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Engineered Nanobodies Targeting Endogenous Digoxin-like Substances Function as Novel Immune Checkpoints Inhibitors and Enhance Immunotherapy against Cancer Metastasis

GSE308031 Mus musculus (mouse) RNA-seq (gene expression) 12 samples Submitted 2025/09/20 Platform GPL24247
At a glance

RNA-seq (gene expression) · mouse · 12 samples

What it is
RNA-seq (gene expression)Gene activity measured by sequencing. The summary does not mention single-cell work, so this is most likely bulk: one average per sample, the usual starting point for comparing groups.
Organism
Mus musculus (mouse)
Samples
12 samples
Data you can get
A processed table (series matrix) and supplementary files from NCBI; the raw sequencing reads from ENA. Which of these hold counts depends on the study, so check the file list.
Summary (written by the submitters)
Digoxin is a well-recognized cardiovascular medication; however, epidemiological investigations have associated its clinical application with an elevated risk of malignancy. To elucidate it, we discovered, for the first time, that therapeutic digoxin can facilitate tumor metastasis by acting as an inducer of immune checkpoints, including PD-1, PD-L1/2, CTLA-4, LAG-3, ICOS, TIM-3, and TIGIT, among others.
Read the full summary ↓
The PD-1/PD-L1 axis plays a pivotal role. Digoxin upregulates PD-L1 production in cancer cells through multiple mechanisms, including digoxin-PKM2 interaction, liquid-liquid phase separation, and Na,K-ATPase degradation. Notably, endogenous digoxin (ED) was significantly upregulated in breast cancer, correlating with increased metastasis. When specific nanobodies (Nbs) were developed to functionally inhibit ED in vivo, pulmonary tumor metastasis was reduced. However, the efficacy of ED-targeting Nbs was limited by endogenous ouabain (EO), another hormone structurally similar to digoxin. To address this, Nbs co-targeting ED and EO demonstrated an increased anti-metastatic effect and a notable reduction in immune checkpoint expression. Furthermore, Nbs simultaneously targeting ED, EO, and PD-1 exhibited significant synergy in suppressing tumor progression with a favorable safety profile. In conclusion, therapeutic digoxin and ED are potent inducers of immune checkpoints. Engineered nanobodies co-targeting cardiotonic hormones (ED, EO) and immune checkpoints present a promising novel approach in the treatment of breast cancer metastasis.
This dataset
Download · which file do I need?

Direct links to NCBI and ENA, no account and no request form. Pick by what you want to do: take the small processed table to look, the supplementary files to re-analyse, or the original raw data to start over.

1 · Just have a look

Processed table

The series matrix: one table of values per sample as the submitters processed it. The smallest download, and it opens in Excel or R. Start here to see what the data look like.

2 · Re-analyse it

Supplementary files

The count tables and other files the authors uploaded. Take only the samples you need, or the whole study as GSE308031_RAW.tar (it can run to tens of gigabytes).

3 · Start from the raw reads

Raw sequencing reads

The original FASTQ files, straight from the sequencer, held at ENA. Large, and you align them yourself, so only choose this if you need to re-process the study from scratch.

What the files usually look like: For RNA-seq the counts are usually a table among the supplementary files (look for 'counts', 'expression', 'FPKM' or 'TPM' in the file name). The series matrix of a sequencing study is sometimes nearly empty.

Also filed as BioProject PRJNA1328618 and SRA study SRP620382. Searching any of these in the dataset finder brings you back here.

Samples in this study

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+ 12 more — browse all 12 samples with per-sample file links →

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