Download 2016 department of medicine research day

2016 DEPARTMENT OF MEDICINE RESEARCH DAY
Title of Poster: The PI3K inhibitor Copanlisib shows activity alone and in combination
with Carfilzomib in multiple myeloma
Presenter: Sarah Larson, M.D.
Division: Hematology-Oncology
☒ Faculty ☐ Fellow ☐ Resident ☐ Post-doc Research Fellow ☐ Graduate Student ☐ Medical Student ☐Other
Principal Investigator/Mentor: Sarah Larson, M.D. Co-Investigators: Andrae Vandross, MD, Erka Von Euw, PhD,
Dylan Conklin, Mao Yu Peng PhD, Zoe Fuchs B.S., Monica Meade M.D, Neil O'brien PhD, Dennis J. Slamon, MD, PhD
Thematic Poster Category:
Pharmacology and Drug Development
Abstract
Background: Copanlisib (BAY 80-6946) is a PI3K inhibitor with preferential activity for the alpha and
delta isoforms. In multiple myeloma, the alpha isoform is of particular importance. Here we present
data showing activity of copanlisib as a single agent as well as in combination with carfilzomib.
Methods: 3 sensitive cell lines (NCI-H929, MM.1S, L-363) and 3 resistant lines (AmO-1, JJN3, COLO677) were identified for our studies of a total of 21 that were initially screened. Each cell line was
evaluated with apoptosis and cell senescence assays. The doses and time points used in these assays
were: Copanlisib at 50nM and 100nM at 72 hours; carfilzomib at 2 nM and 20 nM at 96 hours. Cell
cycle analysis was done with propidium iodide staining. Apoptosis assay done with annexin V FITC
staining. Both assays were analyzed using FACS. Β-galactosidase activity was measured in cell
senescence assays after 96-hour treatment. In the combination experiments, excess over highest
single agent statistics(EOHSA) were used evaluate for potentiation. Pre and post treatment reverse
phase protein array (RPPA) were obtained and confirmed with western blot analysis.
Results: Apoptosis and cell cycle arrest were observed in sensitive cell lines treated with single agent
copanlisib but not resistant lines. Results of cell senescence assays confirmed that apoptosis is the
mechanism of cell death in proliferation studies. Low levels of pS6 were seen in pretreated, sensitive
cell lines. The levels of pS6 were reduced further with copanlisib treatment in sensitive lines only. This
finding, as well as downstream PDCD4 and eIF4A effects were confirmed with western blot analysis.
EOHSA statistics were notable for potentiation carfilzomib/copanlisib combination. These combination
treatments also showed decrease in pS6 expression in sensitive cell lines and not resistant lines.
Discussion: Copanlisib alone, as well as in combination with carfilzomib shows activity in human
multiple myeloma cell lines. These preclinical results are encouraging for use of dual therapy in
patients with relapsed and refractory multiple myeloma. Additionally, pS6 levels may be an important
biomarker to identify those that would benefit from therapy as well as monitoring response to dual
therapy.