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)
| Status: | Completed |
|---|---|
| Conditions: | Blood Cancer, Hematology |
| Therapuetic Areas: | Hematology, Oncology |
| Healthy: | No |
| Age Range: | 18 - 100 |
| Updated: | 2/8/2017 |
| Start Date: | September 2010 |
| End Date: | October 2012 |
Prospective Pilot Study of Bone Marrow and Peripheral Blood Samples From Acute Myeloid Leukemia (AML) Patients to Characterize the Biologic Heterogeneity of the Disease Using Single Cell Network Profiling (SCNP)
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.
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.
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.
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.
Inclusion Criteria:
- Newly diagnosed, relapsed, or refractory (post induction therapy) AML patients age >
18
- For relapsed AML patients, previous treatment regimens received do not limit their
eligibility to this study
- Patients enrolled will have no limitation as to the type of treatment they receive
for their disease.
- Patient is able to give consent
Exclusion Criteria:
- AML-M3 patients
- AML patients age < 18
- AML patients in clinical remission
- AML patients who will not be able to receive diagnostic blood and marrow work up for
any reason
- Patients who received allogeneic stem cell transplantation or autologous stem cell
transplantation
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