Fondazione Ginevra Caltagirone is funding preclinical testing of new drugs inhibitors of pediatric AML

Fondazione Ginevra Caltagirone is funding preclinical testing of new drugs inhibitors of pediatric AML

Fondazione Ginevra Caltagirone has granted over the past three years, projects pediatric presented thanks to Fondazione Umberto Veronesi, a major contribution to the achievement of important results for the treatment of acute myeloid leukemia children.
Strong the important results achieved in this field, has decided with great pleasure for the year 2015/2016 a new loan of 45,000 euros for the continuation of the Research Project of the Dott. Riccardo Masetti entitled:
PRECLINICAL TESTING OF NEW DRUGS inhibitors THE PATHWAY OF HEDGEGHOG IN CELL LEUKEMIA Acute myeloid PEDIATRIC A BAD PROGNOSIS.


The massive sequencing of AML pediatric performed at the laboratory of pediatric hematology and oncology with funding obtained from Fondazione Ginevra Caltagirone has allowed us to define a new subgroup of AML represented by 20% of patients characterized by the anomaly cytogenetic CBFA2T3-GLIS2. Preliminary studies have shown that this subgroup of AML has a peculiar expression profile compared to the other LAM with other genetic abnormalities. In addition it was found that these patients have particularly poor prognosis and a higher risk of recurrence.
Therefore the need of new drugs target highly effective and specific for this subset of leukemias that makes the execution of the preclinical study of fundamental importance for this subset of AML at high risk for which the therapeutic alternatives in addition to conventional chemotherapy are much reduced.


They have been conducted preliminary experiments, in particular was tested the efficacy of a number of drugs inhibitors of the pathway of Hedgehog on a series of cell lines of LAM referred to a positive genetic lesion CBFA2T3-GLIS2 of interest. The first results are very promising. In fact one of these drugs, the GANT61, has demonstrated incredible effectiveness and specificity towards the positive cell line cytogenetic abnormality in question. The drug exposure GANT61 causes cell line mutation of GLIS2 high mortality and apoptosis even at low concentration, instead resulting in ineffective lines LAM negative for the genetic lesion GLIS2.
Although these preliminary results it can be said that we have all the credentials and privileges to take this drug trial that could represent a breakthrough in the treatment of this subgroup AML pediatric high risk for which to date there are no specific therapies.
Main objective of the project is the testing of inhibitors of the Hedgehog pathway with assays of cell viability, cell cycle analysis and apoptosis in cell lines but above all on leukemic blasts of patients with and without chromosomal aberration concerned after drug exposure.
Then you analyze the expression pathway downstream of GLIS2 before and after drug treatment and determine the total amount of protein GLIS2 active before and after treatment to determine or confirm whether the drug determines a direct inhibition of the binding of protein GLIS2 DNA by Chromatin Immunoprecipitation.
Then you will make an in vivo study in mice in which they were previously inoculated leukemia cells carrying the mutation, we will be conducted essays cell viability, cell cycle analysis and apoptosis in cell lines and blasts of patients carry the mutation. The results will be compared with cell lines and blasts from patients with AML negative.
During the second semester, we will instead be carried out first experiments of gene expression and chromatin Immunoprecipitazion after drug treatment to determine if the drug GANT61 determines a sub-expression of genes regulated by GLIS2 who are directly involved in the process of leukemogenesis.


This pharmacological study could have important clinical implications. In fact, as mentioned earlier patients with LAM and the positive genetic lesion CBFA2T3-GLIS2 have a particularly poor prognosis and increased risk of recurrence. Moreover, the treatment of the disease with conventional chemotherapy involves cytotoxic effects that are partially responsible for the therapeutic failure. Provide a clinical information and data on the specificity of a drug into the chromosome aberration test, this allows you to set a possible targeted biological therapy in these patients by reducing the limiting toxicity of conventional chemotherapy. Many drugs target tyrosine kinase inhibitors such as new generation, monoclonal antibodies, inducing cell differentiation and apoptosis in fact are not commonly used in first-line therapy for lack of information about the biology of the disease.
The execution of this study may help the clinician detailed information about the effectiveness and specificity of the drug in the subject of this preclinical testing that would allow you to set customized treatment protocols can induce complete remission, limiting the cytotoxic effects and lower the probability recurrence. Test new drugs and targeted molecular means testing a new therapy model so-called "intelligent" because it comes precisely at the origin of leukemogenesis with the ultimate aim of improving overall survival of pediatric patients with AML poor prognosis.
This study is in a sense the end of a path of research in recent years, thanks to the sensibility and to the help of the Fondazione Ginevra Caltagirone has made a significant contribution to the literature of pediatric acute myeloid leukemia. The pre-clinical trials of a new drug for this specific cytogenetic abnormality would be able to treat more effective and less toxic than a subgroup of AML that is characterized by a particularly poor prognosis and so today the only effective therapeutic option is the transplantation of bone marrow.