Potential Role of CD9 and Implication of Motility Process in Pathogenesis of TEL/ALM1-positive ALL Relapses (LAL TEL/ALM1 and CD9).
Condition: Acute Lymphoblastic Leukemia (ALL)
MeSH terms: Precursor Cell Lymphoblastic Leukemia-Lymphoma; Recurrence
Study type: Interventional

Summary: 
      Down regulation of CD9 in TEL/AML1-positive ALL is addressed in motility assays to explore
      its role in B-ALL pathogenesis and its potential implication in relapses (and prognosis).
    

Detailed description: 
      1. Assess of the impact of CD9 expression level on motility assays (migration and adhesion)
           We have initiated motility assays (fibronectin adhesion experiments and CXCL12
           chemoattracted migration tests with modified Boyden chamber technique) using the CD9
           positive TEL/AML1-positive cell line REH and the CD9 negative cell line RAJI (wild or
           transfected with CD9 cDNA). Data will be analyzed in combination with blocking
           antibodies and chemical antagonist according to the level of CD9 (transcript and
           protein) and of CXCR4. Protein quantifications will be performed by flow cytometry and
           Western Blot. Interactions will be explored by confocal microscopy and biological
           pathways by immunoblot.

           Adhesion results will be validated on patient samples of B-ALL.

        2. Post-transcriptional regulation of CD9 in TEL/AML1-positive ALL To identify miRNAs that
           are potentially deregulated in TEL/AML1-positive acute lymphoblastic leukaemia and
           especially to screen for CD9 -targeted miRNAs, we will use a TaqMan ®MicroRNA Arrays
           approach allowing the simultaneous measurement of about 760 human miRNA.

      Small RNA will be extracted from bone marrow samples of twenty childhood B-ALL to screen
      miRNAs which are differentially expressed between CD9-positive and CD9-negative ALL and
      further compared with miRNAs which were predicted to target CD9 in databases. Validation of
      the selection will be performed by single Q-PCR for selected miRNAs using a novel cohort of
      ten bone marrow samples. Transfection assays and luciferase assays will be further realized
      to confirm that the differential miRNAs really target and affect CD9 expression .
    

Intervention name(s): Impact of CD9 expression level on motility assays; Post-transcriptional regulation of CD9
Intervention type(s): Other; Other
Intervention description(s): 1) Assess of the impact of CD9 expression level on motility assays (migration and adhesion) We have initiated motility assays (fibronectin adhesion experiments and CXCL12 chemoattracted migration tests with modified Boyden chamber technique) using the CD9 positive TEL/AML1-positive cell line REH and the CD9 negative cell line RAJI (wild or transfected with CD9 cDNA). Data will be analyzed in combination with blocking antibodies and chemical antagonist according to the level of CD9 (transcript and protein) and of CXCR4. Protein quantifications will be performed by flow cytometry and Western Blot. Interactions will be explored by confocal microscopy and biological pathways by immunoblot.
Adhesion results will be validated on patient samples of B-ALL.; 2) Post-transcriptional regulation of CD9 in TEL/AML1-positive ALL To identify miRNAs that are potentially deregulated in TEL/AML1-positive acute lymphoblastic leukaemia and especially to screen for CD9 -targeted miRNAs, we will use a TaqMan ®MicroRNA Arrays approach allowing the simultaneous measurement of about 760 human miRNA.
Small RNA will be extracted from bone marrow samples of twenty childhood B-ALL to screen miRNAs which are differentially expressed between CD9-positive and CD9-negative ALL and further compared with miRNAs which were predicted to target CD9 in databases. Validation of the selection will be performed by single Q-PCR for selected miRNAs using a novel cohort of ten bone marrow samples.
Other intervention name(s): Not Apllicable; Not Apllicable
Intervention model: Single Group Assignment
Masking: None (Open Label)
Primary purpose: Other
Keywords: TEL/ALM1_positive ALL relapses