bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prognostic Significance of Telomere Length in Chronic Lymphocytic Leukemia Patients in Early Stage Disease,
Authors: Fabris Sonia , Hoxha Mirjam , Agnelli Luca , Dioni Laura , Bollati Valentina , Ciceri Gabriella , Mosca Laura , Barbieri Marzia , Cutrona Giovanna , Matis Serena , Gentile Massimo , Deliliers Giorgio Lambertenghi , Cortelezzi Agostino , Morabito Fortunato , Ferrarini Manlio , Baccarelli Andrea , Neri Antonino , Bertazzi Pier Alberto 
Published: October 2019

Abstract
Abstract 3890
Chronic lymphocytic leukemia (CLL) is a genetically heterogeneous disease with a variable outcome. The identification of factors that could predict the clinical course of early-stage CLL represents a crucial objective. Although previous studies indicated that telomere length may be a useful independent prognostic factor in the risk stratification of CLL, limited information has been reported in asymptomatic early stage patients (Binet stage A). We investigate the association of telomere length with the major biological and cytogenetic markers known to predict clinical outcome in CLL. The global DNA methylation levels of Alu and LINE sequences, was also investigated. Correlation with disease progression, measured as the time elapsed from diagnosis to first treatment, was evaluated. We measured relative telomere length (RTL) by real-time PCR in a panel of highly purified (>90%) peripheral mononuclear CD19+ cells from 7 healthy donors and 77 untreated CLLs. All the cases were characterized by FISH for the most frequent chromosomal aberrations (Fabris et al. GCC, 2008). Molecular markers including mutation status of the heavy chain variable regions of immunoglobulin genes (IGVH), the expression of the 70-kd zeta-chain T-cell receptor-associated protein kinase (ZAP-70) and CD38 cell surface antigen protocols were previously reported (Cutrona et al., Haematologica, 2008). A quantitative bisulfite-PCR Pyrosequencing method was used to evaluate methylation of Alu and LINE-1. We found a significantly lower RTL values in CLLs (median RTL=0.4 IQR 0.3–0.6) as compared with controls (median RTL=1.0 IQR 0.9–1.3) (P <0.001). A progressive and significant RTL decrease in low (13q- and normal karyotype), intermediate (+12) and high (11q- and 17p-) cytogenetic risk categories (P for trend =0.008) was observed. Patients with IGVH mutated genes had longer telomeres than patients with unmutated genes (P <0.001). No significant association between telomere length and either CD38 or ZAP-70 expression was found. Telomere shortening was significantly correlated with hypomethylation of Alu (P =0.048) and LINE-1 (P =0.001), indicating a contribution to chromosome instability. Finally, follow-up analysis showed a significantly higher risk of starting treatment for patients with shorter telomeres (P =0.0037). Our results extended previous evidence that telomere length could be used as marker for the identification of CLLs with a different prognostic risk.

Disclosures:
No relevant conflicts of interest to declare.
An International Study of High Cut-off Hemodialysis for the Management of Myeloma Kidney,
Authors: Hutchison Colin A , Bevins Anne , Mead Graham , Cook Mark 
Published: October 2019

Abstract
Abstract 3974

Background:
An early reduction in serum free light chains (FLC) improves outcomes for patients with myeloma kidney. The purpose of this study was to assess how high cut-off (HCO) dialysers are being used internationally to achieve this target.

Methods:
Data was collected for 54 patients, from 18 centers in 10 countries. Clinical presentation and treatment parameters, including details of removal of FLCs with high cut off hemodialysis and chemotherapy treatment, were captured using a web-based collection database.

Demographics:
All patients were Caucasian, median age of 65 years (range 43–81). Median biochemistry at presentation was: GFR of 8mls/min/1.73m2 (1–27). creatinine 633.5μmol (168–2263); calcium 2.3mmol/L (0.91–3.83); albumin 34g/L (14–46) and β2M 9.45mg/L (0–55.7). Baseline monoclonal κ and λ FLCs levels were: 5070mg/L (range 2250–20200) and 4200mg/L (range 300–13300), respectively. Of the patients who received a renal biopsy, 81% had myeloma kidney as their primary diagnosis. For initial chemotherapy prescriptions: 78% received bortezomib and 34% received thalidomide. 68.75% of the patients were treated with the Theralite HCO dialyzer; the other 31.25% received treatment with the HCO1100 dialyzer. There was a total of 626 HCO dialysis sessions recorded, with each patient receiving on average 13 session each (median, range 3–35) treatments per patient. Of the 54 patients, only 3 were treated with HDF.

Results:
73.2% of patients demonstrated a decreased serum FLC level over the course of treatment, with 86.7% of these patients demonstrated decreased levels by day 12. The median FLC reduction achieved was 72.96% (15.09–99.62%) by day 12 and 93.03% (40.23–99.96%) by the last dialysis treatment. There was no difference in the percentage FLC reduction achieved between bortezomib (median: total 94.09%, day 12 79.14%) and thalidomide (median: total 77.65%, day 12 66.51%) treatment groups (total p=0.179, day 12 p=0.300). FLC removal was significantly increased in patients receiving Theralite treatment over those receiving HCO1100 at both day 12 (p=0.030) and by the end of treatment (p=0.031) Dialysis independence occurred in 68.2% of patients, median time 32 days (10–249). Patients who became independent of dialysis had significantly greater reductions in serum FLC by day 12 (p=0.032). No significant adverse events related to the study device were reported.

Conclusion:
Reducing FLC in patients with myeloma kidney is associated with increased levels of dialysis independence. This study adds further evidence that a combination of chemotherapy and FLC removal by HCO hemodialysis improves patient outcomes following acute kidney injury secondary to multiple myeloma.

Disclosures:
Hutchison: The Binding Site: Honoraria. Bevins:The Binding Site Group Ltd: Employment. Mead:The Binding Site Group Ltd: Employment.
Lentiviral Transduction of Patient Derived Leukaemic Blasts Allowing In Vivo Bioluminescent Monitoring in An NSG Model of Leukaemia Stem Cell Maintenance,
Authors: Bomken Simon , Buechler Lars , Rehe Klaus , Ponthan Frida , Blair Helen , Heidenreich Olaf , Vormoor Josef 
Published: October 2019

Abstract
Abstract 4000
Serial transplantation of patient derived acute lymphoblastic leukaemia (ALL) blasts continues to contribute to our understanding of the biology of leukaemia stem cells (LSC). Refinement of techniques, and in particular intrafemoral injection and development of the highly immunocompromised NOD/scid IL2Rγ null (NSG) mouse have demonstrated B precursor ALL propagating cells to be both common and present in diverse immunophenotypes. These studies must now be complemented by interrogation of the biological pathways underpinning leukaemia stem cell behaviour and clonal propagation in ALL. We have developed a lentiviral based approach to such studies, using the transfer vector pSLIEW, encoding both enhanced green fluorescent protein (EGFP) and firefly luciferase (luc).
We have recently replaced a bone marrow stromal feeder based transduction protocol with a feeder free protocol, removing the risk of co-transduction of feeder cells. Using the feeder free protocol, we have achieved transduction of primary (n=4) and primograft (n=3) material with between 13.0 and 51.4% eGFP positive cells.
Transplantation of transduced cells by intrafemoral injection into NSG mice resulted in engraftment and disease dissemination. This process was monitored using an IVIS Spectrum bioluminescence imaging system. This technique demonstrated progression of disease to the contralateral femur, spleen, CNS and vertebrae. Disease progression was also monitored by serial bone marrow punctures and 5-colour flow cytometry, which demonstrated no immunophenotypic bias amongst the transduced cells. Flow cytometry of harvested bone marrow and spleen showed between 5.5% and 10.2% eGFP positive cells representing only a moderate decrease from 26.3% eGFP positivity at initial transplantation. This confirms the relative resistance of the SFFV promoter to silencing, making this approach suitable for serial transplantation.
Harvested bone marrow and splenic cells were re-transplanted at 5.5 × 103 – 1.0 × 104 SLIEW+ cells per mouse (total 1 × 105 cells transplanted). Bioluminescent imaging has shown engraftment and dissemination of leukaemia within five weeks, confirming transduction of the leukaemia stem cell compartment.
Further development of the pSLIEW vector to include shRNA sequences now offers the potential for functional studies using patient derived material, transduced with a single lentivector construct and serially engrafted in the NSG assay for leukaemic stem cell maintenance. We believe that this approach will allow us to investigate the genetic programmes underpinning leukaemia stem cell self-renewal.

Disclosures:
No relevant conflicts of interest to declare.
Mitochondrial DNA Minisatellites Showed Higher Informativeness and Sensitivity Than the Nuclear DNA Markers for the Quantitative Determination of Chimerism After Allogeneic Stem Cell Transplantation,
Authors: Kim Hye Ran , Won Eun-Jeong , Choi Hyun-Jung , Kim Hwan-Young , Moon James , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook , Shin Myung-Geun 
Published: October 2019

Abstract
Abstract 4004

Background:
Mitochondrial DNA (mtDNA) is widely used in forensic identification and anthropologic studies on account of its abundance resulting in preferential amplification, sequencing and inherent variability. We developed mtDNA markers to monitor donor cell engraftment after allogeneic stem cell transplantation(SCT), then compared with nuclear short tandem repeat (STR) markers. Patients and methods: The mtDNA control regions and six mtDNA minisatellites (mtMS) (303 poly C, 16184 poly C, 514 (CA) repeat, 3566 poly C, 12385 poly C and 12418 poly A) from the total DNA samples of 215 cases (donor, recipient and after allogeneic SCT) were amplified using the designated specific primers and PCR. The results were compared with those from the six short tandem repeat (STR) markers (D12S391, D18S51, F13A1, HUM RENA-4, HUM FABP2 and Amelogenin). Results: Polymorphisms in the mtDNA control region identify an informative marker in 88% (189 cases) of all cases. Among the six mtMS markers, the informativeness of 303 poly C and 16184 poly C mtMS was 63% and 67% respectively. A combination of direct sequencing through the mtDNA control region, 303poly C and 16184 poly C mtMS could completely distinguish the donor cells from the recipient cells. The results from a typical mixing experiment to determine the sensitivity revealed a detection limit (DL) of the gene scan analysis in a mtDNA mixture to be visible at 1% heteroplasmy in 303 poly C mtMS marker. However, the DL from D12S391 in the same mixing experiment was 5–10% heteroplasmy. Conclusions: mtMS markers, especially 303 poly C and 16184 poly C markers, can provide a sensitive, accurate and quantitative determination of mixed chimerism after a SCT.

Disclosures:
No relevant conflicts of interest to declare.
Selective Targeting of B Cells with Mesenchymal Stromal Cells Treatment Is An Efficacious Strategy for Cutaneous Cgvhd,
Authors: Lai Peilong , Weng Jianyu , Luo Chenwei , Guo Rong , Lu Zesheng , Geng Suxia , Du Xin , Wu Suijing 
Published: October 2019

Abstract
Abstract 4086

Background:
Cutaneous chronic graft versus host disease (cGVHD) is an increasingly common complication in long term survivors of hematopoietic stem cell transplantation, but currently available therapies have demonstrated limited efficacy. Furthermore, Mesenchymal Stromal Cells (MSCs) have been reported to be effective in various immune-mediated disease models, but their therapeutic potentials versus cutaneous cGVHD have not been determined.

Methods:
We enrolled 23 patients suffering from cutaneous cGVHD who had failed to respond to conventional immunosuppressive therapy or relapsed after reduction of prednisone. They received intravenous in vitro expanded bone marrow (BM)-derived MSCs. The median follow-up period was 90 days (range 60–146 days).The dermal manifestations of cGVHD were monitored, and a score was given to the cutaneous response by the physician. B lymphocyte subsets and ratio of CD4/CD8 were detected by flow cytometry before the first and after the last MSC infusion. The plasmatic levels of TNF-α, sICAM-1 and Th1 or Th2 factors, including IL-2 and IL-4, were measured by enzyme-linked immunosorbent assay (ELISA).

Results:
Nineteen patients (82.61%) had skin symptom abatement after MSCs treatment with the skin scores improved according to the NIH criteria. Most patients had healing of skin ulceration, regression on skin findings and increased flexibility of involved joint. Steroid sparing or discontinuation of immunosuppressive medications was possible in all of the patients. Clinical improvement was accompanied by increasing levels of CD19+CD5+ B cells and elevated ratio of CD4/CD8. In addition, after the last MSCs treatment, the patients had significantly higher levels of IL-2 compared to before the first MSC infusion, while they had lower levels of TNF-α, sICAM-1 and IL-4.

Conclusion:
MSCs infusion represents an effective treatment option for cutaneous chronic GVHD. This is of high interest in patients suffering from cutanous cGVHD resistant to conventional immunosuppressive therapy. MSCs alter allo-reactivity by affecting CD19+CD5+ B cells, possibly normalize the ratio of CD4/CD8 and Th1/Th2 and decreased the level of proinflammatory cytokine TNF-α and proadhesive cytokine sICAM-1. Our study demonstrates CD19+CD5+ B cells could be a possible target for therapeutic intervention in cutaneous cGVHD.

Disclosures:
No relevant conflicts of interest to declare.
Conditioning with Treosulfan and Fludarabine for Patients with Refractory or Relapsed Non-Hodgkin Lymphoma (NHL),
Authors: Schmitt Michael , Trenschel Rudolf , Sayer Herbert G. , Schneider Catarina , Glass Aenne , Hilgendorf Inken , Treschl Anne , Junghanss Christian , Borchert Kersten , Koenigsmann Michael , Casper Jochen , Beelen Dietrich , Freund Mathias , Kahl Christoph 
Published: October 2019

Abstract
Abstract 4106

Background:
Treatment of refractory or relapsed non-Hodgkin lymphoma (NHL) remains challenging. Here, 88 patients with refractory or relapsed NHL received treosulfan and fludarabine as a reduced intensity conditioning for allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Material and Methods:
73/88 intensely pre-treated patients experienced a relapse (R) with 18/88 early relapses (ER < 6 months from the last chemotherapy). At the time of allo-HSCT, 26 patients were in complete (CR) and 43 in partial remission (PR), twelve patients had progressive disease (PD) and seven stable disease (SD). 47 patients received an autologous graft followed by allo-HSCT.

Results:
After allo-HSCT, 69 of 88 patients were in CR, seven patients in PR, resulting in an overall response rate of 86.4% (76/88). Thirty-three patients achieved a CR from PR as well as six patients from PD and five from SD. 43/88 (49 %) patients were alive at the end of follow-up. Patients undergoing directly allo-HSCT without preceding auto-HSCT showed a better disease-free survival (DFS, p =.038) with a trend (p =.077) for better overall survival (OS). Patients with ER showed an OS of.35 ±.12 after three and seven years. Chronic graft-versus-host disease (cGvHD) had a positive impact on both OS and DFS (for limited cGVHD versus no cGvHD p =.002 and.004, respectively).

Conclusion:
Allogeneic stem cell transplantation following conditioning with treosulfan and fludarabine constitutes a good therapeutical option for patients with refractory or relapsed NHL and should be considered early in the course of salvage treatment.

Disclosures:
Schmitt: Medac: Participation in Conferences. Sayer:Medac: Travel grant to ASH 2009. Koenigsmann:Medac: Research Funding. Casper:Medac: Participation in Conferences. Beelen:Medac: Participation in Conferences. Freund:Medac: Honoraria, Research Funding.
The Favorable Results of Allo-HSCT in TKI-Resistant CML,
Authors: Markiewicz Miroslaw , Dzierzak-Mietla Monika , Wisniewska-Piaty Katarzyna , Frankiewicz Andrzej , Koclega Anna , Zielinska Patrycja , Wieczorkiewicz-Kabut Agata , Sobczyk-Kruszelnicka Malgorzata , Bialas Krzysztof , Kyrcz-Krzemien Slawomira 
Published: October 2019

Abstract
Abstract 4145

Introduction:
Tyrosine kinase inhibitors (TKI) have revolutionized the treatment of chronic myeloid leukemia (CML). However, for patients who fail TKI or progress to advanced phase disease, allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains the only therapeutic option. The rationale of this study was to evaluate results of allo-HSCT in CML patients who have failed TKI treatment. Patients and Methods: 48 CML pts aged 33 (19–52) years who failed previous treatment with TKI (imatinib-37 pts, imatinib+dasatinib-5, imatinib+dasatinib+nilotinib- 3, dasatinib-2, imatinib+nilotinib-1) received allo-HSCT from HLA-matched siblings (15) or from 10/10 (21) and 9/10 (12) HLA-antigens-Matched Unrelated Donors (MUD) in Hematology and BMT Center in Katowice, Poland, from 10.2002 to 03.2011. The myeloablative preparative regimen consisted of treosulfan 3×14 g/m2 plus fludarabine 5×30 mg/m2 (30 pts) or busulfan 4×4 mg/kg plus cyclophosphamide 4×30 mg/kg (18 pts). Standard GVHD prophylaxis consisted of cyclosporine-A, methotrexate and pre-transplant ATG or thymoglobulin in allo-HSCT from MUD. Source of cells was bone marrow (27 pts) or peripheral blood (21 pts) with median 2.6 or 6.7 x10(6)CD34+cells/kg, respectively. Results: All pts engrafted. 100% donor chimerism has been achieved in 40 (83.3%) pts, 97–99% in 4 pts, progressing mixed chimerism was observed in 4 pts. The 5-year estimated overall survival rate was 79%. 9 pts died 9 (1–21) months following allo-HSCT due to infection (5), GVHD (3) or relapse (1). Bio-molecular relapse or progressing mixed chimerism was observed in 6 pts and was treated with 3 to 5 donor lymphocyte infusions (3 pts), post-transplant imatinib (3 pts) or nilotinib (1 pt). Acute GVHD grade I, II, III and IV was observed in 15, 13, 2 and 1 pt (serious aGVHD grade III-IV in 3 pts (6.25%) only); limited and extensive chronic GVHD in 17 (35.4%) and 9 (18.75%) pts, respectively. Other complications in survivors included CMV reactivation (12), hemorrhagic cystitis (3), H. zoster (2), P. jiroveci (1) and cataract requiring surgery (1). 39 pts (81.25%) are alive 44 (3–92) months post-transplant. Conclusions: Allo-HSCT with myeloablative treo/flu or bu/cy conditioning is a feasible and effective curative therapy in TKI-resistant CML. The alternative therapy with allo-HSCT may overcome TKI resistance, providing long-term remission or cure from CML in patients who failed TKI treatment.

Disclosures:
No relevant conflicts of interest to declare.
Development of the Functional Assessment of Cancer Therapy-Multiple Myeloma (FACT-MM) Scale and Validation in the Eastern Cooperative Oncology Group Trial E1A05,
Authors: Weiss Matthias , Jacobus Susanna , Wagner Lynne I , Cella David , Katz Michael S , Rajkumar Vincent , Fonseca Rafael 
Published: October 2019

Abstract
Abstract 4184

Introduction:
FACT-MM was developed with the aim to create a disease-specific, patient-reported outcomes (PRO) measure as part of the FACT measurement system to assess multiple myeloma (MM)-related symptoms. Literature review identified 52 MM specific symptoms and concerns. 13 MM expert clinicians rated these 52 items on relevance to health-related quality of life (HRQL) for MM patients and added 11 items for comprehensive PRO assessment in MM. These 63 candidate items were rated by 13 MM patients recruited through the International Myeloma Foundation website. Disease-related symptoms and concerns were described and provided by patients through free-text comments. Information from both the MM expert clinician and MM patient surveys including free-text items was analyzed and 14 highest ranked items were selected for the FACT-MM. The FACT-MM subscale (score 0–56), the FACT-G physical and functional well-being subscales (score 0–28), and the FACT-Neurotoxicity (FACT-Ntx) subscale (score 0–44) was administered to 48 E1A05 participants to assess disease and treatment-related symptoms of patients with newly diagnosed MM receiving 8 cycles of VRd versus Vd. Methods: In this study, the first three of seven sequential assessments were evaluated: baseline, cycle 5 and end of treatment (after cycle 8 or early diconstinuation). Instruments were scored as per the FACT scoring guidelines. Descriptive statistics were provided for each timepoint and changes in scores from baseline. The Wilcoxon rank sum test was used to assess differences in scores by ISS stage and ECOG PS. Pearson correlation coefficients were obtained between the scores. Cronbach's alpha was used to evaluate internal consistency of the FACT-MM. Results: At baseline and end of treatment, 46 and 41 patients, respectively, completed assessments. The FACT-MM demonstrated good to very good internal consistency (Cronbach alpha 0.79 – 0.89). The FACT-MM subscale was significantly correlated with the FACT physical and functional well-being subscales at baseline (r = 0.72), cycle 5 (r = 0.64; r = 0.49) and end of treatment (r = 0.72; r = 0.66). All scores declined modestly by cycle 5 and end of treatment from baseline. While sample size was limited, there appeared association with ECOG PS with higher scores for patients with PS 0 status. Conclusions: The 14-item FACT-MM scale is feasible for use to measure myeloma-related symptoms and has demonstrated acceptable psychometric properties based on findings from E1A05, an ECOG myeloma trial.

Disclosures:
Cella: Novartis: Research Funding. Fonseca:Consulting:Genzyme, Medtronic, BMS, Amgen, Otsuka, Celgene, Intellikine, Lilly Research Support: Cylene, Onyz, Celgene: Consultancy, Research Funding.