Pre-existing Kinase Domain Mutations in Ph-positive Leukemias

The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Know the risks and potential benefits of clinical studies and talk to your health care provider before participating. Read our disclaimer for details. Identifier: NCT02643888
Recruitment Status : Not yet recruiting
First Posted : December 31, 2015
Last Update Posted : October 10, 2017
Qendra Spitalore Universitare NeneTereza, Albania
Hanusch-Krankenhaus, Wien, Austria
Information provided by (Responsible Party):
Prof DDr Thomas Lion, St. Anna Kinderkrebsforschung

Brief Summary:
The main task of this study includes analyses of the BCR-ABL1 (breakpoint cluster region/Abelson) gene and mutations in the BCR-ABL1 tyrosine kinase domain within flow-sorted stem cells from the bone marrow and/or peripheral blood specimens. To assess the germline configuration of the patient, DNA from fingernail clippings will be investigated.

Condition or disease

Detailed Description:

Background Single or multiple point mutations in the tyrosine kinase domain (TKD) of the BCR-ABL1 fusion gene in chronic myeloid leukemia (CML) and Ph-chromosome positive acute lymphoblastic leukemia represent the most important known mechanism of resistance to tyrosine kinase inhibitors (TKIs). It is conceivable that pre-existing point mutations in CML stem cells attributable to the genomic instability conferred by the BCR-ABL1 fusion protein give rise to the outgrowth of resistant subclones and onset of treatment-insensitive disease under the selection pressure of TKI therapy. Early detection of such subclones may therefore be of prognostic and therapeutic relevance. The recently published immunophenotype of CML stem cells (Hermann et al. Blood 2014), and our recent report on an NGS (next-generation sequencing)-based method facilitating sensitive detection and quantitative monitoring of BCR-ABL1 subclones carrying single or compound mutations (Kastner et al. European Journal of Cancer 2014) facilitate the screening for clinically relevant mutant subclones in stem cells or early progenitor cells.

Hypothesis Detection of mutant subclones within the stem cell or early progenitor compartments at diagnosis or early into therapy of Ph-positive leukemias, and monitoring of their proliferation kinetics, permit early prediction of resistant disease under the ongoing TKI treatment.

Experimental approach Bone marrow (BM) and peripheral blood (PB) samples will be collected upon informed consent from patients with Ph-positive leukemia at diagnosis (BM+PB), and subsequently at 3-month intervals (PB;BM upon availability) during the first year of therapy based on or including TKIs. Isolation of CD (cluster of differentiation) 34+ cells carrying additional phenotypic markers characterizing stem cells or early progenitor cells (CD38-/CD25+/CD26+ in CML, CD19 in Ph-ALL) will be performed by flow sorting. The entire BCR-ABL1 tyrosine kinase domain (TKD) will be amplified from cDNA (complementary DNA) or specific exons of interest from DNA by established protocols, and bidirectional sequencing will be performed by ultra-deep sequencing using NGS. Mutant subclones identified at diagnosis or after debulking of most of the treatment-sensitive leukemic burden early into treatment (3 month time point), will be monitored by NGS until month 12 of therapy. BCR-ABL1 transcripts will be monitored according to the International Scale (IS) in parallel. DNA isolated from fingernail clippings will be used as germline control for the ABL1 TKD. The testing will be performed in a blinded fashion to prevent treatment adjustments according to experimental data.

Study Type : Observational
Estimated Enrollment : 30 participants
Observational Model: Cohort
Time Perspective: Retrospective
Official Title: Identification of Pre-existing Kinase Domain Mutations in Subclones of Ph-positive Leukemias
Study Start Date : December 2016
Estimated Primary Completion Date : December 2018
Estimated Study Completion Date : December 2019

Resource links provided by the National Library of Medicine

MedlinePlus related topics: Leukemia

Primary Outcome Measures :
  1. Leukemic stem/progenitor cells defined by a specific marker profile will be isolated from BM/PB by flow sorting. Screening for and monitoring of BCR-ABL1 TKD mutant subclones at the cDNA/DNA levels within the 1st year of TKI therapy will be done by NGS. [ Time Frame: 3 years ]

Biospecimen Retention:   Samples With DNA
DNA samples from CML patients obtained from bone marrow/peripheral blood and fingernail clippings

Information from the National Library of Medicine

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Ages Eligible for Study:   Child, Adult, Older Adult
Sexes Eligible for Study:   All
Accepts Healthy Volunteers:   No
Sampling Method:   Non-Probability Sample
Study Population
The study plan includes 30 adult patients with Ph-positive leukemia.

Inclusion Criteria:

  • Established diagnosis of Ph-positive leukemia

Exclusion Criteria:

Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its identifier (NCT number): NCT02643888

Contact: Sandra Preuner-Stix, MSc. 0043140470 ext 4880

Sponsors and Collaborators
St. Anna Kinderkrebsforschung
Qendra Spitalore Universitare NeneTereza, Albania
Hanusch-Krankenhaus, Wien, Austria
Principal Investigator: Thomas Lion, MD PhD Prof Children´s Cancer Research Institute