bluuebunny/crossref_metadata_2025 huggingface dataset:
Improving Hematopoietic Stem Cell Based Gene Therapy By Targeting CD133+ Cells
Authors: Goebel Benjamin , Brendel Christian , Abriss Daniela , Kneissl Sabrina , Brugman Martijn , Buchholz Christian J. , Serve Hubert , Grez Manuel
Published: October 2019

Introduction

Generally, CD34+ cells are used for genetic modification in gene therapy trials. CD34+ cells consist of a heterogeneous cell population with mostly limited long-term repopulating capabilities, resulting in low long-term engraftment levels in particular in those diseases in which gene modified cells lack a proliferative advantage over non-modified cells. Therefore, modifications in gene transfer vectors and gene transfer strategies are required to improve long-term clinical benefit in gene therapy patients. One particular attractive approach to solve this problem is the improvement of HSC based gene transfer by specifically targeting cells with long-term engraftment capabilities.

Material and Methods

We constructed lentiviral gene transfer vectors (LV) specifically targeting CD133+ cells, a cell population with recognized long-term repopulating capabilities. Targeting is achieved by pseudotyping with engineered measles virus (MV) envelope proteins. The MV glycoprotein hemagglutinin, responsible for receptor recognition, is blinded for its native receptors and displays a single-chain antibody specific for CD133 (CD133-LV). These vectors were compared to VSV-pseudotyped lentiviral vectors in in vitro and in vivocompetitive repopulation assays using mobilized peripheral blood CD34+ cells.

Results

Superior transduction of isolated human hematopoietic stem cell populations (CD34+CD38- or CD34+CD133+ cells) compared to progenitor cell populations (CD34+CD38+ or CD34+CD133-) could be shown using the newly developed CD133-LV. Transduction of total CD34+ cells with CD133-LV vectors resulted in stable gene expression and gene marked cells expanded in vitro, while the number of VSV-G-LV transduced CD34+ cells declined over time. Competitive repopulation experiments in NSG mice showed a significantly improved engraftment of CD133-LV transduced HSCs. At ∼12 weeks post-transplantation gene marked hematopoiesis was dominated by the progeny of CD133-LV transduced cells in 42 out of 52 transplanted animals in the bone marrow and 39 out of 45 transplanted animals in the spleen, respectively. Consistent with this data we could show that stem cell content in the CD133-LV transduced population is about five times higher compared to the VSV-transduced population using a limiting dilution competitive repopulation assay (LDA-CRU). Experiments showing proof of principle for the application of this technology for the correction of Chronic Granulomatous Disease (XCGD) using patient derived CD34+ cells are currently ongoing.

Discussion

In conclusions this new strategy may be promising to achieve improved long-term engraftment in patients treated by gene therapy.

Disclosures:

No relevant conflicts of interest to declare.
PI3K/AKT Pathway Is Activated By MYD88 L265P and Use Of PI3K-Delta Inhibitors Induces Robust Tumor Cell Killing In Waldenstrom’s Macroglobulinemia
Authors: Yang Guang , Liu Xia , Zhou Yangsheng , Xu Lian , Cao Yang , Manning Robert , Patterson Christopher , Tripsas Christina K , Hunter Zachary , Buhrlage Sara , Gray Nathanael S , Treon Steven Peter
Published: October 2019

Background

MYD88 L265P is a somatic mutation present in >90% of Waldenstrom's Macroglobulinemia (WM) patients. The downstream signaling pathways that promote malignant cell growth and survival remain to be fully clarified but include IRAK 1/4 and BTK (Treon et al, NEJM 2012; Yang et al, Blood 2013).

Methods

To further clarify the downstream signaling associated with MYD88 L265P in WM cells, we employed Phospho Explorer Antibody Arrays in MYD88 L265P expressing BCWM.1 and MCWL-1 WM cells following lentiviral mediated knockdown of MYD88, or over-expression of MYD88 wild type or L265P; or the use of MYD88 homodimerization inhibitor that block MYD88 signaling. Arrays were scanned by Axon GenePix Microarray Scanner and data analyzed by Ingenuity Pathway Analysis. Western blot analysis was performed using total and phospho-specific antibodies in WM cells. CellTiter-Glo® Luminescent cell viability assay (Promega) was used to assess cell survival following treatment with the PI3K-delta inhibitors, CAL101 and PIK-294 (Selleck Chemicals).

Results

Ingenuity pathway analysis demonstrated significant enrichment for B-Cell Receptor, PI3K/AKT, LPS-stimulated MAPK and ERK/MAPK signaling following MYD88 modulation in both BCWM.1 and MWCL-1 WM cells. Signal proteins that demonstrated the greatest changes in phosphorylation following MYD88 modulation are shown in Table 1. Western blot analysis confirmed the activation of ATK as downstream of PI3K in WM cells. Use of a MYD88 inhibitor significantly decreased phosphorylation of ATK-pS473 and AKT-pY308 in both BCWM.1 and MCWL-1 WM cells. Cell viability analysis demonstrated robust tumor cell killing (EC50 30-50nM) in BCWM.1 and MWCL-1 WM cells following treatment with both PI3K-delta inhibitors (CAL-101 and PIK-294).

Conclusion

In addition to activation of NF-kB through IRAK and BTK signaling, MYD88 L265P promotes survival of WM cells by activation of PI3K/AKT signaling. Inhibition of PI3K-delta is associated with robust killing of WM cells. These studies provide the framework for the investigation of PI3K-delta inhibitors in WM.

Disclosures:

No relevant conflicts of interest to declare.
Argonaute2 promotes myeloma angiogenesis by deregulation of microRNAs
Authors: Lijuan Chen , Jiaren Xu , Wu Shuang , Wang Rong , Xiaoyan Qu , Wenjun Yu , Li Jianyong
Published: October 2019

Dysregulation of miRNAs expression contributes to cancer cell proliferation, apoptosis and angiogenesis. Angiogenesis is a hallmark of multiple myeloma development and progression. Argonaute 2 (AGO2) protein, the core component of RNA-induced silencing complex (RISC), can directly bind miRNAs and mediate target mRNAs degradation. Previous study showed that AGO2 knockdown suppressed the growth and tube formation of HUVECs. In current study, the supernatant of AGO2 over-expression MM lines could induce HUVECs migration and accelerated tube formation. Conversely, the supernatant of AGO2 knockdown MM lines could suppress cell migration and tube formation of HUVECs. Moreover, CAM assay also demonstrated AGO2 can drive neovessel formation of MM in vivo. Using miRNAs microarray, we observed that 25 miRNAs were up-regulated and 14 miRNAs were down-regulated by AGO2 protein. Among these AGO2-associated miRNAs, most Let-7 family and two miR-17-92 cluster members (miR-17a and miR-92-1), known as pro-angiogenetic miRNAs, were the dominant positively regulated miRNAs by AGO2, and the anti-angiogenetic miRNAs, such as miR-145 and miR-361 were inversely regulated by AGO2 protein, which play crucial role in AGO2 mediating angiogenesis.

Disclosures:

No relevant conflicts of interest to declare.
Elevated Neutrophil: Lymphocyte Ratio Does Not Predict Survival In Multiple Myeloma Patients
Authors: Atallah Jean Paul , Azab Basem , Vivekanandarajah Abhirami , Naboush Ali , Sater Houssein Abdul , Farhan Mohammad , Patel Nileshkumar J , Elkosseifi Myriam , Fares Elias , Boumitri Christine , Zaraket Vera , Saad Marc , Hassan Samer , Odaimi Marcel , Terjanian Terenig
Published: October 2019

Background

Cancer associated inflammation is one of the key determinants of outcome in patients with cancer. An elevated neutrophil: lymphocyte ratio (NLR) has been identified as a predictor of worse survival in patients with various solid tumors compared to hematologic malignancies but no reports yet examined its impact on multiple myeloma. The aim of this study was to examine the prognostic value of an elevated NLR in multiple myeloma.

Methods

We had approval by our institutional review board to collect the data on patients diagnosed with multiple myeloma at Staten Island University Hospital between year 2000 and 2012 identified from our local cancer database. Data on demographics, conventional prognostic markers, laboratory analyzes including blood count results, and histopathology were collected and analyzed. A cox proportional survival analysis was carried out to assess the relationship between NLR and mortality. NLR was assessed as a continuous variable as well as categorical variable (quartile 0.5-1.5, 1.6-2.2, 2.3-3.8, and 3.9-22.3).

Results

A total of 96 patients were identified with a median age at diagnosis of 70 (IQR of 61 to 79) years. The median neutrophil count was 3.5 (2.5—5.1) x 10-9/liters, median lymphocyte count 1.5 (1.05-2.4) × 10-9/liters, while the NLR was 2.28 (1.53-3.88). The median overall survival was 147.5 weeks, IQR (88.5-320). NLR did not prove to be a significant predictor of death as a continuous variable (0.95 (0.85-1.06), p =0.35). Furthermore, there was no significant difference in survival with any of the quartiles of NLR. Compare to lowest quartile of NLR, Hazards ratio for the consecutive quartiles were 1.25 (0.56-2.79, p 0.55), 1.36 (0.61-3.04, p=0.45) and 0.89 (0.36-2.22, p=0.80).

Conclusion

NLR does not appear to offer useful predictive ability for outcome and survival in multiple myeloma patients. Our study is limited with small sample size, further studies are needed to validate our results.

Disclosures:

No relevant conflicts of interest to declare.
The Presence Of Spine Bone Plasmacytomas At The Time Of Multiple Myeloma (MM) Diagnosis Confers Poor Prognosis
Authors: Kyrtsonis Marie-Christine , Nikolaou Eftychia , Maltezas Dimitrios , Sarris Katerina , Koulieris Efstathios , Iliakis Theodoros , Vyniou Nora , Vardounioti Ioanna , Karali Vassiliki , Pessach Ilias , Bartzis Vassiliki , Tzenou Tatiana , Dimou Maria , Vassilakopoulos Theodoros P. , Angelopoulou Maria K. , Tsaftaridis Panagiotis , Katodritou Irene , Panayiotidis Panayiotis
Published: October 2019

The negative prognostic consequences of extramedullary plasmacytomas in the context of symptomatic MM is well known. On the contrary, the clinical impact of bone plasmacytomas (BP) has not been extensively studied, although their presence is one of Duries’ major MM diagnostic criteria.

The aim of the present study was to study eventual differences between MM patients presenting at diagnosis with or without BP.

Patients and Methods

Two hundred and twelve symptomatic MM patients were studied from diagnosis to last follow-up. Their median age was 66 years and 60% were men. Twenty-three percent, 27% and 50% of patients were in ISS stage 1, 2 and 3 respectively; MM type was IgG in 59% , IgA in 26%, light chain only in 12%, IgD in 1,5% and non- or oligo-secretory in 1,5%. All patients required treatment and BP ones were additionally given radiotherapy. The median follow-up time of the whole cohort was 46,4 months.

Results

Forty patients (19%) had a BP at the time of their diagnosis. BP was located in the spine (BP-S) in 19 out of 40 and in other sites (BP-O: skull, pelvis, long bones, and ribs) in 21 patients. BP patients had equivalent demographics, stage and MM type as the others. Non-BP patients tended to have a better survival than BP ones but the difference was not statistically significant (p=0.1). However, BP-S patients had a significantly worse outcome than BP-O patients (24±6 versus 48±11 months, p<0.00001), and indeed worse than non-BP patients. It is worthwhile to outline that 17 out of 19 BP-S patients had received new drugs (thalidomide, lenalidomide, Bortezomib) during their course while 8 of them underwent high dose treatment with autologous stem-cell transplantation. In addition, the ISS staging has absolutely no prognostic impact in these patients (p=0.5), on the contrary to the whole cohort (p=0.03).

In conclusion, MM patients presenting spine plasmacytoma at diagnosis constitute a distinct subgroup with adverse outcome that is difficultly predictable, given that the current staging system does not separate them. Further confirmation of this finding in larger cohorts is needed, as well as exploration of the biologic background.

Disclosures:

No relevant conflicts of interest to declare.
Once-Weekly Subcutaneous Bortezomib Plus Oral Cyclophosphamide and Dexamethasone Therapy For Relapsed Or Refractory Multiple Myeloma
Authors: Takamatsu Yasushi , Imamura Yutaka , Nakazato Tomonori , Uike Naokuni , Miyashita Kaname , Saburi Yoshio , Harada Naoki , Ota Shuichi , Imamura Rie , Miyamoto Toshihiro
Published: October 2019

Background

We previously reported that twice-weekly intravenous bortezomib plus doxorubicin and intermediate-dose dexamethasone (iPAD) therapy induced a high complete response (CR) rate of 30% and prolonged remission duration of median 12.1 months in patients with relapsed or refractory myeloma. However, sensory neuropathy was observed in 78% of the patients with grade 3 neuropathy in 22%. To reduce the toxicity, we conducted once-weekly subcutaneous bortezomib therapy given in combination with oral cyclophosphamide and dexamethasone (sVCD).

Methods

This was a phase 2 multicenter study conducted by the Kyushu Hematology Organization for Treatment Study Group (K-HOT). sVCD regimen consists of bortezomib 1.3 mg/m2 administered subcutaneously, and cyclophosphamide 300 mg/m2 and dexamethasone 40 mg given orally on days 1, 8, 15 and 22. The treatment was repeated at a 5-week interval for 6 cycles. The primary endpoint was to determine the complete response rate. The toxicities including sensory neuropathy and injection site reaction were analyzed as the secondary endpoint. The study was approved by the Institutional Review Board at each participating center and was registered in University hospital Medical Information Network (UMIN000006490). All patients provided a written-informed consent.

Results

Thirty-one patients with median age of 69 (range, 43-87) were entered into this study. The numbers of prior chemotherapy line were 1 in 39%, 2 in 23%, and more than 3 in 39% of the patients. Bortezomib, lenalidomide and thalidomide had been given in 52%, 26% and 16%, respectively. CR was achieved in 16%, partial response (PR) in 26%, and minimal response (MR) in 29%, resulting in overall response rate of 71%. Grade 3/4 neutropenia was observed in 35%/3%, thrombocytopenia in 16%/13%, and anemia in 45%/0% of the patients. Grade 3 to 4 non-hematological toxicities were uncommon; the highest incidence of grade 3 events were rash, hyponatremia and hyperglycemia observed in 6%, while no grade 4 events were reported. Grade 3 sensory neuropathy developed in 3% and grade 2 in 19%. Injection site reactions were observed in 39%, but all of them were grade 1.

Conclusion

sVCD therapy containing bortezomib administered subcutaneously in once-weekly schedule successfully reduced the incidence and severity of sensory neuropathy with moderate response rate.

Disclosures:

Miyamoto: Kyushu University Hospital: Employment.
Description Of The Hematological Toxicity Of Different Regimens Using Bortezomib In Multiple Myeloma (CyborD, Vel-Dex and VMP)
Authors: Duquette Dominic , Laroche Vincent
Published: October 2019

Introduction

  The use of bortezomib has evolved since the APEX study published in 2005. It remains, however, the only study providing a detailed account of the changes in hematological parameters in response to bortezomib-based chemotherapy. The APEX trial enrolled multiple myeloma patients in relapse.  In first-line transplant eligible multiple myeloma patients, we use bortezomib twice weekly with low-dose dexamethasone (VelDex) or bortezomib once weekly with dexamethasone and cyclophosphamide (CyBorD). For patients non-eligible to transplant, we use bortezomib once or twice weekly with melphalan and prednisone (VMP).  The goal of this study is to describe changes in platelet counts during these three types of bortezomib-based induction therapys.

Methods.

  We conducted a retrospective chart review to examine the complete blood count (CBC) results during treatment with VelDex, CyBorD or VMP. Neutrophil count, hemoglobin and platelet count were measured before every bortezomib infusion. Patients not receiving the anticipated protocol or less than 3 cycles of VelDex or CyBorD or 6 cycles of VMP were excluded from this review.

Results

  30 patients were included in this review (VelDex = 11, CyBorD =10, VMP = 9) Results are showed in the following graphs. The use of twice weekly bortezomib is associated with a more pronounced drop in platelets.  CyborD once weekly is associated with less severe drop in platelets.

Conclusion

  These findings show that once weekly bortezomib is associated with less severe thrombocytopenia.  This suggests that the blood monitoring could be less frequent than at each dose of bortezomib.  A biweekly monitoring of blood counts could reduce significantly blood tests and waiting time for patients.

Disclosures:

Duquette: Janssen: Consultancy, Honoraria. Off Label Use: CyborD is a widely accepted regimen but has not been accepted by the FDA.
PTEN Regulated BCRP/ABCG2 and Side Population Through PI3K/Akt Pathway In Chronic Myeloid Leukemia
Authors: Huang Fang-Fang , Zhang Li , Yu Yan-Hui , Chen Fang-ping , Zeng Hui , Niu Ting , Wu Deng-shu
Published: October 2019

A small population of cancer stem cells named as “side population (SP)” has been demonstrated to be responsible for many solid tumor maintenance. However, the role of SP in leukemic pathogenesis is still controversial. The resistance of leukemic stem cells in response to targeted therapies such as tyrosine kinase inhibitors (TKIs) results in therapeutic failure or refractory/relapsed disease in chronic myeloid leukemia (CML). The drug pump, ABCG2, is well-known as a specific marker of SP and could be controlled by several pathways including PI3K/Akt. Our data demonstrated that compared to wild type K562 cells, the higher percentage of ABCG2+ cells corresponded to the higher SP fraction in K562/ABCG2 (ABCG2 overexpressing) and K562/IMR (resistance to imatinib) cells, which exhibited enhanced drug resistance along with downregulated PTEN and activated p-Akt. It could be abrogated by both PI3K inhibitor LY294002 and mTOR inhibitor rapamycin. Moreover, in CML patients at accelerated phase/blastic phase (AP/BP), increased SP phenotype rather than ABCG2 expression accompanied with loss of PTEN protein and up-regulation of p-Akt expression was observed. These results suggested that expression of ABCG2 and fraction of SP may be regulated by PTEN through PI3K/Akt pathway, which will be the potentially effective strategy for targeting CML stem cells.

Disclosures:

No relevant conflicts of interest to declare.
Post-Transplant Immune Monitoring In Patients Receiving Allogeneic Stem Cell Transplantation
Authors: Park Silvia , Jung Chul Won , Jang Jun Ho , Kang Eun Suk , Kim Kihyun
Published: October 2019

Introduction

There are still substantial morbidity and mortality caused by insufficient immunologic recovery after allo-HSCT. In this context, we attempt to evaluate the clinical relevance of immune monitoring in allo-HSCT recipients.

Method

Fifty five patients who underwent allo-HSCT between 2008 and 2012 were included. Peripheral blood samples were drawn from recipients before transplant, and on 4, 8, 12, 24, 36 and 48 weeks after transplant. Each blood samples were analyzed by multi-color flow cytometry for determining lymphocyte subsets. MNC were separated from blood specimen, and analyzed for the quantitation of Treg with the use of real-time PCR. We also examined T cell derived IFN-r by using in vitro culture, intracellular staining, and flow cytometry analysis.

Results

The median age was 43, and AML was the most common reason for transplantation (49.1%). Grade II or more aGVHD occurred in 36.4% of cases, and 49.1% exhibited moderate or severe cGVHD. The differences in the proportion (%) and the absolute number (/uL) of CD4+, CD8+ cells, CD4+ derived IFN-r (%), CD8+ derived IFN-r (%), and Treg (%) between the groups (Gr. II or more aGVHD (+) vs (-); moderate or severe cGVHD (+) vs (-)) were compared by Two sample t-test. Patients with Gr. II or more aGVHD showed decreased CD4+ count at 4, 8 and 12 weeks, but showed rather higher CD8+ count at 8 weeks after transplant. T-cell secretion function assessed by IFN-r (%), and Treg (%) was similar between two groups within 12 weeks after transplant. In case of cGVHD, both CD4+ and CD8+ count tended to be higher in patients with moderate or severe cGVHD, and the trends lasted for up to 48 weeks from allo-HSCT. Treg (%) was almost consistently lower throughout the period in these patients. There were 12 relapses within follow up period (median 36.1 months), and higher slope of post-transplant increase in CD8+ count and CD8 derived IFN-r were identified as protective factors for disease relapse.

Conclusion

In view of the results so far achieved, slow recovery of CD8 count and function might be associated with disease relapse. However, this is still a preliminary data, and warrants further evaluation.

Disclosures:

No relevant conflicts of interest to declare.
Allogeneic Stem Cell Transplantation With Reduced Intensity Conditioning Regimen In Patients With Relapsed Hodgkin´s Lymphoma
Authors: Rivas Maria Marta , Berro Mariano , Yantorno Sebastian , Prates Maria Virginia , Milone Jorge H , Basquiera Ana Lisa , Berreta Adriana , Garcia Juan , Remaggi Guillermina , Rolon Juliana Martinez , Arbelbide Jorge , Makiya Monica , Dibar Eduardo , Requejo Alejandro , Julio Feldman Leonardo , Milovic Vera , Foncuberta Maria Cecilia , Burgos Rubén , Kusminsky Gustavo Daniel
Published: October 2019

Introduction

Hodgkin´s Lymphoma (HL) is a highly curable disease. However, there are still patients with primary refractory disease or who relapse after first-line treatment, or even after high-dose chemotherapy with hematopoietic cell rescue. Allogeneic stem cell transplant (ASCT) is therapeutic for this patients.

Objective

To analyze the experience with relapsed HL patients that received ASCT with reduced intensity conditioning (RIC)regimen in 8 Argentine Medical Centers.

Design and Population

We performed a retrospective multicenter analysis from data obtained from medical records. Fifty-four patients with relapsed HL who received ASCT had a median age of 26years. The relationship between male / female was 1/1. Only 3 patients (5.5%) at the time of transplant had a performance status> 1 according to ECOG. Ninety-six percent of the patients had received previously autologous transplant. Most patients 43 (80%) received an identical sibling donor transplant. All patients receiving unrelated donor transplants had in vivo lymphocyte depletion as prophylaxis of graft versus host disease. Forty-three patients (79.6%) received as a conditioning regimen Fludarabine + Melphalan. The disease status at transplant was: complete remission (CR) 33%, partial remission (PR) 54%, stable disease / progressed (SD / PD) 13%.

Results

With a median follow up of 2.7 years, actuarial overall survival (OS) at 1 and 5 years was 65% and 20% respectively and disease free survival (DFS) at 1 and 5 years was 35 % and 18% respectively. The incidence of acute GVHD grade II-IV was 31%. Patients in CR at the time of transplant showed significant differences compared with those who were not in CR in DFS (1-5 years 52-27% vs 19-14%, p=0.01), OS (1-5 years 76-38% vs 59-13%, p=0.02) and non relapsed mortality (NRM) (1-5 years 6-12% vs 34-39%, p=0.04). Age, PS, the use Fludarabine + Melphalan as conditioning regimen, unrelated donor, aGVHD, were not variables that modified the overall survival and disease-free survival.

Conclusion

The ASCT with RIC regimen is a feasible therapeutic option in patients with HL, especially in patients who can achieve CR. The low rate of DFS is still an issue in this setting, may be new drugs may help in optimizing pretransplant response status to improve patients’ outcome.

Disclosures:

No relevant conflicts of interest to declare.
Sequential R-CHOP and R-FM Chemotherapy Followed By Autologous Stem Cell Transplantation Results In High Rates Of Long Term Remission In Advanced Follicular Lymphoma
Authors: Derenzini Enrico , Stefoni Vittorio , Casadei Beatrice , Pellegrini Cinzia , Broccoli Alessandro , Fanti Stefano , Gandolfi Letizia , Stefani Giulia , Argnani Lisa , Zinzani Pier Luigi
Published: October 2019

Introduction

Autologous stem cell transplantation (ASCT) is a potentially curative treatment option for relapsed Follicular Lymphoma (FL) patients, but to date available data do not support the use of ASCT as first line consolidation, given the lack of overall survival (OS) advantage compared to standard therapy. R-CHOP (Rituximab-Cyclophosphamide, Vincristine, Doxorubicin, Prednisone) and R-FM (Rituximab, Fludarabine, Mitoxantrone), have comparable efficacy and are widely used as first and second line combinations. The best way to sequence the available therapies in FL is still undefined. Here we show the long term results of a phase II trial of sequential chemotherapy alternating CHOP and FM plus Rituximab followed by ASCT in patients with stage III-IV and/or bulky FL either at disease onset or first relapse, conducted in our Institution from 2002 to 2008.

Methods

Patients at diagnosis or first relapse were treated in sequence with R-CHOP for 4 cycles, Endoxan 7g/m2 followed by hematopoietic stem cell harvest, R-FM for 4 cycles and ASCT. The ASCT conditioning schedule was BEAM (BCNU, ARA-C, Etoposide, Melphalan) in all cases.

Results

24 patients were enrolled, 12 pts were male. Median age was 44 years. One patient did not undergo ASCT for insufficient left ventricular ejection fraction and was excluded from the analysis. 13 patients were treated upfront whereas 10 patients at first relapse. After a median follow-up of 10 years, progression free survival (PFS) and OS in the whole study cohort were respectively 65% and 87%, with a complete response (CR) rate after the completion of sequential treatment of 100%. PFS and OS for patients treated at disease relapse were 60 and 70% (4 relapses, 3 deaths). Remarkably PFS and OS for the 13 patients treated upfront was 70% and 100% (4 relapses). To date no secondary malignancies were observed.

Conclusions

Sequential treatment alternating standard R-CHOP and R-FM followed by ASCT results in impressive long term PFS and OS rates, both in first line and at relapse. These data represent the proof of principle of a sequential therapy containing alternating alkylating agents and purine analogs followed by ASCT in FL.

Disclosures:

No relevant conflicts of interest to declare.
Use Of Unrelated Donors In Elderly Patients (age >60 years) Undergoing Reduced-Intensity Conditioning Hematopoietic Stem Cell Transplantation For Hematologic Malignancies
Authors: Jean El-Cheikh , Raynier Devillier , Sabine Fürst , Roberto Crocchiolo , Angela Granata , Catherine Faucher , Bilal Mohty , Samia Harbi , Reda Bouabdallah , Norbert Vey , Luca Castagna , Christian Chabannon , Blaise Didier
Published: October 2019

Objectives

The lower morbidity and mortality of reduced-intensity conditioning (RIC) regimens have allowed allogeneic hematopoietic cell transplantation (HCT) in older patients. However, there are only limited data on the feasibility and outcomes of URD HCT in elderly patients.The aim of the study was to compare the outcome in OS and PFS for patients transplanted using unrelated donor (URD) in patients age 60 or older.

Patients and methods

We retrospectively analyzed outcomes in 62 consecutive hematologic malignancy patients aged > or =60 years (median, 62 years; range: 60-70 years) undergoing reduced intensity conditioning regimens (RIC) from URD. In this study, URD was used only when a MRD was not available.

Then we compared the outcome of 17 elderly patients (age >65 years) with 44 younger patients aged between 60 and 65 years. Patient, disease and transplant characteristics are shown in Table 1.

Results

No patients experienced graft rejection. The median HCT comorbidity index score was 2 (range, 0 to 6). With a median follow up of 36 months (range, 5-74), the cumulative incidence of grades II to IV acute GVHD was 28% and of grades III to IV acute GVHD, 13%. At 2 years, the cumulative incidence of chronic GVHD was 27%, progression-free survival (PFS) was 62%, overall survival (OS) was 63%, and relapse was 14%. Non relapse mortality (NRM) was 24% at 2 years.

The cumulative incidence of grade II–IV Acute GVHD was 43% for the younger group and 17% for the older group (P = 0.056). The cumulative incidence of chronic GVHD was not different between the two groups (23% vs. 45% (p=0.3), respectively). Two-year OS and PFS was 57% versus 86% (P = 0.059) and 55% versus 86% (P = 0.03), in the younger and the older group respectively. The 2-year NRM and relapse was 26% versus 14% (P = 0.4) and 19% versus 0% (P = 0.04), in the younger and older group respectively.

Conclusions

This retrospective study suggest that RIC HCT from URD is a safe and effective option for patients aged > or =60 years or older, and in the absence of suitable related donors, well-matched URD may offer a very reasonable alternative, and that does not appear to be associated with a detrimental outcome. However these results are encouraging showing once again that with an adequate selection, age is not a definitive limitation.

Disclosures:

No relevant conflicts of interest to declare.
Is Rituximab Needed With High Dose Chemotherapy and Autologous Stem Cell Transplant When Patients With Non-Hodgkin’s Lymphoma Have Been Exposed To It?
Authors: Gul Zartash , Anderson Stephan , Slone Stacey , Chhabra Saurabh , Monohan Gregory , Howard Dianna , Lawson Amber , Hayslip John
Published: October 2019

Introduction

The value of Rituximab with High dose chemotherapy and autologous stem cell transplant (auto-HCT) has been evaluated in some retrospective studies. It is noted that the addition Rituximab to auto-HCT results in significant improvement in overall (OS) and disease free survival (DFS). However, it is unclear that the addition of Rituximab with auto-HCT would result in significant benefit in patients who have received prior Rituximab based therapy.

Methods

We retrospectively evaluated twenty seven consecutive patients who had B cell non-Hodgkin's lymphoma (NHL) and underwent auto- HCT at Markey cancer center between January 2010 and December 2012. All patients received 375 mg/m2of Rituximab with chemotherapy. Rituximab was not administered with high dose chemotherapy (BEAM: BCNU, Etoposide, Cytarabine and Melphalan) prior to auto-HCT. All patients received GCSF starting day 5 after auto-HCT.

Results

There were 27 patients who underwent auto-HCT for NHL. Median age of the patients was 60 years (36-72 years). Nineteen patients were male and 8 patients were female. Fifteen patients were in complete remission (CR) and the rest were in partial remission (PR). Patients had: Diffuse large B cell lymphoma (DLBCL=11 patients): Mantle cell lymphoma (MCL=13 patients): follicular lymphoma (3 patients). Median CD 34 count of infused cells was 4.14 x 106 cell/kg (2.26- 9.45). Median time to neutrophil recovery was 11 days (9-14). Median lymphocyte count at day 30 after auto HCT was 1130 x 10 6/l (320- 5180). After a median follow up of 7.9 months (3.0- 32.4) eight patients had relapsed. Five patients who had relapsed had refractory disease before auto-HCT. Non -Relapse mortality (NRM) is shown in figure 1. Median overall survival and disease free survival were not reached. Seventy percent patients had not relapsed. We did not find any factors associated with relapse.

Conclusion

We performed a retrospective study to evaluate the impact of Rituximab with high dose chemotherapy and in patients with NHL. Review of literature showed that median 2 year and 5 year OS was 80% and 69% in patients who received Rituximab with auto HCT. Median 2 year DFS was 67%. In our study 70% patients were relapse free at a median follow up of 8 months. We would need a longer follow up to determine the efficacy of Rituximab with auto- HCT in patients with NHL who had prior exposure to Rituximab.

Disclosures:

Off Label Use: The use of Thymoglobulin® for GVHD prevention. Hayslip:Sanofi: Research Funding; Janssen: Research Funding; Pfizer: Research Funding; Celgene: Research Funding.
Assisted Administration Of Subcutaneous 5-Azacytidine to The Patient's Home. Significant Reduction Of Indirect Costs Incurred By Care Recipients and Unpaid Caregivers Together With a Total Adhesion To The Treatment Schedule and Same Safety
Authors: Iuliano Francesco , Iuliano Eleonora , Perricelli Alessia , Pomillo Angelo , Luci Maria , Cervo Gianluca
Published: October 2019

Background and aim

Administration of azacitidine is planned in-patient daily, there is no experience of its administration to the patient's home. The aim of this work was to evaluate the feasibility of a program of home administration of azacitidine able to reducing the cost-of-illness, to increase adherence to treatment while maintaining the same safety of the therapy given in hospital.

Material and methods

Between Jan 2008 and Dec 2012, 22 consecutive  patients, (MDSs, n = 15; CMML, n = 4; AML, n = 3), were enrolled in the study.

The pharmacoeconomic analysis included assessment of direct costs ( hospital inpatient, physician inpatient, physician outpatient, emergency department nursing home care, specialists’ and other health professionals’ care, diagnostic tests, prescription drugs and drug sundries, and medical supplies),indirect costs incurred by care recipients and unpaid caregivers, including time, productivity and travel cost.

Results

Azacitidine 75 mg/m2/day was administered as a subcutaneous injection for 7 consecutive days  every 4 weeks,  Median age of the patients was 71 years (range, 65–83). Median number of courses delivered to each patient was 9 (range, 3–31) Hematologic responses (CR/PR/mCR) were induced in 6 patients (27.0%)  Median number of treatment courses to achieve any response was 2 (range, 1–6) Adverse events were evaluated for the first 6 courses for all patients, for a total of 124 courses. Major adverse events were cytopenia and cytopenia-related infection. Grade 3 or higher neutropenia was 64.4%) but incidence of febrile episode requiring intravenous antibiotics was  8,4% slightly lower than reported from the pivotal clinical study. Grade 3 or higher non-hematologic toxicities were infrequent. Injection site reaction 0.4 and site pain 0% Median follow-up duration of surviving patients was 46.9 months (range, 11.8–55.5). Of the 14 patients  who were RBC transfusion dependent at baseline, 48.0% of these patients became RBC transfusion independent during the treatment period. Adherence to treatment was 100%.

In our experience, despite the high percentage of elderly patients of whom 36% living in rural area, it was possible to give treatment to all patients with 100% adherence. There has been a reduction in direct medical costs due  to less use of hospitalization, a reduction of indirect costs by 63% due to the lower number of working days lost and a drastic reduction of travel costs, with the same efficacy and safety of administration.

Disclosures:

Off Label Use: drug administration at patient home instead of hospital.
Cardiovascular Complications Of Thalassemia Patients In The Iron Chelation Therapy Era
Authors: Tantiworawit Adisak , Tapanya Suebsakul , Phrommitikul Arintaya , Norasetthada Lalita , Chai-adisaksopha Chatree , Rattarittamrong Ekarat , Hantrakool Sasinee , Taechacharn Sucharawee , Nawarawong Weerasak
Published: October 2019

Background

Cardiac complications are the most common cause of death in thalassemia, accounting for up to 71% in the past. Iron chelation therapy is given to patients with iron overload to prevent these complications. The cost effectiveness of iron chelation therapy was arguable.

This study aims to evaluate the prevalence of cardiac complication and the correlation between risk factors in iron chelation therapy era.

Method

This is a cross sectional study from June 2011- May 2012. All thalassemia aged > 15 years old were enrolled. Clinical data and hemoglobin typing were reviewed. Echocardiography and CMR T2*, a technique represent cardiac iron deposition used to evaluate cardiac function, were used to evaluate cardiac complications.

Results

Ninety one patients were enrolled, 63.7% were females, median age of 31 years (16-75). There was 49.5 % homozygous β thalassemia, 31.9% β thalassemia/Hb E disease, 18.7% Hb H disease. Half of patients were transfusion dependent and 63.7% underwent splenectomy. Eighty four percent of patients received iron chelation therapy but few of them got their preferable choice in adequate dosage. Patients with serum ferritin levels more than 1,000 ng/ml. received deferoxamine, deferiprone or deferasirox.

Even with the iron chelation therapy, mean serum ferritin level was still high at 3,820 ng/ml for the whole group.

CMR T2* was more sensitive in detecting cardiac function. The CMR T2* showed shorter signal (≤ 20 msec) in 11.1%. Only 8.2% had impaired ejection fraction <55% by echocardiography. The CMR T2*  ≤ 20 msec was significant correlated with higher maximum ferritin 5,739.14 ng/ml compared to 3,614 ng/ml (p=0.001). Pulmonary hypertension was found 7 patients (12.7%) and 71.42% had underwent splenectomy.

Conclusion

From our study, the CMR T2* is the sensitive method for detecting cardiomyopathy and highly correlated with serum ferritin levels. Splenectomy remains the major risk factor for pulmonary hypertension.

The incidence of cardiac complications has decreased with iron chelation therapy for maintaining acceptable serum ferritin levels but the problem with cardiomyopathy and pulmonary hypertension still exist. Early detection, more sensitive implementation and aggressive iron chelation therapy are necessary to prevent these complications.

The majority of the patients in Thailand which are under universal health-care coverage scheme could not get access to more effective and expensive iron chelator. Regular and adequate chelation plays a major role in the prevention of cardiac complications and the achievement of better quality of life.

Disclosures:

No relevant conflicts of interest to declare.
Bendamustine and Rituximab Achieve a Major Survival Improvement Of Patients With Morbus Waldenström 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 Morbus Waldenström 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 Morbus Waldenström are diagnosed and treated in routine care and whether improvements in survival are achieved.

Methods

A retrospective analysis of all patients with Morbus Waldenström 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

52 patients with a median age of 67 (36-84) were identified. 67.3% were male and 32.7% female. Distribution according to IPSSWM-stage was: 21.2% low risk, 61.5% intermediate risk, 17.3% not evaluable (in most cases due to lacking β2 microglobulin data). 59.6% needed therapy with a median of 3 therapies (1-12). Regimens most frequently applied were: Bendamustine-containig (74.2%), Rituximab-containing (74.2%), Bendamustine+Rituximab-combinations (61.3%) and Chlorambucil-containing (45.2%). 6.5% of patients were treated within a clinical trial. Median overall survival was 12 years (0-19). Median relative survival was 13.6 years. 10-year relative survival was 86%. 10-year relative survival according to IPSSWM-stage was as follows: low risk 92%, intermediate risk 71%. Median overall survival of the patients who never needed any therapy was 11 years compared to 12 years of those patients who needed therapy. Median relative survival for patients who never needed therapy was not reached yet, median relative survival of the patients who needed therapy was 13.6 years.

Conclusions

Systemic treatment of patients with Morbus Waldenström in routine care consists mainly of Bendamustine, Rituximab and Bendamustine+Rituximab-combinations. 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.
GFI1B Mutation Causes a Platelet Function Defect With Reduced Alpha-Granule Content and Abnormal Aggregation Response
Authors: Ward Christopher M , Morel-Kopp Marie-Christine , Chen Qiang , Liang Hai Po , Ng Ashley P. , Roberts Andrew W. , Bahlo Melanie , Stevenson William s
Published: October 2019

GFI1B is a transcription factor important for erythropoiesis and megakaryocyte development, but previously unknown to be associated with human disease. We have identified a family with an autosomal dominant bleeding disorder associated with thrombocytopenia, red cell anisopoikilocytosis and platelet dysfunction. The severity of bleeding is variable with some affected individuals experiencing spontaneous bleeding while other family members only exhibit abnormal bleeding with surgery. All affected individuals had a moderate thrombocytopenia ranging from 72-149 x109/L with an elevated mean platelet volume. Platelet function was perturbed with prolonged collagen/epinephrine closure times on the PFA-100 (294 vs 125 sec, P <0.001). Platelet aggregation studies were abnormal, with an absent collagen response and primary aggregation only with ADP, arachidonic acid and adrenaline. Genetic linkage analysis and massively parallel sequencing were performed on 16 family members to localize the mutation causing the disease phenotype to chromosome 9. A single nucleotide insertion was identified in GFI1B that predicts a frameshift mutation in the fifth zinc finger DNA-binding domain. This mutation alters the transcriptional activity of the protein as determined by luciferase production from the TGFBR3 and GFI1B promoters. Aberrant transcriptional activity was associated with a marked reduction in platelet alpha-granule content as measured by electron microscopy (1.3 vs 3.1 alpha-granules per 1µm2, P<0.001) with similar changes in platelet protein expression. Proteomic analysis of platelet lysates demonstrated altered levels of cytoskeletal proteins, including talin, vinculin and myosin light chain 6. GFI1B mutation represents a novel human bleeding disorder and the described phenotype identifies GFI1B as a critical regulator of platelet number and function.

Disclosures:

No relevant conflicts of interest to declare.
Effects of GLI1 and CTNNB1 Knockdown on Notch Signaling and Proliferation of AML and T-ALL Cell Lines
Authors: Okuhashi Yuki , Takahashi Yusuke , Ohtaka Mika , Shiratori Erika , O Shijyun , Itoh Mai , Tohda Shuji
Published: October 2019

Objective: Although Hedgehog (Hh) and Wnt signaling pathways are known to play a role in cancer cell growth, their specific effects on leukemia cells are not fully understood. Previously we reported that the Hh inhibitor, cyclopamine, and the Wnt inhibitor, quercetin, suppressed the growth of leukemia cells. However, a specific inhibitory role of these chemical compounds on Hh and Wnt signaling could not be confirmed since these compounds also have off-target effects. Therefore the effects of specific inhibition of Hh and Wnt signaling pathways must be investigated in detail. In our previous study, we reported that small interfering RNA (siRNA)-mediated knockdown of NOTCH1 and NOTCH2 suppressed growth of T-ALL cell lines. In this context, the relationship between Hh, Wnt, Notch and mTOR signaling; known to regulate stemness; has not been fully elucidated. To address these questions, the effects of siRNA-mediated knockdown of GLI1 and CTNNB1 (Catenin beta 1), which respectively mediate Hh and Wnt signaling, on cell proliferation as well as Notch and mTOR signaling were studied in AML and T-ALL cell lines.

Methods: Two AML cell lines (NB4 and THP-1) and 2 T-ALL cell lines (DND41 and Jurkat) were used in this study. siRNAs targeting GLI1 (siGL), CTNNB1 (siCT), NOTCH1 (siN1) or control siRNA were transfected into cells by using the pipette tip chamber-based electroporation system. The effects of siGL and siCT transfection on cell proliferation were examined using a colorimetric WST-8 assay and by observing cytospin preparations of the harvested cells. The effects on mRNA and protein expression were examined by quantitative RT-PCR and immunoblotting, respectively.

Results: Transfection with siGL and siCT selectively suppressed the expression of GLI1 and CTNNB1, respectively. siRNA treatment did not lead to any significant effects on cell proliferation or morphology in all tested cell lines. In NB4 cells, GLI1 as well as CTNNB1 knockdown increased the level of NOTCH1, the cleaved NOTCH1 fragment (active form of NOTCH1), HES1, and phosphorylated mTOR protein. In the other cell lines, the expression of these proteins was not significantly altered by siGL and siCT.

Discussion: siRNA-mediated knockdown experiments suggested that Hh and Wnt signaling had insignificant effect on the proliferation of the leukemia cell lines used in this study. We report a novel interaction, namely, the activation of Notch and mTOR signaling by the suppression of Hh and Wnt signaling in NB4 cells. The molecular mechanism and significance of this phenomenon as well as its translation to other cell lines needs further examination. We propose that this finding must be taken into account when Hh- and Wnt-targeted therapy against leukemia are developed.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.