bluuebunny/crossref_metadata_2025 huggingface dataset:
Early Response to a Short Course of Induction Chemotherapy Overcomes the Prognostic Role of IPI in Patients with Aggressive NHL. Preliminary Results of the GISL LA05 Trial.
Authors: Di Renzo Nicola , Luminari Stefano , Montanini Antonella , Petrini Mario , Brugiatelli Maura , Gobbi Paolo G. , Stelitano Caterina , Angrilli Francesco , Musso Maurizio , Baldini Luca , Mazza Patrizio , Pennese Elsa , Federico Massimo
Published: October 2019

The achievement of a clinical response to the first part of induction chemotherapy has been considered for predicting survival in patients with aggressive non Hodgkin lymphoma (NHL). In April 2000, the Gruppo Italiano Studio Linfomi (GISL) started the LA05 trial with the aim of assessing different treatment modalities according to response to 4 initial courses of chemotherapy (CT) assessed according to the international response criteria for NHL. Untreated Patients younger than 65 years with histologically confirmed diagnosis of aggressive NHLs were eligible to the study. All stages and all IPI groups were allowed. After 4 courses of CT patients achieving at least a PR >75% were to be treated with two additional courses of CT (group 1); those achieving PR <75% were randomized to receive either additional 4 courses of the same CT or a modified HDS program followed by autologous stem cell support (group 2); patients with SD or PD were to be assigned to salvage programs (group 3). The study CT regimen was ProMECE-CytaBOM; in July 2005, the protocol was amended allowing the use of other CT regimens, including CHOP and MACOP-B, and recommending the addition of Rituximab for B-cell NHL. D The study is still ongoing and we are not yet able to provide data about the efficacy of HDS in patients of group 2. We present the preliminary results of efficacy of the LA05 trial, in particular focused on the outcome of group 1 patients. As of June 2006, 341 patients were enrolled. Eighty percent of cases had DLBCL, 12% PTCL, 5% GIII FL and 3% MCL. Median age at diagnosis was 49 years (range 18–67). Sixty percent had stage III–IV disease and 38% had a bulky mass. IPI was low (0–1) in 52%, intermediate (2) in 22% and high (3+) in 26% of cases. After 4 courses of CT, 265 patients were evaluable for response; one-hundred and seventy patients (64%) achieved a CR or a PR >75%, 49 (19%) achieved a PR <75% and 46 (17%) had SD or PD. The probability of achieving a CR or a PR >75% was 74%, 62% and 42%, for patients with an initial IPI of 0–1, 2 or 3+. At the end of treatment program, a CR was achieved in 57% of cases and a PR in 14%. After a median follow-up of 21 months (range 1–73), the 2-year OS was 69% (IC 95% 62–75), being 85%, 65% and 25% for group 1, 2 and 3 respectively (p<0.001). Two-year OS was 88%, 78% and 78% for patient in group 1 with and IPI of 0–1, 2 and 3+ respectively (p=NS). The achievement of a CR or a PR >75% after the first part of initial CT overcomes the prognostic role of IPI in patients with aggressive NHL.
Toxicity and Risk Factors Contributing to Delayed Methotrexate (MTX) Elimination in Adult/Elderly Patients (pts) Treated with High-Dose Methotrexate Therapy (HDMTX), a 2-Year, Single Center Survey.
Authors: Schwartz Stefan , Martus Peter , Ludwig Wolf-Dieter , Arnold Renate , Ruhnke Markus , Korfel Agnieszka , Kopitzke David , Auton Tim , Thiel Eckhard
Published: October 2019

HDMTX is a well-established regimen used in a variety of neoplasms, including acute lymphoblastic leukemia (ALL) or lymphoma (NHL). Risk factors contributing to a delayed MTX-elimination and toxicity have been studied in pediatric pts with HDMTX (Relling MV, J Clin Oncol 1994, 12: 1667–72). However, data are scarce on risk factors contributing to delayed MTX elimination in adult pts, on toxicities in elderly pts and from pts treated with HDMTX outside clinical trials. We assessed toxicities and risk factors in all non-pediatric pts treated with HDMTX at the Charité in 2003/04. 141 pts (age 16–81y, median 44y) received 614 HDMTX cycles (median dose MTX/m2: 3000mg, range 320–12000mg) for ALL (39), NHL (38), CNS-NHL (36), sarcoma or other solid tumor (28) within (69) or outside (72) clinical trials. In 99/614 (16%) of HDMTX cycles a 15–85% (median 50%) dose reduction was carried out due to renal dysfunction (58), severe toxicity of prior chemotherapy (18) or other reasons (23). Toxicities in 62/614 (10%) HDMTX cycles caused delays of 1–50 days (median 7 days) of scheduled therapy. 61/614 (10%) HDMTX cycles resulted in an increase of serum creatinine values above the upper limit of normal (ULN) (median of 112μmol/L, range 81–386μmol/L). Delayed MTX-elimination (MTX blood level >150μM 24h, >3μM 36h, >1μM 42h, >0.4μM 48h or >0.1μM 68h) occurred in 156/614 (25%) HDMTX cycles. Grade 3/4 toxicities were recorded in 224 (36%) cycles for leukopenia, 127 (21%) cycles for liver toxicity, 116 (19%) cycles for thrombocytopenia, 88 (14%) cycles for fever with neutropenia and 63 (10%) cycles for mucositis/gastrointestinal toxicity. Presence of a 3rd space and comedications with known interference with MTX elimination or nephrotoxic potential were associated with a delayed MTX elimination (p<0.01). Overweight (BMI≥25kg/m2), urine pH<7 at some time after HDMTX and presence of a 3rd space were associated with an increase of serum creatinine values above ULN (p≤0.05). Pts with delayed MTX elimination or an increase of serum creatinine above ULN were older compared to pts with regular MTX elimination (median age 51y vs. 30y and 54y vs. 35y, p<0.01). Pts receiving HDMTX outside clinical trials more frequently experienced delayed MTX elimination (p<0.01). HDMTX frequently causes grade 3/4 hematotoxicity and increases of serum creatinine. Delayed MTX elimination is associated with advanced age and therapy outside clinical trials. Improved supportive care strategies are needed, especially in pts with advanced age, to ensure predictable MTX-elimination and reduce toxicities associated with HDMTX.
Phase II Study of Enzastaurin in the Treatment of Relapsed/Refractory Mantle Cell Lymphoma.
Authors: Morschhausser Frank , Wolf Martin , Seymour John , Tilly Herve , Pfreundschuh Michael , Kluin-Nelemans Hanneke , Raemaekers John , van’t Veer Mars B. , Milpied Noel , Cartron Guillaume , Pezzutto Antonio , Spencer Andrew , Darstein Christelle , Thornton Donald , Dreyling Martin , Reyes Felix
Published: October 2019

Protein kinase C beta (PKCβ), a pivotal enzyme in B-cell signaling and survival, has been identified as a therapeutic target in B-cell malignancies. Recent studies show that PKCβ is overexpressed in a majority of patients (pts) with mantle cell lymphoma (MCL). Other studies implicate overactivation of the PI3K/AKT pathway in the pathogenesis of MCL. Enzastaurin, an oral serine/threonine kinase inhibitor, suppresses signaling through PKCβ and the PI3K/AKT pathway to induce tumor cell apoptosis, reduce proliferation, and suppress tumor-induced angiogenesis. The primary objective of this phase II, single-arm, multicenter trial was to determine the rate of freedom from progression (FFP) for ≥3 cycles (1 cycle = 28 days). Secondary endpoints included objective response rate (complete + partial) and toxicity. Pts with previously relapsed/refractory MCL, with no more than 4 prior regimens of therapy, received 500 mg enzastaurin orally, once daily, until disease progression or unacceptable toxicity. A total of 60 pts (18 female; 42 male), median age of 66 years (range: 45–85), median international prognostic index of 2 (range: 0–5), and an Eastern Cooperative Oncology Group performance status of 0–2 were enrolled. All pts had CD20 + MCL and failed multiple prior therapies (median = 2, range: 1–5). Most pts had received CHOP-based chemotherapy and/or rituximab. All enrolled pts received at least 1 dose of enzastaurin and were included in the safety and efficacy analysis. Three pts discontinued due to adverse events possibly related to study drug (diarrhea, renal impairment, and syncope). There were no drug-related deaths or grade (Gr) 4 toxicities. No drug-related bone-marrow toxicities, except Gr 3 anemia (1 pt), were reported. There was 1 case each of drug-related Gr 3 diarrhea, dyspnea, vomiting, hypotension, and syncope. Fatigue (Gr ≤2) was the most common toxicity (n = 5). No objective tumor responses were recorded. 22 of 60 enrolled pts (36.7%, 95% CI: 24.5%–48.9%) were FFP for ≥3 cycles. Six of these pts, who had failed prior chemotherapy and/or immunotherapy, achieved stable disease for ≥ 6 months. One pt was FFP for 22 months, and 2 pts are still on treatment for 8+ and 12+ months, respectively. Although no objective tumor responses occurred, 6 pts were FFP for 6 to 22 months. Oral enzastaurin was well tolerated for extended durations, suggesting it may be used as continuous or maintenance therapy after induction therapy and warrants further investigation in MCL.
The VIP-ABVD Regimen Is Not Superior to the CHOP 21 for the Treatment of Non Epidermotropic Peripheral T Cell Lymphoma. Final Results of the “LTP95” Protocol of the GOELAMS.
Authors: Gressin Remy , Peoch Michel , Deconinck Eric , Casassus Philippe , Desablens Bernard , Senecal Delphine , Quitet Philippe , Vilque Jean Pierre , Foussard Charles , Eghbali Houchingue , Jaubert Jerome , Tournilhac Olivier , Lucas Virginie , Delwail Vincent , Choufi Bachera , Thyss Antoine , Huguet Francoise , Maloisel Frederic , Richard Bruno , Milpied Noel-Jean , Colombat Philippe
Published: October 2019

Introduction: There is non consensus today for the treatment of PTCL. The five years OS with the CHOP regimen is estimated to 40%. A five years OS of 65% have been expected with an alternating VIP/ABVD regimen in a cohort ot 58 patients (pts) retrospectively studied by the same institution in Grenoble FRANCE (results presented at the french haematology society in 2000).

Aim of the study and methods: To confirm the superiority of the VIP/ABVD in first line for pts with PTCL. We have compared the 2 years EFS of pts receiving 8x CHOP21 or 6 alternating VIP/ABVD in a random manner. The VIP/ABVD regimen consisted of an alternance of three VIP regimen (VP16 100 mg/sqm D1 to D3; Ifosfamide 1000 mg/sqm D1 to D5 and Cisplatinum 20 mg/sqm D1 to D5) and three ABVD (Adriblastin 50 mg/sqm, Bleomycin 10 mg/sqm, Vinblastin 10 mg/sqm and Deticene 375 mg/sqm D1 and D14). A 40 grays IF irradiation was systematically used for AA stage I-II and on the tumoral sites larger than 5 cm to complete the treatment plan.

Results: 100 pts were included and 88 retained after a centralized pathologic review. 57 pts were diagnosed as PTCLu, 15 LAI, 10 T and 4 NTNB anaplasic, 1 angiocentric and 1 NK type. 45 pts received the CHOP and 43 the VIP/ABVD.

53/88 pts (60%) achieved a CR (34) or PR>75% (19). With a median Fup of 26.5 months EFS and OS are respectively 40% at 2 years and 40% at 4 y. There is no difference between the two groups. Two factors but not the IPI score have significantly influenced the survivals: histologic types (anaplasic vs no anaplasic) and AA stage (I–II vs III–IV).

Conclusion: The VIP/ABVD is not superior to the CHOP regimen for the treatment of PTCL. The combination Ann Arbor stage and the histologic pattern is the best predictive factor of the survivals. New innovative approach are mandatory to improve the prognostic of PTCL.
Imatinib Mesylate (Gleevec®) Is Active in Relapsed/Refractory Waldenstrom’s Macroglobulinemia: Planned Interim Results of WMCTG Clinical Trial 05-140.
Authors: Treon Steven P. , Soumerai Jacob D. , Patterson Christopher J. , Hunter Zachary R. , Villarreal Rose , Ghobrial Irene M.
Published: October 2019

Waldenstrom’s macroglobulinemia is a B-cell disorder characterized by the infiltration of lymphoplasmacytic cells (LPC) in the bone marrow (BM), along with an IgM monoclonal gammopathy. Characteristic of WM is an increased number of mast cells (MC) which are found in association with LPC, and stimulate LPC growth through several TNF-family members including CD40L, APRIL and BLYS (Ann Oncol17:1275; Blood104:917A). As such, we have targeted MC in WM. One important growth and survival factor for MC is stem cell factor (SCF), which signals through CD117. Imatinib mesylate blocks SCF signaling through CD117, and induces apoptosis of WM BM MC and LPC, both of which highly express CD117 (Blood2004; 104:314b). As such, we performed this Phase II study of imatinib mesylate in patients with relapsed and refractory WM. Intended therapy consisted of imatinib mesylate which was initiated at 400 mg po qD over the first month, and subsequently dose escalated to 600 mg po qD for up to 2 years. Dose de-escalation to 300 mg po qD was permitted for toxicity. Thirteen patients were enrolled and are eligible for evaluation at interim analysis. Median age was 64 (range 53–80 years), and median prior therapies was 2 (range 1–5). Nine and four patients had relapsed and refractory disease, respectively. Following a median of 3 months of therapy, median serum IgM levels for all evaluable patients declined from 3190 (range 763–8130 mg/dL) to 2095 (range 695–7340 mg/dL) at best response (p=0.009). On an intent to treat analysis, 6/13 (46.2%) of patients attained a ≥25% decrease in serum IgM. Responses were prompt, and occurred at a median of 2.5 months. Overall, therapy was well tolerated with ≥grade 2 toxicities as follows: anemia (n=4); edema (n=3); hyperglycemia (n=2); leukopenia (n=2); thrombocytopenia (n=1); neutropenia (n=1); and resulted for 4 patients in dose modification (n=4) or treatment cessation (n=3). The interim results of this study demonstrate that imatinib mesylate is an active and well-tolerated salvage therapy for WM.
NF Kappa B as a Therapeutic Target in AML.
Authors: Jenkins Christopher , Pepper Chris , Mills Ken , Burnett Alan
Published: October 2019

NF Kappa B is an inducible transcription factor that regulates the expression of a large number of genes essential for the normal immune and inflammatory responses. However, inappropriate NF Kappa B expression has been implicated in a number of malignancies. In this study, a two fold increase in NF Kappa B1 expression in AML samples, as compared to normal cell populations was shown by Affymetrix gene array. In addition, constitutive activation of the p65 protein component of NF Kappa B was demonstrated by ELISA in 70% of nuclear extracts derived from primary AML samples. In comparison there was no constitutive activation in the normal marrow samples tested. Elevated levels of AML nuclear p65 were inverse correlated with the proportion of cells undergoing spontaneous apoptosis, indicating that increased basal activation of NF Kappa B confers a cytoprotective effect on primary AML blasts. This data strongly suggest that NF Kappa B is a potential therapeutic target in AML. Consequently, the effects of the novel NF Kappa B inhibitor, LC1, were investigated in AML cells in-vitro. Leukemic cells were treated with LC1, at a range of 0.002 – 20mM, and the LC50 values for each sample were determined by MTS assay. The AML cell lines NB4, HL60 and U937 were also evaluated in the same way and demonstrated LC50 values of 2.98, 7.5, and 5.9 μM respectively. K562 cells were remarkably resistant to LC1, and were also shown to have the lowest NF Kappa B expression by Affymetrix gene array. Primary diagnostic AML (n=48) samples were analysed and an LC50 range of between 0.61μM and >100μM was detected with a median of 6.9μM. Annexin V and cell cycle analysis confirmed increased apoptosis as the mode of cell death. A time- and dose-dependent reduction in nuclear p65 NF Kappa B protein levels by LC1 was proven by ELISA. This preceded the induction of apoptosis, demonstrating that the inhibition of nuclear p65 NF Kappa B was not simply a result of cell death. The levels of NF Kappa B expression were correlated with the LC1 in-vitro LC50 response, demonstrating a significant inverse relationship. In addition, the downstream consequences of LC1-induced NF Kappa B inhibition were shown with a decrease in cytokine expression as demonstrated by cytokine array. The potential for synergy between LC1 and the conventional chemotherapeutic agent Ara-C and the proteasome inhibitor Velcade were also measured. Synergy was demonstrated in 60% of cell samples with Ara-C, and 80% with Velcade. This study suggests that LC1 is a potentially useful novel agent for the molecular therapy of acute myeloid leukemia.
The IGF1-R Inhibitor AEW541 Is a Potent Anti-Myeloma Agent That Overcomes Drugs Resistance, and Has Synergistic Activity with Other New Drugs.
Authors: Maiso Patricia , Ocio Enrique M. , Garayoa Mercedes , Pearson Mark A. , Pandiella Atanasio , Miguel J.F. San
Published: October 2019

Multiple myeloma (MM) represents an incurable disease for which development of new therapies is required. Here we report the effect on myeloma cells of AEW541, a new small molecule, belonging to the pyrrolo[2,3-d] pyrimidine class, identified as inhibitor of the IGF-1R in vitro kinase activity. AEW541 showed a potent antimyeloma activity (IC50 <4.5 μM) on MM cell lines both sensitive (MM1S, U266, OPM2, RPMI8226) and resistant (MM1R and U266LR7) to conventional chemotherapeutic agents. In fresh cells from five MM patients a marked antitumor activity was confirmed. AEW541 showed a synergistic effect with dexamethasone and lenalidomide, while it was additive with melphalan and bortezomib. Moreover the triple combination of AEW541, bortezomib and dexamethasone showed even higher anti-MM activity. Gene expression profiles of MM1S cells identified a total of 967 genes to be significantly deregulated (transcriptional changes in gene expression of 2-fold or greater) by treatment with AEW541. The classification of these genes according to functional categories indicated that 3.5% were involved in apoptosis/responses to stress and 13% in the control of cell cycle/proliferation. By Western analyses, we observed that AEW541 affected genes involved in cell cycle and cell death pathways. AEW541 blocked cell cycle progression, and this was accompanied by p27, up-regulation and pRb, CCND1, CCNA and CCNE downregulation. AEW541 induced cell death through an increase in the mitochondrial outer membrane permeability and provoked DNA fragmentation. AEW541 induced apoptosis and at late time points, also activated caspases 8,9 and 3. The pan-caspase inhibitor Z-VAD-FMK only slightly decreased the sensitivity to AEW541. In addition, AEW541 stimulated a caspase-independent pathway, through the release of AIF and Endonuclease G from the mitochondria. It is therefore conceivable that both caspase dependent and independent pathways are activated by AEW541 in MM cells, although the effect of AEW541 on cell cycle arrest is an earlier and more potent event. Finally, AEW541 was able to overcome the protective effect that confers IL-6, IGF-1 and BMSCs to myeloma cells in a dose dependent manner. All these data indicate that AEW541 could be a useful drug for the treatment of MM patients, particularly in combination with other novel agents such as bortezomib or lenalidomide together with dexamethasone.
Pharmacogenetics of Response to Glucocorticosteroids in Adults with Acute Lymphoblastic Leukemia.
Authors: Jamroziak Krzysztof , Balcerczak Ewa , Szemraj Zofia , Salagacka Aleksandra , Grzybowska-Izydorczyk Olga , Cebula Barbara , Szemraj Janusz , Mirowski Marek , Robak Tadeusz
Published: October 2019

Glucocorticoids are crucial components of standard chemotherapy regimens used for the treatment of adult acute lymphoblastic leukemia (ALL). However, little is known on significance of the inherited genetic background for response to steroids in adult ALL. We hypothesized that functional single nucleotide polymorphisms (SNPs) in genes important for steroid pharmacokinetics and pharmacodynamics may contribute to sensitivity or resistance to steroids. In this study we investigated whether analysis of common functional SNP in drug transporter P-glycoprotein gene (MDR1 or ABCB1), phase I cytochrome P450A metabolizing enzymes genes (CYP3A4 and CYP3A5) and glucocorticoid receptor gene NR3C1 can be useful to predict response to induction chemotherapy and survival of adult ALL patients. Sixty-two uniformly treated adult ALL patients were investigated for 6 common SNPs affecting steroid pathway including MDR1 exon 26 C3435T, CYP3A4*1B, CYP3A5*3, CYP3A5*6, NR3C1 bcl I and NR3C1 N363S polymorphisms. Genotypes were identified using direct sequencing or restriction fragment length polymorphism (RFLP) method. In the genotyped cohort 60% patients achieved complete remission after first induction chemotherapy. The analysis of the prognostic impact of the genotyped SNPs on treatment response showed that carriers of the glucocorticoid receptor NR3C1 gene 363S allele had a significantly greater probability of achievement of complete remission as compared to the non-carriers, odds ratio(OR)=1.23, 95% confidence interval (95%CI) = 1.02 −1.47, p=0.042). None of the analyzed polymorphic variants showed significant impact on probability of progression-free and overall survival of the included patients. In conclusion, we found that glucocorticoid receptor NR3C1 gene N363S polymorphism contributes to the response to steroids in patients with adult ALL. Inherited functional polymorphisms in pharmacokinetic pathways of anti-leukemic drugs should be considered as determinants of response potentially useful for individualization of chemotherapy of adult ALL of in the future.
Oxidative Stress in Red Blood Cells, Platelets and Polymorphonuclear Leukocytes from Patients with Myelodysplastic Syndrome.
Authors: Fibach Eitan , Ghoti Hussam , Amer Johnny , Winder Asher , Rachmilewitz Eliezer
Published: October 2019

Myelodysplastic syndrome (MDS) is characterized by refractory cytopenias due to ineffective hematopoiesis. Some patients with severe anemia require multiple blood transfusions and develop iron overload. Consequently, reactive oxygen species (ROS) are generated concomitant with a decrease in cellular antioxidants such as reduced gluthatione (GSH). The generated oxidative stress contributes to cell damage, apoptosis and ineffective hematopoiesis.

Using flow cytometry, we measured the oxidative state of RBC, platelets and PMN in 14 low-risk MDS patients and 25 normal donors. The results indicate that the majority of the patients had higher ROS in RBC (2.79-fold) and platelets (2.91-fold) and lower GSH in their RBC (3.4-fold) and platelets (2.1-fold) than normal (p<0.005). As for PMN, there were no significant differences in ROS, although GSH was significantly (p<0.1) lower in MDS compared with normal donors. The oxidative stress in MDS cells could be ameliorated by a short in vitro treatment with the iron-chelators deferrioxamine and deferiprone, or with the anti-oxidant N-acetylcysteine. These results suggest that the decrease in transfusion requirements with increase in platelet and PMN counts in MDS patients treated with deferrioxamine may be due to indirect antioxidant effect of the iron chelator and suggest that treatment with a combination of iron-chelators and anti-oxidants might be more effective.

ROS generation and GSH content in MDS blood cells ROS generation and GSH content in MDS blood cells Effect of iron chelations and an antioxidant on the oxidative status of MDS cells Effect of iron chelations and an antioxidant on the oxidative status of MDS cells
Targeted Disruption of the Mouse Mitoferrin (Slc25A37) Mitochondrial Solute Carrier Results in Defective Primitive and Definitive Erythropoiesis.
Authors: Paw Barry H. , Gwynn Babette , Langer Nathaniel B. , Shaw George C. , Lambert Amy J. , Boyer Leah A. , Li Liangtao , Stolte Jens , Robledo Raymond F. , Sahr Kenneth E. , Schlaeger Thorsten M. , Hentze Matthias W. , Muckenthaler Martina U. , Kaplan Jerry , Peters Luanne L.
Published: October 2019

We previously described a zebrafish mutant, frascati (frs), which exhibits profound hypochromic anemia and erythroid maturation arrest due to defects in mitochondrial iron uptake. Through positional cloning, we showed that the frs gene encodes a novel member of the vertebrate mitochondrial solute carrier family (SLC25), mitoferrin (mfrn, slc25a37). Mfrn, which is highly expressed in fetal and adult hematopoietic tissues of zebrafish and mouse, functions as the major mitochondrial iron importer essential for heme biosynthesis in vertebrate erythroblasts (Shaw GC, et al. 2006 Nature 440:96–100). To study the function of Mfrn in mammalian organisms, we identified an embryonic stem (ES) cell clone that harbors a gene trap b-geo cassette in intron 1 that inactivates the Mfrn locus. Homozygous disruption of the Mfrn locus results in embryonic lethality at E11.5 from profound anemia due to a failure of primitive erythropoiesis, confirming the requirement of Mfrn in mammalian development . Circumventing the embryonic lethality, we generated Mfrn−/− ES cells to study the role of Mfrn in definitive erythropoiesis by in vitro differentiation of embryoid bodies and mixed chimera assays. Mfrn−/− ES cells were defective in promoting the growth, differentiation, and hemoglobinization of both primitive and definitive erythroblasts by in vitro differentiation of embryoid bodies. In mixed chimera studies, Mfrn−/− ES cells failed to contribute to the erythroid compartment of adult mosaic mice, whereas measurable contribution of Mfrn−/− donor cells could be assayed in the non-erythroid, leukocyte compartment. Transcriptome microarray analysis, using the mouse Affymetrix GeneChip and the custom IronChip, revealed unexpected down-regulation of transcripts for heme-biosynthetic enzymes in Mfrn−/− erythroblasts. The block in protoprophyrin synthesis, as well as mitochondrial heme synthesis, could be partially rescued by the addition of aminolevulinic acid (ALA) to Mfrn−/− erythroblasts in vitro. Our data demonstrate that mitochondrial iron homeostasis, working through the Mfrn iron importer, coordinately regulates the synthetic pathways for porphyrin and heme in developing mammalian erythroblasts.
Hematologic Improvement, Transfusion Independence, and Safety Assessed Using Three Alternative Dosing Schedules of Azacitidine in Patients with Myelodysplastic Syndromes.
Authors: Lyons Roger M. , Cosgriff Thomas , Modi Sanjiv , McIntyre Heidi , Beach C.L. , Backstrom Jay T.
Published: October 2019

Efficacy and safety of azacitidine (Vidaza®), at the FDA-approved dosing schedule of 75 mg/m2/day x 7 days every 28 days, was demonstrated in a phase III CALGB study by Silverman et al (JCO2002; 20:2429) for the treatment of myelodysplastic syndromes (MDS). The objective of this phase II, multicenter, randomized, open-label study in all FAB subtypes of MDS was to study the treatment response and safety of 3 alternative subcutaneous azacitidine dosing schedules, eliminating the need for weekend azacitidine injections. Patients were randomized to either AZA 5-2-2 (75 mg/m2 day x 5 days, followed by 2 days no treatment, followed by 75 mg/m2/day x 2 days), AZA 5-2-5 (50 mg/m2/day x 5 days, followed by 2 days no treatment, followed by 50 mg/m2/day x 5 days) or AZA 5 (75 mg/m2/day x 5 days). After 6 cycles of azacitidine, patients meeting International Working Group MDS response/improvement criteria (Blood2000; 96:3671), defined as complete remission, partial remission, stable disease, or hematologic improvement, were eligible to receive an additional 12 cycles. As of July 1st, a total of 106 patients have been randomized in the AZA 5-2-2 (n=33), AZA 5-2-5 (n=35), and AZA 5 (n=38) treatment arms. Based on FAB classification, most patients have RA (42%, 45/106) or RAEB (30%, 32/106). Of 80 patients who have received ≥2 cycles of treatment, hematologic improvement (major or minor in at least 1 cell line) occurred in 58% (46/80) of the patients (Table). Median time to improvement in the 3 arms ranged between 1 and 3 cycles (AZA 5-2-2: 1.1, AZA 5-2-5: 3.0, AZA 5: 2.6). Of 38 patients who were RBC transfusion dependent at baseline, 27 (71%) became independent (AZA 5-2-2: 59%, 10/17, AZA 5-2-5: 89%, 8/9; AZA 5: 75%, 9/12). The most common grade 3 or 4 events included neutropenia 29%, 28/98 (AZA 5-2-2: 44%, 14/32; AZA 5-2-5: 20%, 6/30, and AZA 5: 22%, 8/36), thrombocytopenia 15%, 15/98 (AZA 5-2-2: 28%, 9/32; AZA 5-2-5: 10%, 3/30, and AZA 5: 8%, 3/36) and anemia 12%, 12/98 (AZA 5-2-2: 16%, 5/32; AZA 5-2-5: 17%, 5/30, and AZA 5: 6%, 2/36). The frequency of patients with a grade 3 or 4 infection was 15%, 15/98 (AZA 5-2-2: 19%, 6/32; AZA 5-2-5: 23%, 7/30; AZA 5: 6%, 2/36) or hemorrhage was 4%, 4/98 (AZA 5-2-2: 6%, 2/32; AZA 5-2-5: 7%, 2/30; AZA 5: 0/36). Updated data will be available at the time of the meeting. Based on preliminary results, these data indicate that the 3 alternative dosing schedules provide clinical benefit (i.e., transfusion independence and hematologic improvement) and are consistent with the FDA-approved 75 mg/m2/day x 7 days dosing results from previous CALGB studies.

Table: Hematologic Improvement Response Rates
Treatment of Myelodysplastic Syndromes with del 5q before the Lenalidomide Era: The GFM Experience.
Authors: Kelaidi Charikleia , Park Sophie , Brechignac Sabine , Mannone Lionel , Vey Norbert , Dombret Herve , Aljassem Lina , Stamatoullas Aspasia , Adès Lionel , Giraudier Stéphane , de Botton Stéphane , Mahé Béatrice , Lepelley Pascale , Picard Françoise , Leroux Geneviève , Daniel Marie-Thérèse , Bouscary Didier , Dreyfus François , Fenaux Pierre
Published: October 2019

Background: MDS with del 5q are characterized by profound anemia, which until the recent introduction of lenalidomide (N Engl J Med2005; 352: 549–57, J Clin Oncol2005;16S:5), was considered generally unresponsive to available treatments. In order to reevaluate the outcome of those patients in the pre-lenalidomide era, we analyzed response of anemia in MDS with del 5q treated with EPO ± G-CSF and thalidomide in previous GFM trials.

Patients: MDS with del 5q included in 419 MDS treated with EPO or Darbepoetin (DAR) ± G-CSF by GFM centers (including 3 successive GFM trials: Blood2004; 104: 321–7; Blood2005;106 suppl 1: 712a; Br J Haematol2006;133: 513–9 and submitted to ASH 2006), and in 134 MDS treated with thalidomide in two successive GFM trials (Br J Haematol2005; 131: 609–18, and submitted to ASH 2006). Patients received at least 30,000 U/w of EPO or 300 mg/w of DAR and doses ranging from 50 to 800 mg/d of Thalidomide during at least 12 weeks.

Results: 48 MDS with del 5q received EPO (or DAR) ± G-CSF, including 30 pts with del 5q alone, 9 with one and 9 with >1 additional cytogenetic abn; 21/48 had marrow blasts ≥5% (7 had >10%). 17 had the “5q- syndrome” according to WHO. Median pre-treatment EPO level was 287 UI/L (range 12–5,665), i.e. significantly more than in non del 5q cases (median 68, p<0.001). 22/48 pts (46%) had erythroid response, including 15 major and 7 minor (11 responses after EPO or DAR alone and 11 after EPO or DAR + G-CSF) vs. 64% in pts without del 5q (p=0.01) (p= 0.066 after adjustment for marrow blasts). The response rate was 52%, 55%, 22% and 33%, respectively in del 5q pts with the 5q- syndrome, one additional cytogenetic abn, >1 additional cytogenetic abn, and marrow blasts ≥5%. Response duration was significantly shorter in MDS with del 5q than in other MDS (mean 12 vs. 24 months, p=0.019) and in pts with 5q- syndrome vs. other MDS with marrow blasts <5% (mean 11vs. 24months, p=0.025). 20 pts with del 5q were treated with thalidomide. 6/20 had an erythroid response (30 %, including 3 major and 3 minor responses) vs. 29% of other MDS (p=NS)

Conclusion: MDS with del 5q with ≤1 additional cytogenetic abn and no excess of marrow blasts may have erythroid response to EPO ± G-CSF but responses are generally very short, while response rates to thalidomide are low. Those results are clearly inferior to results obtained with lenalidomide in MDS with del 5q.
Correlation of Clinical Features with Mutational Status in Children with Juvenile Myelomonocytic Leukemia.
Authors: Yoshida Nao , Yagasaki Hiroshi , Xu Yinyan , Tanaka Makito , Nishio Nobuhiro , Hidaka Hirokazu , Hama Asahito , Takahashi Yoshiyuki , Manabe Atsushi , Kojima Seiji
Published: October 2019

Juvenile myelomonocytic leukemia (JMML) is a rare clonal myeloproliferative disorder that affects young children. It is characterized by specific hypersensitivity to granulocyte-macrophage colony-stimulating factor (GM-CSF) in vitro. Mutations in RAS, NF1, or PTPN11 positioned in the GM-CSF signal transduction pathway, which contribute to myeloid proliferation, have been recognized in the pathogenesis of JMML. Recently a multi-step model for leukemogenesis has been proposed. In this model, the pathogenesis of leukemia requires at least two classes of mutations:primary mutations of genes implicated in cell differentiation such as AML1 and PU.1 andadditional mutations of genes contributing to myeloid proliferation such as FLT3, RAS, and PTPN11.

We hypothesized that in patients with JMML, in addition to known mutations of genes in the GM-CSF pathway involved in myeloid proliferation, potential causative mutations of other classes might be acquired. AML1 encodes a transcription factor that is essential for definitive hematopoiesis, and its mutations have recently been found in adults with acute myeloid leukemia and myelodysplastic syndrome. However, no information is available on the profiles of mutations in these genes and the relationship between these mutations and clinical features of JMML in children. We analyzed mutations of N-RAS, K-RAS, and PTPN11 in 50, and of AML1 in 30 Japanese children with JMML by direct sequencing. Correlation between the mutational status and clinical and laboratory findings, including age at diagnosis, sex, fetal hemoglobin (HbF), platelets count, cytogenetic abnormalities, and hepatomegaly, all which are suggested prognostic factors for JMML, were also assessed. Of the 50 patients with JMML, cytogenetic abnormalities were detected in 14, including 8 with monosomy 7. PTPN11 and N-/K-RAS mutations were found in 24 (48%) and 7 (14%) patients, respectively, and a novel mutation in AML1 was identified in one patient. No simultaneous mutations in these genes were found. Age at diagnosis was older (median 36 vs 11 months, p=0.0005) and HbF level was higher (31.0% vs 5.1%, p=0.033) in patients with the PTPN11 mutation than those with the RAS mutation. No difference was observed between patients with PTPN11 and RAS mutations in sex ratio, white blood cell count, platelets count, and the incidence of cytogenetic abnormalities and hepatomegaly. Our results suggest that the clinical outcome of patients with PTPN11 might differ from those with RAS mutations because a higher level of HbF and older age have been reported to be poor prognostic factors. In one patient with JMML, we identified a novel mutation in the AML1 gene that belongs to the class of genes contributing to cell differentiation instead of the class of genes in the GM-CSF pathway involved in myeloid proliferation.
FIP1L1-PDGFRalpha Positive Hypereosinophilic Syndrome (HES). The Response to Imatinib (IM) Is Durable. A Report of 21 Patients with a Follow-Up of 12 to 67 Months.
Authors: Rondoni Michela , Ottaviani Emanuela , Piccaluga Pier Paolo , Paolini Stefania , Merante Serena , Cilloni Daniela , Malagola Michele , Palmisano Michela , Grafone Tiziana , Messa Francesca , Buccisano Francesco , Tiribelli Mario , Pane Fabrizio , Saglio Giuseppe , Martinelli Giovanni , Baccarani Michele
Published: October 2019

An interstitial deletion in the long arm of chromosome 4 leads to the formation of a novel fusion gene (FIP1L1-PDGFRalpha) coding for a constitutively activated form of PDGFRalpha. The fusion gene has been identified only in patients with the hypereosinophilic syndrome and has become a marker of clonal HES. The PDGFRalpha transcript can be neutralized by several protein tyrosine kinase inhibitors (PTKI). Recently, imatinib mesylate (IM) has been shown to induce remissions (Cools J, N Engl J Med. 2003; 348:1199–200, Pardanani A, Blood2003;102:3093–6, Cortes J, Blood2003; 102:4714–6) but remission duration is still undefined. The aim of this study was to evaluate the duration of IM in FIP1L1-PDGFRalpha positive HES patients. Hypereosinophilic syndrome was defined according to Chusid criteria (Blood, 1994; 83: 2759–2779). The FIP1L1-PDGFRalpha transcript was identified and monitored every six months on bone marrow cells using a nested retrospective reverse transcriptase polymerase chain reaction (RT-PCR), as reported by Cools J (N Engl J Med.2003; 348:1199–200). Twenty-one FIP1L1-PDGFRalpha positive HES patients have been enrolled in a prospective national study of treatment with IM at a daily dose of 400 mg. All patients were male. Median age was 48 years ( range 25–72). All 21 patients achieved a complete hematologic response (CHR) in less than one month. Currently, the median follow-up of all 21 patients is 28 months ( range 13– 67), with 13 patients with a follow-up longer than 2 years. The longest follow-up are 42, 46 and 67 months. All these patients remained in complete hematologic remission and all became PCR negative and have remained PCR negative as of today. All patients are continuing on imatinib, but in several cases the dose has been reduced to 300, 200 or 100 mg daily. No patients developed heart failure.

We conclude that in FIP1L1-PDGFRalpha positive HES patients the therapeutic effect of IM at doses ranging from 400 to 100 mg daily is sustained over time, up to five years, without detectable adverse effects.
Risk of Recurrent Venous Thromboembolism Reduced by High Density Lipoproteins.
Authors: Eichinger Sabine , Pecheniuk Natalie M. , Hron Gregor , Deguchi Hiroshi , Kyrle Paul A. , Griffin John H.
Published: October 2019

Venous thrombosis is clinically distinct from arterial thrombosis, with notable differences in thrombus appearance, pathogenic mechanisms, and therapeutic approaches. High-density lipoprotein (HDL) protects against arterial atherothrombosis, but it is not known if HDL protects against recurrent venous thromboembolism. We hypothesized that HDL protects against recurrent venous thrombosis because of its multiple antithrombotic and anti-inflammatory actions. These protective activities include down-regulation of thrombin generation by acting as an anticoagulant cofactor for activated protein C/protein S, enhancement of protective endothelial nitric oxide synthase activity, reduction of leukocyte adhesion to endothelium, and anti-apoptotic effects on endothelium. To test our hypothesis, we prospectively studied 772 patients who had a first episode of spontaneous venous thromboembolism. During an average follow-up observation period of 48 months we recorded the end point of objectively documented, symptomatic recurrent venous thromboembolism. Patients with a genetic deficiency of a plasma coagulation inhibitor deficiency, with a lupus anticoagulant, or with cancer as well as those who required long-term antithrombotic treatment or were on statin therapy were excluded. The relationship between plasma lipoprotein parameters and recurrence of venous thrombosis was evaluated. Plasma apolipoproteins AI and B were measured by immunoassays for all subjects. Using NMR, we determined the levels of ten major lipoprotein subclasses in plasma for 99 patients with venous thrombosis recurrence and for 297 matched patients without recurrence. Recurrent venous thromboembolism developed in 100 of the 772 patients (12.8%). Patients with recurrence compared to those without recurrence had lower mean (±SD) plasma levels of apolipoprotein AI (1.12±0.22 vs. 1.23±0.27 mg/ml, P<0.001) but similar apolipoprotein B levels. The relative risk of recurrence was 0.87 (95% CI, 0.80 to 0.94) for each increase of 0.1 mg/ml in plasma apolipoprotein AI. For patients with apolipoprotein AI above the 67th percentile of the study population values compared with those with lower levels, the relative risk of recurrence was 0.51 (95% CI 0.32–0.83). Comparisons of plasma lipoprotein particle levels from subjects with recurrence versus those without recurrence gave similar results showing that HDL particle concentrations were lower in subjects with recurrence. Patients with high levels of apolipoprotein AI, HDL-cholesterol, and large HDL particles had decreased risk of recurrent venous thromboembolism. In summary, these novel findings that HDL appears to protect against recurrent venous thrombosis may have implications for testing HDL parameters to predict risk and may imply that lipid-altering drugs that increase HDL might reduce the risk for first or recurrent venous thrombotic events.
LBH589, a Novel Deacetylase Inhibitor (DACi), Treatment of Patients with Cutaneous T-Cell Lymphoma (CTCL). Skin Gene Expression Profiles in the First 24 Hours Related to Clinical Response Following Therapy.
Authors: Prince Miles , George D.J. , Johnstone R. , McCormack C. , Ellis L. , Williams-Truax R. , Atadja P. , Zhao C. , Dugan M. , Culver K.
Published: October 2019

Background: LBH589 is a novel DACi in Phase I trials. Pre-clinical studies have demonstrated that DACi alter gene expression and other DACi have induced disease regression in CTCL. Indeed, CTCL is an ideal disease to assess variation in tumor gene expression over time following drug administration. In this study we evaluated the safety and activity of LBH589 in CTCL and examined changes in tumor gene expression in the first 24 hours following oral LBH589.

Methods: Pts with advanced-stage CTCL, who had progressed following prior systemic therapy were entered into the oral DLT dose level 30 mg M,W,F cohort (n=1), the subsequent MTD dose level 20 mg M,W, F weekly (n=9). LBH589 was continued until disease progression or unacceptable toxicity. Intensive cardiac monitoring was performed. Six pts had 3 mm punch biopsies from CTCL-involved skin lesions at 0, 4, 8 and 24 h after administration, which were subjected to gene expression profiling using Affymetrix U133 plus 2.0 GeneChips with 47,000 probesets. Alteration in gene expression patterns was confirmed by QRT-PCR of selected genes. Individual gene expression analysis is underway, utilizing set enrichment analysis to elucidate the functional categories which correlate with degree of patient response.

Results: 10 pts are currently evaluable for response. 2 of the pts attained a complete response (CR), 4 attained a partial response (PR), 1 achieved stable disease (SD) with ongoing improvement, and 2 progressed on treatment (PD). (RR = 6/10; 60%). Microarray data on 5 pts demonstrated distinct gene expression response profiles between pts. Individual gene expression within patient tumors varied over the timepoints in the first 24 hours following treatment. To demonstrate effects of LBH589 as an epigenetic modulator, global changes in gene expression patterns in responding versus progressing patients have been delineated. In addition, functional categories of genes which correlate with degree of patient response have been identified.

Conclusions: LBH589 induces CR’s in CTCL pts. Preliminary microarray analysis of tumor samples have identified distinct gene expression profiles.
Zanolimumab, a Fully Human Monoclonal Antibody: Early Results of an Ongoing Clinical Trial in Patients with CD4+ Mycosis Fungoides (MF) Type CTCL (Stage IB-IVB) Who Are Refractory or Intolerant to Targretin and One Other Standard Therapy.
Authors: Duvic Madeleine , Kim Youn , Korman Neil J. , Boh Erin , Lerner Adam , Heffernan Michael P. , Hymes Kenneth B. , Pacheco Theresa , Elmets Craig A. , Lisby Steen , Baadsgaard Ole
Published: October 2019

Zanolimumab (previously referred to as HuMax-CD4) is a fully human monoclonal IgG1k antibody, targeting the CD4 molecule on T-cells. It exhibits cytotoxic and anti-proliferative effects and has previously shown efficacy in cutaneous T-cell lymphoma. We report the early safety and peripheral CD4+ cell depletion results from the open-label, dose escalation part of a US phase III efficacy study. So far, 21 patients have been recruited and the results from the 4 mg/kg dose group (9 patients) and 8 mg/kg dose group (6 patients) are available. Zanolimumab was administered iv, once weekly for 12 weeks. In total, 2 Serious Adverse Events have been reported. Of these, 1 case of large cell transformation in a patient with large cells present prior to inclusion was judged possibly related to treatment by investigator. No increase in toxicity was seen upon dose escalation. Marked depletion of CD4+ T-cells was observed after just 1 infusion of zanolimumab at both dose levels. One week following the last dose, the median (range) peripheral blood CD4+ T-cell count compared to baseline values was decreased from 831 per μL (167–1928) to 24 per μL (4–141) in 4 mg/kg dose-group and from 638 per μL (182–1024) to 9 per μL (4–19) in the 8 mg/kg dose-group. These initial data indicate that zanolimumab has an acceptable tolerability profile at the doses tested and results in a rapid and pronounced decrease in peripheral CD4+ counts in MF CTCL patients.
Novel Treatment of Burkitt Lymphoma with Dose-Adjusted EPOCH-Rituximab: Preliminary Results Showing Excellent Outcome.
Authors: Dunleavy Kieron , Pittaluga Stefania , Janik John , Grant Nicole , Shovlin Margaret , Little Richard , Yarchoan Robert , Steinberg Seth , Jaffe Elaine S. , Wilson Wyndham H.
Published: October 2019

Burkitt Lymphoma (BL) is a highly proliferative lymphoma with a propensity to present in extranodal sites. Standard treatment for BL involves intensive, multi-agent chemotherapy that is highly effective but associated with considerable toxicity and treatment-related mortality, particularly in older patients. Based on the observation that DA-EPOCH overcomes the adverse effect of high proliferation in diffuse large B-cell lymphomas, we hypothesized it may be effective in BL. We have undertaken a study of DA-EPOCH-Rituximab in untreated BL in an attempt to maintain a high cure rate with low treatment related toxicity. Eligible patients had untreated BL and could be HIV positive or negative. HIV negative patients (n=13) received 6 cycles of DA-EPOCH-R as previously described (Blood99: 2685, 2002). Patients with HIV-associated BL (n=4) received 3–6 cycles of DA-EPOCH-R for 1 cycle beyond CR for a minimum of 3 cycles. All patients received intrathecal methotrexate prophylaxis and therapy was administered in the outpatient clinic where possible. Characteristics of 17 patients include median age (range) 27 (18–66) and ECOG PS 1 (1–3); stage III/IV 9 (53%); LDH > N 9 (53%); male sex 13 (76%) and; extranodal sites 11 (65%). All 11 (65%) patients with extranodal disease had abdominal or pelvic involvement at diagnosis. No patients had central nervous system disease on presentation. Response is CR/CRu in 100% of patients. One patient received consolidative radiotherapy to a single site of residual disease. At a median potential follow-up of 28 months, OS and PFS are both 100% and EFS is 92.3%. Significant toxicities include fever/neutropenia in 16%, grade 4 neutropenia in 47% and grade 3/4 thrombocytopenia in 22% of cycles. There were no treatment related deaths and no tumor lysis syndrome. In conclusion, DA-EPOCH-R appears to be highly effective with relatively low toxicity compared to standard intensive high-dose regimens for BL. Infusional chemotherapy may reduce or eliminate the risk of tumor lysis syndrome. Accrual continues.

Figure Figure