ASH 2019: Characterizing Molecular Drivers of Myeloma Cells Via CRISPR Activation
Posted: Friday, December 20, 2019
Through the use of gain-of-function studies, researchers were able to validate the relevance of genes implicated in multiple myeloma development and identify new candidate regulators of multiple myeloma cells. Huihui Tang, PhD, of Harvard Medical School, Boston, presented the work he completed with colleagues at the 2019 American Society of Hematology (ASH) Annual Meeting & Exposition in Orlando, Florida (Abstract 313).
“These results provide novel insights into the biology of multiple myeloma cells and may have therapeutic implications towards suppressing the ability of multiple myeloma cells to transition to states of advanced biological aggressiveness,” concluded the authors.
The team of researchers applied CRISPR technology to activate genes in four different multiple myeloma cell lines. Cells were transduced to express transcriptional activators and the Calabrese genome-scale single guide RNA (sgRNA) library. Next-generation sequencing was then applied to quantify the abundance of sgRNA at baseline and at various time points. Rank aggregation algorithms were used to identify genes that had statistically significant enrichment or depletion of sgRNAs, which reflected positive and negative effects of gain-of-function genes on cell survival and proliferation.
Positive regulators of multiple myeloma cell growth were identified, such as the transcription factors POU2F2, POU2AF1, IRF4, and MYC; growth factor signaling mediators IGF1R, IRS1; and Ras family members like KRAS. Among the positive regulators identified, not all were essential for multiple myeloma survival or proliferation in CRISPR knockout studies within the same multiple myeloma cell lines. Researchers then focused on understanding the mechanistic basis of the role of POU2AF1 as a driver of multiple myeloma growth. They found a cluster of more than 50 genes that had not yet been studied in multiple myeloma that showed a distinct enrichment for transcriptional regulators not required for growth of cells in CRISPR knockout studies.
Disclosure: For full disclosures of the study authors, visit ash.confex.com.
