bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Age in the Outcome of Patients with Chronic Myeloid Leukemia in Late Chronic Phase: Clinical and Molecular Results of a Phase II Study of the GIMEMA CML Working Party.
Authors: Martinelli Giovanni , Palandri Francesca , Iacobucci Ilaria , Bassi Simona , Castagnetti Fausto , Amabile Marilina , Cilloni Daniela , Poerio Angela , Soverini Simona , Rege-Cambrin Giovanna , Iuliano Franco , Alimena Giuliana , Latagliata Roberto , Testoni Nicoletta , Pane Fabrizio , Saglio Giuseppe , Rosti Gianantonio , Baccarani Michele
Published: April 2020

To assess the effect of age on response and compliance we performed a sub-analysis within a phase II trial of the GIMEMA CMLWorking Party (CML/002/STI571). Since the WHO cut-off age to define an older patient is 65 years, we identified: 226/284 (80%) younger patients (below 65 years) and 58/284 (20%) older patients (above 65 years) before starting imatinib. Responses (hematologic and cytogenetic) were lower in older age group but progression free survival and overall survival probabilities (median observation time 3 years) were the same. Moreover, among complete cytogenetic responders, no differences were found in the level of molecular response between the 2 age groups. As probably expected, older patients experienced more adverse events (AEs), both hematologic and non-hematologic: this worsen compliance, however, did not prevent a long term outcome similar to younger ones. CML
Dasatinib (SPRYCEL®, BMS-354825) Phase-I/II Study of Patients with Chronic Myeloid Leukemia (CML) Resistant or Intolerant to Imatinib: Results of the CA180031 Study in Japan (Phase I Portion).
Authors: Sakamaki Hisashi , Fujisawa Shin , Tobinai Kensei , Taniwaki Masafumi , Miyawaki Shuichi , Jinnai Itsuro , Utsunomiya Atae , Tanimoto Mistune , Morishima Yasuo , Okada Masaya , Miyazaki Yasushi , Uoshima Nobuhiko , Seriu Taku , Ohno Ryuzo
Published: April 2020

Dasatinib (SPRYCEL®, formerly BMS-354825) is a potent, orally active, multi-targeted kinase inhibitor active against BCR-ABL and SRC family kinases. Studies outside Japan have demonstrated that dasatinib is highly effective in overcoming resistance and intolerance to imatinib (im), inducing durable cytogenetic and hematologic responses in this CML patient population. In establishing the dose for non-Japanese patients, the maximum tolerated dose failed to be reached and the 70-mg-twice-daily (BID) dose was determined to provide the optimal benefit-risk profile. Based on clinical experience to date, dasatinib has the potential to have a major impact on the current treatment paradigm for patients with CML resistant or intolerant to prior therapy, including im. To assess the safety and efficacy of dasatinib in Japanese patients, a phase-I/II study was conducted in patients with all phases of im-resistant or -intolerant CML. In the phase-I portion of this study, dasatinib was administered at one of three escalating dose levels for a period of 4 weeks: 50 mg, 70 mg, or 90 mg BID. As of July 2006, 15 eligible patients have been enrolled and treated. Six evaluable patients (4F, 2M; median age 43 y [range 27–56]; 4 im-resistant, 2 im-intolerant) were treated at the 50 mg BID dose level and one dose-limiting toxicity (DLT) was observed (grade 4 thrombocytopenia). Two patients experienced transient grade 3 ALT elevations but as treatment was uninterrupted, these were not considered DLTs. A further 6 evaluable patients (6M; median age 42 y [range 27–55]; 3 im-resistant, 3 im-intolerant) were treated at the 70 mg BID dose level and one DLT was observed (grade 4 thrombocytopenia). Three patients have recently been enrolled at the 90 mg BID dose level. As the safety of the 70 mg BID dose was established in the phase-I portion of the study, the use of this dose is being expanded into the phase-II element, which plans to evaluate cytogenetic response in chronic phase CML patients at 24 weeks, and hematologic response rate in accelerated- or blast-phase CML and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL) patients at 12 weeks. Accrual to the phase-II portion of this study is ongoing. Updated data including baseline mutational analyses will be presented at the meeting.
Synergism between Anti-Leukemia Therapeutics and Isoprenoid Pathway Inhibitors in K562 Leukemia Cells.
Authors: Askeland Eric J. , Wiemer Andrew J. , Hohl Raymond J.
Published: April 2020

Chronic myeloid leukemia (CML) is a consequence of the BCR-ABL oncogene which mediates a phosphorylation cascade of downstream effectors including those of the Ras family. Drugs that target the isoprenoid pathway, such as the farnesyl transferase (FT) inhibitors have shown promise as novel anti-cancer therapies for some hematologic malignancies. Leukemias are treated with specific inhibitors, like imatinib mesylate in the case of CML, and less-specific chemotherapeutic drugs such as the pyrimidine analog cytarabine (ara-C). We systematically evaluated combinations of these and other agents with isoprenoid pathway inhibitors for synergistic anti-leukemia effects. Toxicity was measured in the BCR-ABL-expressing K562 cells using an MTT assay, and drug synergy was assessed using a modified isobologram method. Cell cycle distribution, Annexin-V apoptosis detection, and Western blot analyses were performed on cells treated with synergistic drug combinations. Imatinib mesylate exhibited significant synergy with the squalene synthase inhibitor zaragozic acid, but not with inhibitors for HMG coenzyme A reductase (lovastatin), FPP synthase (zoledronic acid), GGPP synthase (digeranyl bisphosphonate), and FT (alpha-hydroxyfarnesyl phosphonate). Western blot analysis revealed a moderate decrease in Ras protein levels upon treatment with zaragozic acid. This zaragozic acid-mediated decrease in Ras protein may explain the synergy seen between the drugs since imatinib mesylate also affects Ras by decreasing its phosphorylation through inhibition of the BCR-ABL protein. Apoptosis detection and cell cycle distribution did not yield insight into the mechanism underlying this synergy. Ara-C displayed synergy with digeranyl bisphosphonate in the MTT assay. Subsequent cell cycle analysis showed a concentration-dependent G0–G1 arrest by digeranyl bisphosphonate which was enhanced by ara-C. Annexin-V/propidium iodide staining revealed increased cellular apoptosis in response to either digeranyl bisphosphonate or ara-C with synergistic apoptosis to combination treatments. Western blot analysis for anti-apoptotic proteins showed a decrease in Mcl-1 and Bcl-XL protein levels in the presence of digeranyl bisphosphonate. Ara-C has been shown to alter pro-apoptotic and anti-apoptotic proteins in a manner that induces apoptosis. These modifications in pro- and anti-apoptotic proteins and alterations in cell cycling may constitute a mechanism for the synergy seen between digeranyl bisphosphonate and ara-C. Discovery of these synergistic combinations is important because translation of these cell culture results to in vivo testing and human clinical trials may enhance treatment efficacy and diversify available treatment options. Future work will focus on further determining the mechanisms for these synergistic interactions, finding new synergistic combinations, and applying these regimens to in vivo models and clinical trials.
Clonal Chromosomal Abnormalities in Philadelphia Negative Cells during Dasatinib Treatment in Five Patients with Chronic Myeloid Leukemia.
Authors: Dubruille Viviane , Talmant Pascaline , Loiseau Herve Avet , Moreau Philippe , Harousseau Jean-Luc
Published: April 2020

The emergence of clonal chromosomal abnormalities in Philadelphia negative cells was reported during Imatinib treatment in several patients with chronic myeloid leukemia (CML) in different phases (chronic phase, accelerated phase and blastic crisis). Recently, new tyrosine kinase inhibitors were used in treatment of Imatinib-resistant or Imatinib-intolerant chronic myeloid leukemia. We describe 5 patients with chromosomal abnormalities in Philadelphia negative cells during treatment with Dasatinib. At the beginning of treatment, 4 patients were in chronic phase, 1 patient was in accelerated phase. All patients were in cytogenetic relapse. Additional cytogenetic abnormalities were described in Philadelphia positive cells only in one case (accelerated phase). Clonal cytogenetic abnormalities (4 trisomy 8, 1 monosomy 7) appeared in Philadelphia negative cells during Dasatinib treatment (after 6 months in 1 patient, 9 months in 4 patients).At this time, 4 patients (Imatinib- resistant) received Dasatinib at the dose of 40 mg twice daily (dose reduction due to hematological toxicity); 1 patient (Imatinib cutaneous intolerance in chronic phase) received 140 mg daily. Two patients were in complete cytogenetic response, 2 patients were in partial cytogenetic response (10% and 8.5% Philadelphia positive cells) and obtained a complete cytogenetic response 3 months later, and 1 patient was in minor cytogenetic response. For 2 patients, the presence of trisomy 8 was transient, for 3 patients chromosomal abnormalities were persistent; none showed myelodysplastic changes. At present, all patients are still treated by Dasatinib with the same dosage. Cytogenetic abnormalities occur in Philadelphia negative cells in a fraction of patients with CML treated with Dasatinib. Philadelphia negative abnormal clone can persist despite complete cytogenetic response achievement. Their incidence, etiology and prognosis remain to be clarified and with a short follow-up, no clear clinical consequences can be identified.
Blast Cell Reduction in Elderly Patients with Acute Myeloid Leukemia Secondary to Myelodysplastic Syndrome Treated with Methylprednisolone.
Authors: Suzuki Kei , Sekine Takao , Ohishi Kohshi , Katayama Naoyuki
Published: April 2020

Treatment of elderly patients with acute myeloid leukemia (AML) is challenging, because they are often resistant to chemotherapy and not eligible for standard myelosuppressive chemotherapy. Here we report three elderly patients (above 70 years of age) with AML secondary to myelodysplastic syndrome and a poor performance status (PS 4), in whom methylprednisolone (mPSL; 125 mg/body) resulted in a decrease in blast cells in the peripheral blood (PB). Although this reduction was transient, the blast counts could be controlled by intermittent or daily treatment with mPSL, without significant adverse effects. We examined the effects of mPSL on leukemic blasts (CD34+CD33+CD13+CD4+) in detail in one patient and found that the blast count in the peripheral blood rapidly decreased from 14.2 × 109/l to 0.3 × 109/l for 18 hours after mPSL administration. The neutrophil counts gradually increased. Annexin V assays revealed that the percentage of annexin V+PI−cells among the blast cells increased from 4% to 20% at 9 hours after mPSL treatment, indicating that mPSL decreased the number of blasts in part through induction of apoptosis. In in vitro liquid cultures, mPSL inhibited the survival of blast cells from the patient in a dose-dependent manner. Furthermore, mPSL suppressed the growth of leukemic progenitors in semisolid cultures. Although our experience is limited to these three patients, our findings suggest that mPSL may offer therapeutic benefits for a subset of elderly patients with AML evolved from myelodysplastic syndrome.
The Extrinsic Apoptotic Pathway Is Implicated in Myelodysplastic Syndromes through the Fas/Fas-L/FADD System.
Authors: Ribeiro Patricia , Pereira Herminia , Costa Isabel , Sousa Maria H. , Conduto Antonio , Pocas Isabel , Viegas Paula , Costa Maria M. , Goncalves Cidalia , Rodrigues Antonio S. , Sousa Aida B.
Published: April 2020

The myelodysplastic syndromes (MDS) are characterized by ineffective hematopoiesis, peripheral blood cytopenias, increased apoptosis and possible transformation into acute myeloblastic leukemia (AML). MDS are considered “clonal” stem cell disorders but the precise mechanism is yet unknown. Lately there has been some evidence implicating the extrinsic apoptosis pathway in the pathogenesis of these diseases. The present study included 30 cases (2001/03) classified according to WHO: 6 RA, 1 RARS, 7 RAEB type I, 3 RAEB type II, 2 RCMD, 2 MDS/MDP, and 2 aplastic anemias, 3 AML secondary to MDS and 4 control cases with normal histology. We assessed the expression of TNF-related-apoptosis-inducing ligand (TRAIL) and its receptors, of Fas, Fas-L, Fas-associated-death-domain (FADD), caspase-8, caspase-3 and DNA fragmentation factor (DFFP) by RT-PCR and by immunohistochemical staining. Results were correlated with the expression of CD34, HLA-DR15, CD15, CD20, CD68 and CD117. Patients were stratified according to the international prognostic scoring system (IPSS). Fas, Fas-L and FADD were overexpressed in low risk IPSS categories and in RA and RARS, but gradually lost their expression in more advanced stages and were not expressed in RAEB II and AML cases. This expression was inversely correlated with the expression of caspase-8 and DFFP. We observed that CD34+ and HLA-DR15+ samples were positively correlated with the expression of FADD, caspase-8 and DFFP. TRAIL was positive in normal histology and AML cases but its expression was diminished in all other samples. Our data suggest that the extrinsic apoptotic pathway may be implicated in MDS through Fas, Fas-L and FADD. RAEB II may be grouped with AML, as it shows a lesser degree of apoptosis than lower risk MDS categories. A better understanding of these diseases will allow for a better and more individualized management. This work was supported by a grant from the Terry-Fox Foundation.
A Pilot Study To Test the Efficacy of a Combination of Gleevec with Thalidomide in Patients with Idiopathic Primary Myelofibrosis, Myelofibrosis with Myeloid Metaplasia and Myelodysplastic Syndromes Who Present with Myelofibrosis.
Authors: Batus Marta , Shammo Jamile M. , Venugopal Parameswaran , Gregory Stephanie A. , Raza Azra
Published: April 2020

Purpose: Prompted by the finding that Gleevec can reduce marrow fibrosis in patients with CML who present with myelofibrosis, we sought to evaluate the safety and efficacy of a novel combination of thalidomide and imatinib mesylate (Gleevec®) for treatment of patients with myelofibrosis with myeloid metaplasia and myelodysplastic syndrome who presented with Grade 3/4 myelofibrosis. The rationale for this combination is that the anti-angiogenic and anti-TNF effects of thalidomide may be potentiated by the anti-TGF-b, anti-PDGF effects of Gleevec to reduce marrow fibrosis in this group of patients.

Methods: We enrolled and treated 19 patients with MDS, (RA: 9, RAEB: 9, CMMoL:1) with IPSS score (Low: 3, Int-1: 11, Int-2: 4, High: 1) fulfilling the inclusion criteria. 7 patients had a normal karyotype, 2 patients with del 5q alone, 3 patients with del 5 q with other abnormalities, 2 patients with complex karyotype, and 5 with others. In this study we treated patients with Thalidomide starting at a dose of 100 mg per day escalating to 400 mg per day and Gleevec at 600 mg per day. Treatment was continued for one year or until disease progression. Response to treatment was assessed at 16 weeks of treatment according to the International Working Group (IWG) criteria, and dose modifications were employed according to set criteria per protocol.

Results: Three of nineteen patients responded, with overall response rate of 15.8%. Two patients (10.5%) had complete response. One patient (RAEB-I, del 5q and trisomy 8) achieved CR at 16 weeks and continued to be in CR for 9 months with a follow up bone marrow biopsy showing no overt evidence of the previously diagnosed myelodysplastic syndrome by morphology or flow cytometry. The second patient (RAEB-II, normal karyotype) achieved CR at 1 year of treatment, however developed grade 3 peripheral neuropathy and subsequently, Thalidomide was held; this patient continues to be in CR todate while on Gleevec monotherapy. Finally, one patient (5.3%) had a partial response at 16 weeks for 5 months. Four patients died during treatment due to progressive disease; all other patients did not respond. Most patients tolerated that treatment well, three patients developed grade 3 neutropenia, one patient developed grade 3 pulmonary toxicity, and one patient developed grade 3 peripheral neuropathy, these adverse events were managed by dose reduction/interruption.

Conclusion: Thalidomide and Gleevec is a relatively well-tolerated regimen that can be considered for treatment of a small subset of patients with myelofibrosis and MDS. The finding that the combination led to reduction of bone marrow fibrosis in responding patients is intriguing and warrants further investigation.
HLA-DRB1 Allele Frequency and TNF Polymorphism in Patients with Myelodysplastic Syndromes (MDS).
Authors: Chang C.K. , Gooley T. , Petersdorf E.W. , Scott B.L. , Hansen J.A. , Joachim H.
Published: April 2020

MDS, a heterogeneous group of hematopoietic stem cell disorders, are associated with dysregulated cytokine profiles in marrow and blood plasma, including upregulation of tumor necrosis factor alpha (TNF-alpha). Other data suggest that patients with MDS who carry the human leukocyte antigen (HLA)-DRB1*1501 allele are more likely to respond to immunosuppressive therapy. The genes for HLA-DR and TNF are in close linkage on human chromosome 6. Preliminary data had shown a higher than expected frequency of single nucleotide polymorphism (SNP) at position -238 of the TNF gene. We wanted to determine, therefore, the strength of linkage disequilibrium between a given TNF polymorphism and HLA-DR alleles. We analyzed HLA-DR allele frequencies in 1,040 patients with MDS, in comparison to 2,474 patients with chronic myeloid leukemia (CML); 1,422 and 160 of these patients, respectively, had TNF polymorphism data (AA vs. AC vs. CC alleles) for SNP 308 and 863 available. Significant differences in frequency were observed for HLA-DRB1*0401 (9.1% vs. 6.9%; p=0.03), *0701 (3.8% vs. 1.3%; p<0.0001), *1302 (2.7% vs. 4.3%; p=0.03), and *1501 (13.6% vs. 9.9%; p=0.0009). When determining the association of a given TNF SNP with the presence of two, one, or no copy of a given HLA-DR allele, TNF SNP 863 showed a significant association with HLA-DRB1*0101 (p=0.003), *0301 (p<0.0001), *0401 (p=0.0002), and *1101 (p=0.02); HLA-DRB1*0301 was also associated with TNF SNP 308 (p<0.0001). There was no significant correlation of either one of these two TNF SNPs with diagnosis. However, when only MDS patients were considered, there was a suggestion of a correlation of SNP 308 and HLA-DR*1501 (p=0.11); among patients with the TNF genotype AA or AC only 3 of 24 (12%) carried at least one DRB1*1501 allele, compared to 20 of 56 (36%) among those with the CC genotype. While TNF SNPs remain to be typed in a large proportion of MDS patients, results suggest a higher frequency of some HLA-DR alleles, in particular *0701 and *1501, in patients with MDS than seen in CML, and a correlation of TNF SNPs, in particular, TNF 308 CC with HLA-DRB1*1501. Thus, these data are consistent with an association of DRB1 *1501 and TNF alleles, and it will be of interest to identify the pattern of TNF polymorphism in HLA-DRB1*1501 negative patients, Additional studies are needed to further determine potential haplotypes that are associated with MDS and possible correlations with disease course and treatment responses.
Frequency of the JAK2 Mutation and Its Usefulness as a Marker for Treatment Response and Disease Progression in Korean Patients with Chronic Myeloproliferative Disorders.
Authors: Shin Myung-Geun , Kim Hyeoung Joon , Kim Hye-Ran , Lee Sun-Young , Lee Il-Kwon , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook
Published: April 2020

Background: The recent discovery of a single point mutation in the JAK2 gene (V617F) is a major advance in our understanding of the pathogenesis of BCR/ABL-negative chronic myeloproliferative disorders (CMPD). The frequency of the JAK2 V617F mutation in CMPD differs according to methods and research groups. We investigated the frequency of JAK2 mutations in Korean CMPD patients and demonstrated their usefulness as a new molecular marker for treatment response and disease progression in BCR/ABL-negative CMPD using quantitative real time PCR and pyrosequencing.

Methods: Seventy-eight patients with BCR/ABL-negative CMPD comprising 42 cases of essential thrombocythemia (ET), 26 of polycythemia vera (PV), 7 with idiopathic myelofibrosis (MF), and 3 unclassifiable (UC) CMPD were enrolled in this study. A 364-bp PCR product containing the JAK2 V617F mutation was sequenced in both directions from total bone marrow cells. Restriction enzyme-based assessment with BsaXI was also used to search for JAK2 mutations. A quantitative real-time PCR-based allelic discrimination assay and pyrosequencing (Pyrosequencer PSQ96) were used to quantify the JAK2 V617F mutation status.

Results: A single JAK2 V617F point mutation was identified in 20 (77%) of 26 patients with PV, 12 (26%) of 42 with ET, 7 (100%) of 7 with MF, and 2 (67%) of 3 with UC. The V617F mutation was present in over half of Korean patients with BCR/ABL-negative CMPD, giving an overall frequency of 53%. The proportion of mutant alleles ranged from 36 to 100% in the real-time PCR and pyrosequencing analysis. Patients with MF had a higher percentage of JAK2 mutant alleles than patients with ET (MF>PV>ET). PV patients (n=8) with a good response (phlebotomy only) had a relatively low proportion (58.9±20.1%) of the JAK2 mutant, while patients (n=9) showing a poor response (additional chemotherapy) initially had a higher proportion (76.6±19.0%; p=0.04185).

Conclusion: The frequency of the JAK2 V617F mutation in Korean patients with PV was lower than that in reports from Western countries, although the overall frequency in Korean patients with BCR/ABL-negative CMPD was similar to previous reports. The results also suggest that the JAK2 mutation (quantification of JAK2 mutant alleles) can be used as a new marker for treatment response and disease progression in BCR/ABL-negative CMPD using quantitative real-time PCR or pyrosequencing.
Dose-Modified Oral Chemotherapy for AIDS-Related Non-Hodgkin’s Lymphoma (AR-NHL) in East Africa.
Authors: Orem Jackson , Mwanda O.W. , Fu P. , Banura C. , Kakembo J. , Ness A. , Johnson J. , Subbiah V. , Bako J. , Black J. , Katongole-Mbidde E. , Remick S.C.
Published: April 2020

Background: Dose-modified chemotherapy for AR-NHL in the pre-HAART era has been shown to be equally efficacious and less myelotoxic [N Engl J Med1997;336:1641 (mBACOD); J Clin Oncol2001;19:2171 (mCHOP)]. In resource-constrained settings, intravenous chemotherapy and supportive care of the AIDS / cancer patient are challenging (J Natl Cancer Inst2002;94:718).

Methods: We embarked on a pilot feasibility trial of dose-modified oral chemotherapy [lomustine 50 mg/m2 D1 (C#1 only); VP-16 100 mg/m2 D1–3; and cyclophosphamide / procarbazine 50 mg/m2 each D22–26 at 6-week intervals (1 cycle) for 2 cycles] in HIV-infected patients with biopsy-proven AR-NHL in East Africa.

Results: A total of 49 pts (21 Uganda; 28 Kenya) were treated on study. The majority of pts were female (59%) with median age 39 yrs (range 18–64); poor PS (2 or 3) − 63%; high grade lymphoma (69%); advanced stage (III or IV) − 69%; and B symptoms (79%). At study entry median CD4 count was 200/μL and HIV-1 viral load 97,973 copies/ml. Eighteen pts (36%) had access to ARV. A total of 78 cycles of therapy were administered to 49 pts (median 2; range 0.5–2). The regimen was well tolerated. There were 4 episodes of febrile neutropenia and 3 treatment-related deaths (6% mortality rate). Overall objective response rate is 77% (CR/uCR 56%); median survival 12.3 months; and 18 patients remain alive as of 7/11/06.

Conclusions: Dose-modified oral chemotherapy is efficacious, has comparable outcome to that in the US in pre-HAART era, an acceptable safety profile, and is pragmatic in the resource-limited setting. Further investigation of the oral regimen vs. mCHOP is warranted.
Denileukin Diftitox (Ontak™) Therapy for Patients with Systemic Mastocytosis.
Authors: Quintas-Cardama Alfonso , Kantarjian Hagop , Giles Francis , Cortes Jorge , Verstovsek Srdan
Published: April 2020

Systemic mastocytosis (SM) is characterized by abnormal proliferation and accumulation of neoplastic mast cells in various tissues. Patients (pts) with SM are empirically treated with recombinant interferon-alpha (IFN-α) or cladribine when cytoreduction is required. However, treatment with these agents has been associated with poor response rates, poor tolerance, and/or short response duration, and the prognosis of pts with SM is unchanged. This underscores the urgent need of new therapeutic approaches. Atypical expression of CD25 (the alpha-subunit of the interleukin-2 [IL-2] receptor), on the membrane of neoplastic mast cells represents an attractive therapeutic target. Denileukin diftitox (Ontak™) is an engineered fusion protein containing the active domain of the diphtheria toxin and the full-length sequence of IL-2, thus targeting IL-2 receptor-expressing cells. Upon internalization, protein synthesis is inhibited and cytotoxicity ensues. We designed a pilot trial for SM pts that either had aggressive form, or indolent form with uncontrolled symptoms despite supportive care measures, in which Ontak™ was initially administered at 9 ∝g/kg/day intravenously on days 1 through 5 every 21 days. This dose was increased after 3 cycles to 18 ∝g/kg/day in pts with no response and no significant toxicity. Therapy was discontinued in pts who showed no response after 6 cycles. Response was evaluated following guidelines proposed by Valent et al. (Leuk Res. 25;603–625, 2001). Thus far, a total of 7 symptomatic pts have been treated, 5 with aggressive and 2 with indolent SM. Median age was 58 years (range, 41 to 65), time from diagnosis to Ontak™ therapy 64 mo (range, 1 to 158), Hb 11.8 gr/dL (range, 9.2 to 13), WBC 7.9 ×109/L, (range, 2.5 to 16.9), platelet count 275 ×109/L (range, 54 to 759), eosinophil 2.3% (range, 0% to 6.1%). Five patients were previously treated with imatinib mesylate and one with cladribine; one pt had not received any prior therapies. One pt, who had undergone splenectomy, had hepatomegaly and 1 other had splenomegaly prior to start of Ontak™ therapy. In all patients malignant mast cells expressed CD25. A total of 33 cycles were administered. The median number of cycles was 5.5 (range 3 to 6). No significant responses have been observed among 7 pts treated. No significant difference was found between pre- and post- Ontak™ therapy in the percentage of bone marrow mast cells, and only one patient had decreased serum tryptase level at the end of the therapy (by 33%). No responses in C-findings were noted in patients with aggressive disease. However, symptoms related to SM improved in 3 patients and were eliminated in one. Therapy with Ontak™ was well tolerated with all toxicities being grade 1–2, without any grade 3–4. In summary, therapy with Ontak™ is very well tolerated but has no appreciable activity in pts with SM at the dose and schedule used in this study.
The Novel Tyrosine Kinase Inhibitor (TKI) Exel-0862 Potently Induces Apoptosis in Human FIP1L1-PDGFR-α-Expressing Cells through Caspase-Dependent Cleavage of MCL-1.
Authors: Pan Jingpan , Quintas-Cardama Alfonso , Kantarjian Hagop , Lamb Peter , Tefferi Ayalew , Cortes Jorge , Giles Francis , Verstovsek Srdan
Published: April 2020

The FIP1L1-PDGFR-α fusion kinase induces hypereosinophilic syndrome (HES) in a subset of patients (pts). Imatinib is a potent inhibitor of PDGFR-α and has been associated with durable hematologic responses in pts with HES. However, development of kinase domain mutations may hamper TKI activity, underscoring the need for novel agents to prevent or overcome resistance. We evaluated the efficacy of the novel TKI EXEL-0862 in FIP1L1-PDGFR-α-expressing cells. In 72-hour MTS assay, EXEL-0862 inhibited EOL-1 cell proliferation with an IC50 of 1.0 nM and markedly decreased phosphorylation of STAT3 and Erk1/2, suggesting inhibition of downstream targets of PDGFR-α. More important, EXEL-0862 induced dose-dependent inhibition of PDGFR-α phosphorylation in peripheral blood mononuclear cells from a pt with HES. Exposure of EOL-1 cells to EXEL-0862 10 nM for 24 hours caused apoptosis as evidenced by flow cytometry (annexin-V/PI staining) and transmission electron microscopy. This was coupled with time-dependent cleavage of caspases-3, caspase-9, and PARP and with mitochondrial damage as reflected by cytochrome c translocation into the cytosol, and a diminished mitochondrial transmembrane potential. Of note, apoptotic features were not evident after 16 hours of EXEL-0862 exposure. Immunoblots showed marked downregulation of Mcl-1, but not of Bcl-2, Bcl-XL, or Bax. To elucidate the mechanism of Mcl-1 cleavage, EOL-1 cells were exposed to 10 nM EXEL-0862 for 6 hours followed by cycloheximide (5 μg/ml), a potent translation inhibitor, which resulted in marked Mcl-1 turnover. Treatment with the proteasome inhibitor MG-132 failed to abrogate EXEL-0862-induced Mcl-1 cleavage, suggesting proteasome-independent degradation. Rather, a decrease of pro-form Mcl-1 (p42) and increase of the cleaved Mcl-1 peptide form of Mcl-1 (p28) coincided with caspase-3 activation. Pretreatment with 20 μM pan-caspase inhibitor Z-VAD abolished EXEL-0862-induced Mcl-1 cleavage, pointing to a caspase--dependent mechanism. Collectively, our results establish EXEL-0862 as a solid candidate for the targeted treatment of pts with FIP1L1-PDGFR-α-positive HES.
Effect of Tyrosine Kinase Inhibitor INNO-406 on Human Mast Cells Bearing Mutated c-KIT Tyrosine Kinase.
Authors: Pan Jingxuan , Kantarjian Hagop , Cortes Jorge , Giles Francis , Verstovsek Srdan
Published: April 2020

Systemic mastocytosis (SM) is characterized by a clonal accumulation of mast cells (MC) in bone marrow and other tissues. Malignant MC have abnormal spindle shape, express aberrant surface markers, and carry, in most cases, a mutation involving codon 816 of the c-KIT gene (D816V). This mutation results in a constitutively activated c-kit receptor associated tyrosine kinase and is responsible for disease progression. Agents that affect mutated c-kit may have clinical benefit in SM, but there is none clinically available. INNO-406 is a novel multi-targeted tyrosine kinase inhibitor, previously shown to inhibit the growth of cells expressing c-kit by inhibiting its phosphorylation (Kimura, Blood, 106:3948–54, 2005). We investigated the effects of INNO-406 on mast cells with mutated c-KIT, and compared it to that of imatinib mesylate, tyrosine kinase inhibitor effective against selected c-KIT mutants. HMC-1560 cells carrying juxtamembrane domain c-KIT mutation in codon 560, and HMC-1560, 816 cells carrying both codon 560 mutation and tyrosine kinase domain c-KIT mutation in codon 816, were exposed to increasing concentrations (0.02 to 5 μM) of INNO-406 or imatinib in 72-hour cell proliferation assay. Both INNO-406 and imatinib effectively inhibited the growth of HMC-1560 cells, with IC50 of 45 and 75nM, respectively. Neither drug was effective against HMC-1560, 816 cells at the doses tested. INNO-406 effectively induced apoptosis in HMC-1560, but not in HMC-1560, 816 cells, as judged by the Annexin V flow cytometry test and by PARP specific cleavage seen on western blotting. In addition, INNO-406 was effective in inhibiting phosphorylation of c-kit in HMC-1560 cells in nM range. Our results suggest similar potency of INNO-406 and imatinib in mast cells with mutated codon 560 (found in gastrointestinal stromal tumor), but no activity against cells with mutated codon 816 c-KIT (found in SM).
Interferon Therapy Is Predictive Factor for Improved Survival in Patients with Essential Thrombocythemia (ET).
Authors: Jabbour Elias , Harb Antoine , Giles Francis , Cortes Jorge , Pierce Sherry , Shan Jianqin , Estrov Zeev , Kantarjian Hagop , Verstovsek Srdan
Published: April 2020

Essential thrombocythemia (ET) is a Philadelphia chromosome-negative Myeloproliferative Disease characterized by a relatively indolent clinical course, a predisposition to thrombotic and bleeding events, and a delayed occurrence of disease transformation into myelofibrosis (MF) or acute myelogenous leukemia (AML) in selected patients (pts). Current therapy for ET has not been shown to affect in a positive way either survival or risk of disease transformation.

We retrospectively assessed 158 pts with ET seen at our institution between November 1983 and February 2006, and evaluated their long-term outcome, risk of disease complications and transformation, and prognostic factors for both survival and disease complications. Median age was 51 years (range, 17–83), 59% were females. The median time from diagnosis to MD Anderson visit was 5 years (range 0–21) and from MD Anderson visit median follow up was 1.5 years (range 0–19). Fifty-two % of pts had received therapy before MD Anderson visit. At presentation 53% of the pts were symptomatic, including fatigue in 32%, weight loss in 8%, headache in 7%, and night sweats in 5%; splenomegaly was present in 18 pts (11%); median hemoglobin was 13.3 g/dL (range 7.1–16.6), white blood cell count 9.4 × 109/L (range 1.9–21.9), and platelets 909 × 109/L (range, 153–1000 × 109/L); karyotype was diploid in 129 of 139 evaluable pts (93%).

During the follow up 34 pts (22%) have never received any cytoreductive therapy, while 66 pts (42%) received more than one therapy. Therapy included hydroxyurea in 71 (45%), anagrelide in 47 (30%), and interferon/pegylated interferon in 67 pts (42%). Thrombotic and bleeding events occurred in, respectively, 33 (21%; 22 arterial and 11 venous thrombosis) and 29 (18%) pts. Transformation to MF occurred in 7 (4%) pts after a median of 73 months (range 5–159). At tranformation the karyotype was diploid in all but one pt (+1, der (1;17)(q10;q10), +8). Presently 135 pts (85%) are alive. The 5-year and 10-year overall survivals from MD Anderson visit were, respectively, 88.5% and 77.3%. The median survival has not been reached. The multivariate analysis of patients’ characteristics at presentation to and since MD Anderson visit identified interferon therapy as favorable predictive factor for survival (unlike other therapies). In contrast, this therapy constitutes an independent risk for a thrombotic event. No predictive factor for transformation has been identified.

We conclude that ET is a good prognosis disease with a low rate of transformation into more aggressive disease, and that interferon therapy may improve the outcome. Patients were referred to MD Anderson at different time in the natural history of their disease, and the evaluation of probable impact of interferon therapy on patients’ survival in ET should be verified in newly diagnosed patients.
Delta (9)-Tetrahydrocannabinol Contributes to the Development of Kaposi’s Sarcoma.
Authors: Zhang Xuefeng , Wang Jian Feng , Kunos George , Groopman Jerome E.
Published: April 2020

Kaposi’s sarcoma-associated herpesvirus (KSHV), a DNA tumor virus, appears to be necessary but not sufficient for the development of Kaposi’s sarcoma (KS). This is highlighted by the observation that most KSHV infections are asymptomatic, and more than 95% of persons who become infected never develop KS or other KSHV-related cancers. In addition to KSHV, a variety of factors, including HIV-1 Tat protein, may contribute to the pathogenesis of KS. These cofactors usually induce immunosuppression, enhance susceptibility to infection, modulate the viral life cycle, and/or stimulate angiogenesis and tumorigenesis either individually or in combination.

Delta (9)-Tetrahydrocannabinol (THC), the major active component of marijuana, has been prescribed for medicinal use and has also been reported to induce immunosuppression. To date, contradictory observations have been made about its effects on viral infection/replication and on tumor cell growth. This prompted us to study the biological effect of Δ9-THC on KSHV, and obtain more evidence to assess the potential risks versus benefits of THC administration among KSHV-positive populations. We found an increased efficiency of KSHV infection (40% vs 15%) in human dermal microvascular endothelial cells (HMVEC) in the presence of low concentrations of Δ9-THC (10–100 nM). THC enhanced endocytosis and cell-cell adhesion by 30–60%, which may account for this increase in infection efficiency. Using real-time DNA PCR technology, the number of copies of the KSHV latent gene, LANA-1, was quantitated and correlated with the viral load in endothelial cells. We found that Δ9-THC increased the viral load 20–100 fold in KSHV-infected endothelial cells as compared to treatment with the vehicle control. We then quantitated expression of the KSHV lytic switch gene ORF50 (RTA) by real-time RT-PCR, in the presence or absence of Δ9-THC. ORF50 expression was shown to be up-regulated by Δ9-THC, leading to increased KSHV loads. We also observed that comparable concentrations of Δ9-THC up-regulated vGPCR expression, thereby fostering endothelial transformation and inducing significantly more colony-formation (1.5 fold) in soft agar. Our results indicate that Δ9-THC may enhance KSHV infection, transmission, replication, and viral-mediated endothelial transformation. Thus, THC administration may put individuals at greater risk for the development and growth of Kaposi’s sarcoma.
Imatinib Mesylate in the Treatment of Systemic Mastocytosis with Wild-Type c-Kit Codon 816.
Authors: Hsieh Fred H. , Lichtin Alan E. , Katz Hary T. , Lubell Susan , Vasas Dorothy , Gibbons Aridth , Sharma Pankaj , Haque S.J.
Published: April 2020

Mastocytosis is a heterogeneous group of disorders characterized by the abnormal growth and accumulation of mast cells in one or more organs. Somatic mutations in the c-kit gene are associated with >80% of mastocytosis cases with the most common mutation identified as c-kit D816V. Imatinib mesylate is a potent inhibitor of c-kit tyrosine kinase activity, though the D816V mutation has been demonstrated to confer imatinib resistance both in vitro and in vivo. In this study we sought to determine the safety and efficacy of imatinib in patients with systemic mastocytosis who had no identified mutation that conferred imatinib resistance. We screened 24 patients and identified 5 patients who met study criteria, which included systemic mastocytosis by WHO consensus criteria (Valent et al, Leuk Res, 2003), impaired organ function to justify cytoreductive therapy, and wild-type c-kit exon 17 (including codon 816) DNA sequence analysis from peripheral blood, bone marrow, or lesional tissue specimens. In all cases patients were negative for the FIP1L1-PDGFRA fusion kinase. Four patients consented to the study and were treated with 400 mg of imatinib orally once daily in a dose-escalation protocol up to 800 mg daily for six months. Of the four patients entering the study, two had systemic mastocytosis with an associated hematologic non-mast cell disorder (in both cases CMML) and two had aggressive systemic mastocytosis. At the end of the six month treatment period all patients were assessed to have a partial response. Serum tryptase had fallen between 19–44% and skin lesions diminished in all patients with cutaneous involvement but did not resolve. All subjects reported subjective symptom improvement with regards to pruritis, flushing, GI symptoms, and other constitutional manifestations. However, bone marrow examination performed after the trial period when available demonstrated persistent mast cell infiltration. In two patients cutaneous mastocytosis lesions progressed within 6 months after discontinuation of imatinib. Imatinib was generally well-tolerated, though dose-dependent toxicity was noted in all patients and especially in patients > 65 years of age, and included edema, nausea, muscle cramps, conjunctivitis, cough, fatigue, and anemia.

Imatinib mesylate is safe and may have some efficacy in treating patients with systemic mastocytosis who lack imatinib-resistant mutations in c-kit.
Epidermal Growth Factor Is an Independent Prognostic Factor in Patients with Chronic Lymphocytic Leukemia.
Authors: Zhang Zhong , O’Brien Susan , Keating Michael , Jilani Iman , Kantarjian Hagop , Estrov Zeev , Ferrajoli Alessandra , Giles Francis , Albitar Maher
Published: April 2020

Epidermal growth factor (EGF) is a polypeptide expressed in various epithelial tissues and organs. EGF is believed to play an important role in cell development, differentiation, growth, apoptosis, and epithelial regeneration by the activation of signal transduction pathways that include PI3K/AKT, RAS/ERK, and JAK/STAT. These pathways may lead to both pro- and anti-apoptotic proteins. While hematopoietic cells do not express a significant amount of the EGF receptor (EGFR), bone marrow stromal cell and fibroblast expression of EGFR and EGF is important in the support of mesenchymal stem cell growth and colony formation. Since stromal cells play a major role in hematologic diseases, we evaluated the level of EGF in the plasma of 120 patients with chronic lymphocytic leukemia (CLL). EGF levels were significantly lower in CLL patients (median, 86.7; range, 2.2–500 pg/mL) than in 22 normal controls (median, 151.7; range, 35.1–389.1 pg/mL) (P = 0.02). EGF levels did not correlate significantly with β2-microglobulin (β2-M) level or IgVH mutation status, but correlated negatively with Rai stage (P = 0.0001). EGF level correlated positively with platelet count (R = 0.53, P <0.001) and negatively with WBC count (R = −0.32, P <0.001) as well as percent lymphocytes in bone marrow (R = −0.35, P <0.01). Using univariate Cox proportional hazards models, we demonstrated a negative correlation with survival when EGF was used as a continuous variable (P <0.001). The negative predictive value of EGF was independent of Rai stage, β2-M and thrombopoietin (TPO) level, IgVH mutation status, and prior therapy. EGF level as a continuous variable remained a strong predictor of survival (P <0.01) even when Rai stage, IgVH mutation status, β2-M and TPO levels, and prior therapy were included in the model. When dichotomized according to the median EGF level (86.7 pg/mL), patients with higher levels had longer survival (P <0.001). These data suggest that EGF levels are a powerful and independent prognostic factor in CLL and represent a simple and reliable approach to stratify patients for therapy. Further studies are needed to understand the biological basis of the role of the EGF in CLL to determine whether it reflects the general well-being of the host or the health of stromal elements in the bone marrow.

Figure Figure
Genomic Aberrations in Chronic Lymphocytic Leukemia and Small Lymphocytic Lymphoma by Absolute Lymphocyte Count: Analysis of 1694 Patients.
Authors: Tsimberidou Apostolia-Maria , McLaughlin Peter , O’Brien Susan , Wen Sijin , Wierda William G. , Manning John T. , Abruzzo Lynne V. , Lerner Susan , Hess Mark A. , Kantarjian Hagop M. , Freireich Emil J. , Keating Michael J.
Published: April 2020

Introduction: Genomic aberrations provide information for the pathogenesis and clinical outcomes of patients with chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL). We assessed whether genomic aberrations in CLL/SLL differ by absolute lymphocyte count (ALC) and evaluated their prognostic significance in our series of patients with CLL/SLL.

Methods: We reviewed the medical records of patients with CLL/SLL who presented to The University of Texas M. D. Anderson Cancer Center from 1985 to 2005. Patients were assessed by conventional bone marrow cytogenetics and/or fluorescence in situ hybridization (FISH) analysis using probes for trisomy 12, ATM, LAMP1, D13S319, and P53 genes.

Results: Cytogenetics and/or FISH analysis was performed in 1694 of 2189 consecutive patients with CLL/SLL. Patients with ALC < 5 × 109/L had lower rates of genomic aberrations by cytogenetic and/or FISH analysis than did those with ALC ≥ 5 × 109/L (25% vs. 37%, p=0.02). Deletion 17p +/− other abnormalities and 6q del +/− other abnormalities were each associated with shorter survival compared with other genomic aberrations or normal karyotype (p<0.0001 for both results). The survival rates by genomic aberration are shown in the Figure [insert figure here]. In a multivariate analysis of 23 clinical and laboratory factors, deletion 17p or 6q +/− other genomic aberrations (p<0.0001) was the strongest independent predictor of shorter survival. In patients who required therapy for CLL/SLL, independent factors predicting response were hemoglobin levels > 11 g/dL (p<0.0001), age < 60 years (p=0.005), absence of 17p deletion (p=0.048), and absence of 6q deletion (p=0.03). In multivariate analysis, pretreatment parameters that remained independently significant for longer failure-free survival (FFS) were age < 60 years (p<0.0001), absence of 17p del +/− other abnormalities (p<0.0001), and hemoglobin levels > 11 g/dL (p=0.018).

Conclusions: Patients with CLL/SLL and ALC < 5 × 109/L had lower rates of genomic aberrations by cytogenetic and/or FISH analysis (p=0.02), possibly because of less bone marrow infiltration in the lower-ALC group. Deletion 17p or 6q, with or without other genomic aberrations was the strongest independent adverse prognostic factor for survival in CLL/SLL.

Figure Figure
The Assessment of Anti HSP70 Antibodies in Patients with Chronic Lymphocytic Leukaemia.
Authors: Piszcz Jaroslaw , Kloczko Janusz , Rudy Agnieszka , Blusiewicz Eliza , Lemancewicz Dorota , Mazgajska-Barczyk Katarzyna
Published: April 2020

Background: Chronic lymphocytic leukaemia (CLL) is a one of the disease in which processes of proliferation and apoptosis are seriously altered. It has been intensely studied in the recent years in order to understand the mechanisms of neoplastic development. Hsps or heat shock proteins are molecular chaperones involved in a number of cellular functions in stress conditions. Feng et al. (2002) have reported that heat-stressed apoptotic 12B1-D1 leukaemia cells (BCR-ABL(+)) express HSP70 on their surface. It has also been suggested that an increased surface expression of heat shock proteins on apoptotic tumour cells results in the generation of potent antitumour T-cell responses. Anti Hsp70 antibodies are known to play a role in immunological and neoplastic processes. However their significance in CLL has not been well documented.

Aims: The aim of our study was the assessment of the anti Hsp70 antibody concentration in the patients with CLL.

Material and methods: We assessed 20 peripheral blood samples from the patients with newly diagnosed CLL. (aged 46–74), including 11 males and 9 females. A group of healthy age matched subjects were used as a control group. The patients were in A-C stages of CLL according to Binet scale. Quantitative determination of anti-human Hsp70 antibodies in the serum was done using commercial test (anti Hsp70 Elisa Kits, Stressgen). The results are presented as mean ± SEM. Statistical analysis was done using Shapiro-Wilk, Mann-Whitney and Spearman’s tests.

Results: The levels of anti HSP70 antibodies were significantly lower in the group of patients with CLL in comparison to healthy controls (141,99±74,26 ng/ml vs.298,36±57,06 ng/ml; p< 0,01). In our analysis there was no association between the levels of antibodies and the stage of the disease. There were no correlations between the levels of anti Hsp antibodies and other parametres such as age, gender and some prognostic factors (LDH, β2-microglobulin, and lymphocyte doubling time) in the studied group.

Conclusions: The significantly lower concentrations of anti HSP70 antibodies in CLL patients suggest that these molecules may play a role in the biology of the disease. The encouraging results from our study suggest that it might be interesting to assess their level depending on the response to the applied treatment. Further studies are required to elucidate the role of these proteins in leukaemic conditions.