Extended Data Fig. 5: Longitudinal RNA sequencing of CTCs during digoxin treatment. | Nature Medicine

Extended Data Fig. 5: Longitudinal RNA sequencing of CTCs during digoxin treatment.

From: Digoxin for reduction of circulating tumor cell cluster size in metastatic breast cancer: a proof-of-concept trial

Extended Data Fig. 5

(a) The diagram shows the clinical timeline of patient nr. 5, including treatment history (boxes), radiological assessment (dashed lines) and serum tumor marker (CA 15-3) (grey). (b) Workflow for molecular analysis, including microfluidic separation and capture, robotic micromanipulation, pooling of circulating tumor cells (CTCs) into individual tubes and RNA sequencing. (c) Heatmap displaying differentially expressed genes (DESeq Wald test adjusted P value < 0.05) between CTC pools obtained prior to digoxin intake (n = 5 pre-digoxin pools) and post-digoxin intake (n = 2 post-digoxin pools). Values are displayed as gene-scaled (z-score) log2 counts per million mapped reads after normalization. (d) Enriched pathways among downregulated (top) and upregulated (bottom) genes of CTCs post-digoxin intake (over-representation analysis adjusted P value < 0.05). Shown is a reduced list of ontologies after simplification using semantic similarity. TP, treatment pause; PD, progressive disease; SD, stable disease; TAM, tamoxifen. Panels (a) and (b) were created with BioRender (https://biorender.com).

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