bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bone Turnover In Multiple Myeloma: Impact Of Bortezomib-Based Induction Therapy, High-Dose Melphalan, and Lenalidomide Consolidation On Osteoblast and Osteoclast Function Within The GMMG-MM5 Trial
Authors: Seckinger Anja , Meißner Tobias , Bertsch Uta , Salwender Hans , Dürig Jan , Hänel Mathias , Blau Igor W. , Scheid Christof , Weisel Katja , Klein Bernard , Goldschmidt Hartmut , Hose Dirk 
Published: October 2019

Abstract

Introduction
In multiple myeloma, initially, there are increased numbers of osteoclasts showing increased activity, but bone formation by osteoblasts is keeping step. In later stages, parts of the bone remodeling compartments are disrupted by the interaction with myeloma cells leading to increased bone resorption which can no longer be compensated (myeloma bone disease, uncoupling of bone formation and bone resorption). Lenalidomide and bortezomib have been shown to target both, myeloma cells and the microenvironment: lenalidomide inhibits osteoclastogenesis, bortezomib is also able to stimulate osteoblast differentiation leading to increased bone formation. Aim of this study is to evaluate the impact of bortezomib-based induction treatment, high-dose therapy, and lenalidomide consolidation on alterations of bone turnover, i.e. surrogates of osteoblast- (osteocalcin, OC) and osteoclast- (collagen type I fragments, CTX-I) function, and their induction by myeloma cells (DKK1-level).

Methods
Serum was collected during routine sampling within the GMMG-MM5 trial (EudraCT 2010-019173-16), and levels of CTX-I, OC, and DKK1 were assessed by ELISA in triplicates using commercially available assays according to the manufacturer’s instructions (RnD Systems and Immunodiagnostic Systems). The following time points were assessed: at inclusion (n=365), after induction therapy with either PAd (n=88) or VCD (n=84), stem cell mobilization using CAD (n=69), high-dose melphalan (n=92), and 2 months lenalidomide consolidation (n=92). Up to now, serum samples of 69 patients were measured sequentially at five time points in line with the GMMG-MM5 trial. DKK1 levels were correlated with the expression in CD138-purified myeloma cells (Affymetrix microarrays, n=365).

Results
Prior to treatment, CTX-I levels are increased, those of OC decreased compared to healthy donors (uncoupled bone turnover). DKK1 protein levels are increased and correlate with DKK1-expression in myeloma cells. After induction therapy, osteoclast activity (CTX-I) is decreased below normal values. PAd unlike VCD further decreases osteoblast activity (OC-levels); DKK1-levels are normalized. Subsequent treatment further decreases DKK1-levels below normal values and blocks osteoclast function. After 2 months lenalidomide consolidation, no normalization of osteoblast activity is found.

Conclusion
The main impact on bone turnover by bortezomib-based induction treatment is a reduction of osteoclast activity alongside a decrease in DKK1-levels. During the reported period, no normalization of decreased osteoblast function was observed.

Disclosures:
Seckinger: Novartis Pharma: Research Funding. Goldschmidt:Novartis Pharma: Research Funding. Hose:Novartis Pharma: Research Funding.
Responsiveness Of Cytogenetically Discrete Human Myeloma Cell Lines To Lenalidomide: Lack Of Correlation With Cereblon and Interferon Regulatory Factor 4 Expression Levels
Authors: Greenberg Alexandra J , Walters Denise K , Arendt Bonnie Kaye , Tschumper Renee C. , Kumar Shaji K. , Rajkumar S. Vincent , Jelinek Diane F. 
Published: October 2019

Abstract
The introduction of novel immunomodulatory drugs (IMiDs), such as lenalidomide, has drastically improved the survival of patients with multiple myeloma (MM). While it has been shown that patients with specific cytogenetic subtypes, namely t(4;14), have the best outcomes when treated with bortezomib-based regimens, the relationship between cytogenetic subtypes and response to IMiDs remains unclear. Using DNA synthesis and apoptosis assays, we investigated the relationship between cytogenetic subtype and lenalidomide response in a representative panel of human myeloma cell lines (HMCLs). Additionally, we examined HMCL protein expression levels of the lenalidomide target cereblon (CRBN) and its putative downstream target interferon regulatory factor 4 (IRF4), both of which have previously been shown to be predictive of lenalidomide response in HMCLs. Our results reveal that response to lenalidomide did not correlate with specific cytogenetic translocations. There were, however, distinct groups of lenalidomide-responsive and non-responsive HMCLs, as defined by inhibition of cellular proliferation; notably, all of the hyperdiploid HMCLs fell into the latter category. Repeated dosing of lenalidomide significantly lowered the IC50 of the responsive HMCL ALMC-1 (IC50 = 2.6 μM versus 0.005 μM, p=0.004), but did not have an effect on the IC50 of the non-responsive HMCL DP-6 (p>0.05). Moreover, no association was found between lenalidomide responsiveness and CRBN or IRF4 expression levels. In summary, our data indicate that lenalidomide sensitivity is independent of cytogenetic subtype in HMCLs. Additionally, while CRBN and IRF-4 have been shown to be associated with response to lenalidomide in patients, these findings do not translate back to HMCLs, which could be attributable to factors present in the bone marrow microenvironment. These results suggest that the development of a new treatment paradigm based on cytogenetic subtype of MM alone may be insufficient, and that additional markers may need to be incorporated. Furthermore, the disconnect between our findings of expression of CRBN and IRF-4 in lenalidomide-treated HMCLs and patient data highlights the need for further investigation into the role of the bone marrow microenvironment on the regulation of these previously demonstrated markers of lenalidomide response.

Disclosures:
No relevant conflicts of interest to declare.
The Choice Of Regimens Based On Bortezomib For Patients With Newly Diagnosed Multiple Myeloma
Authors: Yang Li , He Jingsong , Han Xiaoyan , Zheng Gaofeng , Zheng Weiyan , Wei Guoqing , Wu Wenjun , Ye Xiujin , Shi Jimin , Xie Wanzhuo , Li Li , Zhang Jie , Huang Weijia , Zhao Yi , Huang He , Zhang Xuejin , Fu Jiaping , Cai Zhen 
Published: October 2019

Abstract
Bortezomib significantly improved response rates in the treatment of patients with multiple myeloma (MM). However, the process of selecting an optimal bortezomib based regimen as the initial therapy of MM remains ambiguous due to a lack of modern clinical trials demonstrating the efficacy of various bortezomib based treatments. Here, we report four bortezomib-based regimens for treatment of MM patients from three hematological treatment centers in China.

Methods
Newly diagnosed MM patients in three hematological centers in China between February 1, 2006 and May 31, 2013 were treated with combination therapies including bortezomib plus dexamethasone (PD), or triple combinations of PD with adriamycin (PAD), cyclophosphamide (PCD), and thalidomide (PTD) every 28 days.

Results
The overall response rate (≥ partial response, ORR) of all the 215 eligible patients was 90.2%. The ORR for PCD, PAD, PTD and PD were 97.4%, 93.2%, 85.3% and 77.8% respectively, while the effects with VGPR or better were 63.7%, 62.7%, 44.2% and 37.8% respectively. The effect of ORR, VGPR and CR/nCR for PCD regimen was significantly better than PD scheme (P = 0.009, 0.011, 0.005 ). The median PFS of all the patients was 29.0 months with significant differences observed between groups (P =0.047). The median OS of all the patients was not reached, but triple combinations of PD with adriamycin (PAD), cyclophosphamide (PCD), and thalidomide (PTD) were more efficient in treatment of MM patients compared to PD (P =0.005). The frequently observed toxicities were neutropenia, thrombocytopenia, fatigue, infection, herpes zoster, and peripheral neuropathy. Incidence of peripheral neuropathy (PN) in PTD group was significantly higher than other three groups, especially grade 2-3 PN. Treatment with anti-viral agent acyclovir significantly reduced the incidence of herpes zoster.

Conclusions
Our study demonstrated that bortezomib-based regimens were active and well-tolerated in the Chinese MM patients, and triple combinations of PD with adriamycin (PAD), cyclophosphamide (PCD), and thalidomide (PTD) were more efficient for treatment of MM patient, and the patients received PCD or PAD demonstrated significant higher ORR compared to PD.

Disclosures:
No relevant conflicts of interest to declare.
Allogeneic Stem Cell Transplantation - A Long-Term Curative Approach In Patients ≥ 60 Years With Advanced Disease AML/MDS, - The Freiburg Experience Of 250 Consecutive Patients
Authors: Bertz Hartmut , Lübbert Michael , Ohneberg Kristin , Wäsch Ralph , Zeiser Robert , Marks Reinhard , Finke Jürgen 
Published: October 2019

Abstract
Since the introduction of reduced-toxicity conditioning prior to allogeneic hematopoietic stem cell transplantation (alloSCT) we transplanted from 1999 to 2012, 250 consecutive patients (pts) with myeloid malignancies (AML, MDS) aged ≥ 60 years (yrs).
The 144 male and 106 female pts with a median age of 66 yrs (range 60-77) were transplanted for de novo AML (n=95), s/tAML (n=104) and MDS (n=51) with 89% unfavorable cytogenetics (CALGB). Since 2004 pts received a prospective fitness assessment (Deschler et al., Haematologica 2013). In 74% the donor was matched/mismatched unrelated and in 26% related. Only 16% were transplanted in CR1/2, 84 % with advanced or untreated disease. The conditioning regimen was the FBM protocol (fludarabine, carmustine, melphalan; Bertz et al., JCO 2003) in 98%, and 97% of the pts received PBSC. For GVHD prophylaxis in 91% a combination of cyclosporine plus alemtuzumab or ATG-F™ was used.
At day +30, 94% of the pts had achieved CR by standard measures. With a median follow up of 57 months (3-157) 37% of the pts are alive; main causes of death were relapse (n=62), infection (n=35) and age-related diseases (n=13). The probability of OS/DFS was at 1yr 61%/49%, at 2 yrs 49%/41% and at 5 yrs 37%/34%, respectively. The probability for NRM at 1 yr is 24%. Nineteen known prognostic factors for outcome were evaluated: e.g. patient and donor age, graft size, days between diagnosis and alloHCT, CMV, early/advanced disease, cytogenetics, Sorror and Gratwohl score, donor type, HLA-identity. In the multivariate analysis a better OS (factors with p<0.1; table) was seen with a matched donor; a better DFS with a related donor, and high CD34+ graft content; in contrast, a mismatched donor is a risk factor for reduced DFS.TableMultivariate analysis of prognostic factors* for OS and DFSvariablevalueHazard Ratio95% CI lower limit95% CI upper limitP valueOverall survivalRemission at alloHCTadvanced1.370.862.160.1825HLA mismatchyes1.401.011.960.0463HCT-CI (Sorror)>= 21.311.011.960.1007Peripheral blood blastsyes1.210.841.760.3034Disease-free survivalRemission at alloHCTadvanced1.290.722.300.3946Donorrelated0.640.430.950.0258HLA mismatchyes1.440.992.090.0561CD34+ cells> median0.760.551.040.0867Bone marrow blasts> 5%1.210.781.880.3915*in univariate analysis p<0.157 (AIC criterion; Sauerbrei W, 1999 Applied Statistics,48:313-329.70.)
In conclusion, this unique large cohort of older pts with AML/MDS with mainly advanced disease and unfavorable cytogenetics shows a high feasibility, safety and efficacy of alloHCT after the FBM protocol. AML/MDS pts in their 7th and 8th decade of life fit for transplant should be evaluated for alloHCT as very important long-term curative option.

Disclosures:
No relevant conflicts of interest to declare.
Allogeneic Hematopoietic Stem Cell Transplantation From Haploidentical Donor After Reduced Intensity Conditioning Regimen Allows Promising Disease Control In Patients With Hematological Diseases
Authors: Devillier Raynier , Castagna Luca , Crocchiolo Roberto , Furst Sabine , Harbi Samia , El Cheikh Jean , Granata Angela , Mohty Bilal , Faucher Catherine , Blaise Didier 
Published: October 2019

Abstract

Introduction
Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) from haploidentical donors (HID) as alternative donor is currently increasing for patients with hematological diseases. In this setting, post Allo-HSCT high dose Cyclophosphamide (Cy) allows the use of T-repleted graft from HID by decreasing early alloreactive T-cell, leading to low incidences of graft-versus-host disease (GVHD) and non-relapse mortality. However, the efficacy of this procedure is still a matter of debate. The aims of this study were to evaluate the efficacy of HID Allo-HSCT in a comparison with our historical control cohort of patients transplanted from matched related or unrelated donors.

Patients and methods
We report here the experience of HID Allo-HSCT prepared by reduced intensity conditioning (RIC) regimen and post infusion Cy (HID group; n=60) and compared outcome to that of historical control cohort of patients with hematological diseases (n=220) who underwent RIC Allo-HSCT from matched related donors (MRD group; n=110) or unrelated donors (UD group; n=110). The control cohort received RIC regimen based on Fludarabine (150 mg/m2 total dose over 5 days), intravenous Busulfan (6.4 mg/kg total dose over 2 days) and antithymocyte globulin (ATG; Thymoglobulin; 5 mg/kg total dose over 2 days). To better compare the results between these two heterogeneous cohorts of patients including different hematological diseases, we used the disease risk index (DRI) score recently published (Armand at al., Blood, 2012).
Patient characteristics are illustrated in the Table 1. Although HID and MRD/UD groups were significantly different regarding age and graft source, they presented with similar disease risk profile when classified according to the DRI and comorbidities according to the HCT-CI.

Results
In the HID group, day-100 acute GVHD, overall incidence of chronic GVHD and 2-year NRM were 21%, 9% and 15% respectively. Regarding the disease control, with a median follow up of 17 months (range: 5-50), the 2-year cumulative incidence of relapse (CIR) was 23%, which is similar to the MRD and UD control groups (2-year CIR: MRD 31%; UD: 18%). The 2-year PFS and OS were 62% and 68% respectively in the HID group, 52% and 66% respectively in the MRD group and 55% and 56% respectively in the UD group.

Conclusions
These results suggest that RIC Allo-HSCT from HID allows promising anti-tumor effect that is comparable to our previous experience of Allo-HSCT from MRD or UD. We also confirm the low toxicity profile after HID Allo-HSCT. Taken together, these observations suggest that HID is a valuable alternative for patients with hematological diseases but no available MRD or UD.

Disclosures:
No relevant conflicts of interest to declare.
Lower Relapse and Better PFS Among Chemosensitive Patients Undergoing Allogeneic Transplantation By Haploidentical Compared With HLA-Identical DONOR: Results On A Cohort Of 94 Patients With Hodgkin’S Lymphoma
Authors: Castagna Luca , Crocchiolo Roberto , Bramanti Stefania , Furst Sabine , Sarina Barbara , El Cheikh Jean , Granata Angela , Devillier Raynier , Garciaz Sylvain , Bouabdallah Reda , Santoro Armando , Blaise Didier 
Published: October 2019

Abstract

Introduction
haploidentical stem cell transplantation (SCT) represents an alternative option for those patients lacking a HLA-identical donor. Patients with relapsed or refractory Hodgkin's lymphoma may benefit from allogeneic SCT and, in particular, patients without a HLA-identical donor may receive SCT from a haploidentical one. The aim of this retrospective analysis is to compare outcome of HL patients undergoing allogeneic SCT from haploidentical vs. HLA-id. Donor.

Methods
patients with diagnosis of HL who received allogeneic SCT at Istituto Clinico Humanitas (Milan, Italy) and Institut Paoli-Calmettes (Marseille, France) were included in the analysis. Outcome was estimated according to SC source (HLA-id. vs. haplo) and disease status at transplant: CR, PR or SP/PD.

Results
a total of 58 and 36 patients received SCT from a HLA-id. and haploidentical donor, respectively. SCT were performed from February 1999 to December 2012. Ninety-two patients received reduced-intensity conditioning; all but one patient in haplo group received post-transplant cyclophosphamide as GVHD prophylaxis. Median follow-up was 63 months (range: 6-160) in HLA-id. group and 20 months (range: 6-50) in haplo group. In HLA-id. group, 2-year PFS was 62%, 33% and 17% among patients in CR, PR or SD/PD before allogeneic SCT, respectively; 2-year PFS in haplo patients was 86%, 53% and 0% among CR, PR and SD/PD patients. Results are shown in Table 1. Notably, a lower number of relapse events were observed in chemosensitive patients in haplo group compared with HLA-id.: 1/21 vs. 7/28 among CR patients and 2/9 vs. 10/18 among those in PR (see Table 1); this translates into better PFS after haplo-SCT. Conversely, outcome of refractory patients confirmed to be poor in both groups. At last follow-up, among the 30 chemosensitive patients receiving haplo-SCT, last relapse occurred at day +392 post-SCT.

Conclusions
present analysis showed better PFS in haplo group among high-risk, chemosensitive (CR + PR) patients compared with HLA-id. and better OS among patients who were in PR before SCT; this was due to lower relapse rate among chemosensitive patients after haplo-SCT vs. HLA-id. In conclusion, haplo-SCT showed high antitumor activity, leading to promising outcome of chemosensitive patients compared with HLA-id transplants. Present data may challenge donor choice in this subset of patients, who may benefit from graft-versus-lymphoma effect, potentially enhanced by haplo disparity.

Disclosures:
No relevant conflicts of interest to declare.
Platelet-Like-Particles Sheared From Myosin-II-Inhibited Megakaryocytes Highlights The Elevated Thrombocrit Of May-Hegglin Anomaly
Authors: Spinler Kyle R , Shin Jae-Won , Discher Dennis E 
Published: October 2019

Abstract
Megakaryocytes (MKs) in the marrow extend projections into blood flow and generate platelets under shear. Understanding MK differentiation and platelet production is of broad clinical importance and extends a need to augment platelet numbers in patients. Reversible but sustained inhibition of non-muscle myosin-II (NMM-II) with the drug blebbistatin increases MK polyploidization, proplatelet formation, and membrane flexibility, thereby increasing platelet generation under shear. Using a cone and plate rheometer to apply fluid shear to drug-treated MKs in bulk, platelet-like-particles (PLPs) that are collagen-I responsive can be generated with intermediate shear. The MKs naturally down-regulate NMM-IIA activity through phosphorylation of S1943, but this site proves shear sensitive, consistent with results for human platelets. Using micropipette aspiration of MKs, inhibition of NMM-IIA is found necessary to generate CD41+ fragments that approximate the size of human platelets. Localization of NMM-IIA to the fragments is modulated by S1943 as seen by unique distribution patterns resulting from specific S1943 mutations that can be abrogated by addition of blebbistatin. The approach is extended to clinically relevant mutations associated with May-Hegglin anomaly (MHA) co-expressed with wild type protein to mimic heterozygotes. As with blebbistatin inhibition of myosin, May-Hegglin mutants result in a higher frequency of fragmentation during micropipette aspiration, indicating a dominant negative effect. Immunofluorescence documents abnormal myosin aggregation in cells transfected with May-Hegglin myosin mutations compared to wild type constructs. Finally, peripheral blood from a patient with a D1414N May-Hegglin mutation is cultured to produce megakaryocytes used to support both the micropipette and immunofluorescence results. These findings reveal a phospho-switch in NMM-II, from inactive to active in the terminal stages of platelet-poiesis, and that proper myosin activity is critical to fragment size and number. Disruption of normal activity enhances fragment generation suggesting a novel mechanism in MHA: in particular, MHA thrombocytopenia results in an increased thrombocrit due to abnormally large platelets, which overcompensates for the reduction in platelet number.

Disclosures:
No relevant conflicts of interest to declare.
Morphological Distinction Unravels Mechanisms Of Platelet Biogenesis From Bone Marrow Megakaryocytes
Authors: Nishimura Satoshi , Eto Koji , Nagai Ryozo 
Published: October 2019

Abstract
Blood platelets are generated in the bone marrow (BM) from their precursors, megakaryocytes (MK). Although we know that MKs produce platelets throughout life, precisely how platelets are produced in vivo remains uncertain, largely because of the rarity of MKs in the BM and the lack an adequate visualization technique. In the present study, we were able to visualize MK dynamics leading to platelet release in living animals at high resolution.
To clearly understand the nature of thrombopoiesis in BM MKs, we optimized an in vivo imaging technique based on two-photon microscopy that enabled us to visualize living BM in CAG- enhanced green fluorescent protein (eGFP) mice. By visualizing living bone marrow in vivo, we observed that two modes (fragmentation and proplatelet formation) can be ongoing simultaneously in the same mouse. We observed that these two modes detectable by different morphological behavior can be ongoing simultaneously in the same BM of mouse, and are regulated by specific cytokines. Short proplatelets from megakaryocytes predominated at steady state, and more elongated proplatelets were accelerated by thrombopoietin (TPO) with responding to chronic platelet needs including recovery form BM transplantations. In contrast, acute platelet needs by blood loss, 5-FU administration or pritoneal acute inflammation increased cytoplasmic fragmentation following rapid ‘rupture’. Observed two modes are both dependent on tubulin reorganization on platelet biogenesis. In addition, platelet increase at acute phase is independent of proliferation by MK progenitors and this factor might exert apoptosis machinery on already reserved mature type of MKs.
This humoral factor was identified by combination of in vitro screening systems and in vivo MK visualization analysis. Factor serum levels were reduced independently of the thrombopoietin level in human subjects with low platelet counts. It thus appears the cytokine balance dynamically regulates the mode of thrombopoiesis and the cellular programming of MKs. Thus, these novel factor may be a novel therapeutic target in thrombocytopenic situations, especially when associated with acute loss of platelets or when platelet transfusion is limited or unsuccessful.

Disclosures:
No relevant conflicts of interest to declare.