ALBERT

Description: The purification of multipotent mesenchymal stromal cell (MSC) from sub-cutaneous adipose tissue (SCAT) represents a promising approach for several clinical applications. The isolation procedures have been applied in humans only to fairly large amount (5–300 g) of harvested SCAT (Zuk PA et al., 2001; Van Harmelen V et al., 2004; Meyerrose TE et al., 2007). Thus, we tested whether it might be possible to isolate MSC from far smaller tissue volumes. SCAT specimens from healthy donors which underwent to plastic surgery operations (n=5; mean weight ± SD=116.3±91.9 mg) were processed with a protocol modified from Zuk et al. 2001. The released cells were filtered, centrifuged and seeded with expansion medium. Solely small, undifferentiated and adherent proliferating fibroblasts were soon evident in the cultures and cells reached the first confluence within 12.6±5.1 days. After 40 days of expansion, the SCAT-MSC displayed a robust proliferative potential, with a cell count of 1.32±0.53x108. FACS analyses showed that SCAT-MSC were positive (〉95%) for CD90, CD105, CD73 and negative (

Description: Background: Dasatinib (SPRYCEL®, formerly BMS-354825) is a novel, oral, multitargeted kinase inhibitor of BCR-ABL, SRC, and other kinases that is approximately 300 times more potent than Imatinib in vitro. Dasatinib has been shown to be effective and safe in pts with CML resistant or intolerant to Imatinib, As might be expected, older patients experienced more adverse events, both hematologic and non-hematologic with the standard dosage of 100 mg once daily. Data regarding tolerability in elderly pts are scanty The aim of this study is to test if escalated dasatinib dose up to 60 mg once daily is as effective as standard dose of 100 mg QD with a better toxicity profile in elderly patients with chronic myeloid leukemia in late chronic phase resistant to or intolerant of Imatinib. Patients and Methods: As of July 2007, 4 eligible patients have been enrolled and treated :4F ; median age 77 y [range 73–82]; 1 imatinib-resistant, 3 imatinib-intolerant). Dasatinib was given at 20 mg once daily (QD) starting dose with dose escalation to 40 mg QD or 60 mg QD in pts tolerant or lacking response. Complete blood counts were obtained weekly for the first 12 weeks; bone marrow cytology and cytogenetics every 1 months, and molecular monitoring of BCR-ABL transcript levels by real-time qPCR every 12 weeks. The primary endpoint was toxicity. Median time from diagnosis of CML was 62 months (range 36 – 88). Prior therapy included interferon-alpha in 1/4. 50 percent of pts had 400 mg of prior imatinib. 25% had 600 mg. 100 % of pts received imatinib for 〉3 yrs. Best response to prior imatinib therapy was a CHR in 100, and partial (PCyR) cytogenetic responses in 75 % of pts. No BCR-ABL baseline mutations were found. Results: The analyses with a median follow-up of 12 months (7–12) months show hematologic toxicity (grade 3 thrombocytopenia and leukopenia) at 60 mg OD in 2 out 4 pts. Dose interruptions occurred in 1/4 pts at 60 mg One pts was intolerant to dasatinib and switched to HU Patients received an average daily dose of 40 mg/day (range 20–60 mg). Non-hematologic toxicity consisted mainly of grade 1 diarrhea, headache 2 patients experienced grade II muscolo-skeletal toxicity and fatigue at 60 mg/day.) 100% pts had a CHR, and 50% a PCyR. Conclusions: Dasatinib demonstrated substantial hematologic activity in elderly patients with late CP-CML. Despite the maximum dose reached was 60 mg QD only the 75% of pts were able to continued the treatment. because of toxicity. The novel TKIs therapies constitute an important therapeutic challenge in this particularly subset of patients when compared with younger patients

Description: High-dose prophylaxis for severe hemophilia A [25–40 factor (F) VIII Units (u) kg−1 on alternate days] reduces arthropathy, but does so at great cost. (NEJM2007; 357: 535–544). Less intensive prophylaxis regimens might be cost-effective while reducing the need for indwelling venous access devices and their attendant complications. Tailored prophylaxis starts with FVIII, 50 u kg−1 weekly (Step1). If bleeding frequency is unacceptable (4 joint/soft tissue bleeds or 5 bleeds into a joint on a step of protocol), or if a target joint develops (≥3 bleeds into a single index joint-ankle, elbow or knee) over a consecutive 3-month period the dose of FVIII is escalated to 30 u kg−1 twice weekly (Step 2); a third escalation to 25 u kg−1 on alternate days if bleeding frequency remains unacceptable (Step3). Objectives: To assess the degree of joint damage in the 6 index joints for subjects receiving tailored prophylaxis. Methods: We studied an inception cohort of boys ages 1–2.5 years with severe hemophilia A (

Description: Abstract 3741 Poster Board III-677 LNH98.5 was the first randomized study with the addition of rituximab to CHOP in patients with diffuse large B-cell lymphoma. 399 patients were randomized, 197 in CHOP arm and 202 in R-CHOP arm. Patients were aged between 60 and 80 years (median 70 years), had disease stage II to IV, and no contra-indication to one of the drugs. 60% had poor risk lymphoma according to IPI. Response to treatment and early survival analyses were previously presented with 2 years and 5 years median follow-up (NEJM 2002;346:235 and JCO 2005;23:4117). With a median follow-up of 10 years, median age of surviving patients is 78 years, oldest patient being 91 years old. Only 4 patients were lost for follow-up, defined as not seen during the last 18 months, at 5, 7, 8, and 8 years. No event was observed in 105 of the 399 patients, 37 (19%) in CHOP arm and 68 (34%) in R-CHOP arm. Relapse was observed in 73 (59%) and 51 (34%) of CR patients, and death without progression in 16 and 33 patients, respectively. Death occurred in 71% and 56% of the patients, most of them from disease progression but 21 and 20 cancers were observed, representing half of the deaths without progression. Most frequent cancers were colon and lung; two MDS were observed in CHOP arm and one AML in R-CHOP arm. One patient with CHOP presented a multiple myeloma 10 years after DLBCL. During the last 3 years, 10 additional patients relapsed, 4 in CHOP arm and 6 in R-CHOP arm, these late relapses representing 4% of CR patients. Median overall survival was 37 months in CHOP arm and 7 y 9 m in R-CHOP arm with 10-y survival of 28% and 43%, respectively (p

Description: BACGROUND: Acquired aplastic anemia (AA) is thought to be an immune-mediated disease. Immunosuppressive therapy (IST) has been the treatment of choice for patients who did not have suitable donors. Previously, we had published promising results of IST for children with acquired AA. In the study, overall survival rate (OS) at 4 years was 83% in patients with vSAA and 92% in those with SAA/nonSAA. OBJECTIVES: Here we report follow-up results focusing on pediatric patients with relapse and clonal diseases, MDS/AML and PNH. PATIENTS and TREATMENT: From 1992 to 1997, 119 newly diagnosed children with acquired AA (median age; 9) entered AA-92 study. 50 vSAA patients were treated with ATG+CyA+mPSL+danazole +G-CSF, 36 SAA and 28 nonSAA patients were treated with ATG+CyA+mPSL+danazole +/−G-CSF. Complete remission (CR) was defined as a neutrophil count 〉1.5x10^9/L, a platelet count 〉100xx10^9/L, and a hemoblobin level of 〉11 g/dl. Partial response (PR) was defined as a neutrophil count 〉0.5x10^9/L, a platelet count 〉20x10^9/L, and a hemoglobin level of 〉8.0 g/dl. Relapse was indicated by the return of the PB counts to levels meeting the definition of SAA and/or the requirement for blood transfusion. Response rate was 71% at 6 months in vSAA patients, 65% in SAA/nonSAA patients, respectively. There was no statistical difference in 6-month response rate between G-CSF +/− treatments. No patient responded after 6 months. Therefore, 75 responders and 29 non-responders at 6 months were analyzed their OS, relapse rate (RR), and treatment-failure-free survival (TFFS). The median observation time of surviving patients is 118 months, ranging from 75 to 168 months. RESULTS: Among 119 patients, 38 patients received BMT and 17 patients died during an observation period. The OS was 81.9+−3.8% at 10 years, but has not reached plateau. Of the 75 6mo-responders, 23 patients relapsed and the RR was 33.6+−6.0% at 10 years. TFFS of the 75 was 67.4+−5.5%. Fourteen patients received 2nd ATG therapy and 5 of them responded. Eleven of the 23 patients with relapse received alternative donor BMT and 8 are alive. 10y-OS of these relapsed patients was 73.4+−9.4%. Nineteen of the 29 6mo-non-responders received alternative donor BMT and 15 are alive. There is no statistically significant difference in 10y-OS between the responders and the non-responders (89.1+−3.7% vs. 75.0+−8.2%, p=0.09). New clonal abnormalities appeared in 9 of 119 patients (10.0+−3.2%KM probability): monosomy 7(3 patients), trisomy 8(3 patients), trisomy 9, trisomy11, del (13)(1 patient each). There is no statistically difference in 10y-OS, RR, and incidence of clonal abnormality between randomized G-CSF+/− treatments. Clinical PNH with symptomatic hemolysis developed in 3 of the responder patients, 96, 105 and 138 months after the AA diagnosis, respectively. Among 65 surviving responders, 33(51%) have CR and 26(40%) PR at last follow-up time. CONCLUSIONS: Our data demonstrate that IST is effective for children with acquired AA, but relapse are common. Effective 2nd line treatment should be developed. PNH developed even in pediatric AA patients during long-term observation.

Description: Despite the mature results of the new intensive treatments, ABVD is still considered the standard therapy of the HL. Here we report the 2-year results of a phase 2 study which explore the possibility to ameliorate the performance of this (g)old schedule. Primary End-Points: CR and early cardio-toxicity. Secondary End-Points: survival (FFP,OS) and late cardio-toxicity. 23/25 CR (CI 90%) to validate the study (80% standard vs 90% DD-DI ABVD, CI +/− 10% one tail test, 5% sig stat level). Modifications of the standard (st) ABVD and strategy concepts are summarized as follow: Each patient received a total of 6 cycles without RT Adryamicin (ADM) was escalated from 50 to 70 mg/m2 in the cycles 1,2,3,4. The cumulative dose of ADM was 380 mg/m2. The intercycle period was shortened from 28 to 21 days for all 6 cycles; the 4 drugs were delivered at day 1 and 11 of each cycle G-CSF was given from d4 to d8, and from d14 to d18 of each cycle The therapy program was driven by dynamic indicators such as interim-PET and CT fusion images. Normalisation of PET images at the end of 2nd cycles was indicator of early CR, while the persistence of PET+ lesion(s) at the end of the 4th cycle was indicator of failure and consequently the treatment was shift to a high-dose CT/CD34+ rescue program. RT was eliminated. At the end of 6th cycle, PET- residual masses were monitored by PET/TC fusion images. An historical group of 94 HL pts treated with 6–8 cycles of st-ABVD +/− RT was used to compare response, survival and toxicity data. Compared to st-ABVD the theoretical relative dose-intensity (RDI) values of drugs were 1.86 for ADM, 1.33 for DTIC, Bleomycin and Vinblastine, and 1.46 for the entire schedule. Administered RDIs of the schedule (delivered cycles = 184) ranged between 1.10 to 1.55 (median value 1.39). Results: from June 2004 to August 2006 thirty-nine pts were enrolled (tab1). As the 8th August 2006 thirty-five out of 39 pts have performed the interim-PET-TC fusion images and thirty-two completed the treatment: 100% of valuable pts obtained the early CR (35/35) and CR (32/32). All pts are alive and free of disease (tab2 and fig.1). Cardiac functions were monitored by ECG, Ecography, MUGA scan, pro-BNP, and troponin T. Toxicity was mild and similar to standard ABVD. Dose-dense and dose-intense ABVD is feasible, well tolerated and highly active in newly diagnosed patients with advanced/intermediated Hodgkin’s lymphoma: six cycles without RT seem superior to 6–8 cycles of ABVD +/− RT (fig 1) and may be a promising program for optimal long-term results. tab1 presentation features Features total 39 100% Early unfavourable 9 23% Advanced 30 77% Age 〉 45 –yr 4 10% Bulky 19 44% Stage IV 13 33% B symptoms 25 64% Extranodal 9 23% VES 〉 50 mm 20 51% LDH Ratio 〉 1 15 38% IPS ≥ 3 10 26% N. Sites〉3 28 72% Male 13 33% tab.2 Response and Survival N. of patients percent *n°. of valuable pts as 8 August 2006 Early CR 35/35* 100% CR 32/32* 100% FFP-2yr 32/32* 100% OS-2yr 32/32* 100% Freedom From Progression Freedom From Progression

Description: Introduction: We previously reported that plerixafor + G-CSF was as safe as and more effective than placebo + G-CSF in mobilizing hematopoietic stem cell for autologous transplant in patients with multiple myeloma (MM) through 100 days follow-up (DiPersio ASH 2007). We report herein the 12 months data. Methods: This is a phase 3, multi-center, randomized (1:1), double-blind, placebocontrolled study. Multiple myeloma patients requiring an autologous hematopoietic stem cell transplant, in first or second complete or partial remission were eligible. Patients received G-CSF (10μg/kg/day) subcutaneously for up to 8 days. Beginning on evening of Day 4 and continuing daily for up to 4 days, patients received either plerixafor (240μg/kg) or placebo subcutaneously. Starting on Day 5, patients began daily apheresis for up to 4 days or until ≥ 6 x 106 CD34+cells/kg were collected. We report herein the 12 months graft durability and hematology data. Results: As reported previously, the primary endpoint of collecting ≥ 6 x 106 CD34+ cells/kg in ≤ 2 days of apheresis was met in 106/148 (71.6%) patients in the plerixafor group and 53/154 (34.4%) patients in the placebo group, p

Description: Abstract 3356 Poster Board III-244 Background: ALL accounts for approximately 70-83% of all childhood leukemia. High risk ALL is a big problem in our country and allogeneic related HSCT has proven to be a curative treatment for high risk patients with ALL. Patients and Methods: We analyzed the clinical data of 51 children (27 boys and 24 girls) with ALL undergoing related BMT/PSCT from Edgardo Rebagliati Hospital between 1996 and 2008. Survival probabilities were analyzed by Kaplan and Meier method. Outcome of different subgroups was compared by the log-rank test. Results: The recipients median age at transplantation was 10.7 years (range, 1 - 18 years). The disease status at conditioning were 1st CR (n=33), 2nd CR or more (n=18). The median interval from diagnosis to BMT was 14 months (range, 4 - 93 months). We have used two conditioning regimen: TBI/CTX/VP16 (TBI-1200Gy/CTX-120 mg/kg/ VP16-40mg/kg) in 34 patients (66.7%) or TBI/CTX 16 (31.4%) and 1 patient (2%) standard BuCy conditioning regimen. Graft-versus-host disease (GVHD) prophylaxis was methotrexate and cyclosporin. The median infused CD34 cell number was 6.21×106/kg (range 1.61-14.9×106/kg). The resource of hematopoietic cell was bone marrow 33.3% (17 patients), peripheral blood, 64.7% (33 patients) and both 2% (1 patient). Acute GVHD occurred in 10 (19.6%) patients, and chronic GVHD occurred in 15 (29.4%) patients. Twelve year overall survival (OS) is 59.8% ± 8.7% (n= 51) and leukemia-free survival probabilities (DFS) were 62.9% ± 8.2%. Patients with 1st CR and 2nd CR had a 5 year EFS of 60.1% and 67.3% (log-rank test, P 0.92), respectively. The mortality non related to transplant was 29.5%; 10 patients (19.6%) due to progressive disease and five patients (9.9%) by sepsis. Conclusions: Allo-SCT is well tolerated in high risk ALL children with prolonged OS and DFS. Not found statistical significance on OS and DFS between the conditioning regimen used, between status pre-transplant (1st CR vs 2nd CR) and we found statistical significance in favor to PB as a source of SC on DFS. Disclosures: No relevant conflicts of interest to declare.

Description: Radioimmunotherapy (RIT) options for T-cell non-Hodgkin lymphomas (T-NHLs) are limited. We evaluated anti-CD45-RIT in human (h) and murine (m) T-NHL. CD45 was highly expressed on hT-NHL patient samples (median, 2.3 × 105 antigen-binding capacity units/cell) and hT-NHL cell lines (3.4 × 105 CD45 antigen-binding capacity units/cell). Biodistribution studies in hTNHL xenografts showed that 131I-labeled BC8 (anti-hCD45) delivered 154% (P = .01) and 237% (P = .002) more radioiodine to tumor sites over control antibodies at 24 hours and 48 hours, respectively. Importantly, tumor sites targeted with 131I-BC8 exhibited 2.5-fold (P = .05), 3.0-fold (P = .007), and 3.6-fold (P = .07) higher 131I retention over the nontarget organs of lungs, liver, and kidneys, respectively (24 hours). Because the clinical use of anti-hCD45 would target both T-NHL and other hematolymphoid tissues, we evaluated the ability of anti-mCD45 to target mT-NHL. mT-NHL grafts targeted with anti-mCD45 correspondingly retained 5.3 (P 〈 .001), 5.4 (P 〈 .001), and 8.7 (P 〈 .001) times the radioactivity in tumor sites compared with nontarget organs of lung, liver, and kidney. 131I-labeled BC8 therapy yielded improved complete remission rates (75% vs 0%, P 〈 .001) and progression-free survivals (median, 23 days vs 4.5 days, P 〈 .001) compared with controls. These data indicate that the high CD45 expression of T-NHL allows reliable tumor targeting and disease control supporting anti-CD45 RIT for T-NHL patients.

Description: In an attempt to improve outcomes for patients with acute myeloid leukemia (AML) after allogeneic hematopoietic cell transplantation (HCT), we conducted a phase 1/2 study in which targeted irradiation delivered by 131I–anti-CD45 antibody was combined with targeted busulfan (BU; area-under-curve, 600-900 ng/mL) and cyclophosphamide (CY; 120 mg/kg). Fifty-two (88%) of 59 patients receiving a trace 131I-labeled dose of 0.5 mg/kg anti-CD45 murine antibody had higher estimated absorbed radiation in bone marrow and spleen than in any other organ. Forty-six patients were treated with 102 to 298 mCi (3774-11 026 MBq) 131I, delivering an estimated 5.3 to 19 (mean, 11.3) Gy to marrow, 17-72 (mean, 29.7) Gy to spleen, and 3.5 Gy (n = 4) to 5.25 Gy (n = 42) to the liver. The estimated 3-year nonrelapse mortality and disease-free survival (DFS) were 21% and 61%, respectively. These results were compared with those from 509 similar International Bone Marrow Transplant Registry patients who underwent transplantation using BU/CY alone. After adjusting for differences in age and cytogenetics risk, the hazard of mortality among all antibody-treated patients was 0.65 times that of the Registry patients (95% CI 0.39-1.08; P = .09). The addition of targeted hematopoietic irradiation to conventional BU/CY is feasible and well tolerated, and phase 2 results are sufficiently encouraging to warrant further study.