12th Congress of the European Hematology ... - Haematologica

tested across a wide range of human cancers namely in hematologic
malignancies. Besides preliminary results from clinical trials demonstrate
enzyme target inhibition, a favorable toxicity profile and promising efficacy,
with this study we hope to contribute to determine their mechanism
of action and the role of combination with other agents, to define
their place in the therapeutic arsenal of hematologic disorders, namely
in ALLs. For this purpose CEM cells were incubate in absence and presence
of an FTIs, α-Hydroxyfarnesylphosphonic acid (α-HFPA) in different
concentrations (rangin from 10 nM to 300 µM), as single agent and/or
with combination with Doxorrubicin (DOX) and/or MG262, a proteasome
inhibitor during 72 h. Cell growth and viability were evaluated by
Trypan blue exclusion. Cell death was performed by morphological studies
using optic microscopy and by flow cytometry (FC), using annexin-
V and/or propidium iodine incorporation. The proteins related with
apoptotic cell death, BAX, BAD e BCL-2 were evaluated by FC using
monoclonal antibodies. As a marker of FT inhibition we evaluate, by FC,
the levels of lamin A/C before and after α-HFPA administration. To
determine the inhibition of RAS-MAPK pathway we analysed by FC
the levels of ciclin D1. Our results show that α-HFPA induces a decrease
in cell density and viability, as single agent, in a dose and time dependent
manner (IC50 ≥300 µM), inducing cell death by apoptosis. However,
the anti-proliferative effect seems to be higher than the cytotoxic
one, witch may be related with Ras-MAPK pathway inhibition once we
detected a decrease in ciclin D1 levels. The observed decrease in lamin
A/C levels confirm the farnesyltransferase inhibition and may also translate
the inhibition of Ras-MAPK pathway. This study suggests that FTIs
may be a usefull therapeutic approach in ALLs as single agent. However,
the future of treatment for leukemia resides in defining the molecular
pathways underlying the pathogenesis of this disease and in further
elucidating the pharmacogenetic factors of the host.
This work was supported by GAI
1132
SURVIVIN AND VEGF MRNA EXPRESSION IN ELDERLY PATIENTS
WITH DE NOVO OR SECONDARY AML
A. Tsiga, A. Avgitidou, E. Ioannidou-Papayannaki, E. Vlachaki, F. Klonizakis,
A. Kalogeridis, S. Haralambidou-Vranitsa, I. Klonizakis
AUTH, Hippokration Hospital, THESSALONIKI, Greece
Background. Several biologic features of AML differ between older and
younger individuals. Older patients often express multidrug resistance
phenotype and cytogenetic abnormalities, which may be responsible in
large part for the poor outcomes observed in older-aged subgroups. Traditional
cytotoxic chemotherapy is associated with a low complete
response rate and a high treatment-related mortality in older patients,
which explains in part the poorer outcomes in individuals over 60 years
of age. For that reason, treatment for the elderly AML patients should
be based on biologic and prognostic factors. Research into the pathophysiology
of AML has revealed an abundance of intracellular signalling
events that govern angiogenesis and apoptosis of the malignant cell.
VEGF plays an important role in angiogenesis by acting as a potent inducer
of vascular permeability and serving as an endothelial cell mitogen.
Furthermore, the anti-apoptotic properties of VEGF during angiogenesis
are primarily mediated by the induced expression of Survivin. Survivin
is the smallest member of the inhibitory of apoptosis protein family
(IAP family) and a putative oncogene that is aberrantly expressed in
cancer cells. Survivin is also highly expressed in neoangiogenesis. The
AIM of this study is to examine the mRNA expression of Survivin and
VEGF in elderly patients with de novo or secondary AML. Methods. Total
RNA was isolated from peripheral blood cells of 16 elderly AML patients
(median age 73.8±8.7 years, 10 men and 6 women) and 16 individuals,
of the same age, with normal haemopoiesis. Seven patients were diagnosed
with de novo AML, while 9 had secondary AML, derived from
MDS. All of our patients had excess of peripheral blasts (range 60-80%).
Real Time Semi-Quantitative RT-PCR assay was performed in order to
detect the mRNA levels of Survivin and VEGF. Abl was used as a reference
gene. The regulation of the target genes was estimated as an expression
rate. Results. Survivin and VEGF were both up-regulated in all AML
patients that were examined [4.5 fold (p>0.05) and 2.5 fold (p>0.05),
respectively]. More specifically, Survivin was up-regulated by the factor
2.8 (p>0.05), in patients with de novo AML and by the factor 6.8 (pAML patients. VEGF mRNA expression was 1.5 times
(p>0.05) higher in de novo AML patients and 3.2 times (pAML patients. The mRNA expression of both genes in the elderly
patients with secondary AML is statistically significant. Conclusion.
Our data and the findings of other authors, suggest that Survivin and
VEGF may play an important role in the pathophysiology of hematopoi-
12 th Congress of the European Hematology Association
etic malignancies and the progression of leukemia. Antisense approaches,
which could be used to target Survivin or VEGF or inhibit their common
pathway, would be expected to be useful for the treatment of AML,
as well as carry limited toxicity for normal cells. These approaches could
be therefore, more tolerable by elderly patients. Further studies are needed
in order to determine whether Survivin and VEGF could be used as
prognostic markers, minimal residual disease (MRD) indicators or promising
cancer therapeutic targets.
1133
INTRATHECAL LIPOSOMAL CYTARABINE AS THERAPY OF LYMPHOMATOUS MENINGITIS
N. Cascavilla, C. Bodenizza, A.M. Carella, M. Dell’Olio, A.P. Falcone,
M.M. Greco, A. La Sala, S. Mantuano, L. Melillo, E. Merla, M. Nobile,
G. Sanpaolo, P. Scalzulli
Hematology „CSS„ Hospital, SAN GIOVANNI ROTONDO, Italy
Background and aims. The Lymphomatous Meningitis (LM) represents
an event with a generally poor prognosis and a median survival of few
months in untreated patients. Its occurrence is tightly related to the histological
type, the response to the therapy and the introduction of a
proper prophylaxis in the treatment plan. Currently, the systemic therapy
with high doses of Methotrexate (HD-MTX) (3 g/sqm) followed by
radiotherapy (WBRT: 45 Gy) represents the elective treatment. The role
of the intratecal chemotherapy (IT-CHT) isn’t well defined. Recently, an
important role both in the prophylaxis and in the therapy of LM is played
by the intratecal administration of Cytarabine in liposomal formulation
(DepoCyte): randomized studies have shown a significant better effectiveness
and a reduced toxicity of liposomal formulation treatment with
respect to the traditional one. Methods and results. In this work 11 cases
of LM treated with systemic chemotherapy and DepoCyte (50 mg IT
every 15 days x 6 times) are shown. Male/Female ratio is 7/4; average
age is 41 years (range 24-78). In all patients neurological symptoms were
present; lymphomatous cells were identified in the liquor of 6 patients
and Central Nervous System (CNS) localizations were detected by NMR
in 9 patients. The 6 CSF-positive cases were heterogeneous, as follows:
1) B-lineage CD10 + ALL meningeal relapse (after two marrow relapses),
already undertaken to allogeneic staminal cells transplantation, treated
only with DepoCyte; 2) Lymphoblastic T Lymphoma meningeal localization,
with mediastinic mass, treated with DepoCyte associated to
systemic chemotherapy (LSA2L2 Protocol) and autologous stem cell tranplantation;
3) DLBCL meningeal localization treated with DepoCyte
associated to systemic chemotherapy (R-CHOP Protocol); 4) LM at diagnosis
in a 78 years old patient with inadequate compliance to systemic
therapy, treated with DepoCyte; 5) Mantle Cell Lymphoma meningeal
(and systemic) relapse treated with DepoCyte associated to systemic
chemotherapy (Codox-M Protocol); 6) Acute Myeloid Leukemia M3
FAB with meningeal relapse during manintenance therapy treated with
DepoCyte and HAM + RT protocol. In patients 1) to 4) the C.R. was
obtained and they are still alive; only the latter patient, namely, case 5),
died for disease progression. The patient 6) is still in treatment. The
remaining 5 CSF-negative cases had been diagnosed as DLBCL (3 at
diagnosis and 2 at relapse) CNS solitary localizations. In all cases the systemic
therapy with L-VAMP Protocol (modified with HD-MTX) was
associated to IT-CHT with DepoCyte. 4 patients had a good response
to treatment and 3 are still alive and in C.R. Conclusions. Overall,
DepoCyte treatment was shown to be mostly effective, well tolerated
by all patients and devoid of undesired side effects. The association with
various systemic chemotherapy protocols had been demonstrated to be
suitable and endowed with synergic effects. Studies with higher number
of patients might validate the effectiveness of DepoCyte also in
those CSF-negative cases with solitary CNS localization.
1134
USE OF INTRATHECAL LIPOSOMAL CYTARABINE DRAMATICALLY IMPROVED
NEUROLYMPHOMATOSIS IN WALDENSTROM MACROGLOBINEMIA
A. Marfaing-Koka, S. Ovidiu, A. Notar, C. Pignon, L. Chaiba
Hopital Antoine Beclere, CLAMART, France
Neurolymphomatosis, a rare complication of Waldenström’s macroglobinemia,
is characterized by meningeal and root nerve infiltration by
lymphoplasmacytic cells. Combined intravenous and intrathecal
chemotherapy including methotrexate and cytosine arabinoside is usually
effective but poorly tolerated. We report a 61 year-old patient with
waldenström‘s macroglobinemia who presented with rapidly progressive
distal leg weakness. He had been previously treated by six courses
of COP associated with good partial response (IgM serum level decrased
haematologica/the hematology journal | 2007; 92(s1) | 417