bluuebunny/crossref_metadata_2025 huggingface dataset:
Significance of Laminar Air Flow Room for the Reverse Isolation in Neutropenic Cancer Patients.
Authors: Lee Ji Hye , Kim Hae Sook , Yoon Hwi-Joong , Lee Kun Soo
Published: October 2019

Infection is one of the most important causes of death in cancer patients. So many physicians make every effort to control the infection, especially in neutropenic cancer patients. The aim of this study is to find out the role of HEPA filter equipped laminar air flow room reverse isolation for the management of chemotherapy induced febrile neutropenic children with cancer. We evaluated febrile neutropenic patients following chemotherapy from January 2003 to April 2006 at the Department of Pediatrics, Kyungpook National University Hospital, Daegu, Korea. They were promptly managed by antibiotics and antifungal agents and if possible, they were isolated in the aseptic room. They were allocated to three groups of standard ward care, isolation after onset of fever and isolation before onset of fever. Profiles of infection, clinical courses and survival rate were compared among three groups. One hundred and nine episodes of febrile neutropenia from thirty eight cancer patients were observed. Twenty nine were boys and nine were girls with their median age were 5.5 years. The diagnoses included acute leukemia (36.8%), malignant lymphoma (15.8%), and other solid tumor (47.4%). Fifty five episodes were included to standard ward care, forty four episodes to isolation after onset of fever and thirteen episodes to isolation before onset of fever. We found out that one in thirteen episodes of isolation before onset of fever (8%) and twenty six in ninety six episodes in other groups (27%) were microbiologically or clinically defined infections. Fifty six episodes recovered and only one died of infectious cause both isolation after and before onset of fever group, especially all recovered in isolation before onset of fever group. Forty eight episodes recovered and four died in general ward care group, but there was no statistical difference among three groups (p=0.93). Age, sexual difference, underlying disease, absolute neutrophil count (ANC), duration of ANC recovery and incidence of disseminated intravascular coagulation were no difference among three groups. In early isolated group, duration of fever and antibiotics medication were significantly shorter (p=0.002, 0.009) and CRP level was lower than other two groups (p=0.04). Reverse isolation in laminar air flow room for neutropenic cancer children before onset of fever affect the durations of fever and antibiotics treatment with beneficial effect.
Assessment of Quality of Life, Psychological Distress, and Alexithymia in Patients with Cutaneous Lymphoma.
Authors: Sampogna Francesca , Frontani Marina , Baliva Giannandrea , Lombardo Giuseppe , Di Pietro D Stat Cristina , Alvetreti Gabriele , Tabolli Stefano , Abeni Damiano , Russo Giandomenico
Published: October 2019

Cutaneous lymphoma is a visible cutaneous malignancy, which especially in the more severe forms may have a profound effect on patients lives. The aim of this study was to evaluate quality of life (QoL), psychological distress, and alexithymia in patients with cutaneous lymphoma referring to a dermatological hospital, and to correlate these results with clinical parameters. QoL was evaluated with the Skindex-29, that measures QoL in skin conditions on three scale (symptoms, emotions, and functioning), and the EORTC QLQ-C30, that assesses the QoL of cancer patients. Presence of minor non-psychotic psychiatric disorders was assessed using the GHQ-12, defining as case patients scoring 4 or more; alexithymia was evaluated using the TAS-20 questionnaire. In our ongoing study, we analysed data from the first 66 patients. Of them, 67% were men, the mean age was 58 years (range: 26–85y), and there were 15 patients (23%) with cutaneous B-cell lymphoma (CTBL), and 51 with T-cell lymphoma (CTCL), 6 of them with Sézary Syndrome. In our population, 31.3% of patients were GHQ-cases, 18.6% had alexithymia, and another 22% possible alexithymia. Dermatology-specific QoL was particularly impaired in the symptoms and the emotions scales. QoL was always significantly more impaired in patients with CTCL than in those with CBCL. Also, the prevalence of GHQ-cases was higher (35% in CTCL vs 13% in CBCL), as well as the prevalence of people with alexithymia (48% vs 20%). Skindex-29 and TAS-20 scores significantly correlated with the T stage of lymphoma (i.e., the higher the stage, the lower the QoL and the higher the prevalence of alexithymia). The evaluation of quality of life and psychological problems in patients with cutaneous lymphomas may help clinicians to better manage the disease and its burden on patients life. Also, the relation between quality of life impairment and clinical variables may give important information on the course of the disease as well as the possible effect of treatment.
Epidemiology of Transfusion Related Acute Lung Injury in e-FIT, the French Hemovigilance Database: 1994 to 2006. A Study of the French Hemovigilance Network.
Authors: Renaudier Philippe , Vo Mai Mai Phuong , Ounnoughene Nadra , Willaert Beatrice , Caldani Cyril
Published: October 2019

Background: Transfusion Related Acute Lung Injury (TRALI) is a life-threatening complication of allogenic blood transfusion, manifested typically by a non-cardiogenic pulmonary edema. However, the magnitude of the risk of TRALI remains unknown at this time, all the more so as a variety of other clinically similar respiratory complications can be associated with transfusion. Hemovigilance had been implemented in France since 1994. All adverse effects (AE) regardless of their severity should be notified on a normalized form to AFSSaPS, the French health products safety agency, as required by the French law.

Objective : to describe TRALI observed in e-FIT, the French hemovigilance database.

Population and methods: AE possibly, probably or definitely associated with transfusion were considered for inclusion. As the item “ TRALI” was explicitly present since 09/01/2001, we screened the database from 07/01/1994 to 09/01/2001 with “ pulmonary edema AND fluid overload excluded” , or with free comments including the words “TRALI ” , “ anti-granulocyte antibodies ”, “ non-cardiogenic pulmonary edema ” , “ white lung syndrome ” . The subsequent AE were considered for inclusion only if TRALI was the final diagnostic.

Results: From 07/01/1994 to 06/01/2006, 82,436 AE were registered in e-FIT, corresponding to about 30,000,000 labile blood products (LBP) transfused. 117 AE fulfilled the above criteria (3.9/1,000,000 LBP transfused). There were 59 men and 58 women, age = 0 to 86 (median 58); 34 men and 21 women were 60 years old or more. 107 patients presented with pulmonary symptoms, 93 with dyspnea, 63 with chills and/or hyperthermia and 53 with a typical pulmonary edema. 13 TRALI were fatal, 85 were life-threatening and 19 were benign. 42 patients received only packed red cells, 4 only fresh frozen plasma and 40 only platelet concentrates ; among the 31 remainders, 14 received platelets concentrates associated with an other LBP.

Conclusion:

Although the initial French database was not adapted to detect TRALI cases until 2001, the dynamics of notification is of interest ; TRALI is more frequent in elderly patients, as noted in other reports ; Platelets concentrates remains the more frequently LBP involved, but packed red cells also contribute to nearly half of the TRALI reported ; Besides the classical life-threatening presentation, a benign form of TRALI seems to exist ; A national working party is implementing new ways to define and investigate TRALI, which will provide more accurate information on TRALI in France.
The Anti-Apoptotic Role of the Unfolded Protein Response in Bcr-Abl Positive Leukemia Cells.
Authors: Tanimura Atsuko , Yujiri Toshiaki , Tanaka Yoshinori , Tanizawa Yukio
Published: October 2019

Accumulation of unfolded or misfolded proteins within the endoplasmic reticulum triggers the unfolded protein response (UPR). Evidence from several studies suggests that the UPR is activated in various tumors and might play a crucial role in tumor growth. However, the role of the UPR in leukemia remains unclear. Therefore, to define the role of the UPR in leukemogenesis, we used p210 Bcr-Abl-expressing 32D myeloid cell lines (p210 32D). The mRNA expression of UPR-related genes, namely, a spliced form of X-box DNA-binding protein (XBP1) and glucose regulated protein 78 (GRP78), was increased in p210 32D. Luciferase assay indicated that p210 Bcr-Abl induced high levels of the transcriptional activity of the UPR element. Moreover, levels of the phosphorylated eIF2 alpha protein increased in p210 32D. Inhibition of the UPR using IRE1 and ATF6 dominant-negative mutants diminished the ability of Bcr-Abl to protect the cells from etoposide- and imatinib-induced apoptosis, but had no effect on the proliferation of p210 Bcr-Abl-transformed cells. We also evaluated the expression of UPR-related genes in primary leukemic cells from Ph chromosome-positive cells by real-time RT-PCR; it was found that these cells showed higher expression levels than the control cells. Taken together, these results for the first time suggested that UPR is a downstream target of the Bcr-Abl oncoprotein, and it plays the anti-apoptotic role of Bcr-Abl in Ph chromosome-positive leukemia cells.
Efficacy and Tolerability of Pregabalin in Decreasing Pain Associated with High Dose of Imatinib in Newly Diagnosed Philadelphia Chromosome-Positive Chronic Phase CML.
Authors: Iuliano Francesco , Maio Massimo DI , Infusino Stefania , Perricelli Alessia , Pomillo Angelo , Cilloni Daniela , Saglio Giuseppe
Published: October 2019

Background Recently published data suggest that high doses of imatinib mesylate (IHD) (800 mg/daily) may be more effective than standard dose (400 mg/daily) in newly diagnosed Philadelphia chromosome-positive chronic phase CML. Furthermore escalating the dose of imatinib to 800 mg per day can overcome some cases of resistance to imatinib but tolerability of high-dose imatinib continues to be an issue.

Aims To evaluate the efficacy and safety of pregabalin dosed twice daily (BID) for relief of musculoskletal and neuropathic pain associated with IHD.

Methods 13 patients in chronic phase (CP) were enrolled because complained of severe, persistent pain, requiring nonoppioid analgesics. Patients were asked to quantify their pain using a visual analogue scale (VAS) (numeric range 0–10). QoL assessment was based on Therapy Impact Questionnaire. At baseline, the median pain-rate was 5 (range 3–8). Pts received pregabalin in doses ranging from 75 to 300 mg twice daily for 12 weeks.

Results Despite a short study duration, a significant reduction was seen in weekly pain score (p < 0.0001), as well as VAS score (p < 0.0001) allowed pts to receive the scheduled dose of imatinib. Pregabalin was well tolerated, and the most common adverse event was only somnolence. The short study duration precluded the assessment of longer term safety issues such as weight gain.

Conclusions Pregabalin was safe and effective in decreasing pain associated with high dose of imatinib, and also improved mood, sleep disturbance, and quality of life.
Examination of Point Mutations within the ABL Kinase Domain of Bcr-Abl Fusion Gene in Chinese Patients with Chronic Myeloid Leukemia Who Develop Imatinib Resistance.
Authors: Yang Dongguang , Zhang Ri , Gang Honglin , Gu Jian , Cen Jian Nong
Published: October 2019

Objective: To investigate the point mutations within the ABL kinase domain (KD) of Bcr-Abl fusion gene in Chinese patients with chronic myeloid leukemia who develop IM resistance and investigate the mutation.

Method: We collect a total of 17 bone marrow samples obtained from 11 patients who showed hematology resistance(n=7) or cytogenetic refractoriness(n=4). A long semi-nest PCR method was used to amplify the ABL KD of the Bcr-Abl fusion gene. After two cycles of PCR reaction, we get a fragment including 863 bases, purified and sequenced it.

Result: The ABL point mutation was detected in 45.45% of patients. The mutation rate of hematology resistance is 57.14% and the mutation rate of cytogenetic refractoriness is 25%. In total, we find three point mutation in all of our patients tested, ie, G250E(2), E255K(1) and T315I(2). In patients who develop hematology resistance, the earliest time to detect mutation is 4 months later after the therapy of IM and the longest time to detect mutation is 9 months later after the therapy of IM. In patients who develop cytogenetic refractoriness, the time to detect mutation is 12 months later after the therapy of IM .For the patients whose samples are available, no single mutation happens before IM thearpy.

Conclusion: There is high frequency of point mutations clustered within the adenosine triphosphate-binding region of Bcr-Abl fusion gene in Chinese patients with chronic myeloid leukemia, and the mutation is related to IM therapy. It’s good for patients to switch to another therapeutic strategy when the mutations were detected earlier.

Figure Figure
Expressions of G-CSF Receptor in Tumor Cells Might Stimulate the Proliferation of Malignant Cells Upon Treatment of G-CSF.
Authors: Kim Tae Young , Moon Hee Won , Oh Bora , Hwang Sang Mee , Ku Ja-Lok , Lee Dong Soon
Published: October 2019

Abstract 1009

Poster Board I-31

We previously reported that the increased expression of G-CSFR in AML1/ETO+ AML cell, suggesting that cells with G-CSFR expression might easily proliferate in response to therapeutic G-CSF. To confirm the association of AML/ETO gene and increased expression of G-CSFR, we transfected AML1/ETO specific small interfering RNAs (siRNAs) to AML/ETO+ cell line and G-CSFR expression was decreased on transfection by siRNA AML1/ETO gene suggesting a strong association between AML1/ETO gene and G-CSFR up-regulation. To assess the effect of G-CSF on G-CSR expressing leukemic cells, we treated the most commonly used two forms of G-CSF on various leukemic cell lines. Both forms of G-CSF significantly stimulated the cell proliferation (40-80% increment compared to control) and the differentiation of AML1/ETO+ leukemic cells. Western blot showed marked increased signals of JAK2/STAT3 after treatment of G-CSF on AML/ETO+ Kasumi-1 cell line, while there was no increased signals in AML/ETO- CTV-1 cell line. To search the malignant tumors with increased G-CSFR, we screened the expressions of G-CSFR mRNA in 32 kinds of solid tumor cell lines and % kinds of leukemic cell lines, using real-time PCR. Among 32 kinds of solid tumor cell lines, 3 cell lines [hepatoblastoma (HepG2), squamous cell carcinoma of larynx (SNU-899), and breast carcinoma cell line (MCF 10A)] expressed G-CSFR mRNA and amount of G-CSFR expression of HepG2 cell was comparable to Kasumi-1 cell lines. In summary, association of G-CSFR and AML/ETO gene is strongly suggested. Expression of G-CSFR in some tumor cells is suspected, requiring pre-screening of G-CSFR expression before treatment of G-CSF. The present study shows that therapeutic G-CSF might stimulate the proliferation and differentiation of malignant cells with G-CSFR expression (AML/ETO+ AML cell and solid tumor cell lines) and we suggest the prescreening of G-CSFR expression primary tumor cells before treatment of G-CSF.

Disclosures:

No relevant conflicts of interest to declare.
The Use of Purine Analogues Does Not Aggravate Infectious Complications During Induction and Consolidation Therapy of Acute Myeloid Leukemia.
Authors: Seweryn Marek , Wojnar Jerzy , Kata Dariusz , Kyrcz-Krzemien Slawomira
Published: October 2019

Abstract 1011

Poster Board I-33

Background:

Addition of purine analogues to standard induction therapy of acute myeloid leukemia (AML) had previously been demonstrated to increase complete remission rate. The aim of this study was to analyze whether the use of cladribine or fludarabine during induction and consolidation increases the risk of infectious complications.

Material and methods:

118 AML patients, included in two consecutive randomized trials between 1999-2006 in a single centre were analyzed. Induction therapy consisted of daunorubicin + cytarabine (DA-7, n=53) alone or in combination with cladribine or fludarabine (DAC-7 + DAF-7, n=65 ). Consolidation included one course of high-dose AraC + mitoxantrone and one course of high-dose AraC +/- purine analogues. A median age was 45(17-58) years and 48(20-60) years for patients treated with and without purine analogues, respectively.

Results:

The frequency of neutropenic fever as well as microbiologically documented bacterial, fungal and viral infections during induction and consolidation did not differ between two compared groups - receiving or not purine analogues. Time to infection occurrence and infection duration were similar in both study groups.

During induction and both consolidation treatments significant lower values of lymphocytosis were observed in the group of patients treated with purine analogues.

There was a slight tendency to increased rate of mucositis for patients treated with purine analogues (60% vs. 44.3%, p=0.07) during induction treatment, while infections affecting skin and soft tissues were significant frequent for patients treated without purine analogues (43.3% vs. 18%, p=0.03) during second consolidation treatment (high dose AraC).

The usage of intravenous anti-infectious medications (antibiotics, antifungal, antiviral) and periods of hospitalization did not differ between two groups in this study.

Conclusions:

We conclude that the use of purine analogues, either cladribine or fludarabine along with conventional induction and consolidation therapy does not aggreviate infectious complications in adults with AML.

Disclosures:

No relevant conflicts of interest to declare.
PI3K Signaling Pathway Activation Predicts Class I PI3K Inhibitor GDC-0941 Sensitivity in AML.
Authors: Ma Xiaoju Max , Du Changchun , Sun Laura , Shi Xiaoyan , Friedman Lori , Dornan David , Ebens Allen J.
Published: October 2019

Abstract 1057

Poster Board I-79

The PI3K-Akt signal transduction pathway plays a key role in the pathogenesis of many human cancers. In AML malignancy, deregulation of upstream receptor tyrosine kinases such as FLT3, c-Kit, and c-FMS or mutations in K-Ras, N-Ras, B-Raf and CEBPA genes lead to activation of PI3K-Akt signaling to promote cell survival and cell growth. A highly selective Class I PI3K inhibitor, GDC-0941, provides exciting therapeutic opportunities for targeting this pathway in AML. Here we show that GDC-0941 significantly inhibits the viability of the majority of a large panel of AML cell lines tested in vitro (80% or 19/24) at a concentration of < 1 uM. Because not all AML cell lines responded to GDC-0941, we show that PI3K-Akt pathway activation, evidenced by basal pAkt level, can serve as a potential predictive biomarker for GDC-0941 in AML in that sensitive cell lines displayed higher level of pAkt relative to resistant cell lines. Consistently, GDC-0941 treatment leads to decreased pAkt, and therefore the down-regulation of this important pro-survival signaling. Our further analysis shows that GDC-0941 treatment can induce apoptosis and/or cell cycle arrest. We also obtained fresh AML tumor samples to test whether GDC-0941 can similarly induces apoptosis in blast cells and showed that GDC-0941 treatment results in a down-regulation of pAkt level and increased apoptosis. Other PD biomarkers such as phospho-BAD level and Bim expression are both consistent with the observed apoptotic responses. Furthermore, the mammalian target of rapamycin complex 1 (mTORC1) inhibitor, rapamycin, synergizes with GDC-0941 to produce an increased amount of apoptosis in several AML cell lines tested. This is likely due to the fact that long-term treatment with rapamycin induces the sensitivity of the PI3K –Akt signaling pathway by releasing the negative feedback loop of mTORC1-S6K-IRS1/2 module. Importantly, in some AML cell lines we observe synergies between GDC-0941 and AraC. Interestingly, while AraC alone does not induce apoptosis in AML cell lines with PTEN loss or mutation, the synergy between AraC and GDC-0941 comes from increased apoptotic response, suggesting that GDC-0941 can synergize with chemo agents that induces S/G2 cell cycle arrest. Together, our preclinical data suggest that GDC-0941 may be used as a targeted therapy in AML patients as a single agent or in combination with other chemotherapies in clinic.

Disclosures:

Ma: Genentech Inc.: Employment. Du:Genentech, Inc.: Employment, Equity Ownership. Sun:Genentech Inc.: Employment. Shi:Genentech, Inc.: Employment, Equity Ownership. Friedman:Genentech Inc.: Employment. Dornan:Genentech, Inc.: Employment, Equity Ownership. Ebens:Genentech, Inc.: Employment, Equity Ownership, Patents & Royalties.
Erosion of Telomeric Single-Stranded Overhang in Patients with Aplastic Anemia Carrying Telomerase Mutations.
Authors: Calado Rodrigo T. , Regal Joshua A. , Kajigaya Sachiko , Young Neal S
Published: October 2019

Abstract 1079

Poster Board I-101

Loss-of-function mutations in telomerase complex genes reduce telomerase activity and shorten overall telomere length in leukocytes, and they can clinically manifest as bone marrow failure (aplastic anemia and dyskeratosis congenita), familial pulmonary fibrosis, and hepatic cirrhosis. The double-stranded tandem telomeric TTAGGG repeats are followed by a 3' G-rich single-stranded overhang, a crucial structural component responsible for protective t-loop formation. We investigated the length of telomeric overhangs in 25 healthy individuals from 0 to 76 years, 16 patients with aplastic anemia, and 13 of their immediate relatives using a non-denaturing in-gel method and the telomere-oligonucleotide ligation assay (T-OLA). Among healthy controls, overall telomeric length of leukocytes shortened as a function of age. It was longest in umbilical cord blood samples and eroded as a function of a third-order polynomial until 76 years (R2=0.9517; Spearman=0.90; P<0.0001). However, the lengths of single-stranded overhangs were constant with age, as determined by both non-denaturing and the T-OLA methods. In contrast, most patients with marrow failure carrying a telomerase gene mutation showed marked erosion of telomeric overhangs associated with critically short telomeres (P<0.05). Seven telomerase complex-mutant aplastic patients had overhang lengths below the 5% percentile established for healthy volunteers. In aplastic patients with normal genotypes, normal overall telomere lengths, and who effectively responded to immunosuppressive therapy, telomeric overhangs were maintained and were all within the range established for control individuals. In conclusion, telomeric overhang erosion does not participate in physiological aging but eroded telomeric overhangs and abnormal telomere structure appear in pathologic shortening of telomeres, especially caused by loss-of-function telomerase mutations. Disrupted telomere structure caused by short telomeric overhangs may contribute to the mechanisms of abnormal hematopoietic compartment senescence and chromosomal instability in human bone marrow failure.

Disclosures:

No relevant conflicts of interest to declare.
ABCG2 C.421C>A Is Associated with Higher Trough Imatinib Plasma Levels in Patients with Chronic Myeloid Leukemia.
Authors: Takahashi Naoto , Miura Masatomo , Scott Stuart A , Sawada Kenichi
Published: October 2019

Abstract 110

[Background] Despite the excellent efficacy of imatinib for the treatment of chronic myeloid leukemia (CML), trough imatinib plasma levels can vary widely among patients. This may be due, in part, to inter-individual variation in imatinib metabolism and drug transport efficacy. To investigate the role of genetic variation in the pharmacokinetics of imatinib, we analyzed common single nucleotide polymorphisms within important imatinib pathway genes including ABCG2 (BCRP), ABCB1 (MDR1), ABCC2 (MRP2), CYP3A5, and SLC22A1 (OCT1) in 67 CML patients treated with imatinib. In addition, trough imatinib plasma levels were determined using high-performance liquid chromatography-tandem mass spectrometry. [Results] Distinct imatinib pharmacokinetics were identified in association with ABCG2 c.421C>A (p.Q141K; rs2231142) genotype. Specifically, the presence of the variant c.421A allele was significantly (p=0.024) associated with higher imatinib concentrations [median Cmin/Dose 2.70 (range: 1.50-8.30) ng/ml/mg; n=25] compared to patients with the wild-type ABCG2 (c.421C/C) genotype [median Cmin/Dose 2.27 (range: 0.37-5.30) ng/ml/mg; n=42]. ABCG2 is an efflux transporter for many xenobiotics, including imatinib, and is expressed at high levels in the human liver. Previous studies indicate that c.421A causes a 40% reduction in imatinib transport in vitro when compared to the wild-type genotype. Our data suggest that CML patients with ABCG2 c.421A allele may have deficient ABCG2 activity in vivo, resulting in reduced hepatic excretion of imatinib. Of note, although less common among Africans and individuals of European decent, the ABCG2 c.421C>A allele occurs at a high frequency in the Japanese (0.311) and Han Chinese (0.289) populations. [Conclusion] The association of ABCG2 c.421C>A with imatinib pharmacokinetics may explain why some Japanese CML patients administered less than 400 mg/day of imatinib have clinically sufficient trough imatinib plasma levels. Prospective studies are warranted to confirm the association between ABCG2 genotype and imatinib pharmacokinetics in large patient populations.

Disclosures:

No relevant conflicts of interest to declare.
OCT-1 Protein Expression Detected by Flow Cytometry Provided Valuable Information in CML Patients Treated with Imatinib MesylateÃ,Â FHigher hOCT-1 Expression Predict Better Outcome.
Authors: Zhu Huanling , Zhang Qiurong , Jiang Nenggang , Liu Ting
Published: October 2019

Abstract 1117

Poster Board I-139

Objective

The human organic cation transporter-1(hOCT-1) is the major active influx protein responsible for the transport of imatinib into cells. The functional activity of the hOCT-1 protein using 14-C detected by others demonstrated a link between CML molecular response and hOCT-1 activity. However, 14-C labeled detection is not convenient in routine clinical practice. Hence, we use flow cytometry to detect hOCT-1 protein expression level in CML patient and try to find some relation between hOCT-1 expression and imatinib response.

Subjects and methods

In this study, 64 CML CP patients and 31 healthy donors were enrolled. Totally, there are 78 patient' peripheral blood (PB) or bone marrow (BM) samples and 31 donor PB samples were measured. The hOCT-1 protein expression levels were detected by indirect immunofluorescent flow cytometry. The hOCT-1 levels were expressed as mean fluorescent intensity (MFI). In avoided to systematic error, lymphocytes which had little hOCT-1 expression were used as internal negative control.

Results

‡@ Assessing PB hOCT-1 expressing in patients with donors, hOCT-1 level is higher in healthy donors than in CML patient (mean±standard deviation 9.11±6.04,5.60±3.74,P=0.005). ‡AThe hOCT-1 level was compared with molecular response in patients. Of 39 patients achieved optimal molecular response, the hOCT-1 level was 6.49±3.83, versus 3.86±2.91 in 20 patients with non-optimal response(P=0.009). Comparing 39 optimal responders with 16 sub-optimal responders, hOCT-1 level were 6.49±3.83, versus 3.98±1.23(P=0.025. ‡B Assessing CML stages with hOCT-1 expression, there is no significant difference in chronic stage and advanced stage(5.93±3.87, 3.49±1.64, P=0.085).

Conclusions

hOCT-1 expression level measured by flow cytometry is very convenient and clinically available. The hOCT-1 expression level can be an important predictor in CML patients treated with imatinib mesylate.

Disclosures

No relevant conflicts of interest to declare.
Large Granular Lymphocyte (LGL) Expansions Comprising Oligoclonal T Cell or NK Cell Populations in Dasatinib Treated Patients Are Associated with HLA-A*0201, CMV Reactivation and Enhanced Anti-Leukemic Control.
Authors: Mustjoki Satu , Kreutzman Anna , Dix Carolin , Price David A , Ladell Kristin , Ekblom Marja , Stenke Leif , Einsele Hermann , Porkka Kimmo , Seggewiss Ruth
Published: October 2019

Abstract 1123

Poster Board I-145

Introduction

Immunosuppressive effects of the second generation tyrosine kinase inhibitor dasatinib (Sprycel®) on T cells and NK cells have been described in vitro. In contrast, in some CML or Ph+ ALL patients receiving dasatinib, the development of a chronic, oscillating lymphocytosis has been observed in vivo. We previously showed that dasatinib-induced lymphocytosis typically comprises of oligoclonal NK or CD8+ cytotoxic T cell large granular lymphocyte (LGL) expansions and was associated with HLA-A*0201, CMV reactivation and enhanced, long-lasting therapy responses in patients with advanced leukemia.

Patients and Methods

To further elucidate the mechanisms underlying LGL expansions during dasatinib, we analyzed T cell and NK cell effector functions directly ex vivo. Specifically, we evaluated activation, proliferation, cytotoxic activity, cytokine secretion, degranulation, KIR expression profile and apoptosis/necrosis susceptibility.

Results

We observed decreased NK cell cytotoxic activity against the cell line K562 only in patients with NK cell expansions, potentially due to exhaustion based on excessive in vivo proliferation. This reduced lytic ability was restored after pre-treatment with IL-2 for 18h. No correlation between functionality and KIR expression profile was found. Furthermore, patients with LGL expansions exhibited elevated IP10/RANTES levels in the plasma compared to non-LGL patients and healthy controls. Strikingly, within the CD8+ T cell population specific for the CMV epitope NLVPMVATV (pp65, residues 495-503) restricted by HLA-A*0201, we observed distinct CD8high and CD8low fractions in patients with LGL expansions (n=5) but not in patients without LGL expansions (n=4) or in healthy controls (n=3). The CD8high subpopulation was more resistant to inhibition by dasatinib compared to other cell fractions; in addition, dasatinib effectively abrogated down-regulation of both TCR and CD8 in the CD8high subpopulation in vitro.

Conclusions

Thus, we hypothesize that CMV reactivation is intimately linked to the observed LGL expansions in dasatinib-treated patients. The expanded cytotoxic cells may target both CMV-infected and leukemic cells by virtue of epitope sharing and thus explain the enhanced anti-leukemic control we observed. Further investigations based on the application of polychromatic flow cytometry and analysis of T cell clonality are underway to establish the nature of this association in more detail. Dasatinib and derivatives may prove to be useful in modulating anti-leukemic/anti-host immune responses in vivo.

Disclosures

Mustjoki: BMS: Honoraria. Ekblom:BMS: Honoraria. Porkka:Novartis: Honoraria, Research Funding; BMS: Honoraria, Research Funding. Seggewiss:BMS: Honoraria.
Fludarabine, Cyclophosphamide, Anti-Thymocyte Globulin, with or without “ 2Gy TBI, for Alternative Donor Transplants in Acquired Aplastic Anemia (SAA): A Report From the EBMT-SAA Working Party.
Authors: Bacigalupo Andrea , Socie' Gerard , Lanino Edoardo , Prete Arcangelo , Locatelli Franco , Locasciulli Anna , Cesaro Simone , Shimoni Avicahi , Marsh Judith , Brune Mats L , Passweg Jakob R.
Published: October 2019

Abstract 1207

Poster Board I-229

Patients.

Data from 100 patients with acquired severe aplastic anemia (SAA) undergoing an alternative donor transplant, were analyzed. Patients were prepared with a combination of fludarabine (30 mg/m 2×4), cyclophosphamide (300 mg/m 2×4), antithymocyte globulin (3.75 mg/lgx4) (FCA) (n=52, median age 13 years), or FCA supplemented with low dose (2 Gy) total body irradiation (FCA-TBI), but with a lower dose of ATG (total 7.5 mg/kg) (n=48, median age 27 years). The donor was unrelated (n?87) or a one antigen mismatched family donor (n=13).

GvHD.

Acute graft versus host disease (GvHD) grade III-IV was seen in 13% and 7% ; extensive chronic GvHD was recorded in 1 FCA patient and in 4 patients receiving FCA-TBI.

Graft failure.

Rejection/graft failure was seen in 17 patients, equally distributed in the two groups: 9/17 patients survive long term, 3 with autologous recovery, and 6 after a second transplant.. As to predictors of graft failure, patients with a longer interval from diagnosis to transplant (> 2 years) had a trend for a higher risk of GF (22%) as compared to patients grafted <1 year (12%) or between 1-2 years (14%) (p=0.3).

Chimerism data:

within 100 days from BMT the average donor chimerism was 93% and 84% in the FCA and FCA-TBI regimens. Beyond day +100 the average was 89% and 80%.

Survival.

With a median follow up of 1204 days, the overall actuarial 5 year survival is 75%, respectively 73% for the FCA and 79% for the FCA-TBI. The interval diagnosis-transplant (<1 year, 1-2 years and >2 years) was a significant predictor of survival in univariate and multivariate analysis : actuarial survival was 87%, 86%, 58% respectively (p=0.004). Older age and HLA mismatch was a significant predictor of survival in the FCA group, but not in the FCA-TBI patients.

Causes of death.

Twentythree patients died, 13/52 in the FCA group and 10/48 in the FCA-TBI group (p=ns): major causes of death were rejection (n=7), post-transplant-lymphoproliferative-disease (n=4) and GvHD (n=4).

Conclusions.

This study confirms a significantly improved outcome of alternative donor transplants in SAA patients, and suggests that best results are achieved if the transplant is performed within 2 years from diagnosis. FCA seems appropriate only for children with well matched donors, whereas FCA-TBI is preferable in adults, especially when the donor is HLA mismatched. Complications such as rejection and EBV reactivation need to be addressed with modifications of the transplant regimens.

Disclosures:

No relevant conflicts of interest to declare.
Durable Remission with Salvage Autotransplants in Patients with Multiple Myeloma.
Authors: Shah Nina , Ahmed Khawaja Fraz , Qureshi Sofia , Shah Jatin , Orlowski Robert Z , Weber Donna M , Hosing Chitra , Popat Uday , Alousi Amin , Champlin Richard E , Giralt Sergio , Qazilbash Muzaffar
Published: October 2019

Abstract 1227

Poster Board I-249

Background

In comparison with single autologous hematopoietic stem cell transplantation (auto HCT), tandem autologous HCT has resulted in longer event-free and overall survival in randomized trials for patients with newly diagnosed multiple myeloma (MM). Most myeloma patients, however, only receive a single auto HCT. Many of these patients are eligible for a second auto HCT as salvage at the time of relapse. We evaluated the outcome of salvage auto HCT for MM patients treated at our institution.

Methods

We performed a retrospective chart review and identified 62 MM patients (38 males, 24 females) who received a second auto HCT as salvage between 1/3/1992 and 11/4/2008.. Preparative regimen was high-dose melphalan alone or in combination with other chemotherapy agents, including busulfan, topotecan and bortezomib. Three patients received a combination of thiotepa, busulfan and cyclophosphamide.

Results

Median interval between the first and salvage auto HCT was 21 months (range 2-81). Median age at salvage HCT was 55 years (37-73) and median prior treatment regimens were 4 (range 2-16). Twelve patients had chromosomal abnormalities on conventional cytogenetic studies. Patients received a median CD34 cell dose of 4 ×106 / kg (range 2.3-11.2). Fourteen patients (22%) experienced grade 3 or higher toxicity after the salvage auto HCT. Two patients died within 100 days with a TRM of 3%. Median time to neutrophil engraftment was 10 days (8-38). Responses after salvage auto HCT were as follows: CR+ near CR 15%, PR 48%, with an overall response rate of 63%. Twenty-seven (44%) patients received post auto HCT maintenance therapy. Median follow-up from salvage HCT was 25 months. Kaplan-Meier estimates of median progression-free survival and overall survival (OS) were 15.5 and 43.3 months, respectively. Median time to progression after the first and salvage auto HCT was 20 and 12 months, respectively, with total remission duration of 32 months from two HCTs. Median OS from the time of diagnosis was 72 months, comparable to reported results with tandem auto HCT. At last follow up, 20 patients were alive and in remission.

Conclusions

In selected MM patients a second auto HCT for salvage therapy is well tolerated with acceptable toxicity. The combined remission duration and overall survival are comparable to outcomes with tandem autotransplants.

Disclosures

Qazilbash: Cephalon: Speakers Bureau.
Abnormal Free Light Chain Ratios in Chronic Lymphocytic Leukemia: A New Prognostic Factor?.
Authors: Matschke Johannes , Eisele Lewin , Sellmann Ludger , Duehrsen Ulrich , Duerig Jan , Nückel Holger
Published: October 2019

Abstract 1237

Poster Board I-259

Introduction

Free light chains (FLC) have prognostic significance in monoclonal gammopathy of undetermined significance, solitary plasmocytoma of bone, smouldering myeloma, multiple myeloma, Waldenstroms macroglobulinaemia and AL amyloidosis. Although monoclonal protein secretion is a typical feature of plasma cell dyscrasias, it can also be detected in other B cell malignancies including chronic lymphocytic leukemia (CLL). Recent data suggest a significant correlation between abnormal ratio of FLC and outcome. Therefore, we investigated FLC in a large cohort of 120 patients in order to assess the role of FLC in CLL.

Methods and Results

Plasma samples which had been previously cryopreserved and collected at the time before the initiation of therapy or six months after finishing therapy were used. The levels of FLC were assessed using nephelometric immunoassays (The Binding Site) and quantified nephelometrically with the BNII analyser. A normal FLC range (κlγ) was defined as 0.26-1.65. Moreover, in all cases we evaluated the M band on immunofixation (IF).

Abnormal FLC ratios were found in 71 patients (59%) whereas the IF was positive in only 32 cases (27%). In 48 cases the FLC ratio was positive while IF was negative and in only 9 cases the IF was positive while the FLC ratio was normal. In total, 23 patients had both a positive IF and an abnormal FLC ratio.

Patients with an abnormal FLC ratio for γ had a significantly shorter treatment-free survival (TFS) than patients with an abnormal ratio for κ or with a normal FLC ratio (median TFS: 34 versus 78 versus 109 months, p=0.042). Evaluation of several disease characteristics in association with FLC of the patients' B-CLL cells showed no significant differences for FLC in the different risk groups (ZAP-70 status, CD38 status, cytogenetics and Binet stage) suggesting no correlation of the FLC with these already established adverse prognostic factors.

Conclusion

FLC can be detected in a substantial fraction of patients with CLL and the FLC technique improves detection of M-proteins. Moreover, an abnormal FLC ratio is associated with worse outcome, particularly those with a low abnormal FLC ratio. Evaluation of the prognostic significance of abnormal FLC in a larger cohort is currently under way. This data will be presented at the meeting. Future studies are warranted to elucidate the role of FLC as biomarkers of disease and as a prognostic factor for response.

Disclosures

No relevant conflicts of interest to declare.
The Inverse Correlation of Regulatory T Cells and Time to Initial Treatment in Chronic Lymphocytic B-Cell Leukemia.
Authors: Weiss Lukas , Melchardt Thomas , Grabmer Christoph , Egle Alexander , Tinhofer Inge , Greil Richard
Published: October 2019

Abstract 1238

Poster Board I-260

Regulatory T cells (Treg) are thought to be potent suppressors of anti-tumor immunity and increased Treg frequencies are correlated with poor prognosis in patients suffering from various tumor types. In the current study we analysed Treg (CD3+CD4+CD25+CD127−) in the peripheral blood of 101 previously untreated chronic lymphocytic B-cell leukemia (B-CLL) patients as well as 170 healthy volunteers using flow cytometry. Statistical analysis was used for correlation with clinical data. Treg of B-CLL patients expressed high numbers of FOXP3 and efficiently suppressed T cell proliferation. No significant difference in relative Treg numbers between B-CLL patients and healthy controls could be observed, but Treg numbers showed a significant negative correlation with time to initial treatment. Defining a cut-off of 9.75%, patients with higher Treg showed significantly shorter time to initial treatment (median 4.4 yr, 95% CI 2.2 – 6.6 yr) when compared to the lower Treg group (median not reached at 4.25 yr) (logrank P=0.001). Further subdivision of patients into quartiles according to Treg numbers showed a clear dose-dependency (logrank P=0.027). Treg numbers had significant prognostic power for time to initial treatment in univariate (P=0.001) as well as multivariate (P=0.037) Cox-regression analysis including Rai stage, IgVH mutational status, chromosomal aberrations and CD38 expression. In conclusion, higher Treg numbers are significantly and independently correlated with shorter time to initial treatment in B-CLL in a dose-dependent fashion and show significant prognostic power for time to initial treatment. These results provide evidence for the role of Treg in the clinical course of B-CLL.

Disclosures

No relevant conflicts of interest to declare.
Increased Interferon-αa Signaling During Megakaryocyte Ontogeny: Implications for the Spontaneous Resolution of Down Syndrome Transient Myeloproliferative Disorder.
Authors: Wieland Karen , Woo Andrew , Akie Thomas , Cantor Alan B.
Published: October 2019

Abstract 1272

Poster Board I-294

About ten percent of infants with Down syndrome (DS) are born with a transient myeloproliferative disorder (DS-TMD), which spontaneously resolves within the first few months of life. However, the basis for this resolution remains unknown. Acquired mutations leading to exclusive production of a short isoform of the transcription factor GATA-1 (GATA-1s) occur in all cases of DS-TMD, and knock-in mice that exclusively produce GATA-1s have hyperproliferation of megakaryocytes during early fetal liver hematopoiesis, but not during later developmental stages. In this study, we found striking upregulation of the interferon-αa (IFN-αa) receptor and multiple IFN-αa responsive genes, including Ifi203, Ifi205, Irf-1, Irf-8, and Ifitm6, in immunophenotypically isolated megakaryocyte progenitor cells (MkPs) from bone marrow versus embryonic day 13.5 (e13.5) fetal liver of wild type mice. These differences were confirmed at the protein level in megakaryocytes by in situ immunohistochemistry. Addition of IFN-αa to GATA-1s containing e13.5 fetal liver MkPs reduces their hyperproliferation in vitro in a dose-dependent manner. Conversely, injection of neutralizing IFN-αa/β antibodies, but not control IgG, into adult GATA-1s mice markedly increases the percentage of bone marrow CD41+ cells and morphologically recognizable megakaryocytes, in contrast to wild type mice. We propose that increases in IFN-αa signaling during megakaryocyte ontogeny may account for the developmental stage-specific effects of GATA-1s on megakaryocyte hyperproliferation, and possibly the spontaneous resolution of DS-TMD. Interestingly, the genes encoding the IFN-αa/β receptor are located on human chromosome 21 and are expressed at 1.6 times that in trisomy versus disomy 21 cells. We speculate that increased interferon alpha signaling during embryogenesis may be the basis for the strong selective pressure for GATA-1s producing mutations in trisomy 21 fetuses in the first place.

Disclosures

No relevant conflicts of interest to declare.
The PAF Complex Synergizes with MLL Fusion Proteins at .Hox Loci to Promote Leukemogenesis.
Authors: Muntean Andrew G. , Tan Jiaying , Basrur Venkatesha , Elenitoba-Johnson Kojo S.J. , Hess Jay
Published: October 2019

Abstract 1277

Poster Board I-299

Mixed lineage leukemia (MLL) is a histone H3 lysine 4 methyltransferase that is required to maintain a normal hematopoietic stem cell compartment. MLL functions to maintain expression of HOX genes as well as the HOX co-factor MEIS1, which play significant roles in regulating hematopoiesis. MLL is involved in chromosomal translocations with up to sixty different partners in both acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL). HOXA9 and MEIS1, are directly regulated by MLL fusion proteins and are crucial for MLL fusion protein mediated transformation. The deregulated expression of target genes in AML is dependent on specific protein-protein interactions and functional domains of MLL. For example, the tumor suppressor Menin bridges LEDGF to the extreme N-terminus of MLL and both of these interactions are necessary for transformation. Furthermore, a DNA methyltransferase homology region (CxxC domain) of MLL is essential for binding to non-methylated CpG islands and MLL-fusion protein oncogenesis. We have found that sequences downstream of the CxxC domain, termed the RD2 region, that interact with the Polymerase Associated Factor (PAF) complex are also required for MLL fusion protein mediated transformation. The PAF complex interacts with RNA polymerase II and is required for H2B mono-ubiquitination and subsequent histone H3K4 and H3K79 methylation. Together the PAF complex has been shown to be involved in transcriptional initiation, elongation and termination. Interaction of MLL with the PAF complex is mediated through direct contacts with two subunits: Ctr9 and PAF1. The PAF complex synergizes with MLL-AF9 to augment transcriptional activation of the Hoxa9 promoter. Furthermore, MLL fusion proteins recruit high levels of the PAF complex to the Hoxa9 promoter. Importantly, deletions of the MLL RD2 region that abolish interactions with the PAF complex eliminate MLL-AF9 mediated transformation of mouse bone marrow cells. Transcription of PAF components is dramatically downregulated during differentiation of hematopoietic cells, consistent with recent data showing a requirement for the PAF complex to maintain an embryonic stem cell phenotype. Knock down and transplantation experiments are underway to further define how the PAF complex regulates normal MLL function and cooperates with MLL fusion proteins to promote leukemogenesis.

Disclosures

No relevant conflicts of interest to declare.