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Functional profiling of live melanoma samples using a novel automated platform.
Functional profiling of live melanoma samples using a novel automated platform.
- Source :
-
PloS one [PLoS One] 2012; Vol. 7 (12), pp. e52760. Date of Electronic Publication: 2012 Dec 28. - Publication Year :
- 2012
-
Abstract
- Aims: This proof-of-concept study was designed to determine if functional, pharmacodynamic profiles relevant to targeted therapy could be derived from live human melanoma samples using a novel automated platform.<br />Methods: A series of 13 melanoma cell lines was briefly exposed to a BRAF inhibitor (PLX-4720) on a platform employing automated fluidics for sample processing. Levels of the phosphoprotein p-ERK in the mitogen-activated protein kinase (MAPK) pathway from treated and untreated sample aliquots were determined using a bead-based immunoassay. Comparison of these levels provided a determination of the pharmacodynamic effect of the drug on the MAPK pathway. A similar ex vivo analysis was performed on fine needle aspiration (FNA) biopsy samples from four murine xenograft models of metastatic melanoma, as well as 12 FNA samples from patients with metastatic melanoma.<br />Results: Melanoma cell lines with known sensitivity to BRAF inhibitors displayed marked suppression of the MAPK pathway in this system, while most BRAF inhibitor-resistant cell lines showed intact MAPK pathway activity despite exposure to a BRAF inhibitor (PLX-4720). FNA samples from melanoma xenografts showed comparable ex vivo MAPK activity as their respective cell lines in this system. FNA samples from patients with metastatic melanoma successfully yielded three categories of functional profiles including: MAPK pathway suppression; MAPK pathway reactivation; MAPK pathway stimulation. These profiles correlated with the anticipated MAPK activity, based on the known BRAF mutation status, as well as observed clinical responses to BRAF inhibitor therapy.<br />Conclusion: Pharmacodynamic information regarding the ex vivo effect of BRAF inhibitors on the MAPK pathway in live human melanoma samples can be reproducibly determined using a novel automated platform. Such information may be useful in preclinical and clinical drug development, as well as predicting response to targeted therapy in individual patients.
- Subjects :
- Animals
Antineoplastic Agents pharmacokinetics
Automation, Laboratory
Cell Line, Tumor
Drug Screening Assays, Antitumor
Female
Humans
Indoles pharmacokinetics
Inhibitory Concentration 50
Melanoma drug therapy
Melanoma pathology
Mice
Mitogen-Activated Protein Kinases antagonists & inhibitors
Molecular Targeted Therapy
Sulfonamides pharmacokinetics
Xenograft Model Antitumor Assays
Antineoplastic Agents pharmacology
Indoles pharmacology
MAP Kinase Signaling System drug effects
Melanoma metabolism
Mitogen-Activated Protein Kinases metabolism
Proto-Oncogene Proteins B-raf antagonists & inhibitors
Sulfonamides pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 7
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 23285177
- Full Text :
- https://doi.org/10.1371/journal.pone.0052760