BD Rhapsody™ Single-Cell Analysis System Workflow: From Sample to Multimodal Single-Cell Sequencing Data.

Advancements in single-cell sequencing have revolutionized our understanding of complex biological systems such as the immune system and allowed us to overcome limitations in various disciplines of life science research such as oncology, developmental biology, or neurobiology (Perkel, Nature 595. https://www.nature.com/articles/d41586-021-01994-w , 2021).The BD Rhapsody™ Single-Cell Analysis System enables us to capture multimodal information from thousands of single cells in parallel ("Multiomics") covering mRNA expression levels, protein expression levels, the immune repertoire for T-cell receptors (TCR) and B-cell receptors (BCR), and the identification of antigen-specific T cells and B cells using dCODE Dextramer® (RiO) from Immudex. The system utilizes microwell-based cartridges that allow to capture a broad range of single cells and an imaging device for sample quality control and workflow quality control (including viability and multiplets). The power of Multiomics relies on simultaneously measuring several aspects of single cells, including gene expression and protein abundance, using next generation sequencing (NGS) as a single readout.Here we describe the complete BD Rhapsody™ Single-Cell Analysis System from the sample preparation including different options for the antibody and/or dCODE Dextramer® staining through to the data analysis.For updated protocols, guides, and technical bulletins, please visit the BD Scomix page: https://scomix.bd.com/hc/en-us or the BDB webpage: https://www.bdbiosciences.com/en-eu .

Deregulated MYC expression induces dependence upon AMPK-related kinase 5.

Deregulated expression of the MYC oncoprotein contributes to the genesis of many human tumours, yet strategies to exploit this for a rational tumour therapy are scarce. MYC promotes cell growth and proliferation, and alters cellular metabolism to enhance the provision of precursors for phospholipids and cellular macromolecules. Here we show in human and murine cell lines that oncogenic levels of MYC establish a dependence on AMPK-related kinase 5 (ARK5; also known as NUAK1) for maintaining metabolic homeostasis and for cell survival. ARK5 is an upstream regulator of AMPK and limits protein synthesis via inhibition of the mammalian target of rapamycin 1 (mTORC1) signalling pathway. ARK5 also maintains expression of mitochondrial respiratory chain complexes and respiratory capacity, which is required for efficient glutamine metabolism. Inhibition of ARK5 leads to a collapse of cellular ATP levels in cells expressing deregulated MYC, inducing multiple pro-apoptotic responses as a secondary consequence. Depletion of ARK5 prolongs survival in MYC-driven mouse models of hepatocellular carcinoma, demonstrating that targeting cellular energy homeostasis is a valid therapeutic strategy to eliminate tumour cells that express deregulated MYC.