bluuebunny/crossref_metadata_2025_split huggingface dataset:
Excellent Long-Term Survival of Patients with Steroid-Refractory and Steroid-Dependent Acute Graft-Versus-Host Disease after Extracorporeal Photochemotherapy.
Authors: Greinix Hildegard T. , Knobler Robert M. , Worel Nina , Mitterbauer Margit , Schulenburg Axel , Schneeberger Armin , Karlhofer Franz , Hoecker Paul , Leitner Gerda , Rosenmayr Agathe , Kalhs Peter 
Published: April 2020

Abstract
Despite posttransplantation immunosuppressive therapy, acute graft-versus-host disease (GVHD) remains a major cause of sickness and death. Second-line therapies for steroid-refractory acute GVHD have been used with limited success. Extracorporeal exposure of peripheral blood mononuclear cells to the photosensitizing agent 8-methoxypsoralen and UV-A radiation (ECP) has been shown to be effective in the treatment of selected diseases mediated by T cells. We have reviewed the responses and long-term outcome of 59 hematopoietic stem cell transplant (HSCT) patients treated from 1996 to 2003 with ECP for steroid-refractory acute GVHD, defined as progression or no improvement of acute GVHD after a minimum of 4 (range, 4–49, median 17) days of treatment with prednisone (n=37) or steroid-dependent acute GVHD, defined as flare-up of GVHD during prednisone taper (n=22). Patients received HSCT from 17 related and 42 unrelated donors. In 28 cases an HLA-mismatch between recipient and donor was present. Prior to ECP, grade III–IV GVHD was observed in 23 patients (39%) and grade II GVHD in 36 (61%). Organs involved included skin in 97% of patients, liver in 39%, and GI tract in 17%. Treatment consisted of ECP on two consecutive days per week (=1 cycle) for a median of 7 (range, 1–45) cycles administered within a median of 3 (range, 0.5–31) months in addition to cyclosporine A and prednisone. Three months after initiation of ECP complete resolution of GVHD was achieved in 82% of patients with cutaneous, 61% with liver, and 61% with gut involvement. Complete responses were obtained in 86% of patients with grade II, 55% of patients with grade III, and 30% of patients with grade IV acute GVHD. Probability of transplant-related mortality (TRM) at 4 years after HSCT is 15% in patients with complete response to ECP compared to 88% in patients not responding completely. After a median follow-up of 46 (range, 9–45) months since discontinuation of ECP, 28 (47%) patients are alive including 22 without chronic GVHD. Probability of survival (OS) at 4 years after HSCT is 59% in patients with complete response to ECP compared to 11% in patients not responding completely. Besides response to ECP only organ involvement and grade of GVHD at start of ECP, and ability to timely taper steroids during ECP had a significant impact on both TRM and OS. Thus, ECP is an effective adjunct therapy for acute steroid-refractory and steroid-dependent GVHD. Our long-term results demonstrate durability of responses without adverse events.
IL6 Protection of Myeloid Leukemic Cells Against P53 Induced Apoptosis Is Mediated by Activation of P13kinase/AKT and Stabilization of C-FLIP.
Authors: Vesely Diana L. , Hoffman Barbara , Liebermann Dan A. 
Published: April 2020

Abstract
Myeloid leukemic cells (M1) proliferate continuously in culture unless induced by Interleukin-6 (IL6) to undergo a terminal differentiation program into macrophages, followed by apoptosis. M1 cells lack the tumor suppressor p53, which is a critical determinant of the cellular decision to either growth arrest and repair DNA damage or to undergo apoptosis. Activation of a temperature sensitive p53 protein (p53 val) at the permissive temperature in M1 cells results in rapid apoptosis. IL6 treatment blocks this p53-mediated apoptosis. Towards understanding the basis for this p53-mediated apoptosis and its abrogation by IL6, we have shown that at the permissive temperature p53 activates the pro-apoptotic Fas/CD95 pathway by up regulating the Fas/CD95 receptor and cleaving antiapoptotic c-FLIP. On the other hand, antagonistic Fas antibody protects against apoptosis. IL6 decreases Fas/CD95 ligand expression and prevents cleavage of FLIP. Treatment of M1p53ts cells with the specific Akt inhibitor Ly294002 abrogated IL6 protection and resulted in Flip cleavage, suggesting that IL-6 blocks apoptosis by phosphorylating and activating P13kinase/Akt, which in turn promotes FLIP stability. Inhibition of the ERK pro-survival pathway did not abrogate IL6 protection against p53 apoptosis like Akt inhibition caused. Nevertheless M1p53ts-Flip cell lines fail to completely protect against p53 mediated apoptosis, suggesting that FLIP is itself insufficient to protect against apoptosis. Therefore we examined what other proteins may synergize with FLIP to protect from apoptosis, and found that that the pro-survival bcl2 protein family member MCL-1 is strongly up regulated by IL6 in our M1p53 cell line. Generation of M1p53-FLIP-MCL1, M1p53-MCL1, and M1p53 -uncleavable FLIP cell lines is underway to determine whether there is synergy between MCL-1 and FLIP in escape from p53 mediated apoptosis of M1p53 cells. Thus far taken together these data support a model for leukemic progression where cells that acquire the ability to produce an autocrine survival factor, such as IL6, can bypass normal p53 surveillance function by targeting downstream inhibitors of apoptosis such Akt, c-Flip and MCL-1.
Cytokine Production and Hematopoiesis Supporting Activity of Cord Blood Derived Unrestricted Somatic Stem Cells.
Authors: Koegler Gesine , Radke Teja , Lefort Aurelie , Sensken Sandra , Fischer Johannes , Sorg Rudiger Volker , Wernet Peter 
Published: April 2020

Abstract
Particularly within the last several years cord blood (CB) has become a well established hematopoietic source for unrelated transplantation in children. Assuming, however, a threshold of 2x107 nucleated cells (NC)/kg body weight required for transplantation in adults over 70kg, an average of only 25% of the CB units presently stored in the major CB banks contain sufficient NC to safely engraft patients. Thus far the majority of studies have assessed selective cytokine driven expansion of progenitors for transplantation, which however may be associated with exhaustion of stem cells and an increase of GvHD in adult patients. Therefore, we assessed cytokine production and hematopoiesis supporting stromal activity of CB derived unrestricted somatic stem cells (USSC, Koegler et al. JEM2004: 200: 2: 1–13) in comparison to bone marrow mesenchymal stem cells (BMMSC) and hematopoietic progenitor expansion solely driven by recombinant cytokines. Initiation of USSC generation was initiated from fresh (n=325) and cryopreserved CB (n=66). In the presence of H5100 medium/10−7M dexamethasone, USSC cultures from fresh CB were initiated from 57.1% of CB samples (n= 52 out of 91 CB samples), whereas in DMEM-medium/10−7M dexamethasone the frequency was 31.6% (n=74 out of 234 CB samples). The generation frequency from frozen HES separated CB was only 19.5%. Cytokine production by USSC and BMMSC was determined qualitatively by cytokine mRNA expression array analyses or quantitatively by Luminex or ELISA analyses. To analyze hematopoiesis supporting activity, CB CD34+ cells were expanded in co-cultures with USSC, BMMSC or in the presence of Flt3-L, SCF, TPO. Expansion of CD34+ cells, total cells and colony-forming cells (CFC) were determined after 1, 2, 3 and 4 weeks of culture. USSC constitutively produced SCF, LIF, TGF-1ß, M-CSF, GM-CSF, VEGF, IL-1ß, IL-6, IL-8, IL-11, IL-12, IL-15, SDF-1a and HGF. When USSC were stimulated with IL-1ß, G-CSF was released. Production of SCF (p= 0.0104) and LIF (p= 0.046) was significantly higher in USSC compared to BMMSC. At 1, 2, 3 and 4 weeks, respectively co-cultivation of CD34+ cells on the USSC layer resulted in a 14.62±1.1-fold, 110.1±17.9-fold, 151.8±39.7-fold and 183.6±40.4-fold amplification of total cells and in a 30.56±4.4-fold, 101.4±27.5-fold, 64.67±15.8-fold and 29.4±3.1-fold amplification of CFC. Expansion achieved was significantly higher for USSC compared to BMMSC feeder layers. USSC produce functionally significant amounts of hematopoiesis supporting cytokines and are able to expand CD34+ cells from CB in significantly higher amounts compared to BMMSC. Therefore the USSC is a suitable candidate to direct ex-vivo expansion of hematopoietic CB cells for short term reconstitution.
Increased Adipocytogenesis and Hematopoiesis in Long-Term Bone Marrow Cultures from SMAD3−/− Mice.
Authors: Cao Shaonan , Zhou Shuanhu , Epperly Michael W. , Glowacki Julie , Shields Donna , Greenberger Joel S. 
Published: April 2020

Abstract
The abrogation of an intact TGFß/TGFß receptor signal transduction pathway in SMAD3−/− mice is associated with a decreased inflammatory response, and decreased acute and late response to ionizing irradiation damage. We tested the role of SMAD3 in bone marrow cellular capacity for hematopoiesis and in marrow stromal cellular potential for adipocytogenesis. Long-term bone marrow cultures (LTBMCs) were established from SMAD3−/−, +/−, and +/+ mice. Permanent clonal bone marrow stromal cell (MSC) lines were established from each mouse strain. The SMAD3+/+ and SMAD3+/− LTBMCs generated hematopoietic progenitor cells (by CFU-GEMM assay), and maintained active cobblestone island numbers and production of non-adherent cells for 22–23 weeks. In striking contrast, the SMAD3−/− LTBMCs demonstrated a significant increase in longevity of hematopoiesis as measured by each assay for over 34 weeks and non-adherent cells continued to form GEMM beyond 34 weeks. The number of CFU-GEMM per 5x104 cells (d 14) in SMAD3−/− cultures (188 ± 8.7) was 1250% greater (p&lt;0.0001) than in SMAD3+/+ cultures (15.3 ± 2.3). Microscopic inspection at 27 weeks showed that more (16.5%, p&lt;0.001) SMAD3−/− adherent cells contained lipid droplets (6.6% ± 0.7) compared with SMAD3+/+ adherent cells (0.4% ± 0.2). Because of that difference, we examined the regulation of adipocyte differentiation in MSC lines from those mice. First, there was more extensive (37-fold) adipocyte differentiation in SMAD3−/ − than SMAD3+/+ cultures. Second, there was a 7-fold attenuation of TGF-ß inhibition of adipocytogenesis in cells lacking SMAD3. RT-PCR evaluation showed greater expression of the adipocyte gene, ADIPSIN, and absence of its downregulation by TGFß in SMAD3−/− compared to SMAD3+/+. The data suggest that SMAD3 is a critical inhibitor of adipocyte differentiation. The data document pleiotrophic effects of interruption of the TGFß/TGFß receptor signal transduction cascade for both inhibition of hematopoietic support capacity and adipocyte differentiation of the bone marrow microenvironment.
Indian Hedgehog Stimulates Proliferation of Short-Term NOD/SCID-ß2mnull Repopulating Cells on Coculture with Human Stromal Cells.
Authors: Kobune Masayoshi , Kawano Yutaka , Kato Junji , Ito Yoshinori , Chiba Hiroki , Kuribayashi Kageaki , Takimoto Rishu , Matsunaga Takuya , Hamada Hirofumi , Niitsu Yoshiro 
Published: April 2020

Abstract
Hematopoietic stem cells (HSCs) represent a subset of bone marrow cells that are capable of self-renewal and of giving rise to all types of blood cells. However, the mechanisms involved in controlling the differentiation and self-renewal of HSCs remain largely unknown. The Indian hedgehog (Ihh) signal has been shown to play an essential role in inducing hematopoietic tissue during embryogenesis. We investigated the roles of the Ihh in postnatal hematopoiesis using coculture system between CD34+ cells and human stromal cells. Ihh gene transfer into hTERT-stromal cells enhanced the expression of BMP4, angiopoietin-1 and Wnt5A, and their hematopoietic supporting potential was elevated compared with control stromal cells, as indicated by the colony-forming units in culture (CFU-C, 26±2 vs. 59±3-fold of the initial cell number; CFU-Mix, 63±37 vs. 349±116). Engraftments of NOD/SCID-ß2mnull repopulating cells (RCs) at 8 weeks post-transplantation expanded on Ihh-stromal cells were significantly higher compared with control coculture results and engraftments were neutralized by addition of an anti-hedgehog antibody. Limiting dilution analysis indicated that SCID RCs proliferated more efficiently on Ihh-stromal cells than those on control stromal cells. The degree of expansion on Ihh-stromal cells was estimated to be 6.6-fold greater than that of control stromal cells, according to the Poisson predicted frequency. However, engraftment of NOD/SCID-ß2mnull RCs was decreased by 13 weeks post-transplantation of hematopoietic cells that had been expanded on either Ihh-stromal cells or control stromal cells, although engraftment of human hematopoietic cells that were expanded on Ihh-stromal cells is higher than that for cells expanded on control stromal cells (%CD45+ cells: 3.28±0.60 vs. not detected). These results suggest that Ihh-stromal cells are quite potent to support human short-term RCs in NOD/SCID-ß2mnull mice. Thus, hedgehog signaling is considered to play an important role in the hematopoietic microenvironment.
Identification of Tenascin-C as a Key Molecule Determining the Stromal Cell Dependent Erythropoiesis.
Authors: Seki Masanori , Kameoka Junichi , Takahashi Shinichiro , Harigae Hideo , Sasaki Takeshi , Obinata Masuo 
Published: April 2020

Abstract
Stromal cells in the bone marrow provide an optimal microenvironment for hematopoiesis through cell-to-cell interaction as well as cytokine production. We have previously established 33 bone marrow stromal cell lines from SV40 T-antigen transgenic mice, and demonstrated the lineage-specific activity of supporting hematopoiesis (J Cell Physiol1995: 164; 55). Of the 33 cell lines, 27 clones showed stimulatory abilities to support large erythroid colony formation in the presence of erythropoietin, while 6 cell lines did not. Since these activities were not dependent on the amount of produced cytokines, the existence of unknown molecules responsible for the selective activity of erythropoiesis has been implicated. To identify molecules expressed on bone marrow stromal cells which are responsible for determining the stromal cell dependent erythropoiesis, we compared the gene expression profiling by cDNA microarray between cell lines which support erythropoiesis (E+; TBR 9, 184, 31-2) and cell lines which do not (E-; TBR 17, 33, 511). Out of 7226 genes examined, 138 genes were up-regulated 3-fold or more, and 282 genes were down-regulated 3-fold or more, in E+ cell lines. Among these genes, we have selected one of the up-regulated genes, tenascin-C, as a candidate for erythroid progenitor supporting genes, and examined its expression and function in detail. First, we confirmed the expression of tenascin-C by real time quantitative RT-PCR. The expression of tenascin-C adjusted by the expression of GAPDH in the three E+ cell lines were all higher than the three E- cell lines with a mean of 3.6 times. Next, we examined the effect of suppressing tenascin-C expression by small interfering RNA (siRNA) on the erythroid colony formation. siRNAs were synthesized by in vitro transcription method, and were transfected into TBR cell lines by lipofection. At 24 hours after the transfection of tenascin-C siRNA, the expression of tenascin-C was reduced to 10 to 40% of the control siRNA as determined by real time quantitative RT-PCR. The number of erythroid colony dependent on TBR 9 and TBR 184 in the presence of tenascin-C siRNA was significantly lower than that in the presence of control siRNA (13.0±6.5 versus 0.75±0.95 colonies/104 fetal liver cells; p&lt;0.05). These results suggest that tenascin-C plays an important role in the stromal cell dependent erythropoiesis.
Biochemical Characterization and Gene Expression Profiling of Cytarabine Treated Stromal Fibroblasts Reveal Regulatory Mechanisms Affecting Hyaluronan and Heparan Sulfate Proteoglycans.
Authors: Zweegman Sonja , Kessler Floortje L. , Kerkhoven Ron , Heimerikx Mike , Huijgens Peter C. , Drager Angelika M. , van den Born Jacob 
Published: April 2020

Abstract
High dose chemotherapy and radiation, as applied in stem cell transplantation procedures, have been found to impair the hematopoiesis-supportive capacity of bone marrow stroma. However, the molecular mechanisms involved are far from elucidated yet. As bone marrow stromal glycosaminoglycans (GAGs) play an important role in various regulatory steps in hematopoiesis, we examined the effect of chemotherapy on the synthesis and composition of GAGs and gene expression profiles of the hematopoiesis supportive M210B4 fibroblastic stromal cell line. To this end GAGs were metabolically labeled with 3H-glucosamine and 35S-sulfate, and analyzed by conventional biochemical techniques.
Cytarabine treatment resulted in a pronounced increase (1.6±0.3-fold increase) in hyaluronan (HA) and a reduction in the content (47±7% reduction) and in the extent of sulfation (3H-glucosamine to 35S-sulfate ratio increased from 2,1 versus 2,6) of heparan sulfate proteoglycans (HSPG). Gene expression analysis showed a corresponding increase in hyaluronan synthase 1 (Has 1) gene expression and a decrease in hyaluronidase 2 (Hyal 2) gene expression. The decrease in HSPG was found to correspond with downregulation of the Exostosin 1 (Ext 1) gene, encoding a bifunctional glycosyltransferase, required for the biosynthesis of HS. Finally, there was a trend towards decreased expression of the N-deacetylase/N-sulfotransferase-1 (Ndst1) gene, which might explain the decrease in sulfation of HSPG after cytarabine treatment, as deacetylation is required for sulfation of HSPG to occur.
The functional consequence of the decrease in HSPG was investigated by Stromal Derived Factor-1α (SDF-1α) binding. SDF-1α was added to either untreated or treated M210B4 cells and allowed to bind for 90 minutes. After washing, bound SDF-1α was quantified by staining with the anti-SDF-1α antibody K15C. Cytarabine treatment resulted in a 31.1 ± 11.8% decline in K15C binding, indicating a decline in GAG-mediated binding of SDF-1α (n=6, Mann Whitney test, p&lt;0.05). In conclusion, cytarabine treatment alters the expression of genes involved in GAG synthesis and degradation, thereby affecting the amount and structure of stromal GAGs. Accordingly, the chemokine-binding capacity of HSPG was negatively affected. In view of the critical role of SDF-1α in migration of stem cells and its presumed participation in the creation of the stem cell niche, our results suggest that chemotherapy-induced changes in stromal GAGs might affect homing and hematopoiesis. In addition, because of known functional capacities of HA, malignant hematopoiesis as well as tumor growth and invasion may be promoted.
Hodgin’s Lymphoma in Elderly Patients: A Comprehensive Analysis of the German Hodgkin Lymphoma Study Group (GHSG).
Authors: Engert Andreas , Ballova Veronika , Haverkamp Heinz , Pfistner Beate , Diehl Volker 
Published: April 2020

Abstract
Background: With improved prognosis of patients with Hodgkin’s lymphoma (HL), interest increasingly focuses on high-risk groups such as elderly patients. We thus performed a retrospective analysis of the German Hodgkin Lymphoma Study Group (GHSG) database to determine clinical presentation, toxicity of treatment and outcome of elderly HL patients.
Methods: A total of 4251 patients included in the GHSG studies HD5-9 were analysed of whom 372 (8.8%) were 60 years or older and 3879 (91.2%) were younger than 60 years. Patient characteristics, treatment results, toxicity, failure free survival (FFTF) and overall survival (OS) were compared.
Results: The median age of the two cohorts was 65 and 31 years, respectively. Statistically significant differences in patient characteristics were as follows: elderly patients had more frequently mixed cellularity subtype (35% vs. 19%), B-symptoms (50% vs. 43%) and elevated ESR (59% vs. 51%). Less frequently observed in elderly patients were nodular sclerosis subtype (41% vs. 66%), large mediastinal mass (9% vs. 20%), bulky disease (49% vs. 60%) and Karnofski index &lt;8, (11% vs. 3%). Acute toxicity during chemotherapy was higher in elderly patients (87% vs 79%). The most obvious differences were observed for infections (17% vs 6%). The significantly higher rate of infections was due to more severe leukopenia in elderly patients (grade IV: 41% vs 25%). As a result, significantly fewer elderly patients received at least 85% of intended chemotherapy dose (74% vs. 90%). At a median follow-up of 65 months, 38% and 11% of patients respectively died. The survival analysis showed a statistically significant poorer prognosis for elderly patients in terms of 5-year OS (65% vs. 90%), FFTF (60% vs. 79%) and Hodgkin-specific FFTF (73% vs. 82%).
Conclusion: Elderly patients have a poorer risk profile compared to younger HL patients and experience more severe treatment-associated toxicity. Higher mortality during treatment as well as lower dose-intensity are the major factors explaining the poorer overall outcome of elderly Hodgkin lymphoma patients.
Fertility in Male Patients with Hodgkin’s Disease - Results from the German Hodgkin Lymphoma Study Group (GHSG).
Authors: Sieniawski Michal , Josting Andreas , Breuer Kai , Diehl Volker , Engert Andreas 
Published: April 2020

Abstract
Background:
Treatment results in Hodgkin Disease (HD) have improved tremendously over the last two decades. Therefore long term sides effects of therapy as infertility are of growing importance. However, patients (pts) with HD have increased risk for inadequate semen quality even prior to treatment. To investigate the influence of disease and therapy on the fertility status in pts with HD we performed semen and hormone analysis before and after treatment.
Patients and Methods:
All analysis were evaluated in pts with first diagnosis of HD enrolled into trials of the GHSG between 1988 and 2002, in 40 centers in Europe. Pts had no history of chemotherapy or radiotherapy. All analysis were evaluated according to WHO-guidelines.
Results:
We included 243 male pts with a median age of 26,5 years (range 16–57,5 years). At first diagnosis 138 pts were in clinical stage (CS) I/II, 105 in CS III/IV, and systemic symptoms were present in 106 pts. 202 pts underwent fertility screening before therapy: normospermia was diagnosed in 40 (20%) pts and 162 (80%) had inadequate semen quality. Normal values of FSH, LH, and testosterone were found in 140/151 (93%), 99/125 (79%), and 84/101 (83%) of pts respectively. 112 pts underwent at last one fertility screening after therapy. Treatment consisted of chemotherapy alone in 10 pts, in 9 pts of radiotherapy alone and in 93 pts of combined modality. In 40 (36%) pts a recovery of spermatogenesis was observed. Normal values of FSH, LH, and testosterone after treatment were found in 32/63, 40/51, and 36/40 pts respectively. Table 1 shows time of onset of spermatogenesis and normal values of fertility hormones after the end of therapy.
Table 1
1st year 2nd year 3rd year after 3rd year Spermatogenesis (40/112) 7 (18%) 9 (24%) 10 (26%) 14 (32%) Normal FSH (32/63) 8 (25%) 12 (38%) 6 (19%) 6 (19%) Normal LH (40/51) 13 (33%) 16 (40%) 6 (15%) 5 (13%) Normal testosterone (36/40) 17 (47%) 12 (33%) 1 (3%) 6 (17%)
Recovery from azoospermia was observed in 8 (89%) pts treated with radiotherapy, in 1 (10%) pt treated with chemotherapy alone and in 31 (33%) pts treated with combined modality. Recovery rate from azoospermia was lower in pts treated with alkylating agents, BEACOPP chemotherapy, with systemic symptoms and elevated BSG.
Conclusions:
We confirmed that HD pts had inadequate semen quality even prior to treatment. The majority of pts had azoospermia after treatment, but recovery of spermatogenesis was observed, in general 2 years after the end of therapy. Hormone values usually recovered in the first two years. Patient treated with radiotherapy alone recovered in higher frequency. A detailed analysis (including multivariate analyses) will be presented at the annual meeting.