bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of CD95 in EBV infected B-Cells.
Authors: Le Clorennec Christophe , Youlyouz-Marfak Ibtissam , Adriaenssens Eric , Coll Jean , Bordessoule Dominique , Bornkamm Georg W. , Feuillard Jean
Published: October 2019

Expression of CD95 is regulated by various agents such as TNFa, IFNg, or Daunorubicine, able to induce the transcriptional factors NF-kB, STAT1 and p53 respectively. The CD95 promoter is known to harbour binding sites for these factors. These factors are modulated both in expression and activity during EBV infection of B-cells. EBV is known to induce CD95 expression. The objective of this work was to understand the contribution of p53, STAT1 and NF-kB in the regulation of CD95 expression by EBV.

To control the activity of NF-kB, STAT1, p53 and LMP1, we have developed and used various episomal inducible vectors. One of the characteristics of these vectors is that the gene of interest is expressed from a bidirectional doxycycline regulatable promoter that allowing simultaneous expression of truncated NGF receptor, used as a surrogate marker of inducibility. We have subcloned the following cDNAs into these vectors: dominant negative IkBa, active (STAT1a) and dominant inactive natural isoform of STAT1 (STAT1b), p53, a dominant negative mutant of LMP1 (LMP1CT) and LMP1 wild type. After stable transfection of an LCL, induction of the cDNA of interest was performed with doxycycline for 24H. Positive cells for NGFR were purified using magnetic beads and were additionally treated with TNFa, IFNg or Fludarabine for 24H. To control the latency III program, we have used the EREB2-5 cell line harboring an estrogen-regulatable EBNA2 gene. The function of CD95 was studied by quantifying apoptosis induction after CD95 cross linking with a specific antibody.

We found that induction of latency III program in EREB 2-5 cells was associated with an increase of both p53 and CD95 expression.

Inhibition of LMP1 by LMP1CT decreased the expression of CD95. CD95 expression was hardly regulated by p53. Inhibition of both NF-kB and STAT1 decreased CD95 expression. Only IFNg was able to increase expression of CD95. Finally, we show that induction of CD95 expression by EBV render the infected B-cell sensitive to the induction of CD95-mediated apoptosis. Our results suggest that the latencyc III program of EBV sensitize the infected B-cells to CD95-mediated apoptosis via STAT1 and NF-kB activated by LMP1. In the view of the virus/host equilibrium, this process may render EBV-infected B-cells in latency III susceptible to elimination by the immune system.
The SCL Transcription Factor Supports Erythroid Cell Survival and Differentiation Downstream of the Erythropoietin Receptor.
Authors: Lahlil Rachid , Martin Richard , Aplan Peter D. , Begley C. Glenn , Damen Jacqueline E. , Krystal Gerald , Hoang Trang
Published: October 2019

Erythroid cell development critically depends on the SCL/Tal1 transcription factor and on erythropoietin signalling. In the present study, we have taken several approaches to show that the two genes operate within the same pathway to consolidate the erythroid lineage. Signaling through the erythropoietin receptor (EpoR) upregulates SCL protein levels in a clonal cell line (TF-1) in vitro, and in murine fetal liver cells in vivo, when Epor−/− cells were compared to those of wild type littermates at E12.5. In addition, we provide functional evidence for a linear pathway from EpoR to SCL that regulates erythropoiesis. Interfering with SCL induction or SCL function prevents the anti-apoptotic effect of Epo in TF-1 cells and conversely, ectopic SCL expression is sufficient to substitute for Epo to transiently maintain cell survival. In vivo, SCL gain of function complements the cellular defects in Epor−/− embryos to support cell survival and maturation during primitive and definitive erythropoiesis, as assessed by cellular and histological analyses of Epor−/− SCLtg embryos. Moreover, several erythroid specific genes that are decreased in Epor−/− embryos are rescued by the SCL transgene including glycophorinA, bH1 and bmaj globin, providing molecular confirmation of the functional and genetic interaction between Epor and SCL. Conversely, erythropoiesis becomes deficient in compound Epor+/−SCL+/− heterozygote mice, indicating that the genetic interaction between EpoR and SCL is synthetic. Finally, using EpoR mutants that harbour well defined signalling deficiencies, combined with gain and loss of function approaches for specific kinases, we identify MAPK as the major signal transduction pathway downstream of EpoR that upregulates SCL function, necessary for erythroid cell survival and differentiation. Taken together, our observations are consistent with the view that cytokines can influence cell fate by altering the dosage of lineage transcriptional regulators.
Adult T Cell Leukemia/Lymphoma (ATLL) Is a Malignant Counterpart of Regulatory T Cells (Treg).
Authors: Kohno Tomoko , Yamada Yasuaki , Akamatsu Norihiko , Kamihira Shimeru , Tomonaga Masao , Matsuyama Tosifumi
Published: October 2019

Adult T cell leukemia/lymphoma (ATLL) is a lymphoproliferative disorder caused by a retrovirus, human T-lymphotropic virus type 1 (HTLV-1). ATLL is subclassified into four subtypes, smoldering, chronic, acute, and lymphoma. The acute type progresses rapidly and is usually resistant to conventional chemotherapy. In contrast, the chronic type shows an indolent clinical course and the patients survive for several years, even without chemotherapy. Irrespective of the subtypes, however, ATLL patients are in a severely immune-suppressed condition and can easily acquire opportunistic infections such as Pneumocystis Carinii pneumonitis. Suppression of cell-mediated immunity has also been reported in HTLV-1 carriers. Although ATLL cells show the activated helper/inducer T-cell phenotypes, CD4+ and CD25+, they exhibit strong immune-suppressive activity in vitro. The recent notion of CD4+ CD25+ regulatory T cells (Treg) prompted us to investigate the origin of ATLL cells from the standpoint of Treg. Forkhead/winged helix transcription factor (Foxp3) is a functional marker of Treg, which plays a central role in their generation. There are other marker molecules for Treg, including glucocorticoid-induced TNFR family-related protein (GITR) and the chemokine receptors CCR4 and CCR8.

In the present study, we examined primary ATLL cells from 48 patients: 36 patients with acute type and 12 patients with chronic type. We also examined ATLL cell lines, HTLV-1-infected T-cell lines and peripheral blood mononuclear cells (PBMC) from healthy adults as control cells. We used RT-PCR for detection of Foxp3, GITR, CCR4, and CCR8 mRNA expression. Foxp3 and/or GITR mRNA were detected in over 90% of the patients, and 50% of the patients expressed both. There was no difference between subtypes. In contrast, Foxp3 and GITR mRNA were scarcely detected in the PBMC from healthy adults. Furthermore, we confirmed GITR expression at the protein level by flow cytometry. CCR4 and CCR8 mRNA were also detected in almost all ATLL samples, at significantly higher levels than in the normal PBMC. Corresponding to the results of the primary cells, ATLL cell lines and HTLV-1-infected T-cell lines also expressed GITR mRNA, although HTLV-1-negative cell lines, Jurkat and Molt4, completely lack it. Next, we examined whether GITR affects ATLL cell proliferation using a GITR- expressing IL-2-dependent ATLL cell line, KK1. We found that GITR ligand induced proliferation of KK1 cells in an IL-2-negative condition. Thus, these results indicate the Treg origin of ATLL cells and show that GITR expression is possibly involved in the development of ATLL.
Junctional Adhesion Molecule-1 Is Involved in bFGF-Induced Angiogenesis In Vivo.
Authors: Cooke Vesselina G. , Naik Meghna U. , Skarnes William , Naik Ulhas P.
Published: October 2019

Neovascularization is a multistep process that occurs in the body in both physiological and pathological conditions. We have recently shown that Junctional Adhesion Molecule-1 (JAM-1), a member of the Ig superfamily of molecules, is involved in endothelial cell adhesion and migration, leading to angiogenesis. In quiescent endothelial cells, JAM-1 is located at the cell-cell junctions where it forms a complex with integrin αvβ3. Upon treatment of the cells with growth factors, such as bFGF, JAM-1 dissociates from its complex with αvβ3 and redistributes to the cell surface. Blockage of the extracellular domain of JAM-1 inhibits bFGF-induced endothelial cell morphology, proliferation and angiogenesis. Additionally, functional knock-down of JAM-1 using the RNAi technique in endothelial cells showed decreased adhesion and migration of these cells, indicating a possible role for JAM-1 in angiogenesis. In this report, we show that JAM-1 has an important role in bFGF-induced angiogenesis in vivo. Here we present for the first time the generation JAM-1 knock-out mice, using the gene trap strategy. We have successfully confirmed the JAM-1 −/− genotype via Southern, Northern, and Western blot analyses. JAM-1 −/− mice are viable and do not seem to have any external abnormalities, except that they appear to be smaller in size. Retinal fluorescein angiogram revealed no evidence for morphological defects in the vasculature of JAM-1 −/− mice. To evaluate the role of JAM-1 in angiogenesis, we performed an aortic ring assay with both wild type and JAM-1−/− mice. Mouse thoracic aortas were harvested, cross-sectioned into rings of 1-mm thickness, and cultured in a three-dimensional Matrigel supplied with 50 ng/ml bFGF. Vascular sproutings were counted every other day for a period of 7 days at which time they were stained with crystal violet and photographed. Aortic rings from WT mice treated with bFGF showed a 2.8-fold increase in microvessel growth, compared to WT controls with no supplementation of bFGF. In contrast, microvessel sproutings in bFGF treated aortic rings from JAM-1 −/− mice were no more than the vessels in the WT control mice. These results suggest that JAM-1 may be important for bFGF induced angiogenesis. To further confirm the role of JAM-1 in angiogenesis, WT and JAM-1 −/− mice were injected in their flank region with Matrigel containing 80 ng/ml bFGF and 60 U/ml heparin. Two weeks after injection, Matrigel plugs were excised, embedded in paraffin, and the presence of blood vessels was visualized by H&E staining. Matrigel plugs from control WT mice that were not treated with bFGF showed no vascularization, while bFGF supplied Matrigel plugs from WT mice showed a robust vessel growth. Interestingly, bFGF-treated Matrigel plugs form JAM-1−/− mice failed to produce any blood vessels. These ex vivo and in vivo studies using JAM-1−/− mice suggest that JAM-1 has a unique and essential role in bFGF-induced angiogenesis.
Are There Two Distinct Mechanistic Pathways That Explain the Varying Clinical Presentations, Laboratory Findings, and Outcomes of Thienopyridine-Associated Thrombotic Thrombocytopenic Purpura (TTP)?.
Authors: Kim Benjamin , Carson Kenneth R. , McCaffrey Kathryn , Finley Denise , Zakarija Anaadriana , Bandarenko Nicholas , Pandey Dilip K. , Raisch Dennis W. , Kwaan Hau C. , Yarnold Paul R. , Bennett Charles L.
Published: October 2019

Introduction: The antiplatelet agents ticlopidine and clopidogrel are thienopyridine derivatives that are commonly used for secondary prevention of cerebrovascular events, prevention of coronary artery stent thrombosis, and for treatment of acute coronary syndrome. While these chemically related agents are the first and third most common causes of drug-associated TTP respectively, clinical presentations, laboratory findings, and outcomes suggest that there may be two distinct mechanistic pathways.

Methods: Clinical and laboratory data for 60 cases of ticlopidine-associated TTP and 35 cases of clopidogrel associated TTP were reviewed.

Results: Type 1 thienopyridine-associated TTP, characterized by onset of disease > 5 days after drug initiation, occurred in 98% of the ticlopidine patients versus 63% of the clopidogrel patients. Laboratory studies identified severe deficiency of ADAMTS13 activity and the presence of IgG inhibitors in 7 ticlopidine (Tsai, et al.; Annals of Internal Medicine, 2000) and 2 clopidogrel (Bennett, et al.; NEJM, 2000) patients. Survival was 68% in the ticlopidine patients versus 91% in the clopidogrel patients. Type 2 thienopyridine-associated TTP, consisting of TTP onset within 5 days of drug initiation, occurred in 2% of the ticlopidine patients versus 37% of the clopidogrel patients. Laboratory studies failed to identify deficient ADAMTS13 activity or the presence of IgG inhibitors in 1 clopidogrel patient (Zheng, et al.; Blood, 2004). Survival was 0% in the ticlopidine patients versus 38% in the clopidogrel patients.

Conclusion: The proposed type 1 and type 2 thienopyridine-associated TTP may be distinguished on the basis of clinical presentation, ADAMTS13 levels, and outcome. Type 1 thienopyridine-associated TTP—characterized as occurring after > 5 days of thienopyridine treatment, severely deficient ADAMTS13 activity and presence of IgG inhibitors, and high survival rates with plasmapheresis—is more likely to occur in patients receiving ticlopidine. Type 2 thienopyridine-associated TTP—characterized as occurring within 5 days of thienopyridine therapy initiation, normal ADAMTS13 activity and undetectable IgG inhibitors, and poor survival rates despite plasmapheresis—is more likely to occur in patients receiving clopidogrel. Additional confirmatory studies on larger numbers of patients are needed.
Autologous Transplant Event-Free Survival (EFS) Following Failure of CHOP-Rituximab (CHOP-R) for Diffuse Large B-Cell Lymphoma (DLBCL) Is the Same as the EFS Following Failure of CHOP Alone.
Authors: Vose Julie M. , Bierman Philip J. , Lynch James C. , Bociek Gregory , Armitage James O.
Published: October 2019

Several studies have now identified that the addition of rituximab (R) to CHOP chemotherapy has increased the complete response, EFS, and overall survival (OS) for patients with previously untreated DLBCL. However, concerns of increased resistance of DLBCL following failure of CHOP-R and the inability to salvage those patients with high-dose chemotherapy and autologous stem cell transplantation have been raised. In an attempt to address this issue, we evaluated 103 patients with high risk, relapsed, or refractory DLBCL receiving high-dose chemotherapy and autologous stem cell transplantation at our center between 1999 and 2003. Fifty-six (54%) of the patients received CHOP as their initial induction therapy and 47 (46%) received CHOP-R. The patients ranged in age from 20–73 years (median 50). Sixty patients were male and 43 female. Sixty two of the patients were transplanted < 12 months from diagnosis and 41 were transplanted ≥ 12 months from diagnosis. The patients had received a median of 2 prior therapies (range 1–4). The majority (81%) were chemotherapy sensitive at the time of transplantation. The median follow-up of surviving patients is 27 months ( range 3–74). All patients received a BEAM (C) - based regimen [carmustine, etoposide, cytarabine, melphalan or cycylophosphamide] +/− anti-CD20 monoclonal antibodies. There was no difference in the 2-year EFS for patients failing CHOP vs. CHOP-R (60% vs. 68%, p=0.49). In addition, there was no difference in the 2-year overall survival (OS) of patients failing CHOP vs. CHOP-R (72% vs. 68%, p=0.63). In a multivariate analysis, the only factor predicting for an event (relapse or death) post-transplant was having received ≥ 2 prior chemotherapies (Relative Risk (RR) 7.5, p=0.048) and predicting for death from any cause was chemotherapy resistance (RR 2.5, p=0.037) and an increased lactic dehydrogenase at transplant (RR 4.1, p=0.002). The additional use of a monoclonal antibody with the transplant regimen did not significantly impact outcome in this analysis. This study demonstrates that patients with DLBCL failing CHOP-R are able to be salvaged with high-dose chemotherapy and autologous stem cell transplantation and have a similar 2-year EFS and OS as patients being transplanted following CHOP failure.
Hematopoietic Stem Cell Transplant (HSCT) as Primary Treatment for T-Cell Lymphobastic Lymphoma (T-LBL) : An Intention to Treat Analysis.
Authors: Song Kevin W. , Nevill Thomas J. , Gascoyne Randy D. , Toze Cynthia L. , Barnett Michael B. , Forrest Donna L. , Hogge Donna E. , Lavoie Julye C. , Nantel Stephen H. , Shepherd John D. , Smith Clayton A. , Sutherland Heather J. , Voss Nicholas J. , Connors Joseph M.
Published: October 2019

The role of HSCT for T-LBL is not well defined. Most series are subject to referral bias and do not address the true impact of HSCT, especially as primary treatment. Since 1987 all patients in British Columbia with T-LBL have been considered eligible for HSCT as part of primary treatment. Between 6/87–12/03, 27 patients were diagnosed with T-LBL. Patients with > 25% involvement of the bone marrow were excluded.

Characteristics: Age range 18–56 y (median 26); M:F 19:8; Ann Arbor stage I-II 6; III-IV 21; bone marrow involved 7/27; peripheral blood involved 2/27; CNS involved 1/27; LDH elevated 15/25; mediastinal mass 23/27.

Initial treatment: All received an anthracycline-containing regimen similar to CHOP. All achieved at least a partial remission.

HSCT: 23/27 (85%) proceeded to HSCT in first response.

Source of stem cells: 4, related donor (2 marrow, 2 peripheral blood); 19, autologous (15, mafosphamide purged marrow; 1, 4-hydroperoxy-cyclophosphamide purged marrow; 1, unpurged marrow; 1, unpurged peripheral blood; 1, combined unpurged marrow and peripheral blood). Conditioning was cyclophosphamide (150 mg/kg) and TBI (750–1200 cGy) +/− etoposide 1.8 g/m2 in 22 of 23 patients. CNS prophylaxis with intrathecal methotrexate and cytarabine was planned for all patients. All 23 had maintained chemosensitivity at time of HSCT. Time from diagnosis to HSCT was 1.3–6.4 months (median 2.2). Reasons for not proceeding to HSCT 2 refused; 1 decided to wait until relapse; 1 did not have adequate organ function. Median follow-up of living patients was 33 months (range 4–142 months).

Results: Overall survival (OS) and event free survival (EFS) at 3 years for all patients were 79% (95% CI 61–97%) and 75% (95% CI 57–93%); patients who underwent HSCT 85% (95% CI 70–100%) and 81% (95% CI 64–98%). 4 patients relapsed post HSCT at 2.2, 10.8, 11.7 and 40.4 months post HSCT.

Sites of relapse: 2, mediastinum; 1, bone marrow; 1, mediastinum and bone marrow. One patient who received HSCT died of a treatment related complication (interstitial pneumonitis). Significant factors for improved survival post HSCT (univariate): Absence of bone marrow involvement [EFS at 3 years 92% vs 57% (p=0.026)].

Non-significant factors: normal LDH, stage I/II at diagnosis, age < 30 yrs.

Conclusions: Patients with T-cell lymphoblastic lymphoma treated with a CHOP-like regimen achieve adequate control of disease to proceed to HSCT. HSCT as primary treatment results in excellent long-term disease free survival with acceptably low treatment related mortality. Bone marrow involvement at diagnosis, even though < 25%, predicts for a worse outcome.
Front-Line High-Dose Chemotherapy (HDT) Combined with Rituximab for Adults with Aggressive Large B-Cell Lymphoma (DLBCL) : Goelams 074 Trial.
Authors: Milpied Noel-Jean , Lamy Thierry , Casassus Philippe , Deconninck Eric , Gressin Remy , Maisonneuve Herve , Le Maignan Christine , Ramee Jean-Francois , Tournilhac Olivier , Dugay Jacqueline , Legouffe Eric , Delwail Vincent , Colombat Philippe
Published: October 2019

Background: The beneficial effect of adding rituximab to CHOP has been shown for elderly patients with DLBCL (Coiffier et al, N.E.J.M 2002). We have shown that HDT with autologous stem cell transplantation is superior to CHOP in young adults with DLCL ( Milpied et al GOELAMS 072 trial, N.E.J.M 2004). The feasibility of adding rituximab to front-line HDT remains to be established.

Methods: A prospective pilot trial was proposed to patients with DLBCL, with intermediate-high or high adjusted IPI, up to the age of 60 y.o. This program consisted of 2 courses of high-dose CHOP-like regimen, 15 days apart, with rituximab (375/mg/m2) on day 1 of each course, followed by rituximab on d 22, harvest of G-CSF mobilised peripheral blood stem cells on d 28,29, then rituximab on d 36 followed by a course of high-dose methotrexate with cytarabin. For patients who achieved at least a PR after these 3 courses, a BEAM regimen started on d 66 to 80 followed by the infusion of stem cells.

Results: Between 04/2002 and 05/2003, 42 pts gave their informed consent and were included in that trial. Median age was 50 y.o (18–60 y.o), 23 had WHO PS ≥ 2, the LDH level was >N in 41 pts and 38 had stage III or IV disease. The age-adjusted IPI was intermediate-high in 23 and high in 19 pts. The program was completed in 30 pts (71%), 3 pts died of toxicity before the BEAM regimen, 8 pts failed to achieve at least a PR after the first 3 courses and 1 refused the autologous transplant. On an intent-to-treat basis, the response rate at the end of the treatment was CR/Cru=64%, PR=7%, less than PR or progression= 22% and toxic death=7%. No toxic death was directly attributable to the addition of rituximab. With a median FU of 19 m, the KM 2y probability of OS and EFS are 79% and 59% respectively, these figures compare to those achieved without rituximab in the previous trial as shown on the table:

Conclusion : The addition of 4 doses of rituximab to this HDT program is feasible with no unexpected toxicity, allows the harvesting of sufficient numbers of stem cells to support an autologous transplant with a BEAM regimen in responding patients. This treatment is now being randomly prospectively compared with CHOP-14-rituximab in youg adults with DLBCL (Goelams 075 trial).Supported in part by Roche which kindly provided rituximab for that trial and by DRC Nantes programme N°:02/2N 2002.

GOELAMS 072 (IPI 2 only) GOELAMS 074 (IPI 2–3) 2y OS 80% 79% 2y EFS 60% 59%
Autologous Stem Cell Transplantation as First Line Therapy in Peripheral T Cell Lymphomas. Update of a Prospective Multicenter Study.
Authors: Reimer Peter , Ruediger Thomas , Schertlin Tobias , Geissinger Eva , Weissinger Florian , Mueller-Hermelink Hans Konrad , Wilhelm Martin
Published: October 2019

Peripheral T-cell lymphomas (PTCL) represent a heterogeneous group of non-Hodgkin’s lymphomas, which in general show a poor outcome following conventional chemotherapy. Long-term remissions are achieved in only 15 to 35 %. However, the impact of more aggressive therapeutic approaches such as myeloablative therapy with autologous stem cell transplantation (ASCT) as first line therapy is poorly defined mainly due to the lack of prospective PTCL-restricted studies. In 6/00 we initiated the first prospective PTCL-restricted multicenter study of myeloablative radiochemotherapy in primary diagnosed PTCL. The results of the first 30 patients (pts) are in press. We update our data on all pts entering the study.

Study design: Pts < 65 years with PTCL of all subtypes without primary cutaneous lymphoma and ALK1 expressing anaplastic large cell lymphoma were included. Treatment consisted of 4–6 courses of CHOP protocol followed by DexaBEAM or ESHAP regimen and collection of stem cells. Subsequently pts underwent total body irradiation (TBI) and high dose cyclophosphamide chemotherapy (60 mg/kg body weight) with ASCT.

Patient characteristics: From 6/00 to 8/04 65 pts (42 male) with a median age of 50 years were enrolled. Main subtypes were Peripheral T-cell lymphoma not otherwise specified (NOS, n= 26) and Angioimmunioblastic T-cell lymphoma (AILT, n= 19). According to the Ann Arbor classification, 81% of the pts had stage III/IV disease. The International Prognostic Index (IPI) was low/low intermediate in 54% and intermediate high/high in 46% of the pts, respectively.

Results: So far 54 of 65 pts are eligible for evaluation, while the remaining 11 pts are still under therapy. Thirty-three pts could be transplanted (61%). After a median follow up of 10 months after transplantation 22 pts (67%) are in sustained remission and 8 pts (27%) had relapsed. Post transplantation two pts died treatment-related (one secondary AML, one multiorgan failure). Twenty-one pts (39%) did not proceed to ASCT mainly due to progressive disease (n= 16). Treatment-related toxicity was comparable to other high-dose studies in malignant lymphomas.

Conclusion: Our data show feasibility and efficacy of first-line ASCT following myeloablative radiochemotherapy in PTCL. Sustaining remission seems achievable for a majority of pts. However, additional treatment strategies are required to prevent early progression prior myeloablative therapy. Longer follow-up is necessary to confirm long-term remission rate.
Long-Term Follow-Up of High-Dose Chemotherapy Followed by Autologous Stem Cell Transplantation in Peripheral T-Cell Lymphomas at Diagnosis.
Authors: Cortelazzo Sergio , Tarella Corrado , Dodero Anna , Gianni Alessandro Massimo , Francesco Zallio , Zanni Manuela , Rambaldi Alessandro , Barbui Tiziano , Corradini Paolo
Published: October 2019

Background: Limited experience is available on the feasibility and long-term efficacy of high-dose (HD) chemotherapy followed by autologous stem cell transplantation (ASCT) in patients with peripheral T-cell lymphoma (PTCL) at diagnosis.

Patients and methods: Between January 1993 and December 2003, 54 patients (median age 44 years, range 20-61; 66% male) with PTCL with high-risk features were candidate to HD chemotherapy and ASCT as a first-line treatment. Histologic PTCL subtypes included: PTCL not otherwise specified (n=25), ALCL alk-positive (n=17), angioimmunoblastic (n=9), others subtypes (n=4). 44 patients (81%) presented with stage III or IV, 58% had elevated s-LDH and 73% presented with two or more adverse risk factors according to the International Prognostic Index (IPI). Treatment program consisted of: 1) induction and intensification phase with two APO and two DHAP courses than either the sequential administration at 15-20 days intervals of HD-cyclophosphamide, HD-cytarabine and HD-etoposide (n=26) or 8 weeks of MACOP-B followed by HD-cytarabine and mitoxantrone (MAD) (n=29); 2) consolidation phase with HD-mitoxantrone and HD-melphalan (n=12) or BEAM (n=27) followed by ASCT; 3) radiotherapy on bulky sites.

Results: Thirty-nine patients (71%) attained a clinical response (n=32 CR; n=7 PR) and were autografted while 16 patients (29%) did not for the following reasons: progressive disease (n=10), toxicity (n=2), toxic death concomitant with refractory disease (n=2), refusal (n=2). At a median follow-up of 66 months (range, 10-127 months), 27 patients (49%) are alive (25 in CR) and 28 died (51%): 21 for disease progression, 5 for treatment-related-toxicity and 2 for other reasons (n=1 other disease, n=1 car accident). The estimated 2-year TRM was 7 %. Using an intention-to-treat-analysis, the estimated 5 year overall survival (OS) and progression-free survival (PFS) projections were 52% (95% CI, 38% to 66%) and 50% (95% CI, 36% to 64%) respectively. Patients autografted with ALCL alk-pos subtype had a better prognosis compared to other subtypes with 5 year OS of 84% versus 52% (P<0.03) and 5 year PFS of 84% versus 36% (P<0.01), respectively. No statistical difference in PFS or OS was observed between patients treated with either sequential HD therapy or MACOP-B/MAD. Univariate analysis revealed no significant differences in OS and PFS for patients with IPI score <2 versus ≥ 2.

Conclusions: Our study indicates that up-front HD and ASCT: 1) induces a high rate of long term remission in patients with ALCLalk-pos subtype; 2) is of limited benefit in patients with non-ALCLalk-pos subtype whose expected long-term progression free survival was lower than 40%. In this subgroup other treatment options including consolidation with allogeneic transplantation from HLA identical donor, should be evaluated.
Time-Dependent Effect of Non-Hodgkin’s Lymphoma Grade on Disease Free Survival (DSF) of Relapsed/Refractory Patients Treated with High-Dose Chemotherapy (HDC) Plus Autologous Hematopoietic Stem Cell Transplant (AHSCT).
Authors: Fu Pingfu , Van Heeckeren Willem J. , Wadhwa Punit D. , Bajor David J. , Creger Richard J. , Cooper Brenda W. , Laughlin Mary J. , Lazarus Hillard M. , Gerson Stanton L. , Koc Omer N.
Published: October 2019

HDC and AHSCT is an effective treatment option for patients with relapsed or refractory NHL. The outcome of this treatment depends on the histological grade (indolent versus aggressive) with a well documented time dependent difference in temporal relapse pattern. We developed statistical models to describe the time dependent outcome after AHSCT based on histologic grade and other variables. During January 1993 to December 2003, 114 relapsed or refractory NHL patients were treated at single institution using high-dose BCNU 600 mg/m2, etoposide 2400 mg/m2, and cisplatin 200 mg/m2 IV over 5 days and AHSCT. Median age was 53.5 (range: 25–70) years, 78 patients had aggressive NHL and 36 indolent NHL. Immediately before transplant, 75 patients received involved-field radiotherapy (IFR) to sites of prior bulky (> 5 cm) disease. Twenty-seven patients had mobilized stem cell transplant (13 with in vivo purging and 14 with ex-vivo purging). At a median follow up of 33 (range: 3 to 118) months, the estimated 5 year Kaplan-Meier probabilities of overall survival (OS) and DSF were 61% and 51%, respectively. DFS was similar up to about 1.2 years between indolent and aggressive NHL (Figure 1). DFS of the group who had purged stem cells was improved marginally over non-purged group. Cox proportional hazards model was used to adjust for differences in baseline characteristics between grades that might affect DFS. The proportionality assumption of Cox model was tested by adding a time-dependent covariate for each variable. This showed that the proportionality did not hold for lymphoma grade, indicating the relationship between the grade and DFS differed over time. The results of piece-wise Cox model after controlling age, IFR, purging status indicate there was no difference of DFS between the grade groups up to 1.2 years (optimal cut-off point) post-transplantation (p = 0.748). However, the relative risk for experiencing relapse after 1.2 year in patients with indolent compared with patients with aggressive NHL was 4.22 (p = 0.006) with 95% confidence interval (1.5, 11.9). Our results statistically prove the previously described biological differences in outcomes of patients with indolent and aggressive NHL following HDC. The time-dependent effect of lymphoma grade on DFS indicates the need for early (within first year) incorporation of novel therapeutic approaches, such as post-transplant immunotherapy, tandem high-dose therapy and, allogeneic transplant to generate graft-versus-lymphoma effect, in management of patients with indolent NHL undergoing autologous transplantation.

Figure 1: DFS by tumor grade. Figure 1: DFS by tumor grade.
Lack of Prognostic Implications of Induction Response to Total Therapy 2 (TT-2) in Newly Diagnosed Patients with Multiple Myeloma (MM) for Post-Transplant Even-Free Survival (EFS) and Overall Survival (OS).
Authors: Tricot Guido , Crowley John , Stover Julie , Milner Teresa , Fassas Athanasios , Zangari Maurizio , Thertulien Raymond , Anaissie Elias , van Rhee Frits , Talamo Giampalamo , Hollmig Klaus , Barlogie Bart
Published: October 2019

Between October 1998 and February 2004, a total of 668 recently diagnosed myeloma patients were enrolled in TT-2, of whom 566 completed the first and 417 the second autotransplant. Twenty percent of patients were ≥ 65 years of age and 34% had abnormal metaphase cytogenetics; 24% had received a single cycle of chemotherapy prior to enrollment and 32% had a B2 microglobulin level of ≥ 4mg/L. All patients who completed at least the first transplant could be assessed for response to induction therapy consisting of VAD, DCEP, CAD and DCEP given with hematologic growth factor support at 4 to 5 week intervals. We examined whether the quality of induction response had an impact on event-free (EFS) and overall survival (OS). Responses were defined as complete remission (CR) if serum and urine immunofixation were negative with normal bone marrow plasmacytosis in aspirate and biopsy; partial remission (PR) if serum M protein was reduced by ≥ 75% and urine M-protein ≥ 90% with a normal bone marrow plasmacytosis and < PR for all other responses. Superior EFS (p=.04) and OS (p=.06) from first transplant were observed among the 86 CR patients (15%) with a 4-year estimate of EFS of 69% and OS of 83%, compared with 59% and 72%, respectively, for PR patients (N=214) and 52% and 68%, respectively, for < PR patients (N=266) (Figure 1 and 2). With a median follow-up of 3 years from the first transplant, we conclude that, although a stringently-defined CR after induction therapy resulted in a better EFS and OS, there was no difference in outcome between PR and < PR patients, who had virtually super imposable survival curves. These data, as well as a review of the literature (Vesole et al. Blood1994; 84: 950, Alexanian et al. Blood1994, 84: 4278 and Singhal et al. Bone Marrow Transplant2002, 30: 673) provide a strong argument for change in current Medicare policy, which denies autotransplants to myeloma patients who have failed primary induction therapy. Even with intensive induction therapy as applied in TT-2, more than two-thirds of patients achieving < PR prior to the first transplant are projected to be alive at 4 years after their autotransplant.

Figure Figure Figure Figure
Angiotensin-I Converting Enzyme Inhibitors and the Survival of Patients with Multiple Myeloma Treated by Autologous Stem Cell Transplantation.
Authors: Buchler Tomas , Krejci Marta , Svobodnik Adam , Adam Zdenek , Minarik Jiri , Bacovsky Jaroslav , Scudla Vlastimil , Mayer Jiri , Vorlicek Jiri , Hajek Roman
Published: October 2019

Introduction:

The aim of our study was to analyze the impact of angiotensin-I converting enzyme inhibitors (ACEI) on the outcome of patients with multiple myeloma (MM) treated by autologous peripheral blood stem cell transplantation (APBSCT), as it has been reported that low angiotensin-converting enzyme (ACE) activities are associated with poor prognosis in multiple myeloma (MM).

Patients and Methods:

Patients with MM who underwent APBSCT (n=168) were studied retrospectively. Patients taking ACEI alone or in combination with other antihypertensive agents during the APBSCT were allocated to the ACEI group (n=25; 15%). The ACEI group was compared both to all other patients (n=143; 85%) and to patients with hypertension who were taking other or no antihypertensive medication (n=44). The induction chemotherapy consisted of 4 cycles of VAD. For stimulation, cyclophosphamide was given, followed by G-CSF. High-dose melphalan was used as conditioning.

Results:

The ACEI and non-ACEI groups were similar in terms of age, initial levels of albumin, beta2-microglobulin, CRP, LDH, performance status, stage according to IPI and Durie-Salmon criteria, myeloma type, and maintenance treatment. Patients with hypertension (n=69) and patients without hypertension (n=99) had similar overall survival (OS) after APBSCT (65.5 months versus 60.9, respectively; p=0.561), and progression-free survival (PFS) after APBSCT (32.6 months versus 29.6 months; p=0.635). Patients taking ACEI had significantly worse OS after APBSCT (32.7 months) compared to patients not taking ACEI (65.2 months; p=0.033). Among patients with hypertension, OS was significantly shorter in the ACEI group than in the non-ACEI patients (p=0.024). Statistically significant differences were also observed in PFS between hypertensive patients treated by ACEI and other hypertensive patients (19.3 months versus 48.6 months, respectively; p=0.041). Most deaths in both groups were myeloma-related. Renal failure was a main or contributing cause of death in 3 patients from the ACEI group and in 8 patients from the non-ACEI group (p=0.233). Also, there was a trend favoring non-ACEI hypertensive patients in comparison to ACEI hypertensive patients in the number of complete responses after PBSCT (40% versus 17%; p=0.058).

Conclusion:

In this retrospective study we have found that patients with MM and hypertension treated with ACEI at APBSCT have worse survival in comparison to patients with MM receiving other or no antihypertensive drugs. Further research in this area could lead to important insights into the pathophysiology of MM and to the reevaluation of ACE activity as a prognostic factor in MM.
Reduced Risk of Residual Molecular and Hematological Relapse in Patients with Cml after KIRir-Ligand Mismatched Hematpoietic Stem Cell Transplantation.
Authors: Elmaagacli Ahmet H. , Ottinger Hellmut , Michael Koldehoff , Rudolf Peceny , Steckel Nina K. , Trenschel Rudolf , Biersack Harald , Grosse-Wilde Hans , Flasshove Michael , Beelen Dietrich W.
Published: October 2019

We compared the incidence of molecular and hematological relapse in 236 CML patients (pts) after HLA-identical (n=158) [group 1], HLA class I antigen mismatched and KIR-ligand compatible (n=49) [group 2], and HLA class I antigen mismatched and KIR-ligand incompatible (n=29) [group 3] hematopoietic stem cell transplantation. In group 1 133/158 (84%) pts were in first chronic phase of CML, the corresponding figures were 33/49 (67%) pts in group 2, and 19/29 (64%) in group 3 (p<0.05). Accordingly, 25/158 (16%) pts in group 1, 16/49 (33%) pts in group 2, and 10/29 (34%) pts in group 3 were in advanced disease phases of CML (accelerated phase, 2nd chronic phase, or blastic phase).

Stem cell grafts were from sibling donors in 97% of pts in group 1, 37% of pts in group 2, and in 59% of pts in group 3 (p<0.01 group 1 versus groups 2+3). Median age and gender constellations were not different between the three groups (age medians between 37–41 years, range 16–63).

Acute GVHD grade 2–4 occurred in 62/158 (39%) pts in group 1, in 24/49 (49%) pts in group 2, and in 15/29 (52%) pts in group 3 (NS). Estimates of chronic GVHD did not differ significantly between the study groups with 94% + 2% in group 1, 95% + 4% in group 2, and 91% ± 6% in group 3.

Molecular relapse as detected by real-time RT-PCR occurred in 1/29 (3%) pts of group 3 compared to 62/158 (39%) of pts in group 1, and in 11/49 (22%) pts in group 2 (p<0.001). A hematological relapse developed in 20/158 (13%) pts in group 1, in 2/49 (4%) pts in group 2 and in 0/29 (0%) pts in group 3 (p<0.05). Multivariate analysis on the risk molecular relapse included possible influencial covariates like disease stage, age (> 40 years or < 40 years), gender matching of donor and recipient, occurrence of acute and chronic GVHD, HLA matching between donor and recipient, donor source [sibling or unrelated], and graft type [bone marrow or PBSCs]. This analysis confirmed that KIR-mismatches are a strong independent predictor for the occurrence of bcr-abl transcripts after transplantation (p<0.02). The 8-year overall survival estimates were not different between the three groups (67% group 1, 52% group 2 66% group 3). In conclusion, KIR-ligand incompatibility is an important prognostic factor for the occurrence of molecular and hematological relapse after allogeneic transplantation for CML.
High-Dose Imatinib Mesylate Treatment in Patients (Pts) with Previously Untreated Early Chronic Phase (CP) Chronic Myeloid Leukemia (CML).
Authors: Cortes Jorge , Talpaz Moshe , O’Brien Susan , Giles Francis , Beth Rios Mary , Levtak Laurie , Kantarjian Hagop
Published: October 2019

Imatinib has become the treatment of choice for most with CML. The standard dose (SD) for CP CML is 400 mg daily, but pre-clinical and clinical observations suggest that higher doses (HD) may be more effective. We have treated 222 with previously untreated CML in early CP with imatinib in 3 consecutive trials: one using SD imatinib (400 mg/day) (n=50; all entered in April 2001) and 2 subsequent trials using 400 mg twice daily (total dose 800 mg/day) (n= 172; from June 2001 until present). The 2 HD trials had identical inclusion criteria and will be considered together for this analysis. Pts followed for at least 3 months (mo) are evaluable (n=210) for this report (n=49 at 400mg, 161 at 800 mg). The median age was 48 years (range, 15 to 84); platelets were >450 x109/L in 71 pts (34%), 78 (37%) had peripheral blood (PB) blasts, and 11 (5%) had clonal evolution. Sokal risk group classification was good in 128 (61%) pts, intermediate in 61 (29%) pts, and poor in 21 (10%) pts. There was no difference in pre-treatment characteristics between the standard SD and HD groups. The results at 18 months are as follows:

Response % Response p value* 400 mg/day 800 mg/day CR=Complete remission, Molecular Major=BCR-ABL/ABL <0.05%, Molecular CR=BCR-ABL undetectable (confirmed by nested PCR), *p value by log-rank Median follow-up (months) 36 19 Cytogenetic CR 81 96 0.0002 Cytogenetic Major 99 93 0.15 Molecular Major 47 67 0.0007 Molecular CR 8 24 0.02

Four pts treated with SD have transformed (3 to BP, 1 to AP) and 3 (2 to BP, 1 to AP) in the HD groups (p=0.05) (median time to transformation 11 mo, range 3 to 27). Estimated progression-free survival at 12 mo is 92% in the SD group and 99% in the HD group (p=0.42) (p=0.12 for the estimated transformation-free-survival, 94% and 99% for SD and HD at 12 mo). 4 have died (1 in SD and 3 in HD). Extramedullary toxicity was similar in the 2 groups, but myelosuppression was more common with HD, with grade ≥3 anemia, neutropenia and thrombocytopenia occurring in 7%, 39%, and 27% of pts receiving HD, respectively, and 4%, 20% and 12% of pts receiving SD. At 12 mo, the median actual dose for the HD group is still 800mg, with 40/112 (36%) evaluable having required dose reduction. This compares with 7/43 (14%) of those treated with SD. We conclude that high-dose imatinib results in higher rates of complete cytogenetic and molecular remissions, with some increase in myelosuppression.
Mechanisms of Plasmin-Catalyzed Inactivation of the Factor VIII.
Authors: Nogami Keiji , Shima Midori , Matsumoto Tomoko , Nishiya Katsumi , Takeyama Masahiro , Tatsumi Kohei , Tanaka Ichiro , Yoshioka Akira
Published: October 2019

Factor VIII (FVIII) functions as a cofactor for factor IXa in the intrinsic tenase complex. This tenase activity is down-regulated by activated protein C (APC) or factor Xa (FXa). Plasmin, the most potent fibrinolytic protease, inactivates FVIII as well as other clotting factors. However, the mechanism of FVIII inactivation by plasmin is poorly understood. FVIII activity reached to the peak value of ~2-fold increase at 3 min after the addition of plasmin in a one-stage clotting assay. Then, the activity was decreased rapidly and was undetectable within 30 min. This time-dependent reaction was not affected in the presence of von Willebrand factor and phospholipid. The activation of FVIII by plasmin was an ~50% level of that by FXa. The rate constant (min-1) of inactivation of FVIIIa by plasmin possessed ~11.3- and ~2.5-folds greater than those by FXa and APC in the presence of protein S, respectively. SDS-PAGE analysis showed that plasmin cleaved the 90~210-kDa heavy chain of FVIII to 50, 48,45, 40, and 38-kDa fragments via 90-kDa fragment. Using western blot and N-terminal sequence analyses, these fragments derived from the heavy chain were identified as A11-372, A1337-372-A2, A11-336, A2, and A137-336, respectively, by cleavages at Arg372, Arg740, Lys36 and Arg336 in the A1 domain. On the other hand, the 80-kDa light chain was cleaved to 67-kDa fragment via 70-kDa fragment by cleavages at Arg1721 and Arg1689, respectively, consistent with the pattern of cleavage by FXa. However, the cleavage at Arg336 by plasmin was much quicker than that at Arg372, contrast with that by FXa. Furthermore, this cleavage was faster than that by APC, consistent with rapid inactivation of FVIII. In addition, the cleavage at Arg336 of FVIIIa by plasmin was faster than that of isolated A1 or A1/A3-C1–C2 dimer, different with that by FXa. These results demonstrate the importance of cleavage at Arg336 for the mechanism of plasmin-catalyzed FVIII inactivation. Furthermore, this cleavage appears to be selectively modulated by the A2 domain that may interact with plasmin.
Diamond Blackfan Anemia: An RPS19 Transcript Variant Specifically Interacts with Nuclear Proteins.
Authors: Frojmark Anne-Sophie , Badhai Jitendra , Davey Edward J. , Dahl Niklas
Published: October 2019

Diamond Blackfan Anemia (DBA) is a rare congenic anemia characterized by reduced erythrocyte precursors. Mutations in the ribosomal protein S19 (RPS19) gene are found in about 20% of patients, although it is unclear how this relates to the pathophysiology of the disease. Two transcripts have been reported for human RPS19 in the NCBI database (BC000023 and BC018616), which only differ in the length of the 5′UTR. Both transcripts predict translation to the same protein product and we therefore speculate that the extended 5′UTR may be of importance. While the short (569bp) transcript is well studied, the long (873bp) transcript has not been investigated. We determined the size and tissue distribution as a basic characterization of the long RPS19 transcript. Furthermore, we examined proteins interacting with the different RPS19 transcript variants. Two methods were used determine the size of long RPS19 transcript. Ribonuclease protection assays and 5′-RACE indicate a transcript length consistent with the reported sequence with a 5′UTR of 372bp. Interestingly the 5′UTR predicted in the long transcript contains sequence for a previously reported promoter, suggesting a second novel promoter. Tissue distribution was determined using reverse transcriptase PCR on isolated RNA from a panel of tissues including heart, bone marrow, fetal liver, liver, brain, fetal brain, muscle, kidney, uterus, adrenal gland, lungs, placenta, prostate testes, ovary, thymus, colon, lymphocytes and from K562 and HeLa cell lines. Analysis indicated a ubiquitously expression of the long RPS19 transcript. In order to investigate interactions between the RPS19 transcripts and regulatory proteins, a UV cross-linking assay was performed using in vitro transcribed RNA representing both short and long transcripts. A protein of 55 kDa from nuclear fractions prepared from the erythroid cell lines K562, UT-7 and Cos-1 cells was found to bind to the long RPS19 transcript but not to the short RPS19 transcript. Our results provide evidence for an additional long, RPS19 transcript containing an extended 5′UTR that is ubiquitously transcribed and indicates the existence of a second promoter for RPS19. We further show that a nuclear protein, present in two different erythroid cell lines, interacts with the long RPS19 transcript indicating a function for the extended 5′UTR. This data suggests additional regulatory pathways involving RPS19 mRNA which may be of relevance to the understanding of the molecular mechanisms behind DBA.