Multiple Myeloma (MM) is an onco-hematologic disease characterized by high genetic heterogeneity. Evidence derived from literature has shown that MM is the consequence of a "multi-step" pathogenic process that involves several stages. During this process, the disease progressively becomes increasingly aggressive and resistant. The miRNA dysregulation, the aberrant activation of numerous signal transduction pathways (STP) and the reprogramming of common metabolic mechanisms play a key role in the MM tumorigenesis processes.
Therefore, the study of miRNA, of the aberrant STP activation profile and of the altered metabolism represent a challenge for understanding molecular determinant of resistance developed during the history of MM disease.
The aim of this project is to identify, through a combined application of innovative technologies, specific aberrant signals and peculiar metabolic phenotypes in MM cell lines and primary samples collected at diagnosis and/or at relapse/refractoriness disease.
miRNAs modulate regulatory cell pathways by influencing target genes and may serve crucial functions in oncogenesis. miRNA detection and quantification will be performed by real-time quantitative reverse-transcription polymerase-chain reaction.
Proteomic profiles of cellular populations will be analyzed by using the Reverse Phase Protein Array (RPPA), a high-throughput technology capable of processing, with high efficiency and accuracy, simultaneously numerous proteins in their total and activated form using a limited number of cells.
The impact of aberrant signal on cell metabolism will be analyzed by Seahorse XF Analyzer, a new instrument able to simultaneously measure, in real time and on live cells, several metabolic parameters and functions.
This information, along with the identification of aberrant cross-talking pathways, which may develop during the treatments history, will allow the elucidation of mechanisms involved in progression and resistance of MM.
MM is considered an incurable disease, and little is known about the insurgence of resistance mechanisms that cause the failure of the currently adopted therapeutic strategies. Any additional information is thus precious, as it concurs on improving the current knowledge of this deadly disease and helps developing new and more effective therapeutic strategies. miRNAs, STPs and cellular metabolism are among the most promising novel targets that have been in the spotlight of current cancer research, given their role in multiple aspects of cell biology and their relatively easiness to be targeted with specific inhibitors. Thus, we hypothesize that a multi-level integrated approach aimed at the identification of peculiar profiles associated to resistance would be uniquely suited to the task of dissecting the mechanism of resistance insurgence. Indeed, we plan to deploy the cutting-edge, innovative technology owned by our lab for miRNA, proteomic and metabolic analysis, to unravel the crosstalk between these cellular processes and how this contributes to the establishment of a resistant phenotype in MM cells. In fact, RPPA has been successfully applied by our lab and others to elucidate the phosphoproteome changes in the main STPs involved on the major cellular processes as proliferation, survival and differentiation, during the neoplastic transformation. The current golden standard for metabolic analysis on live cells, the Seahorse XF analyzer, has been invaluable in unraveling the metabolic shifts of cancer cells, and can detect how the metabolism of these cells reacts when challenged with external stresses, such as therapeutic drugs. miRNAs profiling is still contributing to provide novel biomarkers in disease progression and aggressiveness. To our knowledge, the above-described integrated approach has never been attempted on MM cells, thus constituting a promising strategy for the generation of reliable data to be used to overcome the therapeutic resistance in this disease.