encouraging but need to be further confirmed.
Clinical Outcomes in Adult T Leukemia/Lymphoma: Report of 55 Cases from Peru.
Authors: Beltrán-Gárate Brady E. , Carrasco-Yalan Antonio A. , Lopéz-Odría Olga M. , Riva-Gonzales Luis , Ríos Hugo , Chumpitáz-Anchiraico Gloria , Falcón-Lisarazo Silvia , Navarro-M Juan , de Mendoza Fernando Hurtado , Castillo-Aguirre Jorge
Published: April 2020

Adult T -cell leukaemia/lymphoma (ATL) is an aggressive disease associated with human T-cell lymphotropic virus type-I (HTLV-I) with heterogeneous clinical presentation and outcomes, described in Southern Japan, Europe, Caribbean and previously on Pacific coast of South America including Peru (EHA 2001, abst. 304). Shimoyama’s ATL classification (BHJ1991:79) includes four types: acute, lymphomatous, chronic and smoldering; recently a new clinical type cutaneous had been described (BJD2005:152). Herein we show our experience in ATL in our institution between October 1997 and May 2005 All our 55 cases shown positivity to HTLV-I Western Blot test; immune-histopathology, blood smears and flow cytometry showed mature T-cell lymphocyte. Median age at diagnosis 61 years old (range 23–84), female/male ratio: 1.2. Thirty-one (56%) patients had ECOG status performance at diagnosis ≥ 2. Clinical types: acute (n=26), lymphomatous (n=22), smouldering (n=2), cutaneous (n=5) with no chronic type observed. Median haemoglobin diagnosis was 12.1 g/dl (range 6.9–17), median albumin was 3.2 g/dl (range 1.8 – 4.9), median beta-2 microglobulin was 4.6 g/dl (range 1.5 – 16.9), median globulin was 2.7 g/dl (range 1.5 – 8.2) and DHL was 796 UI/ml (range 331 – 13000). Twenty-five per cent (9/36) debuted with hypercalemia (acute type=7, lymphomatous=2). Acute ATL showed median leukocytes of 48,900 cell/mm3 (range 6830–259,000). IPI risk score was: low (n=6), low intermediate (n=7), high intermediate (n=16) and high (n=27).

Treatment for acute ATL was based on acute leukaemia and lymphoma regimens without any response but one, interestingly this patient was treated with Fludarabine 25 mg/mt2 day 1–5 each 28 day for 6 cycles and got complete remission. Twenty-one over 24 lymphomatous ATL type were evaluable for treatment response: overall response 38% (9/24) with 26% complete response. Smouldering and cutaneous ATL types received mainly topic treatments. Stratify analysis for IPI, DHL, beta-2 microglobulin, globulin and albumin for acute and non-acute ATL did not show any statistic difference. Median overall survival was: acute type (2.0 months DE 0.2), lymphomatous type (10.2 months DE 3.6), smouldering type (17.2 months) and cutaneous type (36.2 months DE 19.5) (Log Rank 30,76 p=0.0)

ATL persist is a poor prognostic peripheral T-lymphocytic malignancy with bad clinical and therapeutically outcomes mainly in the acute type. Cutaneous type seems to be less aggressive.

Figure Figure
Anti-CD20 Monoclonal Antibody (Rituximab) for Therapy of T-Cell/Histiocyte-Rich Large B-Cell Non-Hodgkin Lymphoma.
Authors: Park EunSil , Lim Jae Joung , Kang Hyoung Jin , Shin Hee Young , Ahn Hyo Seop
Published: April 2020

T-cell/histiocyte-rich large B cell lymphoma (THR-BCL) is morphologic variants and subtypes of diffuse large B cell lymphoma (DLBCL). Four immunomorphologic criteria were applied to distinguish THR-BCL from DLBCL.

a diffuse or vaguely nodular pattern of neoplastic infiltrate, the presence of scattered large atypical lymphoid cells that are of B-cell phenotype, predominance of a reactive background infiltrate, composed of both T-cell and non-epithelioid histiocytes, minimal presence of small B-cells in neoplastic areas.

Bcl-6 protein expression, which was generally accepted markers of germinal center differentiation, was noted more than half of the THR-BCL cases. THR-BCL predominantly affected middle-aged men who present with advanced disease and an unusually high percentage of bone marrow (BM) involvement. Here we report a pediatric case of THR-BCL. 14-year-old boy was presented with both cervical mass which was observed 1 month ago. He complained generalized weakness and chest discomfort had fever. Cervical lymph node and BM biopsy were taken. Laboratory finding was as follows; Hemoglobin 10.4 mg/dL, hematocrit 31 %, leukocyte 6,730/μL, platelet 219,000/μL. Na 128 mmol/L, K 4.6 mmol/L, Cl 95.4 mmol/L, BUN 10 mg/dL, Cr 1.2 mg/dL, LDH 2,108 U/L. Lymph node specimen showed diffuse large B cell lymphoma and numerous histiocytes. Immnuohistochemical staining; CD 20(+), CD 10(−), CD 3(+), CD 68(+). Ann Arbor stage is IVB, international prognostic index is high-intermediate risk. He received induction chemotherapy with prednisolone, vincristine, cyclophosphamide, doxorubicin, rituximab and achieved remission and now he have continued chemotherapy for 7 months in state of complete remission. THR-BCL is a distinct clinicopathologic entity within DLBCL that is characterized by an aggressive behavior and high risk of treatment of failure. That is the reason why delineate this subgroup. Experimental therapeutic strategy was indicated in patients who relapsed after having obtained a complete remission or had partial response after initial chemotherapy.
Histone Deacetylase Inhibitors Induce Growth Inhibition, Cell Cycle Arrest and Apoptosis in Mantle Cell Lymphoma and Reduce VEGF Production.
Authors: Heider Ulrike , Kaiser Martin , Zavrski Ivana , Sterz Jan , Jakob Christian , Fleissner Claudia , Possinger Kurt , Sezer Orhan
Published: April 2020

Mantle cell lymphoma (MCL) is a B cell lymphoma characterized by the translocation t(11;14)(q13;q32) and an overexpression of cyclin D1. Since current therapy regimens are not effective enough, novel treatment strategies are urgently needed. Histone deacetylase (HDAC) inhibitors have emerged as a new group of agents effective in several human malignancies. This study evaluated the effects of HDAC inhibitors on mantle cell lymphoma.

Three human mantle cell lymphoma cell lines (JeKo-1, Hbl-2 and Granta-519) were exposed to different concentrations of the HDAC inhibitors sodium butyrate and SAHA. MTT assays showed a reduced survival of cells after exposure to these agents. After 48 hours of incubation, IC50 of viability was noted at 1.0μM in JeKo-1, at 3.9μM in Hbl-2 and at 4.3μM in Granta-519. For sodium butyrate, IC50 was observed approximately at 1.5mM in JeKo-1, 1.9mM in Hbl-2 and 11.2mM in Granta-519 after 48 hours. Annexin-V staining showed that all of the SAHA and sodium butyrate-treated cell lines underwent specific apoptosis in a time and dose dependent manner. Cell cycle analysis revealed a decreased proportion of cells in S phase and increased proportion of cells in the G0/G1 and/or G2/M phases after treatment with HDAC inhibitors. To evaluate the effects of HDAC inhibitors on cell cycle regulators, the protein levels of p21, p27, and cyclin D1 were analyzed. SAHA increased p21 in all three cell lines. Expression of p27 was enhanced in JeKo-1 and Granta-519, while it remained constant in Hbl-2. Treatment with sodium butyrate increased the expression of p21 in JeKo-1 and Hbl-2, while in Granta-519 no effect was noted. The expression of p27 remained constant in all cell lines after exposure to sodium butyrate. Incubation of cells with both HDAC inhibitors lead to a downregulation of cyclin D1. In addition, we examined the influence of SAHA on the production of the angiogenic cytokine VEGF. Interestingly, treatment with SAHA led to a decrease of VEGF secretion in comparison to untreated cells. Based on these findings, we provide evidence that HDAC inhibitors have antiproliferative effects in mantle cell lymphoma and may represent a future therapeutic option for this disease.
Lack of Lipid Raft Domain Reorganization Following Ligation of CD20 Is Associated with Resistance to Rituximab-Induced Complement-Dependent Cytotoxicity.
Authors: Olejniczak Scott H. , Hernandez-Ilizaliturri Francisco J. , Clements James L. , Czuczman Myron S.
Published: April 2020

The chimeric monoclonal antibody (mAb) rituximab has previously been shown to induce cell death upon binding surface CD20 of normal and malignant B-cells by three distinct and complementary mechanisms:

complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), and direct apoptosis.

The contribution of each of these mechanisms to the clinical efficacy of rituximab observed in lymphoma patients has yet to be fully elucidated. While laboratory and clinical data supports a role for CDC, ADCC, and/or direct signaling in rituximab activity, a better understanding of individual mechanisms and their interaction is necessary to develop strategies to improve rituximab’s biological activity or overcome resistance. Several previous studies have demonstrated that rituximab has the ability to induce lipid raft domain (LRD) polarization and stabilization of CD20 in LRDs. The contribution of LRD re-organization to complement activation following rituximab binding to surface CD20 was studied in rituximab-resistant B-cell NHL cells and MOLT-4 T-cell lymphoma cells transfected with wild-type and truncated forms of CD20. Rituximab-resistant cell lines (RRCL) were generated from well characterized B-cell lymphoma cell lines (Raji, RL, SU-DHL-4) by exposing them to increasing concentrations of rituximab or rituximab plus human serum as a source of complement. Individual clones were then isolated by limiting dilution from RRCL and the parental cell lines from which they were derived. Standard 51Cr-release assays were used to measure CDC-associated cell lysis following exposure of cells to rituximab in the presence of human serum. Following repeated exposure to increasing concentrations of rituximab (alone or in the presence of human serum) most clones were found to be significantly more resistant to rituximab-induced CDC than parental cells from which they were derived. Rituximab-sensitive cells were found to rapidly polarize CD20-containing lipid raft domains following rituximab binding whereas resistant cells demonstrated significant impairment of this process. Upon chemical disruption of lipid rafts in sensitive cells, the ability of rituximab to induce CDC was significantly reduced. Additionally, MOLT-4 T-cells transfected with truncated forms of CD20 were shown to have a reduced ability to polarize into lipid rafts following anti-CD20 binding and to have a significant loss of rituximab-induced CDC. Taken together these data suggest that lipid raft polarization following rituximab binding to wild-type CD20 is a required event in the induction of CDC and that impairment of lipid raft polarization is a potential mechanism contributing to rituximab resistance.
Molecular Characterization of a Novel Subtype of Pediatric Acute Lymphoblastic Leukemia.
Authors: Mullighan Charles G. , Ross Mary E. , Zhou Xiaodong , Dalton James , Behm Fred G. , Shurtleff Sheila A. , Downing James R.
Published: April 2020

Gene expression profiling is a powerful tool to classify and predict subtypes of pediatric acute lymphoblastic leukemia (ALL). In addition to accurately classifying known subtypes of ALL, microarray analysis of 327 cases of ALL at our institution using Affymetrix U95 arrays identified a previously uncharacterized novel subgroup of 14 cases of B-ALL with a gene expression signature distinct from other leukemias, suggesting a unique pathogenesis. These cases lacked a recurring cytogenetic anomaly, showed a high frequency of aberrant expression of CD2, lacked molecular markers of other subtypes of ALL, and had a favorable prognosis. We have used several complementary candidate and genome-wide approaches to identify the molecular basis of this novel subtype. Gene expression profiling using Affymetrix U133A and B arrays was used to refine the signature, and when applied to an additional 36 cases of cytogenetically normal B-ALL, 5 cases were identified that share the novel gene expression profile. Intriguingly, the tyrosine kinase PDGFRA was overexpressed in the majority of the novel cases, with 6 novel cases showing exceedingly high levels of PDGFRA expression, which was confirmed by western blotting and flow cytometry. Unexpectedly, 3 of these cases harbored the FIP1L1-PDGFRA fusion characteristic of idiopathic hypereosinophilic syndrome. However, FISH examination demonstrated that this fusion was present in a minor subclone only. RT-PCR and FISH approaches failed to identify alternative PDGFRA fusions, and PDGFRA sequencing failed to identify evidence of activating mutations. Furthermore, in unstimulated cultures or following PDGF-AA stimulation, PDGFRA-overexpressing ALL blasts were not sensitive to imatinib (Gleevec). Other genes in the gene expression profile with potential roles in leukemogenesis, include include the B-cell receptor signaling mediator BRDG1 and the tyrosine phosphatase PTPRM. Using FISH and genomic sequencing of multiple candidates, no potentially leukemogenic abnormality has been identified. To determine if gene amplification, deletion, or copy-neutral loss of heterozygosity is characteristic of this subtype of ALL, genome-wide array-based comparative genomic hybridization and Affymetrix 100K single nucleotide polymorphism arrays were performed on all cases. Deletion of the CDKN2A/2B locus, and deletion of a region at 8p22–23.1 flanking the SPAG11 locus were each found in multiple cases, however no single unifying abnormality was identified. In summary, we have identified and characterized a novel subtype of B-ALL with unique gene expression signature, high incidence of CD2 and PDGFRA overexpression, and favorable outcome. Although no uniform causal molecular lesion has been identified, FIP1L1-PDGFRA fusion may represent one mechanism of PDGFRA overexpression. Further studies and analysis of additional cases may prove informative.
Histamine Monitoring in Patients with CML during Treatment with Imatinib: Comparison to Monitoring by Cytogenetics and BCR/ABL.
Authors: Agis Hermine , Herndlhofer Susanne , Semper Hans , Pirc-Donoewenater Hendrati , Haas Oskar , Mannhalter Christine , Esterbauer Harald , Sperr Wolfgang R. , Geissler Klaus , Valent Peter
Published: April 2020

The tyrosine kinase inhibitor STI571=imatinib is highly effective in the treatment of chronic myeloid leukemia (CML). Although most patients (pts) show a complete cytogentic response (CCR) to this tyrosine kinase inhibitor, drug resistance may occur. Therefore, monitoring of minimal residual disease (MRD) during imatinib-therapy is of clinical importance. However, most MRD-parameters require special technology and equipment and are expensive. We found that histamine levels are highly upregulated at diagnosis in pts with CML (controls, n=39: median, 55.2 ng/ml, range, 19.1–108 ng/ml vs CML, n=44: median, 5,684.5 ng/ml, range, 181–47,816 ng/ml; p<0.0005) and correlate with the presence of basophils (R=0.87). To define the value of whole blood histamine as a marker of MRD in CML, histamine levels were determined serially by RIA before and during treatment with imatinib in 80 pts with CML (chronic phase, CP, n=70; accelerated phase, AP, n=10). Of the pts with CML-CP, 29 were previously untreated, whereas 41 pts had received interferon-alpha or a bone marrow transplant prior to imatinib. From the 10 pts with CML-AP, 3 had received previous interferon-alpha. Imatinib was given at a dose of 400 mg/day (CML-CP) or 600 mg/day (CML-AP and CP pts who did not respond adequately to 400 mg/day) orally. The rate of complete cytogenetic response (CCR) amounted to 77.6%, the rate of major cytogenetic response was 80%. Blood histamine levels decreased significantly in CML pts during treatment with imatinib, and returned to normal levels in all pts with CCR. In most pts, loss of CCR was accompanied or was preceded by a recurrent increase in histamine as well as an increase in BCR/ABL determined by light cycler-based real time PCR. The pts who did not enter CCR exhibited higher histamine levels and higher levels of BCR/ABL compared to those with continuous CCR (p<0.05). Unexpectedly, whereas the numbers of basophils were found to correlate well with histamine levels during therapy with imatinib (R=0.96), no correlation was found between histamine and Ph+ metaphases (R=0.34) or histamine and percentage of BCR/ABL (R=0.14), suggesting that basophil-histamine is an independent variable. Basophil-histamine should thus be considered as a simple and reliable additional marker to monitor MRD in pts with CML.
Subcutaneous (SC) Homoharringtonine (HHT) for Patients (pts) with Chronic Myelogenous Leukemia (CML) in Chronic Phase (CP) after Imatinib Mesylate Failure.
Authors: Quintas-Cardama Alfonso , Cortes Jorge , Verstovsek Srdan , Laddie Nakia , Estrov Zeev , Kantarjian Hagop
Published: April 2020

HHT is a cephalotaxus alkaloid obtained from an evergreen tree that inhibits the synthesis of proteins leading to apoptosis. HHT by intravenous (IV) administration has demonstrated significant activity in pts with CML after failure to interferon (IFN) therapy, with cytogenetic (CG) responses in 30% of cases. A phase I study of SC HHT previously reported in pts in accelerated or blast phase (before imatinib was available), demonstrated efficacy (5 of 10 responses) and adequate tolerance at the same doses used IV. We thus started a phase II study to determine the activity of SC HHT in pts with CML in late CP after imatinib failure. Therapy with HHT consisted of an initial IV loading dose of 2.5 mg/m2 over 24 h, followed by 1.25 mg/m2 SC twice daily for 14 days every 28 days. Six pts have been treated and 5 are evaluable (1 pt lost to follow-up after 1 cycle). Best response to prior imatinib therapy for a median of 64 weeks (range, 12 to 96) was: complete hematologic response (CHR) in 3 pts and complete CG response (CG CR) in 2, and primary resistance in 1 pt. All 5 evaluable pts had also failed IFN, and other therapies: Ara-C (n=2), farnesyl transferase inhibitors (n=2). At the time HHT was started, all had lost CHR. Median age was 52.5 years (range, 36 to 66), time from diagnosis to treatment 58 months (18 to 164), WBC 24.35 x 109/L (6/4 to 43), platelets 308 x 109/L (147 to 536), and peripheral basophils 4/5% (3% to 17%). Mutations of the BCR-ABL kinase domain were identified in 2 pts (2 mutations each: G250E and Y253H in one, D276G and F359I the other). Pts received a median of 4.5 courses of SC HHT (range, 1 to 5). CHR was achieved in all 5 evaluable pts. Of these, 1 achieved a CG CR and a minor CG response (lowest Ph 80% and 90%, respectively). The pt that achieved CG CR had D286G and F359I mutations before therapy with HHT. Interestingly, this pt developed extramedullary blast disease (bones) that required radiotherapy while remaining in CG CR in peripheral blood and bone marrow. All pts developed myelosuppression and 3 had their HHT dose reduced due to prolonged neutropenia. Grade 3–4 non-hematologic toxicity included neutropenic fever (n=1), and fatigue (n=1). We conclude that SC HHT is well tolerated and may have clinical activity in pts with CML after imatinib failure. This study was terminated because of drug availability from this source; a new trial will be conducted obtaining HHT from a different sponsor.
Imatinib Mesylate in the Treatment of Relapse of Chronic Myeloid Leukemia after Allogeneic Stem Sell Transplantation.
Authors: Zhu Kanger , Zhang Tao , Chen Jie , Zhong Juan
Published: April 2020

Objective: To determine whether imatinib mesylate is effective against chronic myeloid leukemia (CML) that relapses as CML in chronic phase following allogeneic stem cell transplantation (allo-HSCT).

Methods: We studied 5 patients treated with imatinib mesylate for recurrent CML after allo-HSCT. 2 patients relapsed as CML in chronic phase with hematological evidence of relapse, and one of whom was refractory to donor lymphocyte infusion (DLI). The remaining 3 patients relapsed as CML in chronic phase with cytogenetic evidence of relapse. The patients were followed for hematological and cytogenetic response.

Results: all of 5 patients achieved hematological and cytogenetic response within 5 months.The 2 patients with hematological evidence of relapse and minimal level of donor chimerism experienced severe pancytopenia, followed shortly by a hematological recovery, disappearance of BCR-ABL fusion gene positive cells, and conversion of complete donor chimerism. In 3 patients, a hematological and cytogenetic response was associated with chronic graft-versus-host disease. With a median follow-up of 36 (12~46) months, after cessation of imatinib mesylate, all patients are alive and remain in complete cytogenetic remission.

Conclusion: We have shown that patients who relapsed as CML in chronic phase after allo-HSCT showed very good therapeutic responses to imatinib mesylate, which were rapid, durable, with conversion to full donor chimerism and complete disappearance of BCR-ABL fusion gene positive cells, in spite of a prolonged relapse, an apparent loss of donor chimerism, and a failed prior response to DLI.
Pregnancy during Treatment with Imatinib: A Case Report.
Authors: Fogliatto Laura , Brum Caroline , Zago Alethea , Faltoe Thiago , Rodrigues Fabio , Falhauber Gustavo , Leugeur Denise , Silla Lucia
Published: April 2020

CFSA, an 18-year-old woman with a history of weight loss and abdominal discomfort was diagnosed with chronic myelogenous leukemia (CML) in December 2002. The WBC were 440 x 109 /L with 25% neutrophils, 20% bands, 9% metamyelocytes, 9% myelocytes, 8% promyelocytes, 10% basophils, 9% eosinophils, 1% lymphocytes and 8% blasts. The hemoglobin was 9,8 g/dL and platelet count 416 x 109/ L. A bone marrow examination demonstrated granulocytic hyperplasia and the presence of Philadelphia chromosome in 100% of the metaphasis. A diagnosis of CML was made and she was started on hydroxyurea and imatinib 600mg daily. No matched HLA donor was found. There was a partial hematological response. In February 2005 on imatinib (600 mg) she was pregnant and an ultrasound scan showed a viable fetus of 17 weeks. The WBC were 22 x 109 /L with 50% neutrophils, 23% bands, 8% metamyelocytes, 12% myelocytes, 1% promyelocytes, 5% lymphocytes, hemoglobin 11,4 g/dl and platelet count 290 x 109. Imatinib was stopped and she remained off treatment during pregnancy. Cesarean section was performed at 38th week because of preeclampsia (hypertension, headache and scotomas). A healthy girl weighing 2980 g, 46,5 cm in length with Apgar score of 9 was delivered. The infant’s physical examination, WBC, hemoglobin, platelet count and cranial ultrasonography were normal. The baby is presently healthy with no developmental abnormalities. There are few reports on the use of 600 mg a day of imatinib and pregnancy. It is of notice, although probably unrelated, the event of preeclampsia. The effects of imatinib on fetus are unkown and should be considered teratogenic until more studies or cases are reported.
STI 571 Combined with Hematopoetic Stem Cells Transplantation for Chronic Myeloid Leukemia.
Authors: Meng Fan Yi , Sun Jing , Liu Qifa , Xu Dan , Wei Yongqiang , Zhang Yu , Jiang Qianli
Published: April 2020

Objective: to study the efficacy of STI 571 combined with allogenic hematopoietic stem cell transplantation (Allo-HSCT) or autologous peripheral blood stem cell transplantation (APBSCT) in chronic myeloid leukaemia (CML).

Methods: 18 CML patients were divided into 2 groups: group A (14) and group B (4). Patients in group A including 10 in CML accelerating phase or blast crisis phase, 4 in chronic phase. They all underwent Allo-HSCT and the median duration of STI 571 treatment before Allo-HSCT was 25 days (7–60d). 9 had related complete HLA-matched donors and 5 had unrelated complete matched donors, the preparation regimen was TBI+CTX +VP16 or BU/CY ±ATG, routine protocol was used to prevent graft versus host disease (GVHD). Patients of group B were all in chronic phase, the median duration of STI 571 treatment before APBSCT was 5.5 months (4–26d), and bcr/abl detected by FISH was continuously negative. Mobilization protocol was CAE + G-CSF, 3 of 4 underwent TBI+ CY+ VP16 followed by APBSCT.

Results: in group B, PBSC were separated after 5 days of mobilization, and (3.9–9.6)×106/kg Cd34+ cells were obtained, however FISH- bcr/abl positive cells in separation products were higher than in bone marrow cells before separation (0.8% vs 2.8%). After a median follow up of 24 months (18–28 months), 2 cases relapsed, and one remains FISH-bcr/abl negative. In group A, 8 had GVHD, and after a median follow up of 8 months (4–20 months), 2 cases relapsed, 2 died of transplantation- related complications, 1 died of relapse, 9 remained disease free. Hematopoiesis was reconstituted in 8–21 days in patients of group A.

Conclusions: no obvious side effect was observed in STI 571 combined hematopoietic stem cell transplantation in CML patients.
Prognostic Factors for Allotransplantation in CML.
Authors: Copelan Edward C. , Elder Patrick
Published: April 2020

The role of hematopoietic cell transplantation in the treatment of chronic myelogenous leukemia (CML) has changed dramatically as the effectiveness of Gleevec has become clearer. The use of transplantation as first-line therapy and the timing of transplantation in those who do not achieve cytogenetic remission varies between centers. In order to better understand the likelihood of success following transplantation, we have analyzed prognostic factors in 126 patients with chronic phase CML who underwent allogeneic transplantation between 1984 and 2004. Median age was 37 (range 14–66). There were 69 males and 57 females. The interval between diagnosis and transplantation was most closely associated with survival in multivariate Cox analysis (P<.001). Interval was significant when analyzed as a continuous variable; also patients transplanted within 3 months experienced significantly longer survival than those > 3 months and patients less than 1 year from diagnosis had better survival than those more than 1 year from diagnosis. (P<.001). Patients with fully matched sibling donors experienced significantly better survival that those with other donors (P=.01). Treatment with Interferon or Gleevec, age, use of peripheral blood cells vs marrow, and year of transplant were not significant prognostic factors. There is no evidence that age, which is often heavily considered in determining timing of transplantation, is a significant prognostic factor in patients with CML undergoing allogeneic transplantation after busulfan and cyclophosphamide.

Because interval between diagnosis and transplantation significantly affects outcome, allogeneic transplantation should be considered early in patients receiving Gleevec when an adequate cytogenetic response is not achieved or at the first sign of relapse. Earlier identification of failure, e.g. using quantitative PCR for Bcr-Abl, might improve outcome and should be studied.
Treatment of Anemia in Myelodysplastic Syndrome (MDS) with Darbepoetin-Alpha (DPO) and Granulocyte Colony Stimulating Factor (G-CSF).
Authors: Rose Shelonitda S. , Ali Yaqoob , Maffei Barbara , Saidi Parvin
Published: April 2020

MDS is a clonal stem cell disorder characterized by ineffective hematopoiesis and peripheral cytopenias. Anemia is often severe, transfusion dependent and associated with impaired quality of life. Evidence shows that treatment with erythropoietin (EPO) has an overall response rate of 20% and with G-CSF becomes more effective with an erythroid response rate of around 40%. Studies have independently demonstrated a synergistic effect between EPO and G-CSF on the anemia of MDS. DPO, a modified form of EPO with a longer half life has been shown to be effective in correcting anemia in a significant proportion of MDS patients. Synergy between DPO and G-CSF has not been studied. We report the clinical course and changes in the blood counts in a 77 year old male diagnosed with MDS, Refractory Anemia with Excess Blasts Type I (RAEB-I) who received DPO and G-CSF combination treatment. His baseline CBC showed WBC-2.4 ×103/μL, Hb-9.7g/dL, and Plt-106×103/μL. EPO level was 64.4 mU/mL. He was initiated on EPO 20,000 units weekly (QW) which was titrated to 40,000 units QW after one month. His Hb increased to 12 g/dl. For convenience, he was switched from QW EPO injections to DPO 100 μg every two weeks (QOW). This was followed by an initial drop in the Hb to 11g/dL requiring an increase in DPO dose to 200 μg QOW which was followed by stabilization and maintenance of Hb between 10–11 g/dL. After 7 months, Hb dropped to 9 g/dL requiring an increase in DPO to 300 μg QOW. Hb increased to 10 g/dl and remained between 10–10.5 g/dL for 7 months. Hb dropped again to 8.3g/dL. The DPO dose was increased to 300 μg QW, and he was given a course of IV iron for a total of 1 gm. Following transient Hb increase to 9–10g/dL, for few months Hb dropped to 8g/dL and he became transfusion dependent. In addition, he became progressively neutropenic with ANC <500/μL and thrombocytopenic with Plt <50×103/μL. At this point he was initiated on G-CSF 480 μg TIW along with DPO 300 μg QW. One month after the initiation of DPO plus G-CSF his CBC showed WBC 4.0×103/μL, Hb-11.2 g/dL and Plt-92×103/μL. The Plt response was transient and eventually remained stable between 50–60 ×103/μL. He continues to remain transfusion independent after 8 months of combination treatment maintaining Hb between 10–11 g/dL. In addition to having a major erythrocyte and neutrophil response following the initiation of combination treatment of DPO and G-CSF, this patient also had transient Plt response. The course of this patient suggests that the addition of G-CSF to DPO may improve anemia and neutropenia in patients with MDS who do not have a sustained response to DPO alone. In addition it also suggests that in-vivo synergy may exist between DPO and G-CSF for the production of red cells in MDS.
Oral Prednisolone Produces a Durable Response in Pediatric Myelodysplastic Syndromes (MDS).
Authors: Chandy Mammen , George Biju , Mathews Vikram , Viswabandya Auro , Bajel Ashish , Srivastava Alok
Published: April 2020

Treatment options for children with MDS are limited except for allogeneic bone marrow transplantation. We present our experience with the use of long duration corticosteroids in children with MDS. Fifty five children (age< 15 years) with pediatric MDS or myelofibrosis treated between 1991–2004 with corticosteroids were analyzed. All of these patients presented with cytopenia involving one or more lineages and a hypercellular marrow with dysplasia and increased reticulin. Prednisolone was started at 1 mg/kg daily or on alternate days for a period of 1–2 months. If there was no response, steroids were rapidly tapered and stopped. In children who showed a response, the steroids were slowly tapered to alternate day steroids and stopped over a period of 6–12 months. In patients who had relapse of symptoms on tapering, the dose of steroids was increased to previous levels and then tapered at a much slower rate. The median age was 3 years (range: 3 months – 14 years) with 36 males and 19 females. Forty six has pediatric MDS while 10 had pediatric myelofibrosis. The median duration of symptoms prior to starting steroids was 7 months (range: 1–24). Forty one patients (74.5%) showed a response with 31 (56.4%) showing a complete response while 10 patients (18.1%) showed a partial response with improvement in counts and transfusion independence. Fourteen patients (25.5%) showed no improvement and continued to be transfusion dependant. Fourteen of the 41 patients (34%) who responded to steroids relapsed on tapering of steroids but showed response to repeat courses of steroids. The median duration of steroid use in patients who have responded is 12 months (range: 1 – 96). None of the patients had any serious side effects including infections, hypertension or diabetes except one patient who developed disseminated tuberculosis 12 months after stopping steroids. Nine patients have shown progression of disease while 1 patient developed features of SLE 24 months after diagnosis of MDS. At a median follow up of 34 months (range: 4– 120), 32 patients (58%) are alive with 27 (49%) in complete remission [14 off treatment, 13 on treatment] and 5 in partial remission but transfusion independent. Prednisolone induces a durable response in children with MDS. The biological basis of this response needs to be evaluated.
Genes Important for Disease Progression of MDS-Leukemia Revealed by 40K cDNA Microarray.
Authors: Miwa Hiroshi , Teramoto Hidemi , Patel Vyomesh , Letwin Noah , Castellone Maria D. , Imai Norikazu , Shikami Masato , Imamura Akira , Gutkind Silvio , Nitta Masakazu , Lee Norman H.
Published: April 2020

[Background and Objectives] One of the causes of death in patients with myelodysplastic syndrome (MDS) is leukemic transformation. After the indolent phase, which can be treated with chemotherapy, leukemic MDS will progress to a phase of aggressive blast proliferation, which cannot be controlled by chemotherapy. In this study, we would like to know the molecular mechanisms of the disease progression.

[Materials and Methods] Bone Marrow samples were collected from both indolent (at initial diagnosis) and aggressive (after chemotherapies) phases from 4 MDS-leukemia patients. We extracted RNA from each sample and then amplified the RNA, reverse transcribed the amplified RNA to cDNA, and fluorescently labeled before hybridization for the microarray.

[Results and Discussion] Compared to the expression level at indolent phase, several genes including MEK kinase 5 (ASK1), Rho-associated protein kinase 1 (ROCK1), LIM domain kinase 2 (LIMK2) and GTP binding protein Rheb, were commonly up-regulated more than 2-fold at aggressive phase of 4 cases. These results were confirmed by real time RT-PCR. Since multidrug resistance protein was up-regulated at the ratio of around 1.5, more than 2-fold up-regulation in all 4 examined cases was considered to be significant. These molecules can be candidate markers to predict transition from indolent to aggressive phase of the disease, and also can be targets for novel therapy.
A Retrospective Review of Outcomes of Patients with Chronic Myelomoncytic Leukemia Treated with 5-Azacitidine.
Authors: Dhruva Anand , Damon Lloyd E. , Linker Charles , Martin Thomas G. , Navarro Willis H.
Published: April 2020

The chemotherapeutic agent 5-azacytidine (5-AZA) is an active agent in the treatment of myelodysplastic syndrome (MDS). Phase III data with 5-AZA for MDS of all types suggest a response rate (CR+PR) of 23% and hematologic improvement rate of 37% (Silverman, et al, 2002, JCO, 20:2429). We reviewed the outcomes of patients with the MDS subtype chronic myelomonocytic leukemia (CMML) in order to better characterize outcomes for patients with this specific subtype.

Methods: Retrospective analysis of outcomes in 13 patients (median age 67; range 50–82) with CMML, who were drawn from a group of 165 consecutive patients treated with 5-AZA between 1991 and 2004 at UCSF on an NCI Special Exception Program, was performed. All patients signed informed consent to participate in the Expanded Access Protocol and IRB approval at UCSF was obtained to retrospectively review this data. IPSS of patients were 2 LOW, 7 INT-1, 3 INT-2, and 1 HIGH. 11 patients had CMML-1 and 2 patients had CMML-2. Responses were determined using the MDS International Working Group (Cheson et al., 2000 Blood96:3671) criteria.

Results: Three patients (23%) had CR and 6 patients (46%) had HI at 4 months. 9/13 patients had a response at 4 months for an overall response rate of 69% (95% CI 38.6% to 90.0%). Cytogenetic responses were seen in 1 out of 4 evaluable patients. The median AML free survival was 11 months. The median overall survival was 14 months.

Figure Figure

Conclusions: For patients with CMML, 5-AZA provides an effective treatment option with significant improvements in peripheral blood counts, marrow blasts, and transfusion requirements. The overall response rate of 69% correlates well with larger studies in MDS.
Dysregulated Expression of Myeloid Nuclear Differentiation Antigen (MNDA) in MDS: Possible Role in TRAIL-Induced Apoptosis.
Authors: Briggs Robert C. , Shults Keith , Flye Leanne , McClintock-Treep Sara A. , Jagasia Madan H. , Goodman Stacey A. , Boulos Fouad I. , Jacobberger James W. , Stelzer Greg , Head David R.
Published: April 2020

A reduced level of MNDA expression has been reported with gene expression profiling in both sporadic and familial MDS. We undertook studies to evaluate MNDA protein expression in MDS, and its possible functional significance. Patients (n=7) clinical histories, lab data, cytogenetics, and morphology were reviewed blinded to study results to confirm diagnoses. Patients had Refractory Cytopenia with Multilineage Dysplasia (n=6) or Refractory Anemia with Excess Blasts-1 (n=1)(WHO). Patients lacking primary marrow disease (n=5) were used as controls. Samples were assessed with multiparametric flow cytometry (MPFC) for CD45 antigen density, log side scatter, CD34, CD71, and MNDA (using a specific rat monoclonal antibody) with an FC500 (Beckman-Coulter) flow cytometer, and analyzed with Winlist 5.0 and ModFitLT 3.0 (Verity Software) using published methods and ECFI-generated algorithms. Analyses were blinded to clinical results. Immunohistochemistry (IHC) for MNDA was performed using the same antibody and published methods. Both MPFC and IHC demonstrated MNDA expression in MDS predominantly in granulocyte-macrophage (G-M) progenitors, similar to controls. G-M progenitors in MDS demonstrated reduced MNDA protein expression compared to controls, with 3 patterns: uniform reduced MNDA protein, a mix of normal and absent MNDA protein, or a mix of reduced and absent MNDA protein. We interpret the normal expression in some cases as consistent with presence of a mixture of normal and MDS cells, while the mix of reduced and absent expression may represent clonal evolution of MDS. To evaluate possible functional significance of MNDA expression we used stable K562 clones that express ectopic MNDA protein via retroviral transduction. Parental K562 cells lack MNDA expression. Compared to parental K562, ectopic expression of MNDA failed to protect cells from intrinsic path apoptosis induced by genotoxic agents (mechlorethamine, etoposide, cisplatin) or H2O2. Apoptosis was evaluated by morphology, TUNEL assay, trypan blue exclusion, and phosphorylation of Histone H2B at serine 14 (a specific marker of apoptosis). In sharp contrast, ectopic MNDA expression provided marked protection of cells from extrinsic path apoptosis induced by TRAIL (TNF-related apoptosis inducing ligand) exposure. Disordered apoptosis is hypothesized to be a factor in MDS pathogenesis. Our results indicate MNDA promotes viability in myeloid progenitors, and its down-regulation in MDS might relate to the G-M progenitor sensitivity to TRAIL-induced apoptosis that has been reported in MDS.