BAP1 shRNA inhibits cutaneous melanoma cell growth with known germ line BRaf/NRas mutation status. Nikolai Klebanov BS1,2, Grace LaMuraglia BS1, ...
Uveal Melanoma Inhibition with Calcium Channel Blockade BAP1 shRNA inhibits cutaneous melanoma cell growth with known germ line BRaf/NRas mutation status
Wellman Center for Photomedicine
HARVARD
MEDICAL SCHOOL
Nikolai Klebanov 1Wellman
BS1,2,
Grace
Raj Kumar, Michael, Taylor, Ching Ni Njauw, Zhenyu Ji, Benchun Miao, Chelvi Rajadurai and Hensin Tsao 1 1, Anpuchchelvi Department WellmanKim Center for BS Photomedicine, Massachusetts General Hospital,Rajadurai and Dana Farber Cancer MS Institute1 and Harvard Medical School, Boston, MA 1, LaMuraglia BSof Dermatology, , Ji-Soo , Raj Kumar PhD
and Hensin Tsao MD, PhD1
Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, Boston, MA; 2Tufts University School of Medicine, Boston, MA
ABSTRACT
RESULTS
RESULTS
FDA library screening
Background: Uveal malignant melanoma (UMM), the most common primary intraocular tumor, has remarkable metastatic potential and clinical trials of therapies have shown limited benefits. We aimed to identify possible candidate drugs active against UMM and to characterize their biological function in-vitro. Methods: UMM cell line OMM2.3 was subjected to 1953 FDA-approved compounds to rank drugs based on cell viability inhibition. We characterized the function of the top raking drug on UMM and CMM lines in-vitro by a) growth inhibition in 2D/3D culture; b) assays of cell cycle progression, apoptosis, and senescence. OMM2.3 uveal cell line was used for initial screening. 6 UMM and 10 CMM lines were used for 2D cell culture; 2 UMM and 3 CMM lines were used for all subsequent assays. Main outcomes and measures included identification of compound(s) preferentially-active against uveal (than cutaneous) melanoma cells; evidence of cell cycle arrest/apoptosis/senescence effects to suggest possible mechanisms of action.
Rank
Drug Name
1
Fluoxetine hydrochloride
2
Amlodipine
3
Dronedarone hydrochloride
4
Prochlorperazine dimaleate
5
Afatinib
6
Cinacalcet hydrochloride
7
Amlodipine besylate
8
Crizotinib
9
Crystal violet
10
Alexidine hydrochloride
Virtual blot of apoptotic effects on CMM and UMM cell lines
§ Amlodipine ranked as #2 and #7 among 1953 distinct FDA compounds in highthroughput screening § Based on these results, chose amlodipine, verapamil, diltiazem (calcium channel blockers) and JTV-519 (ryanodine receptor blocker) for 2D cell viability screening § Hypothesized that calcium signaling may be an important contributory pathway in uveal melanoma pathogenesis
A.
C.
Table 1. Amlodipine ranked #2 and #7 among n=1953 compounds subjected to screening,
Results: Amlodipine (and as amlodipine besylate) ranked twice among the top 10 2D Cell Viability Screening drugs active against the OMM2.3 uveal melanoma cell line (#2, #8 of n=1953 § Amlodipine and JTV-519 yielded the highest compounds). In 2D cell culture, amlodipine and JTV (in DMSO solvent) were both found selective growth inhibition among the four to have significantly lower half-maximal inhibitory concentration (IC50) in UMM than compounds selected for 2D screening (Figure 3) CMM (p
