1. Identification of a Maturation Plasma Cell Index through a Highly Sensitive Droplet Digital PCR Assay Gene Expression Signature Validation in Newly Diagnosed Multiple Myeloma Patients
- Author
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Marina Martello, Vincenza Solli, Rosalinda Termini, Ajsi Kanapari, Daniel Remondini, Enrica Borsi, Andrea Poletti, Silvia Armuzzi, Barbara Taurisano, Ilaria Vigliotta, Gaia Mazzocchetti, Elena Zamagni, Alessandra Merlotti, Paola Tacchetti, Lucia Pantani, Serena Rocchi, Ilaria Rizzello, Katia Mancuso, Michele Cavo, Carolina Terragna, Martello M., Solli V., Termini R., Kanapari A., Remondini D., Borsi E., Poletti A., Armuzzi S., Taurisano B., Vigliotta I., Mazzocchetti G., Zamagni E., Merlotti A., Tacchetti P., Pantani L., Rocchi S., Rizzello I., Mancuso K., Cavo M., and Terragna C.
- Subjects
digital PCR ,plasma cell maturation ,Plasma Cells ,Organic Chemistry ,differentiation stage ,Reproducibility of Results ,General Medicine ,self-renewal ,Real-Time Polymerase Chain Reaction ,hedgehog signaling ,Catalysis ,Computer Science Applications ,Inorganic Chemistry ,gene expression ,Humans ,RNA ,Hedgehog Proteins ,Prospective Studies ,Physical and Theoretical Chemistry ,Multiple Myeloma ,Transcriptome ,Molecular Biology ,Spectroscopy ,multiple myeloma ,differentiation stages ,Retrospective Studies - Abstract
DNA microarrays and RNA-based sequencing approaches are considered important discovery tools in clinical medicine. However, cross-platform reproducibility studies undertaken so far have highlighted that microarrays are not able to accurately measure gene expression, particularly when they are expressed at low levels. Here, we consider the employment of a digital PCR assay (ddPCR) to validate a gene signature previously identified by gene expression profile. This signature included ten Hedgehog (HH) pathways’ genes able to stratify multiple myeloma (MM) patients according to their self-renewal status. Results show that the designed assay is able to validate gene expression data, both in a retrospective as well as in a prospective cohort. In addition, the plasma cells’ differentiation status determined by ddPCR was further confirmed by other techniques, such as flow cytometry, allowing the identification of patients with immature plasma cells’ phenotype (i.e., expressing CD19+/CD81+ markers) upregulating HH genes, as compared to others, whose plasma cells lose the expression of these markers and were more differentiated. To our knowledge, this is the first technical report of gene expression data validation by ddPCR instead of classical qPCR. This approach permitted the identification of a Maturation Index through the integration of molecular and phenotypic data, able to possibly define upfront the differentiation status of MM patients that would be clinically relevant in the future.
- Published
- 2022