"Nanoranger" characterizes leukemia cells at the single-cell level using long-read sequencing.
Single-cell RNA sequencing (scRNA-seq) has expanded our ability to distinguish cell populations and elucidate their interactions with each other. However, there are still difficulties in distinguishing leukemia cells from normal hematopoiesis based on gene expression alone. Livius Penter, BIH Digital Clinician Scientist Fellow and specialist at the Department of Hematology, Oncology and Tumor Immunology at Charité - Universitätsmedizin Berlin, together with colleagues Mehdi Borji and Adi Nagler, addressed this problem.
Using the long-read sequencing platform from Oxford Nanopore Technologies (ONT), the researchers have developed the nanoranger protocol, which allows genetic information to be read from scRNA-seq cDNA libraries in a targeted and flexible manner. The protocol helps to find somatic mutations more easily and thus identify leukemia cells and their subpopulations. The nanoranger protocol also helps to read out a number of other molecular markers, such as mitochondrial DNA mutations, fusion genes, chimeric antigen receptor (CAR) sequences or alternative splice variants.
The researchers recently published their findings on the nanoranger protocol in Nature Communications. In the paper, they present a number of applications, such as the elucidation of cellular mechanisms associated with response or resistance to an experimental therapy (decitabine and ipilimumab) that was tested in acute myeloid leukemia (AML) as part of a clinical trial. In the future, nanoranger will help to elucidate the development of therapy resistance in hematologic neoplasms. In addition, nanoranger could serve as a tool for clinical hematological diagnostics, allowing comprehensive information on the state of leukemia and immune cells to be obtained at the same time.
Publication
Penter, L., Borji, M., Nagler, A. et al. Integrative genotyping of cancer and immune phenotypes by long-read sequencing. Nat Commun15, 32 (2024). https://doi.org/10.1038/s41467-023-44137-7