bluuebunny/crossref_metadata_2025 huggingface dataset:
Selective Inhibition Of The PI3K-Alpha Isoform Blocks Myeloma Cell Growth and Survival
Authors: Guenther Andreas , Burger Renate , Klapper Wolfram , Staudinger Matthias , Gramatzki Martin
Published: October 2019

Introduction

Both, the phosphoinositide-3kinase (PI3K)/AKT pathway as well as its nutrient-dependent downstream target, the mToR (mammalian target of rapamycin) kinase, are essential for the growth and survival of malignant plasma cells. PI3K exists in four isotypes (α - δ), the role of each of these isotypes in multiple myeloma (MM), however, is not defined. Therefore, we evaluated systematically the in vitroanti-myeloma activity of NVP-BYL719 (Novartis), a newly developed inhibitor highly selective for the PI3K α isoform, and compared it with the activity of other inhibitors targeting the PI3K/AKT/mToR signaling network at different sites. Since inhibition of mToRC1 may lead to PI3K activation by a feedback loop, we looked also at combinations of NVP-BYL719 and rapalogs.

Materials and Methods

An MTS based growth assay was used to measure growth inhibition of NVP-BYL719 in comparison to different inhibitors (the pan-PI3K inhibitors Ly294002 and NVP-BKM120, the dual PI3K/mToR inhibitor NVP-BEZ235, and the allosteric mToR inhibitors rapamycin and everolimus) on six human malignant plasma cell lines and two non-myeloma cell lines, K562 and Raji. Western blot analysis was used to confirm inhibitor activity and specificity. In addition, combinations of PI3K and mToR inhibitors were tested and antagonistic/synergistic activity was calculated with the CalcuSynTM (Biosoft) software.

Results

The selective PI3K alpha inhibitor NVP-BYL719 inhibited myeloma cell growth at a dose level (IC50 < 10 µM in 5/6 cell lines), where non-MM cell lines were not affected. Only cell line U266 was showed moderate sensitivity. The levels of the PI3K isoforms αwere comparable in all tested MM cell , as shown by Western blot. NVP-BYL719 blocked phosphorylation of AKT and of protein S6, an mToR downstream target. Of note, whenever rapamycin was combined with a PI3K inhibitor including NVP-BYL719, synergistic growth inhibition was seen, even in combination with the dual PI3K/mToR inhibitor NVP-BEZ235.

Conclusions

Here, the PI3Kα isoform is reported as being essential for the growth of myeloma cells. This provides the rationale for the therapeutic exploration of such isoform specific PI3K inhibitors in MM allowing dose intensification with less toxicity. Allosteric mToR inhibitors, already active in early clinical trials in MM patients may provide interesting combination partners for these PI3K-inhibitors.

Disclosures:

Guenther: Novartis: Consultancy. Gramatzki:Novartis: Consultancy.
LICs Of APL-Like Leukemia Lead a Step-Wise and Divergent Differentiation Course
Authors: Liu Xiang-Zhen , Dai Jian-Min , Ding Fei , Ding Jing , Chen Juan , Guo Ning , Zhang Wu , Li Jun-ming , Chen Sai-juan , Chen Zhu , Wang Zhen-Yi , Zhu Jiang
Published: October 2019

The differentiation of leukemia-initiating cells (LICs) is generally regarded as a one-way alterative process to self-renewal. However, how differentiation impacts the leukemic stemness is largely unexplored. Here we show that a top tier of undifferentiated (UD), and a medium tier of partially differentiated (PD) LICs coexist within a PML/RARa-expressing mouse leukemia model. PD-LICs not only self-renew or even de-differentiate, but also mature into terminally differentiated (TD) monocytic or granulocytic non-LICs. Notably, PD-LICs but not UD-LICs were partially refractory to the differentiation-inducing effect of ATRA, a cognate ligand of PML/RARa. Nevertheless, the upregulation of monocytic transcriptional factor Irf8, albeit being dampened by RA signaling, relieves the differentiation arrest of most PD-LICs. Thus, our study reveals a sequential and divergent differentiation course of PML/RARa-expressing LICs under the regulation of discrete stage- and lineage- specific mechanisms.

Disclosures:

No relevant conflicts of interest to declare.
Quality Controls Of Immune Regulatory Properties Of Ex-Vivo, GMP-Grade Expanded Mesenchymal Stromal Cells For Clinical Use
Authors: Menard Cedric , Pacelli Luciano , Bassi Giulio , Dulong Joelle , Bezier Isabelle , Zanoncello Jasmina , Bifari Francesco , Ricciardi Mario , Schrezenmeier Hubert , Sensebé Luc , Tarte Karin , Krampera Mauro
Published: October 2019

Aim of CASCADE is to standardize GMP-grade production and clinical use of Mesenchymal Stromal Cells (MSC). Immunological Unit is aimed at setting up and validating a standardized panel of functional assays to fully characterize the immunomodulatory properties of MSC obtained from bone marrow and adipose tissue through different GMP-grade expansion protocols (platelet lysate- vs. fetal calf serum).

Immune cells were isolated using indirect immunomagnetic depletion (purity >96%). MSC were expanded in the same medium used for production and harvested at 70% confluence. Primed MSC (pMSC) were obtained after 48h-treatment with rh-IFNγ and rh-TNFα. MSC or pMSC were cocultured with T, B, NK cells for 4 or 6 days, and proliferation was evaluated by CFDA-SE dilution. T cells were stimulated with αCD3 + αCD28 antibodies; B cells were activated with CD40L, its enhancer, IL-2, CpG 2006, and anti-IgM/IgA/IgG; NK cells were activated with 100 U/ml rhIL-2. Cocultures were performed also with specific molecule inhibitors: L-1MT (IDO), snPP (HO-1), NS-398 (COX2), L-NMMA (iNOS) and anti-IFNγ antibody. For MSC immunogenicity assay, allogeneic T cell proliferation was evaluated at day 5 of culture; in addition, NK cells were activated for 2 days with rh-IL2, and MSC and pMSC were used as target cells.

Inflammatory milieu significantly upregulated MHC class I and II, CD54, CD106, CD40, CD274, CD112, CD155 expression, and downregulated NKG2D ligands and mesenchymal markers (CD73, CD90, CD105). AT-derived MSC expressed less MHC class II, CD200 and CD106 molecules than BM-MSC. MSC coculture inhibited T and NK cell proliferation without inducing apoptosis, and this effect was greater in presence of primed MSC. Only primed MSC were capable of suppressing B cell proliferation. MSC inhibited apoptosis of resting T, B, and NK cells, while inflammatory priming increased their pro-survival activity. Activation of IDO and HO-1 was the main mechanism involved in MSC immune modulation. MSC never promoted allogeneic T cell proliferation; by contrast, IL-2-activated NK cells could efficiently recognize and kill allogenic unprimed MSC, while primed MSC became insensitive to NK cells. Some differences were observed depending on the origin and culture conditions of clinical-grade MSC.

All the experimental protocols to assess MSC inhibitory effects on immune effector cells have been standardized and will be applied for the release of GMP-grade MSC produced inside the CASCADE Consortium.

Disclosures:

No relevant conflicts of interest to declare.
BB02 Improves Therapeutical Effectiveness Of Extracorporeal Photopheresis With 8-MOP In a Murine Model Of Graft-Versus-Host Disease
Authors: Garcia-Bernal David , Blanquer Miguel , del Rio Jose Antonio , Correal Enrique , del Carmen Algueró Maria , Majado Maria Juliana , Moraleda Jose María
Published: October 2019

Introduction

Extracorporeal photopheresis (ECP) is a cell-based immunomodulatory therapy involving the separation of peripheral blood autologous mononuclear cells followed by ex-vivo administration of 8-methoxypsoralen (8-MOP) and UVA radiation before reinfusion. ECP is efficient for the treatment of multiple diseases mediated by unregulated T cell populations, such as cutaneous T cell lymphoma, autoinmune diseases or graft-versus-host disease (GVHD), the major complication after allogeneic bone marrow transplantation. Our aim in the present work was to compare the therapeutic effectiveness of 8-MOP with other two new compounds (BB01 and BB02) in a experimental murine model of GVHD.

Methods

Murine GVHD was induced after transplanting bone marrow cells and splenocytes from donor Balb/c mice into C57Bl6J recipients previously conditioned with a lethal dose of 10 Gy split into two doses of 5 Gy spaced 24 hours apart. To investigate the therapeutic effectiveness of ECP with either 8-MOP, BB01 or BB02, splenocytes from separate cohorts of C57Bl6J with GVHD were isolated 12 days after transplantation, incubated with the different compounds, irradiated with UVA light and injected intravenously once a week for four weeks. Survival after transplantation was monitored daily and clinical GVHD was graded using a previously described score analyzing weight loss, posture (hunching), activity, skin integrity and fur texture (Cooke et al, 1996). Mice of each group were evaluated and graded from 0 to 2 for each criterion, obtaining a clinical index by summation of the five criteria scores (maximum index=10).

Results

Mice treated weekly with BB02 showed a significant higher survival than those treated with 8-MOP (p=0,038), while BB01 had a similar effect to that of 8-MOP. Mice treated with either compound improved their clinical GVHD score compared to untreated mice group, being significantly lower with BB02 than with BB01 and 8-MOP (p=0,023).

Conclusions

BB02 was more efficient than 8-MOP in the reversal of murine GVHD, while BB01 showed the same therapeutic effectiveness than 8-MOP.

Acknowledgments

Work financed by the Spanish Ministry of Science and Innovation (Ref: BFU2010-19599) and the Spanish Net of Cell Therapy (TerCel) from Institute of Health Carlos III.

Disclosures:

No relevant conflicts of interest to declare.
Outcomes After a Sequential Clofarabine/Arac-C Chemotherapy Followed By Reduced-Intensity Conditioning (RIC) and Allogeneic Stem Cell Transplantation (allo-SCT) In Pediatrics Patients With JMML Or Primary/Relapsed Refractory AML: A Pilot Study
Authors: Audrey Grain , Sirvent Anne , Strullu Marion , Mechinaud Francoise , Mohty Mohamad , Chevallier Patrice , Guillaume Thierry , Rialland Fanny
Published: October 2019

Introduction

The FLAMSA sequential treatment with chemotherapy followed by reduced-intensity conditioning (RIC) for allogeneic stem cell transplantation (allo-SCT) has been introduced few years ago for adult refractory acute myeloid leukemia (AML) showing high activity and relatively good survivals in this particular setting (Schmid, Blood, 2006). There is no study at our knowledge reporting the results of the sequential approach in pediatric refractory AML patients. Here we report our own experience in 6 children using a debulking chemotherapy combining clofarabine and Ara-C followed by RIC before allo-SCT.

Patients and Methods

These preliminary results included 3 males and 3 females with a median age of 7 years (range: 4-11). All cases had received a sequential regimen before allo-SCT at the CHU of Nantes (n=5) or at the CHU of Montpellier (n=1) for primary refractory AML (n=1), refractory relapsed AML (n=3), slow responder relapsed AML (n=1) and blastic JMML (n=1). Sequential regimen consisted of 1) clofarabine 30 mg/m²/d days-13 to -9, Ara-C 1g/m²/d days-13 to-9 followed by RIC combining cyclophosphamide 60 mg/kg/d day-5, iv Busulfan 3.2 mg/Kg/d days -4 to -3 and ATG 2.5 mg/Kg/d days -3 to -2 in 5 patients and 2) clofarabine 30 mg/m²/d days-13 to -9, Ara-C 1g/m²/d days-13 to-9 followed by RIC total body irradiation 4 grays day-5, cyclophosphamide 40 mg/Kg/d days -4 to -3, and ATG 2.5 mg/Kg/d days -3 to -2 in 1 patient. One patient received a graft from a sibling donor while the five other patients received a graft from an unrelated donor (10/10 n=3; 9/10 n=2). All patients received PBSC as stem cells graft. Ciclosporine alone was used in case of related donor while ciclosporin+MMF were used in case of an unrelated donor.

Results

Engraftment was observed in 4 patients (67%) and 3 patients out of the 5 refractory cases achieved complete remission (CR) after transplant. The 2 patients who had an autologous reconstitution, relapsed and died rapidly. Considering the 4 patients achieving full engraftment and CR, only one relapsed at day+60 and died of relapse. The three other patients are alive in CR at +12, +35 and +51 months post-transplant.

Conclusion

To our knowledge, this is the first report of a sequential allo-SCT approach for refractory pediatric AML patients. Although the number of patients is limited in our cohort, the results showed here are very encouraging as half of the patients are alive in CR with full engraftment. These results have to be confirmed prospectively.

Disclosures:

No relevant conflicts of interest to declare.
Factors Affecting Unit Predominance In Double Umbilical Cord Blood Transplantation
Authors: Santos Fernanda NC , Hitomi Yokoyama Ana Paula , Kondo Andrea Tiemi , Macedo Erika Maria , Conti Fabiana Mendes , Sakashita Araci M. , Kutner Jose Mauro
Published: October 2019

Background

Umbilical cord blood transplantation is seen as an alternative source of hematopoietic support for patients who lack suitable HLA-matched donors. Disadvantages of this method include the relatively low cellularity, late engraftment and consequently restrictive use to pediatric patients, due to the low number of hematopoietic progenitors. Transplantation of two cord blood units has been seen as a strategy to overcome the low yield, often leading to the long-term establishment of hematopoiesis supported by a single cord blood unit. Predictive factors of the predominant unit are still unclear. Total nucleated cell count, CD34+, CD3+ dose were independent factors associated with unit predominance, although this is still a controversial issue.

Methods

This retrospective study included 21 patients with hematologic malignancies and immunodeficiencies who underwent double unit cord blood transplantation. We analysed HLA mismatch, ABO group, total nucleated cell count (TNC), CD34+ dose and post-thaw viability and their correlation with the prevalent unit. Transplantations were performed from 2007 to 2013. Six patients were excluded due to incomplete information and another three died before engraftment.

Results

Median NC and CD34+ doses were similar between engrafted and non-engrafted units. Nine transplantations (60%) resulted in engraftment of the higher TNC unit. TNC dose in the predominant units varied between 2.02-3.05x107/kg (median dose: 2.651x107/kg), and in the non-engrafted units between 1.69x107/kg-4.39x107/kg (median dose: 2.44x107/kg), which was not statistically significant (p= 0.65). Sixty percent of the transplantations (9 patients) also resulted in engraftment of the unit with higher CD 34+ (median dose: 2.05x106/kg), compared to 40% (6 patients) in which the lower dose unit prevailed (median dose: 1.95 x106), also not statistically significant (p= 0.861). As for ABO, 26% of the units were ABO matched with the recipient, while the majority was HLA mismatched (93%).

Conclusion

Multiple factors related to the unit and patient influence UCB predominance. In the sample studied, no particular variant played a specific role, probably due to the limited sample size. Current evidence suggests that immunologic-related mechanisms (CD34+ and CD3+ dose) are involved, yet further studies are necessary for a better understanding of which factors lead to unit engraftment.

Disclosures:

No relevant conflicts of interest to declare.
Enteral Versus Parenteral Nutritional Support Post Allogeneic Haematopoietic Cell Transplantation – Results Of a Randomized Controlled Trial
Authors: Kennedy Glen A , Andersen Sarah , Banks Merrilyn
Published: October 2019

Background / Aims

Nutritional support post allogeneic haematopoietic progenitor cell transplantation (HPCT) has proven efficacy in preventing malnutrition post-HPCT with resultant improved patient outcomes. However, there is no consensus as to the most efficacious approach to supplemental feeding post-allogeneic HPCT, leading to wide variation in clinical practice with respect to use of enteral (EN) vs. parenteral (PN) feeding, especially in adult units. We aimed to determine the tolerability and efficacy of EN vs.PN in adult patients undertaking allogeneic HPCT.

Methods

Prospective randomized controlled study in adult patients >18yrs of age undertaking allogeneic HPCT with either myeloablative or reduced intensity conditioning (RIC). Informed consent was obtained before commencing conditioning, with patients randomized to receive either EN or PN after commencing HPCT conditioning if they could not maintain adequate oral nutritional intake, defined as maintaining >60% of daily caloric requirements orally. Patients with severe gastro-intestinal toxicity, including severe mucositis, were excluded from randomisation. The primary endpoint was tolerance of route of supplemental nutritional support, defined as <30% of patients needing to change to the alternative route of support for any reason.

Results

In total 38 patients were enrolled onto the study, including 9 patients (24%) undertaking myeloablative and 29 patients (76%) RIC HPCT. Only 19 patients (50%) required nutritional support post-HPCT. Of these 19 patients, only 9 (47%) were able to be randomized between EN (n=5) and PN (n=4), with 10 patients excluded from randomization due to presence of severe gastrointestinal toxicity in 7 (37%), markedly deranged liver function in 1 (5%), and withdrawal of consent in 2 (11%). The 5 patients randomized to EN met on average 74% of their goal nutrition and 100% required changing to PN due to (gastro-intestinal) intolerance. The 4 patients receiving PN met on average 91% of requirements, with none requiring change to EN.

Conclusions

For adult patients undertaking myeloablative or RIC allogeneic HPCT, supplemental feeding with EN commencing at failure to maintain adequate oral nutritional intake is not feasible, due to the presence of significant gastrointestinal toxicity in these patients at this time. Further study is required to determine whether prophylactic nasogastric tube placement would improve the feasibility and tolerance of EN in this patient population.

Disclosures:

No relevant conflicts of interest to declare.
Hypogammaglobulinemia In Children Undergoing SCT: An Immunoglobulin Isotype Class Switch Arrest?
Authors: Lindqvist Henric , Remberger Mats , Sundberg Berit , Uzunel Mehmet , Winiarski Jacek , Sundin Mikael
Published: October 2019

Treatment-requiring hypogammaglobulinemia is common early after allogeneic hematopoietic stem cell transplantation (SCT) in children. Usually, it resolves but in several children it persists, demanding long-term immunoglobulin (Ig) G replacement therapy. By IgG replacement, the morbidity risk may not be reduced, but infectious complications are decreased.

Here we studied clinical events, Ig and immune cell recovery, IgG2m allotypes and dynamics of cytokines important for B-lymphocytes and Ig production in 84 children undergoing SCT. The aim was to identify risk factors for ≥3 months IgG replacement need as well as studying the biology behind the hypogammaglobulinemia seen post-SCT. Clinical data were collected from charts. Interleukin (IL)-4, -5, -6, -7 and -10, BAFF and APRIL were analyzed in patient sera pre- and post-SCT. Donors and patients were IGHG2 genotyped. Uni- and multivariate statistic analyzes were applied.

As many as 48% of the children needed ≥3 months IgG replacement. These were commonly younger (<5 years) and had acute GvHD grade II-IV. However, children with ≥3 months IgG replacement had better overall survival (88 vs. 69%, P=0.036). Significantly lower Ig levels post-SCT, but equal immune cell recovery were seen when comparing the ≥3 vs. <3 months IgG replacement groups. Significantly higher rates of mixed chimerism in CD3 and C33 populations was seen in the <3 months IgG replacement group. IGHG2 genotyping showed no significant frequencies between the groups. Pre-SCT IL-6 and -7 and post-SCT BAFF and APRIL levels were significantly higher in the ≥3 months IgG replacement group. This suggests an unfavorable cytokine milieu for graft-derived immune recovery with possibly later Ig isotype class switch arrest. Age <5 years (OR 8.16, 95% CI 2.62-25.4, P<0.001), acute GvHD grade II-IV (OR 5.61, 95% CI 1.78-17.6, P=0.038) and pre-SCT IL-6 levels >50 pg/mL (OR 8.03, 95% CI 1.26-51.0, P=0.024) were identified as significant risk factors for ≥3 months IgG replacement need.

To conclude, requiring ≥3 months IgG replacement does not need to be unfavorable and could be an effect of deteriorated cytokine signaling resulting in impaired Ig production. Further prospective studies investigating post-SCT hypogammaglobulinemia with and without IgG replacement are warranted.

Disclosures:

No relevant conflicts of interest to declare.
Prolonged Thrombocytopenia Is Associated With Increases Of CD8+ CX3CR1+ Cells In The Bone Marrow After Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Zhang Xiaohui , Wang Guoxiang , Zhu Honghu , Liu Yanrong , Liu Daihong , Xu Lanping , Han Wei , Chen Huan , Chen Yuhong , Wang Feng-Rong , Liu Kaiyan , Huang Xiaojun
Published: October 2019

Prolonged thrombocytopenia is a common complication after allogeneic hematopoietic stem cell transplantation(allo-HSCT),which is associated with a high mortality and poor prognosis. The aim of this study was to assess the impact of the CD8+CX3CR1+ T cells on the development and maturation of megakaryocytes in patients with the prolonged thrombocytopenia after allo-HSCT in order to identify the risk factors related to thrombocytopenia after allo-HSCT.

The changes in CD8+ T cells and their homing receptors CX3CR1, CXCR4 and VLA-4 in bone marrow of patients( N=89) with and without (N=94 ) prolonged thrombocytopenia following allo-HSCT and the impact of activated CD8+ T cells on apoptosis and ploidy of megakaryocytes in vitro ware determined. The percentage of CD8+CX3CR1+ T cells was significantly higher in prolonged thrombocytopenia patients than control (P<0.001).The increase in CD8+ T cells was not observed in peripheral blood. Patients with prolonged thrombocytopenia exhibited a marked increase in the number of low ploidy(≤8) megakaryocytes compared to those without(P<0.05). Depletion of CD8+ T cells increased apoptosis of megakaryocytes (P<0.05), which was corrected by reconstitution of CD8+ T cells (P<0.05). We demonstrated that prolonged thrombocytopenia is associated with increased expression of CX3CR1 and recruitment of CD8+ T cells into the bone marrow. Activated CD8+ T cells suppress apoptosis of megakaryocytes in vitro. Our findings shed light on the prolonged thrombocytopenia is associated with increases of CD8+ CX3CR1+ cells in the bone marrow after allo-HSCT patients.

Disclosures:

No relevant conflicts of interest to declare.
Engraftment Syndrome (ES) After Autologous Stem Cell Transplantation (SCT): Analysis Of Diagnostic Criteria and Risk Factors
Authors: Gerivaz Rita , Costa Fatima , Schuh Juliana , Monteiro Luis P , Ferreira Gilda , Sousa, MD Aida B.
Published: October 2019

ES is increasingly recognized as an important complication of autologous SCT. There is no consensus over diagnostic criteria (leading to variations in reported incidence from 7 to 55%) nor over risk factors. In order to better characterize this syndrome and its potential severity, we reviewed 380 consecutive autologous SCT (in 304 patients [pts]) performed in our center from 1/2006 to 8/2012.

Methods

During this period ES was diagnosed according to the Maiolino criteria (MC): non infectious fever beginning 24h pre-engraftment + pulmonary infiltrates or rash or diarrhea. We retrospectively reviewed all cases according to the Spitzer criteria (SC): 3 major criteria within 96h of engraftment (non infectious fever, rash, pulmonary infiltrates) or 2 major + 2 minor criteria (hepatic and renal disfunction, weight gain, encephalopathy). The following risk factors for ES were evaluated: sex, underlying disease, mobilization regimen (G-CSF with or without chemotherapy), conditioning regimen, number of reinfused CD34+ cells/kg, cryopreservation duration, time to engraftment and bacteremia rate. The following outcomes were analyzed: days on antibiotics, number of platelet transfusions, in-hospital days, and transplant related mortality (TRM).

Results

ES was diagnosed in 29 SCT (8%) in 27 pts (2 tandem SCT had ES in both) by MC. Only 8 of these fullfilled the SC. Most cases (83%) required corticosteroids and improved; 1 death was attributed to ES. By multivariate analysis the single risk factor for ES was a diagnosis of non Hodgkin’s lymphoma as opposed to Hodgkin’s (15% vs 3.7%, p=0.04); none of the remaining risk factors was statistically significant. Pts with ES spent more days on antibiotics (median 12 vs 10, p=0.008) and in the hospital (23 vs 19, p<0.000) and there was a trend to a higher TRM at d+30 (3.7 vs 1.3%) and d+100 (7.4 vs 4.2%).

Conclusions

We were able to confirm in our series that ES prolongs antibiotic duration and hospitalization and may increase the risk of death. Although ES is a complex entity with a difficult diagnosis, its prompt recognition is critical, as timely institution of corticosteroids diminishes the risk of multiorgan failure, avoids unnecessary use of antimicrobials, and reduces hospital days and costs. The SC are more stringent but too insensitive. We believe that, until prospective studies validate clearer diagnostic recommendations, transplant teams are best served by the MC.

Disclosures:

No relevant conflicts of interest to declare.
Imatinib Mesylate Versus Allogeneic Hematopoietic Stem Cell Transplantation For Patients With Chronic Myeloid Leukemia In Chronic Phase
Authors: Xiaoli Liu , Gao Guanlun , Zhou Xuan , Xu Na , Xiao Yajuan , Cao Rui , Yin Changxin , Zhou Hongsheng , Wei Yongqiang , Zhang Yu , Xu Dan , Xu Bing , Feng Ru , Liu Qifa , Sun Jing , Meng Fanyi
Published: October 2019

Background and Objective

Following the introduction of the tyrosine kinase inhibitor (TKI) imatinib in treatment of chronic myeloid leukemia (CML) patients, the allogeneic hematopoietic stem cell transplantation (allo-HSCT) scene in CML has changed dramatically. This retrospective cohort study was designed to compare medical outcomes of Imatinib mesylate and allo-HSCT for patients with CML in chronic phase.

Patients and Methods

From February 2002 to February 2012, 198 patients treated consecutively at the Nanfang Hospital,Southern Medical University were assigned to two groups according to treatment with imatinib or allo-HSCT. One hundred fifteen cases of imatinib group were given imatinib at an initial dose of 400mg daily and the dose was then adjusted according to the patient´s blood and therapy response. All the patients were evaluated for hematologic, cytogenetic and molecular response every 1-3months. Eighty-three cases of allo-HSCT group received myeloablative preconditioning regimen, and methotrexate (MTX) and cyclosporine A (CsA) were used for graft-versus-host disease(GVHD), parts combined with mycophenolate mofetil (MMF) and antihuman thymocyte globulin(ATG). The primary end points of the study were complete cytogenetic response (CCyR), relapse rate, overall survival (OS) and progression-free survival (PFS) after therapy.

Results

In total, 59 (68.9%) patients treated over 12 months achieved a CCyR after 12 months in imatinib group, while 67 (95.7%) patients in allo-HSCT group. The relapse rates were 14.8% (n=17) in imatinib group and 10.8% (n=9) in allo-HSCT group (P=0.456). Ten-year cumulative OS rates were 93.9% in imatinib group and 77.1% in allo-HSCT group(P=0.015) and ten- year cumulative PFS rates of two groups were 86.1% vs.88.0%(P=0.508). For Sokal rating stratified analysis, the ten-year OS rates of two groups were 96.4% vs.68.0% (P = 0.049) for high-risk patients,92.6% vs. 57.1% (P = 0.019) for intermediate-risk patients , while the ten-year PFS rates of two groups were 89.3% vs. 88.0% for high-risk patients (P = 0.942), 70.4% vs. 85.7% for intermediate-risk patients (P = 0.405).The ten-year OS rates and PFS rates were not significant difference for low-risk patients. The cumulative OS rates of two groups were 94.7% vs. 73.5%(P=0.019)for the patients who were not less than 30 years old,and the cumulative PFS rates of two groups were 84.2% vs. 94.1% respectively (P=0.147).

Conclusion

Imatinib mesylate treatment is superior to allogeneic hematopoietic stem cell transplantation for patients with chronic myeloid leukemia in chronic phase.

Disclosures:

No relevant conflicts of interest to declare.
Outcome Of Hematopoietic Cell Transplantation (HCT) In Pediatric Patients With Non-Hodgkin Lymphoma (NHL): Single Institution Results From Saudi Arabia
Authors: Belgaumi Asim F , Sumaili Hassan A , Al-Kofide Amani A , Ayas Mouhab , El-Solh Hassan , Al-Ahmari Ali , Al-Jefri Abdallah , Anas Mohammed , Mohamed Amal , Al-Seraihy Amal
Published: October 2019

Although HCT is an accepted component of the treatment strategy for relapsed/refractory pediatric NHL, only few studies have reported on the outcome for these patients. Most have reported on small numbers of patients, with survivals ranging from 27% to 75%.

Clinical data were retrospectively retrieved for patients with NHL who had undergone HCT. Pre-HCT information, including pathologic diagnosis, response to first- and second-line therapy and pre-HCT disease status were collected, in addition to details of the transplant process and patient and disease outcome.

Between 1996 and 2012, 28 pediatric patients with NHL underwent HCT. Primary diagnosis for these patients included Burkitt lymphoma (n=13), Large B-cell lymphoma (n=4), T-Lymphoblastic lymphoma (n=4), NK/T cell lymphoma/leukemia (n=3), Peripheral T-cell lymphoma (n=2), B-lymphoblastic lymphoma ((n=1) and anaplastic large cell lymphoma (n=1). The median age at HCT was 7.65 years (mean 8.2; range 1-14.3). Twenty had suffered a relapse of their disease, while five had primary progression; three patients with NK/T lymphomas underwent HCT as part of their first-line therapy. Fourteen patients had autologous (autoHCT) and 14 had allogeneic HCT (alloHCT). Among alloHCT, 11 had matched-related grafts while 3 had unrelated umbilical cord blood (UCB) grafts. At the time of HCT, 23 patients were in CR (CR1=7, CR2=15, CR3=1), and 5 had partial responses. HCT conditioning was myeloablative for all patients; in 18 patients, it was TBI-based. Fourteen patients suffered recurrence of their lymphoma post HCT at a median of 1.17 months from HCT (mean 6.2; range 0.63-42); 4 died in CR due to transplant-related toxicity, of these 3 were post alloHCT and one post autoHCT. Three patients have developed secondary malignancies (SMN; 2 post alloHCT and 1 post autoHCT). 10 patients were alive at last follow-up, all of whom were in CR. The 5-year estimated OS from SCT is 38.7%, with and EFS of 26%. There was no difference in 5-year OS or EFS among patients who received alloHCT v. autoHCT (OS 28.6% v. 49%; p=0.53, EFS 14.3% v. 37.5%; p=0.25) and among patients who did or did not receive TBI (OS 33.3% v. 48%; p=0.37, EFS 27.8% v. 18.8%; p=0.66). OS/EFS for patients with Burkitt lymphoma was 23.1%. Of the three patients with NK/T cell lymphoma two remain alive in CR 13.7 and 5.1 years after HCT.

The outcome of relapsed/refractory non-Hodgkin lymphoma of childhood remains suboptimal. In addition to a high post-HCT relapse rate of 50%, HCT-related toxic mortality and SMN contribute to the poor outcome for this cohort of patients.

Disclosures:

No relevant conflicts of interest to declare.
Outcome Of Hematopoietic Cell Transplantation (HCT) In Pediatric Patients With Hodgkin Lymphoma (HL): Single Institution Results From Saudi Arabia
Authors: Sumaili Hassan A , Belgaumi Asim F , Al-Kofide Amani A , Al-Seraihy Amal , El-Solh Hassan , Al-Jefri Abdallah , Al-Ahmari Ali , Mohamed Amal , Anas Mohammed , Ayas Mouhab
Published: October 2019

Most pediatric patients with Hodgkin lymphoma are cured of their disease with standard combined-modality first-line therapy. Those who relapse are subjected to salvage chemo-radiotherapy, and patients who respond often undergo either autologous or allogeneic HCT, with a reported outcome ranging from 40%-60%. Variables affecting the outcome of such patients are not clearly defined.

This study retrospectively reviewed the clinical characteristics and outcome of patients who underwent HCT at our institution.

Between 1995 and 2012, 29 pediatric (age <14 years) patients with HL underwent HCT. This cohort included 24 boys and 5 girls. Their median ages at initial diagnosis and at HCT were 9.85 years (mean 8.85; range 3.6-13.75) and 12.18 years (mean 11.24; range 5.6-14.9), respectively. 28 patients had classic HL (23 nodular sclerosis, 3 mixed cellularity, 1 lymphocyte-depleted, and 1 lymphocyte-rich) and one patient had nodular lymphocyte-predominance HL. Ten had persistent/progressive disease following first line therapy, while 19 had relapsed following achievement of complete response (CR). For these patients median time to relapse from completion of first-line therapy was 16.9 months (mean 20.1; range 1.9-53.1). All patients received salvage chemotherapy and/or radiotherapy prior to HCT; fifteen patients achieved CR, 13 had a partial response and one had progressive disease. Two patients had allogeneic bone marrow (BM) grafts from matched-related donors, while the rest had autologous grafts (16 BM; 10 PBSC; 1 BM+PBSC) following chemotherapy-based myeloablative conditioning. Twelve patients have relapsed/progressed post-HCT at a median of 6.04 months (mean 11.8; range 1.02-71.4). Nine patients have died; eight because of disease progression and one due to sepsis post HCT. Only two patients died within the first 100 days post HCT, giving a Day-100 mortality rate of 6.8%. Two patients who relapsed after HCT were salvaged with chemo/radiotherapy and remain disease free 2.8 and 9.7 years later. The 5-year estimated overall survival (OS) from HCT for the whole cohort is 61.6%, with an event free survival (EFS) of 57.9%. Patients who had persistent/progressive disease at the end first-line therapy or relapsed <6months off therapy had a worse OS and EFS as compared to those who relapsed later (OS 42.9% v. 75.3%, p=0.047 [Taron-Ware]; EFS 41.7% v. 60.8%, p=0.052 [Taron-Ware]).

The outcome of patients with relapsed/refractory HL following HCT is encouraging, as a majority of patients survive free of their lymphoma. Timing of relapse/progression remains an important prognostic factor and patients who fail early may be considered for novel therapeutic approaches.

Disclosures:

No relevant conflicts of interest to declare.
Salvaged Cord Blood Transplantation For 19 Patients With Progressive Hematologic Malignancies
Authors: Zimin Sun , Wen Yao , Changcheng Zheng , Juan Tong , Huilan Liu , Liangquan Geng , Tang Baolin , Xiaoyu Zhu , Kaidi Song , Ding Kaiyang
Published: October 2019

Few clinical studies have investigated the role of salvaged unrelated cord blood transplantation (CBT) for progressive hematologic malignancies. The aim of this report is to identify the potential benefits of unrelated CBT in progressive hematologic malignancies.19 consecutive patients with progressive myeloid and lymphoid malignancies who received salvaged CBT following myeloablative conditioning regimens (12 TBI/CY/Ara-C, 6 Bu / CY / Ara-C, and 1 Bu /CY) from July 2005 to December 2012 were analyzed retrospectively. Of the 19 patients, 6 suffered from acute myeloid leukemia(AML), 5 acute lymphoid leukemia(ALL),：1 acute mixed lineage leukemia (AMLL), 1 myelodysplastic syndrome-refractory anemia with excess blasts(MDS-RAEB), 2 acute myeloid leukemia transformed from myelodysplastic syndrome, and 4 lymphoma, all of them in non-remission (NR) before transplantation. Median age of them were 13（range 6-32）years and median body weight were 45 (range 18–73 ) kg. All patients received 1 CBT unit ≤2 loci HLA-mismatched with the recipient. Infused TNC was 4.07（range 2.76-6.02）×107/kg and CD34+ stem cell 2.08（range 0.99-8.65）×105/kg. All patients were engrafted with neutrophil exceeded 0.5×109/L on median day +17（range 14-37d）and plt counts of＞20×109/L on median day +35 (range 17-70d). 10 patients (52.6%) experienced pre-engraftment syndrome (PES) and 3 (15.8%) patients progressed to acute GVHD. The incidence of Ⅱ-Ⅳ aGVHD and cGVHD were 10.5% and 21.1%. With a median follow up of 10(range 1-58) months，10 cases survived and 2 relapsed．The estimated 2 year overall survival (OS), disease-free survival (DFS) and non-relapse mortality (NRM) rate was 45.6%, 34.2% and 39.8%. Salvaged CBT might be a promising modality for treatment of progressive hematologic malignancies, even with high leukemia burden.

Disclosures:

No relevant conflicts of interest to declare.
The Mechanisms Of Damage To Bone Marrow By Iron Overload In Patients With Immuno-Related Pancytopenia
Authors: Fu Rong , Huang Lei , Shao Zonghong
Published: October 2019

Objective

To investigate the mechanisms of iron overloading bone marrow damage in patients with immuno-related pancytopenia.

Methods

Forty-seven IRP patients (21 iron overloading IRP patients) and 10 normal controls were enrolled in this study. The expression of ROS, apoptosis, Bcl-2 and Caspase-3 of the bone marrow mononuclear cells (BMMNC) were analyzed by flow cytometry. Antioxidants were added to the iron overloading IRP BMMNC, and then the changes were detected by FCM. The number and apoptosis of the T lymphocytes of IRP patients were detected.

Results

The ROS and apoptosis of the BMMNC, myelocytes, erythrocytes and stem cells in the bone marrow were significantly higher than those of non iron overloading IRP patients and normal controls(P<0.05). The expression of Bcl-2 in BMMNC, erythrocytes and stem cells of the iron overloading IRP patients were significantly lower than those of non iron overloading IRP patients (P<0.05). The levels of Caspase-3 in myelocytes, erythrocytes and stem cells of the iron overloading IRP patients were significantly higher than those of non iron overloading IRP patients or normal controls (P<0.05). After being treated with antioxidants, the expression of ROS, Caspase-3 and apoptosis of the iron overloading IRP BMMNC significantly decreased, the opposite of the Bcl-2. The percentage of the CD4+ lymphocytes (40.86±8.74)% and CD4+/CD8+ (1.44±0.36) in PB of the iron overloading IRP patients were significantly higher than those of non iron overloading IRP patients (35.96±7.03)% and (1.13±0.37)and normal controls(28.00±6.73)% and (0.79±0.21) (P<0.05), the opposite of CD8+ lymphocytes (P<0.05). The apoptosis of CD8+ lymphocytes (27.35±10.76)% and the ratio of CD8+ apoptosis /CD4+ apoptosis (2.51±0.80) in BM were significantly higher than those of the non iron overloading IRP patients (15.47±8.99)% and (1.39±0.47) (P<0.05). The apoptosis of the erythrocytes and stem cells coated with auto-antibodies in BM of the iron overloading IRP patients were significantly higher than those of the non iron overloading IRP patients.

Conclusion

Iron overload damaging the bone marrow hematopoiesis might be through the following mechanisms. 1. The increased ROS induced by iron overload affected the expression of Caspase-3 and Bcl-2, which caused the higher BMMNC apoptosis; 2. The abnormal number and ratio of T lymphocytes caused by iron overload aggravate the abnormality of immunity of IRP; 3. Iron overload may increase the damage to erythrocytes and stem cells coated with auto-antibodies.

Disclosures:

No relevant conflicts of interest to declare.
Pulmonary Irradiation Fibrosis Is Preceded By Increased Endothelial Cell Gene Expression
Authors: Kalash Ronny , Au Jeremiah , Epperly Michael W. , Goff Julie P. , Dixon Tracy M. , Franicola Darcy , Greenberger Joel S.
Published: October 2019

Irradiation of the C57BL/6NHsd mouse lung results in pulmonary fibrosis. An acute phase (days 1 to 28) following irradiation recognized by an inflammatory response is followed by a latent phase (day 28 to day 125) where lung appears normal. Fibrosis occurs from day 125 till death characterized by bone marrow stromal cell migration to and proliferation in lung and increased collagen deposition. To investigate the role of endothelial cells, C57BL/6NHsd female mice were irradiated to 20 Gy to the pulmonary cavity shielding the rest of the body. Lung was excised at serial times, endothelial cells separated and tested for expression of endothelial genes vWF, VEGF, FGF1, CCL13 and CTGF; inflammatory genes IL-6; IGFbp7 and genes associated with fibrosis including MnSOD, Nrf2, NfkB, TLR4 and TGF-B. Endothelial cells were separated by using antibodies to CD45, or PECAM and sorting using flow cytometry. RNA was extracted using Triazol and gene expression determined using Real Time Polymerase Chain Reaction (RT-PCR). In irradiated total lung there was significant increase in endothelial cell markers at all times, compared to nonirradiated lung. Transcripts for vWF increased by 474.1 ± 388.5% (p = 0.017) by 2 days after irradiation, 520 ± 55.4% (p < 0.0001) at day 50, and 484.8 ± 26.8% (p < 0.0001) by day 200. VEGF and CTGF also remained elevated. In contrast, total lung MnSOD gene expression increased by 140.6 ± 46.1% at day 2 (p = 0.0185), decreased at day 50 to 17.9 ± 11.2% (p = 0.2808), and increased to 31.5 ± 11.4% by day 200 (p = 0.023). However, purified endothelial cells compared to alveolar cells, showed increased expression of not only the endothelial cell genes but also MnSOD, Nrf2, NfkB, and TGF-B. MnSOD expression in endothelial compared to alveolar cells at day 28 after irradiation was higher at 244.5 ± 7.8 vs 80.5 ± 4.9 (p = 0.0016), at day 150 expression was 140.1 ± 25.0 vs 5.4 ± 1.2 (p < 0.0001) and day 200 expression was 368.5 ± 6.4 vs 51.8 ± 10.3 (p = 0.0007). Lung endothelial cells display critical biomarkers of irradiation injury leading to fibrosis.

Disclosures:

No relevant conflicts of interest to declare.
Bendamustine + Rituximab-Combinations and R-CHOP Achieve a Major Survival Improvement Of Patients With Follicular Lymphoma In Routine Care
Authors: Weide Rudolf , Feiten Stefan , Friesenhahn Vera , Heymanns Jochen , Kleboth Kristina , Thomalla Jörg , van Roye Christoph , Köppler Hubert
Published: October 2019

Introduction

Progress has been made in diagnosis and treatment of patients with follicular lymphoma who receive their treatment within prospective clinical trials. Due to necessary inclusion and exclusion criteria only a very limited number of patients are treated in studies. Therefore results from clinical trials can't be transferred into routine care. Little practice data are available how patients with follicular lymphoma are diagnosed and treated in routine care and whether improvements in survival are achieved.

Methods

A retrospective analysis of all patients with follicular lymphoma who were treated in an oncology group practice in Germany between 1995-2012. Relevant clinical data concerning diagnosis, treatment and survival were transferred from clinical files into a database and analyzed statistically using SPSS and SURVSOFT.

Results

174 patients with a median age of 60 (27-87) were identified. 43.7% were male and 56.3% female. Stage distribution at initial diagnosis was as follows: Stage I 45 patients (25.9%), stage II 23 patients (13.2%), stage III 32 patients (18.4%), stage IV 62 patients (35.6%), in 12 patients (6.9%) initial stage could not be retrieved. 90.8% needed therapy with a median of 2 different therapies (1-12). Regimens most frequently applied were: Rituximab-containing (67.1%), Bendamustine-containig (41.8%), Bendamustine+Rituximab-combinations (39.9%) and R-CHOP (27.8%). 15 patients (9.5%) received radiotherapy only. 13.3% of patients were treated within a clinical trial. 5 and 10 year absolute overall survival was 88.9% and 73.9%. Relative survival after 5 and 10 years was 94.4% and 86.6%. Median overall survival according to stage was 28 years for stage I, median not reached for stage II, 21 years for stage III and 16 years for stage IV. Median relative survival was 17.4 years for stage IV and has not been reached for stages I-III.

Conclusions

Systemic treatment of patients with follicular lymphoma in routine care consisted mainly of Bendamustine+Rituximab-combinations and R-CHOP. Employment of the most active chemoimmunotherapies leads to a marked prolongation of survival compared to historical controls and registry data.

Disclosures:

No relevant conflicts of interest to declare.
HLA Haplotype Frequency Analysis Within India: Pre-Requisite For Bone Marrow Donor Registry and Cord Blood Bank Planning
Authors: Maiers Martin , Halagan Michael , Leahy Nicole , Joshi Sangeeta , Ballal H Sudarshan , Jagannathan Latha , Damodar Sharat , Srinivasan Periathiruvadi , Narayan Saranya , Khattry Navin , Malhotra Pankaj , Shah Sandip A , Rajagopal Raghu , Cereb Nezih , Yang Soo Young , Dedhia Leenam , Parekh Sunil , Mammen Joy J , Daniels Dolly , Weisdorf Daniel J.
Published: October 2019

To better plan recruitment of volunteer adult unrelated donors for a registry and umbilical cord blood donations for cord banks to supply grafts for hematopoietic cell transplantation to serve Indian transplant candidate patients, we analyzed existing HLA typing data to estimate Indian national and regional populations’ HLA haplotype frequencies.

Indian BMT centers, renal transplant centers, adult donor registries and cord blood banks collaborated to collect HLA typing and demographic data.  We have produced a catalogue of the frequency, and regional diversity, of HLA haplotypes in the Indian population. Haplotype frequency analysis was performed on the registry datasets (N=26,239) using a maximum-likelihood method capable of providing high-resolution estimates from typing data with ambiguities.  These frequencies were generated for 20 regional subgroups into cohorts defined by either geographic state or by native language; and the 14 largest groups were used to develop a computational model of match rates.  Patient family typing data was used to set phase for a group of N=485 patients that was used to run searches against the NMDP registry.

The match rates for Indian patients searching the NMDP USA database (over 11 million donors including DKMS) were 39.7% at the 8/8 allele level (A, B, C, DRB1 high-resolution) and 92.2% at the 7/8 level.  Matching models for patients (averaged across the 14 regional subgroups) showed a match rate within India at the current registry sizes of 13.7% at the 8/8 level and 38.2% at the 7/8 level.  Increasing the registry to 1,000,000 would raise this to 55.0% for 8/8 and 84.1% for 7/8.  A cord blood inventory of 50,000 units would provide a match rate of 22.2% at the 6/6 level and 78.9% at the 5/6 level.  Increasing to a cord blood inventory of 250,000 units would increase these match rates to 42.5% at the 6/6 level and 90.3% at the 5/6 level.

These data will define the needs and serve as critical planning data to define and direct donor recruitment goals to establish an effective Donor Registry and cord blood banking facilities able to serve the needs of Indian patients who could benefit from the curative potential of transplantation. Developed data will be shared with the broader scientific community to facilitate future studies of HLA in areas such as disease association, anthropology and transplantation.

Disclosures:

Joshi: Manipal Hospital, Bangalore: Employment. Damodar:NARAYAN HRUDAYALAYA: Employment. Cereb:Histogenetics: Employment. Young Yang:Histogenetics: Employment. Dedhia:Marrow Donor Registry (India): Employment.