Reverse Phase Protein Arrays: Mapping the Path Towards Personalized Medicine

Abstract

Reverse phase protein array (RPPA) technology evolved from the advent of miniaturized immunoassays and gene microarray technology. Reverse phase protein arrays provide either a low throughput or high throughput methodology for quantifying proteins and their post-translationally modified forms in both cellular and non-cellular samples. As the demand for patient tailored therapies increases so does the need for precise and sensitive technology to accurately profile the molecular circuitry driving an individual patient’s disease. RPPAs are currently utilized in clinical trials for profiling and comparing the functional state of protein signaling pathways, either temporally within tumors, between patients, or within the same patients before/after treatment. RPPAs are generally employed for quantifying large numbers of samples on one array, under identical experimental conditions. However, the goal of personalized cancer medicine is to design therapies based on the molecular portrait of a patient’s tumor, which in turn result in more efficacious treatments with less toxicity. Therefore, RPPAs are also being validated for low throughput assays of individual patient samples. This review explores reverse phase protein array technology in the cancer research field, concentrating on its role as a fundamental tool for deciphering protein signaling networks and its emerging role in personalized medicine.

A pre-existing population of ZEB2+ quiescent cells with stemness and mesenchymal features dictate chemoresistance in colorectal cancer

“…Reverse-Phase Protein Array (RPPA) allows the simultaneous evaluation of phosphorylated, cleaved, or unmodified proteins generating comprehensive profiles of pathway activation in different cell or tissue samples [38,39]. RPPA was used to compare PKH26 + and PKH26 − cells isolated ex vivo from CCSCs-derived tumor xenografts in order to obtain a broad picture of signaling pathways modulated in these two populations.…”

Cyclophilin D regulates lifespan and protein expression of aging markers in the brain of mice

“…A novel methodology that quantitates the abundance of hundreds if not thousands of different proteins in biological samples termed Reverse Phase Protein Array (RPPA) is becoming increasingly popular (Gallagher and Espina, 2014), however it still requires the extensive validation of the antibodies used. In our case, not more than four antibodies have been verified to be suitable for RPPA protocols (i.e., that of the RPPA Core facility of the MD Anderson Cancer Center) for the intended proteins.…”

Quantitative proteomic analyses in blood: A window to human health and disease

“…Precision medicine takes into account individual variability in genes, proteins, environment, and lifestyle for each person allowing a more focused and often targeted therapeutic strategy to be developed and implemented, which often reduces side effects and improves efficacy . Reverse phase protein arrays have been applied in clinical trials for various cancers to profile and compare functional states of protein signaling pathways between patients, within tumors, and within the same patients before and after treatment, which facilitates the development of personalized medicine approaches . Dijk et al.…”