MICROFABRICATED INSTRUMENTATION TO MEASURE SPHINGOLIPID SIGNALING IN HUMAN ACUTE MYELOID LEUKEMIA

An innovative platform to measure the activity of the sphingolipid pathway in single cells from
primary, human, acute myeloid leukemia (AML) will be developed. A multidisciplinary group
(chemist, bioengineer, oncologist and computational scientist) with a history of successful
collaborations will pursue the development of engineered microfluidic instrumentation and
supporting hardware using medically relevant probes to answer fundamental questions
regarding heterogeneity in single AML cells. Fluorescent probes to track simultaneously the
three major pathways comprising the ceramide-sphingosine axis in AML cells will be
developed so that a detailed understanding of sphingolipid signaling in the tumor cells is
achieved. Electrophoretic separations within a microfabricated device will be optimized for the
single-cell measurements as a component of the work flow. Automation and integration will
greatly increase throughput to yield a microdevice which is compatible with common clinical
workflows. A powerful attribute of the proposal is that these measurements will be performed
on single cells from primary samples and will avoid the confounding aspects of population-
averaged data yielded by bulk cell assays. Furthermore, by simultaneously tracking all arms of
the sphingolipid pathway, we will identify the strategies that AML cells use to dynamically
reprogram their growth-promoting pathways via sphingolipid signaling during drug treatment.
The proposed microfabricated devices will in the future provide key information concerning the
best treatment option(s) for patients as well yielding an assessment of treatment efficacy to
contribute fundamental data to the emerging field of precision medicine.