Dysregulated apoptosis plays a key role in the pathogenesis and progression of neoplastic disorde... more Dysregulated apoptosis plays a key role in the pathogenesis and progression of neoplastic disorders, allowing tumor cells to survive beyond their normal life-span, and to eventually acquire chemo-radioresistance (Laconi et al., 2000; Pommier et al., 2004). Thus, targeting either the intrinsic or the extrinsic pathways of apoptosis represent attractive therapeutic strategies for restoring apoptosis sensitivity of malignant cells, or activating agonists of apoptosis (Waxman & Schwartz, 2003). Due to the ability of death receptor ligands to induce cell death, there has been considerable interest in the physiological roles and therapeutic potential of these cytokines as anti-cancer agents. Death receptor ligands of the tumor necrosis factor α (TNFα) superfamily are type II transmembrane proteins that signal to target cells upon cell-cell contact, or after protease-mediated release to the extracellular space (Ashkenazi, 2002). Members of this family, including Fas ligand (FasL), TNFα, an...
Background The outcome of Diffuse Large B Cell Lymphoma (DLB-CL) has been definitely improved by ... more Background The outcome of Diffuse Large B Cell Lymphoma (DLB-CL) has been definitely improved by the addition of the anti-CD20 rituximab to conventional chemotherapy. However, despite the advantages offered by rituximab, results are still disappointing in patients presenting with adverse prognostic factors. For these patients, rituximab-supplemented intensive programs with autograft might represent a suitable option. Thus, a prospective multicenter study has been performed using first-line the Rituximab-supplemented high-dose sequential chemotherapy program delivered with multiple autologous peripheral blood progenitor cell (PBPC) support (R-HDS-maps regimen), in DLB-CL patients with score 2 or 3 according to the age-adjusted International Prognostic Index (aaIPI). Preliminary results have been already reported (ASH 2005; Leukemia2007, 21: 1802). Updated results after a prolonged follow-up are here presented. Methods. R-HDS-maps includes: 3 APO courses; sequential administration of ...
We showed recently that the dismal outcome of MCL might be improved in 28 young patients (pts) (&... more We showed recently that the dismal outcome of MCL might be improved in 28 young patients (pts) (<61 years) by an up-front Rituximab supplemented high-dose sequential chemotherapy (R-HDS) supported by stem cell autograft (ASCT) (A.M. Gianni et al, Blood, 2003). Following this encouraging experience we treated other 26 pts…
We previously published the results of a pilot study showing that vaccination with tumor-loaded d... more We previously published the results of a pilot study showing that vaccination with tumor-loaded dendritic cells (DCs) induced both T and B cell response and produced clinical benefit in the absence of toxicity in patients with relapsed, indolent non-Hodgkin lymphoma (iNHL). The purpose of the present short report is to provide a 15-year follow-up of our study and to expand the biomarker analysis previously performed. The long-term follow-up highlighted the absence of particular or delayed toxicity and the benefit of active immunization with DCs loaded with autologous, heat-shocked and UV-C treated tumor cells in relapsed iNHL (5-year and 10-year progression-free survival (PFS) rates: 55.6% and 33.3%, respectively; 10-year overall survival (OS) rate: 83.3%). Female patients experienced a better PFS (p=0.016) and a trend towards a better OS (p=0.185) compared with male patients. Of note, we observed a non-negligible fraction of patients (22%) who experienced a long-lasting complete re...
Triple-negative breast cancer (TNBC) is an aggressive disease with poor prognosis and limited the... more Triple-negative breast cancer (TNBC) is an aggressive disease with poor prognosis and limited therapeutic options. Recent advances in the immunotherapy field have enabled the development of new treatment strategies, among which the use of bispecific antibodies (BsAbs), able to redirect T cells against tumors, has shown promising results. In particular, a BsAb that uses TNF-related apoptosis-inducing ligand receptor 2 (TRAIL-R2) as a target was constructed and demonstrated good results in redirecting CD3+ T cells to kill TRAIL-R2-expressing TNBC cells. In the present study, we investigated whether treatment with selinexor, a selective inhibitor of nuclear export (SINE) targeting exportin-1/chromosome maintenance protein 1 (XPO1/CRM1), could potentiate the antitumor activity of this BsAb. In combination experiments, we found that selinexor-exposed TNBC cells exhibited greater growth inhibition when treated with the TRAIL-R2xCD3 BsAb than that expected by simple additivity. Similarly, ...
Molecular remission (MR) is associated with improved outcome in mantle cell lymphoma (MCL). If MR... more Molecular remission (MR) is associated with improved outcome in mantle cell lymphoma (MCL). If MR is not achieved, patients are at high risk of relapse. We retrospectively describe the molecular and clinical follow-ups of 4 patients with molecular relapses (M-rels) who were treated with rituximab. The 4 patients received rituximab-supplemented, high-dose sequential chemotherapy and autologous stem cell transplantation as induction treatment and achieved clinical remission and MR. M-rel was defined as polymerase chain reaction (PCR) positivity in 2 consecutive samples in the absence of clinical relapse. M-rels occurred at 3, 6, 39, and 52 months and were always confirmed by direct sequencing of the clonal rearrangement. Minimal residual disease was monitored by qualitative and real-time quantitative PCR. All patients received 4 courses of rituximab, with 2 additional infusions if PCR positivity remained. After 4-6 courses of rituximab, all patients re-entered MR. No clinical relapses...
Background: Early interim-PET (PET-2) is the most powerful factor able to predict treatment outco... more Background: Early interim-PET (PET-2) is the most powerful factor able to predict treatment outcome in advanced-stage Hodgkin Lymphoma (HL) patients treated with ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine). The 2-y PFS of PET-2 positive patients is only 12%, but the optimal treatment for this patient subset is still unknown. For this reason in January 2006 a treatment policy was designed by GITIL (Gruppo Italiano Terapie Innovative nei Linfomi) to early intensify chemotherapy with BEACOPP (bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone) for HL patients with a PET-2 positive after 2 ABVD cycles. Patients and methods: 136 HL patients with advanced-stage or intermediate-stage with adverse prognostic factors (more than 3 nodal sites, ESR > 50 mm, sub diaphragmatic presentation, bulky lesion), consecutively admitted to nine GITIL Italian centers were treated with two ABVD courses and re-evaluated with interim-PET. Twenty-...
Dissecting the genetics of classical Hodgkin lymphoma (cHL), a common lymphoma derived from germi... more Dissecting the genetics of classical Hodgkin lymphoma (cHL), a common lymphoma derived from germinal center B (GCB) cells, has remained a difficult task compared to other lymphomas, due to the rarity (usually <5%) of tumor cells (the Hodgkin and Reed/Sternberg - HRS - cells) in the lymph node. Thus, we microdissected 1200-1800 single HRS cells per case from frozen biopsies of 36 patients, along with a similar number of adjacent non-neoplastic cells, and performed in duplicate whole-genome DNA amplification (WGA) followed by independent whole-exome sequencing, thus allowing to control for possible biases introduced by the WGA step. The putative somatic protein-changing mutations identified in a major tumor clone (i.e., ≥20% variant allele frequency) were then validated by targeted deeper sequencing of the same WGA DNA, as well as by ultra-deep sequencing and digital PCR of unamplified whole-biopsy DNA. We found highly recurrent (11/36 patients; 31%), heterozygous, STAT6 missense m...
From January 1986, 205 consecutive HL patients (p) were enrolled in a multicenter observational s... more From January 1986, 205 consecutive HL patients (p) were enrolled in a multicenter observational study to explore the role of HDS followed by ASCT in refractory or relapsed HL (refHL/relHL). Patients and methods: The mean age was 32.4 (11–61), the male/female ratio was 104/101. Limited (I–II) or advanced stage (III–IV) was present in 87 and 118 p, respectively. The first-line chemotherapy was ABVD (93 p), hybrid MOPP/ABVD (8p), alternating MOPP/ABVD (47p), other (37p), two or more chemotherapy lines (20 p). Radiotherapy (involved or extended field) was given in 95 p. 58 p had refHL, and 147 relHL: 130/147 were in first and 17/147 were in 2nd or more than 2nd relapse, respectively. The first relapse occurred before-(e-Rel) or later than (l-Rel) 12 months in 31 and 99 p, respectively. The standard HDS included DHAP (optional), Cyclophosphamide 7 gr./m2, Methotrexate 8 gr/m2, Vincristine 1.4 mg/m2, Etoposide 2 gr/m2. In 58/205 p. Metotrexate was substituted with ARA-C, 4gr/m2 per 4 days...
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is expected to play a key role in... more Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is expected to play a key role in cancer therapy due to its high cancer cell-specificity and potent antitumor activity. We have previously demonstrated that CD34+ cells transduced with an adenovirus encoding the full-length human TRAIL gene (CD34-TRAIL+) exert a potent anti-lymphoma effect in NOD/SCID mice. To investigate the mechanism of action of CD34-TRAIL+ cells, in vivo experiments were perfomed to analyze the tumor homing capacity of CD34-TRAIL+ cells and the effects of transduced cells on tumor vasculature. Tumor homing of CD34-TRAIL+ cells was investigated in NOD/SCID mice bearing subcutaneous lymphoid tumors. Following a single intravenous injection of CD34-TRAIL+ cells (5 x 106), nodules were excised at different time-points and immunostained with an anti-human CD45 antibody. Sections were digitally recorded and the total number of CD45+ cells per tissue section was then counted using the imaging software Imag...
519 Introduction High-dose (hd) therapy with stem cell autograft is an effective treatment for bo... more 519 Introduction High-dose (hd) therapy with stem cell autograft is an effective treatment for both non-Hodgkin's (NHL) and Hodgkin's Lymphoma (HL). However, the occurrence of secondary malignancies, particularly myelodysplastic syndromes/acute leukemias (sMDS/AL), is a critical issue, representing a major cause of failure in patients potentially cured after hd-chemotherapy. Aim of the study To evaluate: frequency-cumulative incidence-risk factors, of both sMDS/AL and solid tumors in a large series of lymphoma patients, treated with the hd-sequential (HDS) chemotherapy approach, followed by peripheral blood progenitor cell (PBPC) autograft. Patients and Methods Data have been collected on 1,347 lymphoma patients treated in the last two decades at 11 Centers, associated to GITIL. Patients received either the original or modified HDS regimens; PBPC were collected after hd-cyclophosphamide, and, in a subgroup, after a 2nd round of mobilization with hd-Ara-C. The series included...
Recent trials have shown that anti-CD20 monoclonal antibody Rituximab may be effectively employed... more Recent trials have shown that anti-CD20 monoclonal antibody Rituximab may be effectively employed in association with high-dose (hd) chemotherapy and peripheral blood progenitor cell (PBPC) autograft in the management of high-risk B-cell lymphoma. Addition of Rituximab has a dual effect: increased tumour cytoreduction and in vivo purging prior to PBPC harvesting. We here report the results of a prospective, multicenter trial evaluating Rituximab-supplemented hd-sequential chemotherapy (R-HDS) as frontline treatment in patients with high-risk Diffuse Large B-Cell Lymphoma (DLBCL). So far, 6 Italian Centres associated to GITIL have participated to the study. Eligibility criteria included: i. biopsy-proven DLBCL, with CD20+ phenotype; ii. no previous cytotoxic treatments; iii. age between 16–60 yrs.; iv. advanced stage disease with 2–3 aaIPI score. The R-HDS regimen includes an initial debulkying (3 APO courses) and then the sequential administration, at 15–20 day intervals, of: i. cyc...
1653 Introduction: A significant proportion of Hodgkin lymphoma (HL) patients refractory to first... more 1653 Introduction: A significant proportion of Hodgkin lymphoma (HL) patients refractory to first-line chemotherapy or relapsing after autologous transplantation are not cured with currently available treatments and require new treatments. The PI3K/AKT and RAF/MEK/ERK pathways are constitutively activated in the majority of HL. These pathways can be targeted using the AKT inhibitor perifosine (Æterna Zentaris GmBH, Germany, EU), and the RAF/MEK/ERK inhibitor sorafenib (Nexavar®, Bayer, Germany, EU). We hypothesized that perifosine in combination with sorafenib might have a therapeutic activity in HL by overcoming the cytoprotective and anti-apoptotic effects of PI3K/Akt and RAF/MEK/ERK pathways. Since preclinical evidence supporting the anti-lymphoma effects of the perifosine/sorafenib combination are still lacking, the present study aimed at investigating in vitro and in vivo the activity and mechanism(s) of action of this two-drug combination. METHODS: Three HL cell lines (HD-MyZ,...
Background: Lymphomas are increasing in frequency and represent the fifth most common cancer diag... more Background: Lymphomas are increasing in frequency and represent the fifth most common cancer diagnosis in the United States. 1D09C3 is a fully human anti-HLA-DR monoclonal antibody (mAb) derived from a human combinatorial antibody library (HuCAL®) that has consistently demonstrated activity against various lymphoid tumors, both in vitro and in vivo (Nat Med2002;8:801–7). Therefore 1D09C3 may offer a novel therapy to patients with B-cell lymphoma who have failed therapy. Two phase I studies are ongoing to determine i) the maximum tolerated dose (MTD) and phase II recommended dose (RD), ii) the pharmacodynamic (pd)- and pharmacokinetic (pk)-profile and iii) the immunogenicity of 1D09C3 given within a bi-weekly and weekly schedule in this patient population. Material and methods: Patients with relapsed/refractory B-cell lymphoproliferative diseases are included in the two studies. 1D09C3 dose escalation is performed using modified Fibonacci increments with a minimum of 3 patients (pts)...
This is an update of the GITMO-IIL trial comparing R-HDS and CHOP-R in high-risk FL <60 years.... more This is an update of the GITMO-IIL trial comparing R-HDS and CHOP-R in high-risk FL <60 years. The whole patient (pt) population is now evaluable for analysis with a median follow-up of 36 months. Eligibility was based on age-adjusted IPI ≥2 (125 pts) or according to the Italian Lymphoma Intergroup score ≥3 (11 pts).136 pts were stratified according to histology (grade I or II 101, grade III 35) and randomized (68 each). Clinical features were: median age 50 yrs. (22–60), stage III–IV 98%, elevated LDH 78%, bulky disease 66%, B symptoms 47%, extranodal disease (other than BM) 45%, ECOG PS >1 47%. R-HDS has been already described (Ladetto et al ASH 2005). The CHOP-R arm consisted of CHOP and Rituximab delivered sequentially as already published (Rambaldi et al Blood 2002). Cross-over was allowed for pts failing CHOP-R. Centralized minimal residual disease (MRD) analysis with the bcl-2/IgH was planned on BM cells. Analysis was “intention to treat”. Toxic deaths were 4 (2 in each...
550 Background: Interim FDG-PET performed after 2 chemotherapy courses (PET-2) is the most powerf... more 550 Background: Interim FDG-PET performed after 2 chemotherapy courses (PET-2) is the most powerful predictor of treatment outcome in advanced-stage, ABVD-treated HL (AAHL) patients (pts). Whether early therapy intensification in PET-2 positive HL pts improves overall treatment efficacy for all AAHL pts compared to standard ABVD is still unproven. Patients and study design: In the HD 0607 clinical trial (ClinicalTRial.gov identifier 00795613), AHL (stage IIB-IVB) pts prospectively enrolled by 26 Italian and one Israelian institution are treated with 2 ABVD courses and a PET-2 performed afterwards. PET-2+ pts are randomized to either BEACOPP escalated (Be) plus BEACOPP baseline (Bb) (4+4 courses) or Be+Bb (4+4) and Rituximab (R). PET-2 negative pts are treated with 4 additional ABVD and, upon CR achievement, randomized to either consolidation radiotherapy (Rxt) on the sites of initial bulky disease or no further treatment. Primary endpoint is the efficacy and feasibility of the overa...
3711 INTRODUCTION: Patients with refractory or relapsed classical Hodgkin Lymphoma (cHL) represen... more 3711 INTRODUCTION: Patients with refractory or relapsed classical Hodgkin Lymphoma (cHL) represent an unmet medical need and would benefit from the development of new therapies. Histone deacetylases (HDACs) and the RAF/MEK/ERK pathway are aberrantly controlled in cHL and influence a broad repertoire of tumor processes, suggesting a rationale for therapeutically targeting these pathways. We targeted these pathways using the HDAC inhibitor Givinostat (Italfarmaco S.p.A., Milan, Italy), and the RAF/MEK/ERK inhibitor Sorafenib (Nexavar, Bayer, Germany, EU) in order to investigate in vitro and in vivo the activity and mechanism(s) of action of this two-drug combination. METHODS: Three cHL cell lines, including HDLM-2, L-540 and HD-MyZ, were used to investigate the effects of Givinostat and Sorafenib, used alone or in combination, by means of in vitro assays evaluating cell growth and cell survival. Additionally, live cell imaging was used to asses the production of reactive oxygen specie...
4547 Background: Knowledge of the expression of molecular drivers and potentially druggable targe... more 4547 Background: Knowledge of the expression of molecular drivers and potentially druggable targets may enhance prognostic classification of metastatic UC. We aimed at assessing the expression of multiple key molecular biomarkers (BMK) by IHC and their potential to enhance prognostic allocation of patients (pts) with UC. Methods: We analyzed formalin-fixed paraffin embedded tumor from pts with UC undergoing first-line CT with MVAC for locally-advanced unresectable (LA, T3-4±N+) and metastatic (M) disease between the years 2000 and 2013. The expression of the following panel of BMKs by IHC were evaluated using conventional protocols: ERCC1, EGFR, HER2, VEGFR-3, PDGFRα, p53, p63. Expression levels were dichotomized as positive (2+,3+) or negative (≤1+). Fisher exact test was used to evaluate the association with response and setting (LA vs M). Cox regression multivariate (MVA) models evaluated the association with PFS and OS of each biomarker, adjusted for recognized prognostic variables (setting [LA vs M],...
Dysregulated apoptosis plays a key role in the pathogenesis and progression of neoplastic disorde... more Dysregulated apoptosis plays a key role in the pathogenesis and progression of neoplastic disorders, allowing tumor cells to survive beyond their normal life-span, and to eventually acquire chemo-radioresistance (Laconi et al., 2000; Pommier et al., 2004). Thus, targeting either the intrinsic or the extrinsic pathways of apoptosis represent attractive therapeutic strategies for restoring apoptosis sensitivity of malignant cells, or activating agonists of apoptosis (Waxman & Schwartz, 2003). Due to the ability of death receptor ligands to induce cell death, there has been considerable interest in the physiological roles and therapeutic potential of these cytokines as anti-cancer agents. Death receptor ligands of the tumor necrosis factor α (TNFα) superfamily are type II transmembrane proteins that signal to target cells upon cell-cell contact, or after protease-mediated release to the extracellular space (Ashkenazi, 2002). Members of this family, including Fas ligand (FasL), TNFα, an...
Background The outcome of Diffuse Large B Cell Lymphoma (DLB-CL) has been definitely improved by ... more Background The outcome of Diffuse Large B Cell Lymphoma (DLB-CL) has been definitely improved by the addition of the anti-CD20 rituximab to conventional chemotherapy. However, despite the advantages offered by rituximab, results are still disappointing in patients presenting with adverse prognostic factors. For these patients, rituximab-supplemented intensive programs with autograft might represent a suitable option. Thus, a prospective multicenter study has been performed using first-line the Rituximab-supplemented high-dose sequential chemotherapy program delivered with multiple autologous peripheral blood progenitor cell (PBPC) support (R-HDS-maps regimen), in DLB-CL patients with score 2 or 3 according to the age-adjusted International Prognostic Index (aaIPI). Preliminary results have been already reported (ASH 2005; Leukemia2007, 21: 1802). Updated results after a prolonged follow-up are here presented. Methods. R-HDS-maps includes: 3 APO courses; sequential administration of ...
We showed recently that the dismal outcome of MCL might be improved in 28 young patients (pts) (&... more We showed recently that the dismal outcome of MCL might be improved in 28 young patients (pts) (<61 years) by an up-front Rituximab supplemented high-dose sequential chemotherapy (R-HDS) supported by stem cell autograft (ASCT) (A.M. Gianni et al, Blood, 2003). Following this encouraging experience we treated other 26 pts…
We previously published the results of a pilot study showing that vaccination with tumor-loaded d... more We previously published the results of a pilot study showing that vaccination with tumor-loaded dendritic cells (DCs) induced both T and B cell response and produced clinical benefit in the absence of toxicity in patients with relapsed, indolent non-Hodgkin lymphoma (iNHL). The purpose of the present short report is to provide a 15-year follow-up of our study and to expand the biomarker analysis previously performed. The long-term follow-up highlighted the absence of particular or delayed toxicity and the benefit of active immunization with DCs loaded with autologous, heat-shocked and UV-C treated tumor cells in relapsed iNHL (5-year and 10-year progression-free survival (PFS) rates: 55.6% and 33.3%, respectively; 10-year overall survival (OS) rate: 83.3%). Female patients experienced a better PFS (p=0.016) and a trend towards a better OS (p=0.185) compared with male patients. Of note, we observed a non-negligible fraction of patients (22%) who experienced a long-lasting complete re...
Triple-negative breast cancer (TNBC) is an aggressive disease with poor prognosis and limited the... more Triple-negative breast cancer (TNBC) is an aggressive disease with poor prognosis and limited therapeutic options. Recent advances in the immunotherapy field have enabled the development of new treatment strategies, among which the use of bispecific antibodies (BsAbs), able to redirect T cells against tumors, has shown promising results. In particular, a BsAb that uses TNF-related apoptosis-inducing ligand receptor 2 (TRAIL-R2) as a target was constructed and demonstrated good results in redirecting CD3+ T cells to kill TRAIL-R2-expressing TNBC cells. In the present study, we investigated whether treatment with selinexor, a selective inhibitor of nuclear export (SINE) targeting exportin-1/chromosome maintenance protein 1 (XPO1/CRM1), could potentiate the antitumor activity of this BsAb. In combination experiments, we found that selinexor-exposed TNBC cells exhibited greater growth inhibition when treated with the TRAIL-R2xCD3 BsAb than that expected by simple additivity. Similarly, ...
Molecular remission (MR) is associated with improved outcome in mantle cell lymphoma (MCL). If MR... more Molecular remission (MR) is associated with improved outcome in mantle cell lymphoma (MCL). If MR is not achieved, patients are at high risk of relapse. We retrospectively describe the molecular and clinical follow-ups of 4 patients with molecular relapses (M-rels) who were treated with rituximab. The 4 patients received rituximab-supplemented, high-dose sequential chemotherapy and autologous stem cell transplantation as induction treatment and achieved clinical remission and MR. M-rel was defined as polymerase chain reaction (PCR) positivity in 2 consecutive samples in the absence of clinical relapse. M-rels occurred at 3, 6, 39, and 52 months and were always confirmed by direct sequencing of the clonal rearrangement. Minimal residual disease was monitored by qualitative and real-time quantitative PCR. All patients received 4 courses of rituximab, with 2 additional infusions if PCR positivity remained. After 4-6 courses of rituximab, all patients re-entered MR. No clinical relapses...
Background: Early interim-PET (PET-2) is the most powerful factor able to predict treatment outco... more Background: Early interim-PET (PET-2) is the most powerful factor able to predict treatment outcome in advanced-stage Hodgkin Lymphoma (HL) patients treated with ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine). The 2-y PFS of PET-2 positive patients is only 12%, but the optimal treatment for this patient subset is still unknown. For this reason in January 2006 a treatment policy was designed by GITIL (Gruppo Italiano Terapie Innovative nei Linfomi) to early intensify chemotherapy with BEACOPP (bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone) for HL patients with a PET-2 positive after 2 ABVD cycles. Patients and methods: 136 HL patients with advanced-stage or intermediate-stage with adverse prognostic factors (more than 3 nodal sites, ESR > 50 mm, sub diaphragmatic presentation, bulky lesion), consecutively admitted to nine GITIL Italian centers were treated with two ABVD courses and re-evaluated with interim-PET. Twenty-...
Dissecting the genetics of classical Hodgkin lymphoma (cHL), a common lymphoma derived from germi... more Dissecting the genetics of classical Hodgkin lymphoma (cHL), a common lymphoma derived from germinal center B (GCB) cells, has remained a difficult task compared to other lymphomas, due to the rarity (usually <5%) of tumor cells (the Hodgkin and Reed/Sternberg - HRS - cells) in the lymph node. Thus, we microdissected 1200-1800 single HRS cells per case from frozen biopsies of 36 patients, along with a similar number of adjacent non-neoplastic cells, and performed in duplicate whole-genome DNA amplification (WGA) followed by independent whole-exome sequencing, thus allowing to control for possible biases introduced by the WGA step. The putative somatic protein-changing mutations identified in a major tumor clone (i.e., ≥20% variant allele frequency) were then validated by targeted deeper sequencing of the same WGA DNA, as well as by ultra-deep sequencing and digital PCR of unamplified whole-biopsy DNA. We found highly recurrent (11/36 patients; 31%), heterozygous, STAT6 missense m...
From January 1986, 205 consecutive HL patients (p) were enrolled in a multicenter observational s... more From January 1986, 205 consecutive HL patients (p) were enrolled in a multicenter observational study to explore the role of HDS followed by ASCT in refractory or relapsed HL (refHL/relHL). Patients and methods: The mean age was 32.4 (11–61), the male/female ratio was 104/101. Limited (I–II) or advanced stage (III–IV) was present in 87 and 118 p, respectively. The first-line chemotherapy was ABVD (93 p), hybrid MOPP/ABVD (8p), alternating MOPP/ABVD (47p), other (37p), two or more chemotherapy lines (20 p). Radiotherapy (involved or extended field) was given in 95 p. 58 p had refHL, and 147 relHL: 130/147 were in first and 17/147 were in 2nd or more than 2nd relapse, respectively. The first relapse occurred before-(e-Rel) or later than (l-Rel) 12 months in 31 and 99 p, respectively. The standard HDS included DHAP (optional), Cyclophosphamide 7 gr./m2, Methotrexate 8 gr/m2, Vincristine 1.4 mg/m2, Etoposide 2 gr/m2. In 58/205 p. Metotrexate was substituted with ARA-C, 4gr/m2 per 4 days...
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is expected to play a key role in... more Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is expected to play a key role in cancer therapy due to its high cancer cell-specificity and potent antitumor activity. We have previously demonstrated that CD34+ cells transduced with an adenovirus encoding the full-length human TRAIL gene (CD34-TRAIL+) exert a potent anti-lymphoma effect in NOD/SCID mice. To investigate the mechanism of action of CD34-TRAIL+ cells, in vivo experiments were perfomed to analyze the tumor homing capacity of CD34-TRAIL+ cells and the effects of transduced cells on tumor vasculature. Tumor homing of CD34-TRAIL+ cells was investigated in NOD/SCID mice bearing subcutaneous lymphoid tumors. Following a single intravenous injection of CD34-TRAIL+ cells (5 x 106), nodules were excised at different time-points and immunostained with an anti-human CD45 antibody. Sections were digitally recorded and the total number of CD45+ cells per tissue section was then counted using the imaging software Imag...
519 Introduction High-dose (hd) therapy with stem cell autograft is an effective treatment for bo... more 519 Introduction High-dose (hd) therapy with stem cell autograft is an effective treatment for both non-Hodgkin's (NHL) and Hodgkin's Lymphoma (HL). However, the occurrence of secondary malignancies, particularly myelodysplastic syndromes/acute leukemias (sMDS/AL), is a critical issue, representing a major cause of failure in patients potentially cured after hd-chemotherapy. Aim of the study To evaluate: frequency-cumulative incidence-risk factors, of both sMDS/AL and solid tumors in a large series of lymphoma patients, treated with the hd-sequential (HDS) chemotherapy approach, followed by peripheral blood progenitor cell (PBPC) autograft. Patients and Methods Data have been collected on 1,347 lymphoma patients treated in the last two decades at 11 Centers, associated to GITIL. Patients received either the original or modified HDS regimens; PBPC were collected after hd-cyclophosphamide, and, in a subgroup, after a 2nd round of mobilization with hd-Ara-C. The series included...
Recent trials have shown that anti-CD20 monoclonal antibody Rituximab may be effectively employed... more Recent trials have shown that anti-CD20 monoclonal antibody Rituximab may be effectively employed in association with high-dose (hd) chemotherapy and peripheral blood progenitor cell (PBPC) autograft in the management of high-risk B-cell lymphoma. Addition of Rituximab has a dual effect: increased tumour cytoreduction and in vivo purging prior to PBPC harvesting. We here report the results of a prospective, multicenter trial evaluating Rituximab-supplemented hd-sequential chemotherapy (R-HDS) as frontline treatment in patients with high-risk Diffuse Large B-Cell Lymphoma (DLBCL). So far, 6 Italian Centres associated to GITIL have participated to the study. Eligibility criteria included: i. biopsy-proven DLBCL, with CD20+ phenotype; ii. no previous cytotoxic treatments; iii. age between 16–60 yrs.; iv. advanced stage disease with 2–3 aaIPI score. The R-HDS regimen includes an initial debulkying (3 APO courses) and then the sequential administration, at 15–20 day intervals, of: i. cyc...
1653 Introduction: A significant proportion of Hodgkin lymphoma (HL) patients refractory to first... more 1653 Introduction: A significant proportion of Hodgkin lymphoma (HL) patients refractory to first-line chemotherapy or relapsing after autologous transplantation are not cured with currently available treatments and require new treatments. The PI3K/AKT and RAF/MEK/ERK pathways are constitutively activated in the majority of HL. These pathways can be targeted using the AKT inhibitor perifosine (Æterna Zentaris GmBH, Germany, EU), and the RAF/MEK/ERK inhibitor sorafenib (Nexavar®, Bayer, Germany, EU). We hypothesized that perifosine in combination with sorafenib might have a therapeutic activity in HL by overcoming the cytoprotective and anti-apoptotic effects of PI3K/Akt and RAF/MEK/ERK pathways. Since preclinical evidence supporting the anti-lymphoma effects of the perifosine/sorafenib combination are still lacking, the present study aimed at investigating in vitro and in vivo the activity and mechanism(s) of action of this two-drug combination. METHODS: Three HL cell lines (HD-MyZ,...
Background: Lymphomas are increasing in frequency and represent the fifth most common cancer diag... more Background: Lymphomas are increasing in frequency and represent the fifth most common cancer diagnosis in the United States. 1D09C3 is a fully human anti-HLA-DR monoclonal antibody (mAb) derived from a human combinatorial antibody library (HuCAL®) that has consistently demonstrated activity against various lymphoid tumors, both in vitro and in vivo (Nat Med2002;8:801–7). Therefore 1D09C3 may offer a novel therapy to patients with B-cell lymphoma who have failed therapy. Two phase I studies are ongoing to determine i) the maximum tolerated dose (MTD) and phase II recommended dose (RD), ii) the pharmacodynamic (pd)- and pharmacokinetic (pk)-profile and iii) the immunogenicity of 1D09C3 given within a bi-weekly and weekly schedule in this patient population. Material and methods: Patients with relapsed/refractory B-cell lymphoproliferative diseases are included in the two studies. 1D09C3 dose escalation is performed using modified Fibonacci increments with a minimum of 3 patients (pts)...
This is an update of the GITMO-IIL trial comparing R-HDS and CHOP-R in high-risk FL <60 years.... more This is an update of the GITMO-IIL trial comparing R-HDS and CHOP-R in high-risk FL <60 years. The whole patient (pt) population is now evaluable for analysis with a median follow-up of 36 months. Eligibility was based on age-adjusted IPI ≥2 (125 pts) or according to the Italian Lymphoma Intergroup score ≥3 (11 pts).136 pts were stratified according to histology (grade I or II 101, grade III 35) and randomized (68 each). Clinical features were: median age 50 yrs. (22–60), stage III–IV 98%, elevated LDH 78%, bulky disease 66%, B symptoms 47%, extranodal disease (other than BM) 45%, ECOG PS >1 47%. R-HDS has been already described (Ladetto et al ASH 2005). The CHOP-R arm consisted of CHOP and Rituximab delivered sequentially as already published (Rambaldi et al Blood 2002). Cross-over was allowed for pts failing CHOP-R. Centralized minimal residual disease (MRD) analysis with the bcl-2/IgH was planned on BM cells. Analysis was “intention to treat”. Toxic deaths were 4 (2 in each...
550 Background: Interim FDG-PET performed after 2 chemotherapy courses (PET-2) is the most powerf... more 550 Background: Interim FDG-PET performed after 2 chemotherapy courses (PET-2) is the most powerful predictor of treatment outcome in advanced-stage, ABVD-treated HL (AAHL) patients (pts). Whether early therapy intensification in PET-2 positive HL pts improves overall treatment efficacy for all AAHL pts compared to standard ABVD is still unproven. Patients and study design: In the HD 0607 clinical trial (ClinicalTRial.gov identifier 00795613), AHL (stage IIB-IVB) pts prospectively enrolled by 26 Italian and one Israelian institution are treated with 2 ABVD courses and a PET-2 performed afterwards. PET-2+ pts are randomized to either BEACOPP escalated (Be) plus BEACOPP baseline (Bb) (4+4 courses) or Be+Bb (4+4) and Rituximab (R). PET-2 negative pts are treated with 4 additional ABVD and, upon CR achievement, randomized to either consolidation radiotherapy (Rxt) on the sites of initial bulky disease or no further treatment. Primary endpoint is the efficacy and feasibility of the overa...
3711 INTRODUCTION: Patients with refractory or relapsed classical Hodgkin Lymphoma (cHL) represen... more 3711 INTRODUCTION: Patients with refractory or relapsed classical Hodgkin Lymphoma (cHL) represent an unmet medical need and would benefit from the development of new therapies. Histone deacetylases (HDACs) and the RAF/MEK/ERK pathway are aberrantly controlled in cHL and influence a broad repertoire of tumor processes, suggesting a rationale for therapeutically targeting these pathways. We targeted these pathways using the HDAC inhibitor Givinostat (Italfarmaco S.p.A., Milan, Italy), and the RAF/MEK/ERK inhibitor Sorafenib (Nexavar, Bayer, Germany, EU) in order to investigate in vitro and in vivo the activity and mechanism(s) of action of this two-drug combination. METHODS: Three cHL cell lines, including HDLM-2, L-540 and HD-MyZ, were used to investigate the effects of Givinostat and Sorafenib, used alone or in combination, by means of in vitro assays evaluating cell growth and cell survival. Additionally, live cell imaging was used to asses the production of reactive oxygen specie...
4547 Background: Knowledge of the expression of molecular drivers and potentially druggable targe... more 4547 Background: Knowledge of the expression of molecular drivers and potentially druggable targets may enhance prognostic classification of metastatic UC. We aimed at assessing the expression of multiple key molecular biomarkers (BMK) by IHC and their potential to enhance prognostic allocation of patients (pts) with UC. Methods: We analyzed formalin-fixed paraffin embedded tumor from pts with UC undergoing first-line CT with MVAC for locally-advanced unresectable (LA, T3-4±N+) and metastatic (M) disease between the years 2000 and 2013. The expression of the following panel of BMKs by IHC were evaluated using conventional protocols: ERCC1, EGFR, HER2, VEGFR-3, PDGFRα, p53, p63. Expression levels were dichotomized as positive (2+,3+) or negative (≤1+). Fisher exact test was used to evaluate the association with response and setting (LA vs M). Cox regression multivariate (MVA) models evaluated the association with PFS and OS of each biomarker, adjusted for recognized prognostic variables (setting [LA vs M],...
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