Francesca Simone

Cryopreserved autologous bone marrow infusion following high dose chemotherapy in patients with acute myeloblastic leukemia in first relapse

Thirteen patients with AML in first relapse were treated with high dose combination chemotherapy followed by cryopreserved autologous bone marrow transplantation (ABMT). The first four patients received the COATA-Roma regimen, consisting of CTX, VCR, CA, 6-TG and ADM; nine additional patients received the BAVC regimen consisting of BCNU, AMSA, VP-16 and CA. A median of 1.6 X 10(8) fractionated nucleated bone marrow cells/kg body weight were reinfused. The median of GM-CFU-C recovered was 4.7 X 10(4)/kg. Out of 13 patients, 10 (76.9%) achieved CR, 3 had profound aplasia and died from hemorrhagic or infectious complications. Of the 10 patients who achieved CR, 1 died after 1 week from heart failure, 5 relapsed respectively 17, 20, 21, 21, 42, weeks after ABMT, 4 are still in CR after 2+, 14+, 17+, and 120+, weeks. Of the 9 patients treated with BAVC regimen, 8(88.8%) achieved CR. Four patients relapsed after a median of 19.7 weeks and 4 are still in complete remission. Of interest is the fact that the second complete remission of one patient is longer than the first one, despite the fact that marrow was not purified by any in vitro treatment. In conclusion we can say that BAVC regimen is highly effective in obtaining second complete remission in patients with AML and prolonged disease free survival can be achieved at least in a small number of cases.

Intermediate-dose cyclophosphamide and granulocyte colony-stimulating factor is a valid alternative to high-dose cyclophosphamide for mobilizing peripheral blood CD34+ cells in patients with multiple myeloma

Peripheral blood stem cells (PBSC) are widely used in the setting of dose-intensive chemotherapies in patients with multiple myeloma (MM). Although the granulocyte colony-stimulating factor (G-CSF), following chemotherapy or not, is considered the standard growth factor for mobilizing PBSC, the optimal chemotherapeutic regimen still remains to be defined. Cyclophosphamide (CTX) is an effective drug in the treatment of MM which is capable of mobilizing PBSC if followed by growth factors, even though administration of high-dose CTX (7 g/m2) results in severe toxicity requiring hospitalization and increasing costs. We have retrospectively analyzed the results obtained in 38 newly diagnosed MM patients treated with 1.2 g/m2 CTX on days 1 and 3 combined with 40 mg dexamethasone daily for 4 days. The results were compared with those obtained in 25 newly diagnosed MM patients treated with 7 g/m2 CTX. A higher number of CD34+ cells/kg was collected during the first leukapheresis and a statistically significant lower consumption of G-CSF was observed following two doses of 1.2 g/m2 CTX compared to the 7 g/m2 CTX dose. The possibility of treating patients with day-hospital regimens, with a satisfactory yield of hematopoietic cells harvested, may have relevant economic implications for treatment strategies in MM patients.

Autologous bone marrow transplantation in acute myeloid leukemia after in-vitro purging with an anti-lacto-N-fucopentaose III antibody and rabbit complement

Two AML patients, whose leukemic clonogenic cells totally reacted to the anti-lactofucopentaose III S4-7 monoclonal antibody (MoAb), underwent autologous bone marrow transplantation, in first complete remission, after in-vitro purging with S4-7 MoAb and complement. After ablative chemotherapy (BAVC regimen) and reinfusion of S4-7 purged cells, regeneration of marrow cells occurred with prompt recovery of granulopoiesis and erythropoiesis. A more delayed platelet recovery was observed. The two patients are in complete remission at 20 and 11 months from ABMT. The results indicate that immunologic purging with S4-7 MoAb is safe and suitable for selected AML patients undergoing ABMT.

High-dose idarubicine, busulphan and melphalan as conditioning for autologous blood stem cell transplantation in multiple myeloma. A feasibility study

Extensive studies have tested the clinical impact of double and triple sequential transplants as front-line therapy in MM, following the suggestion that dose escalation can overcome the marked drug resistance characteristic of this disease, but the superiority of such approaches vsone single transplant has still to be demonstrated. The aim of our study was to evaluate the feasibility and efficacy of high-dose idarubicine intensification of a standard busulphan–melphalan conditioning regimen in MM. Twenty-eight patients (median age 55 years) with sensitive disease received PBSCT after high-dose idarubicine combined with busulphan and melphalan and followed by s.c. rhG-CSF until PMN recovery. The most severe toxicity was represented by oral mucositis which resolved with hemopoietic reconstitution. Overall response and CR rate were 52% and 40%, respectively. Currently, 36 patients are alive and 19 are progression-free a median of 20 months (12–36) from transplant. The 3-year projected probability of progression-free survival for patients transplanted after first-line treatment is 60%. The combination of Ida/Bu/Melph appears a promising alternative regimen for PBSCT in myeloma, with low transplant-related toxicity and fast hematological recovery. Long-term follow-up and a prospective randomized study, now ongoing, will probably clarify whether an idarubicine-intensified regimen will result in superior outcomes to conventional conditioning and even be comparable to a double consecutive transplant program. Bone Marrow Transplantation (2000) 26, 1045–1049.

In vitro pharmacological purging of human bone marrow is enhanced by the use of lonidamine

Lonidamine (LND), previously reported as a useful antitumor substance in combination with physical or chemical agents, has been studied for its capacity in increasing pharmacological elimination in vitro of residual tumor cells from human bone marrow. Different drugs were tested in association with LND against mixtures of human bone marrow and a tumor cell line, clonogenic human leukemic blast progenitors, and normal human bone marrow precursors. The results demonstrated that LND increased the efficacy of anthracycline derivatives (Adriamycin, Mitoxantrone) both on the tumor cell line and on the leukemic blast progenitors, while VP-16 or ASTA-Z 7654 was not affected by the same substance. The toxicity on normal stem cells reflected that of each drug and was not modified by the addition of LND. While a consistent dose-dependent CFU-GM reduction was observed immediately after treatment with the different drugs, a complete recovery was reached after 7 and 14 days of long-term marrow cultures. Because of the low toxicity and the efficacy demonstrated in association with certain agents in increasing tumor cell elimination in vitro, LND could play an important role in in vitro purging prior to autologous bone marrow transplantation.

Acute myelogenous leukemia in first relapse treated with two consecutive autologous bone marrow transplantations: a pilot study

The feasibility and the antileukemic activity of a double sequential autograft has been evaluated in 3 patients affected by AML in first hematological relapse. Bone marrow collection and cryopreservation was performed twice: during first complete remission (CR) and during second CR. At the time of relapse patients underwent first ABMT with BAVC preparative regimen achieving a second CR without any remarkable complications. After 4, 5 and 4 months respectively a second ABMT was performed with a different preparative regimen, consisting of cyclophosphamide and fractionated total body irradiation (Cy + F‐TBI). A delay in platelet recovery was observed after the second procedure as compared to the first, while neutrophils recovery was comparable. 1 patient died in CR (on day +91 after second ABMT) of interstitial pneumonitis. 2 patients are alive and well without evidence of disease after 46 and 53 months of unmaintained second CR. This experience shows the high antieleukemic potential of treatment with a double sequential autograft; 2 relapsed patients in fact are long‐term survivors with a second CR longer than the first.

Effect of Hemopoietic Growth Factors on the Proliferation of Acute Myeloid and Lymphoid Leukemias.

The effect of G-CSF, GM-CSF, IL-1 and IL-3 on the proliferation of acute leukemia cells (evaluated as (3)HTdR uptake) was investigated in short-term liquid cultures and compared with that observed on normal bone marrow (BM) populations enriched for immature cells. In acute myeloid leukemias (AML), a marked leukemic proliferation was induced in 10/18 cases by IL-3, in 9/18 by GM-CSF, in 7/18 by IL-1 and in 4/18 by G-CSF. In acute lymphoid leukemias (ALL), marked stimulation was observed in 7/11 cases with IL-3 and in 5/11 with GM-CSF, whereas IL-1 and G-CSF were ineffective. Both in AML and ALL, the combination of several factors did not result in an additive synergistic effect. Purified normal BM cells responded to all four growth factors and their combinations produced an additive effect on cell proliferation which probably relates to the heterogeneity of the cell populations studied. The effect of a G-CSF, GM-CSF, IL-1 and IL-3 on the proliferation of acute leukemia cells by different growth factors suggests that caution should be exercised in their clinical use in these diseases.

Autologous Bone Marrow Transplantation (ABMT) for Patients with Chronic Granulocytic Leukemia (CGL) in Blastic Phase (BP)

Autologous hemopoietic stem cell infusion after high dose chemo/radiotherapy performed in BP of CGL is able to reinduce II CP in 90% of patients; however the median survival does not last more than 6 months. This report concerns 5 patients with Phl positive CGL in BP treated with BAVC regimen followed by hemopoietic stem cells reinfusion. An intensification program of cyclic chemotherapy has been applied after ABMT with the aim to prolong the survival of these patients. Since 1980 the majority of CGL patients seen at the Institute of Hematology of Rome were subjected to collection and cryopreservation of hemopoietic stem cells from bone marrow (BM) or peripheral blood (PB). Details of the methods have been described elsewhere (1,2). Of the 5 patients presented in this report 4 received stem cells collected from PB and 1 from BM. Four patients were males, median age was 39 years (range 29–41). Therapy during CP consisted of hydroxyurea; no splenectomy had been performed in any patient. Type of transformation was myeloid in 4 cases and mixed in one. BAVC conditioning regimen consisted of BCNU 800 mg/mq day 1; AMSA 150 mg/mq/day on days2–4; VP-16 150 mg/mq/day days 2–4; CA 300 mg/mq/day c.i. days 2–4. Stem cells were reinfused 24 hours after last dose chemotherapy with a median of 11×108(range 3–21) nucleated cells/kg body weight. After complete hematological reconstitution (25–30 days) from autologous infusion an intensification program of cyclic chemotherapy was begun consisting of 3 courses of L-VAMP (VCR 1.5 mg/mq i.v. hour 0,MTX 200mg/mq c.i. hours 1–3,CA 500 mg/mq c.i. hours 3–7, ASNASI 5000 U/mq i.m. hour 24,PDN 40 mg/mq/day days 1–5);3 courses of DAT (DNR 60 mg/mq i.v. day l,CA 60 to 150 mg/mq/8 hours s.c. days 1–5,6-TG 70 mg/mq/8 hours p.o. days 1–5) and 3 courses of HiDAC (CA 1 g/mq/12 hours c.i. in 3 hours on hour 0,12; ASNASI 6000 U/mq i.v. on hour 18).

Possibility of Progenitor Cell Mobilization during the Hematological Recovery following Peripheral Blood Stem Cell Autograft

Twenty-four patients with hematological malignancies were studied during recovery following autografting in order to establish the proportion of patients that show CD34+ cell mobilization and the kinetics of mobilized CD34-positive cells. The patients showed a peak in peripheral blood (PB) CD34+ cells after a median of 14 days (range 12–20) following reinfusion. According to the number of circulating CD34+ cells, two groups could be clearly distinguished: 17 patients (group A) with <10 PB CD34+ cells/µl (median 1.2, range 0–5) and 7 patients (group B) with >10 CD34+ cells/µl (median 51, range 13–123). Compared to group A, patients of group B showed a faster hematological reconstitution of both polymorphonuclear leukocytes >500/µl (12 vs. 15 days) and platelets >50,000/µl (12 vs. 17 days). The expression of the β1 integrin CD49d was similar in the two groups of patients, while a lower expression of the β2 integrin CD11a and a greater expression of the L-selectin CD62L were observed in the PB CD34+ cells of group B patients. Both in the PB and in the BM, the number of CFU-GEMM, CFU- GM, CFU-E and BFU-E of group B was significantly greater than that of group A. However, when the clonogenic potential of a single CD34+ cell was evaluated, no major differences in the number of colonies produced per CD34+ cell were found between the two groups.

In Vitro and In Vivo Inhibitory Effects of Interferon α 2b on CML Granuloerythroid Precursors

Chronic myelogenous leukemia (CML) is a clonal hemopathy in which malignant transformation probably occurs in an early hemopoietic precursor cell. The initial chronic phase is easily treated by chemotherapy, but the fatal course of the disease rapidly evolves into the acute phase that is very difficult to control with conventional drugs.