Prospective Pilot Study of Bone Marrow and Peripheral Blood Samples From AML Patients to Characterize the Biologic Heterogeneity of the Disease Using Single Cell Network Profiling (SCNP)
Condition: Acute Myeloid Leukemia
MeSH terms: Leukemia; Leukemia, Myeloid; Leukemia, Myeloid, Acute
Study type: Observational

Summary: 
      Treatment options and clinical outcomes for acute myeloid leukemia (AML) have not improved
      for more than 40 years. AML patients are still suffering from receiving costly, ineffective
      chemotherapy treatments with very high chances of bad side effects. The purpose of this study
      is to take a look at leukemia cells to see if the investigators can learn what makes them up
      and makes them aggressive and hard to treat. We want to use this information to create new
      treatments that the investigators hope are more effective and less harmful for AML patients.

      Newly diagnosed, relapsed or refractory (post induction therapy) AML patients that are 18
      years of age or older will have bone marrow and blood samples taken for their regular AML
      treatment. When these tests are done during their treatment the investigators will need to
      get some extra blood and bone marrow to do this research. The patients will not be asked to
      have an extra needle stick or bone marrow biopsy to get these samples. The patients will have
      the same number of blood and bone marrow tests whether they participate in this study or not.
      We will only need to get about two teaspoons of blood and two teaspoons of bone marrow each
      time the patient has these tests during their regular AML treatment. The research the
      investigators do with these sample will not decide or change the care the patients get for
      their AML.
    

Detailed description: 
      The proposed study will be an open-label prospective investigation using fresh bone marrow
      and paired peripheral blood samples obtained from patients with AML starting at the time of
      diagnosis and including samplings at times of response assessment (CR and NR) continuing
      through relapse for responders and for other subsequent treatments including salvage therapy
      after resistance/relapse. Approximately 100 AML patients are expected to be enrolled. There
      is no extra marrow and blood sampling taken from patients who enroll into this study because
      we only study their marrow and blood when they undergo such sampling for routine medical
      needs. The SCNP results will not be used to guide or adjust current, or future, treatment for
      patients enrolled in this study and they will not be contacted regarding the study results.

      Fresh whole bone marrow (5-10ml) and paired peripheral blood (5-10ml) will be collected from
      AML patients at participating institutions when these patients need marrow/blood tests prior
      to initiation of therapy, post each induction, post consolidation, at relapse and/or
      refractory, which is according to their standard care. When marrow cannot be obtained it is
      acceptable to collect only peripheral blood from the patient. Patient's demographic
      information, diagnosis, treatment option, outcome will be de-identified by the study PI.
      Samples must be collected in a standard 10 ml green top heparinized vacutainer labeled with
      sample de-identified ID, time, and date of collection. Samples will then be shipped to
      Nodality Inc. via FedEx in ambient temperature shipping kits on the same day. Nodality Inc.
      personnel will process the sample for study within 36 hours from collection. Leukemic blast
      SCNP will be conducted under the supervision of the researchers at Nodality Inc. and at
      facilities owned by Nodality Inc. Samples will be fractionated into bone marrow mononuclear
      cells (BMMC) or peripheral blood mononuclear cells (PBMC) and then aliquoted. All but one of
      these fractionated aliquots will be cryopreserved. The fresh, fractionated aliquot will be
      incubated with cytokines (e.g. interleukins, Flt3L), growth factors (e.g. SCF, GM-CSF and
      G-CSF), chemotherapeutic agents (e.g. cytarabine, ara- C, etoposide), and other modulators.
      Cells will then be fixed, permeabilized, and stained with antibodies that recognize
      extracellular markers (i.e. surface phenotypic markers such as clusters of differentiation,
      drug transporters, and receptors) in conjunction with intracellular activation-state specific
      epitopes (readouts) of designated signaling molecules. Subsequently, cells will be processed
      by multiparametric flow cytometry for SCNP. Blast populations will be defined by using
      combination of surface markers (CD33, CD34, CD38, CD45, CD11b) and approximately 30 different
      signaling nodes (paired modulator/readout e.g. Flt3L → phospho-Erk), depending on sample cell
      number (ideally 5-6 million; minimum 3 million). Signaling readouts will be evaluated in the
      total blast population, as well as within individual subpopulations, defined by CD33, CD34,
      CD 38, CD45, CD11b, and Side Scatter (SSC). Signaling readouts will be analyzed for each
      signaling node (univariate analysis) as well as a combination of nodes when the data set
      allows for multivariate analyses. In the "bridging" assay, cell surface markers and signaling
      readouts will be compared between fresh and cryopreserved AML samples including BMMC and PBMC
      when available in addition to BM samples.
    

Observational model: Other
Time perspective: Prospective
Sampling method: Non-Probability Sample
Study population: 
        Newly diagnosed or relapsed AML patients at the Mary Babb Randolph Cancer Center
      
Biospecimen description: 
      whole blood
    
Keywords: acute myeloid leukemia; single cell network profiling; AML; SCNP