Engineering a spatiotemporal macrophage circuit via STING phase separation to override immune suppression in pancreatic cancer

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies, largely due to its highly immunosuppressive tumor microenvironment (TME), which fuels metastasis and resistance to immunotherapy. Through comprehensive analysis of single-cell RNA sequencing (scRNA-seq) datasets, we identified multiple heterogeneous tumor-associated macrophage (TAMs) subpopulations as key regulators of PDAC progression, which co-express MRC1 and exert their effects by actively suppressing anti-tumor immune responses. To overcome this barrier, we developed a spatiotemporal macrophage reprogramming platform that leverages STING phase separation to reprogram TAM plasticity and reshape the immune landscape. This system, PMMB, integrates a CSF-1R inhibitor and a STING agonist within a macrophage-mimetic nanostructure, enabling sequential, controlled reprogramming of TAMs. By leveraging STING phase separation, PMMB stabilizes TAMs in an anti-tumor CD80⁺ phenotype while preventing excessive in..., Single-cell RNA sequencing of PDAC tumors in mice
To create a subcutaneous xenograft PDAC model, 5×106 KPC cells were administered via injection into the right flank of male C57BL/6 mice. PMMB was injected into the tail vein of mice on days 0, 3, 6, 9, and 12. On Day 14, the tumors from the mice were collected for scRNA-seq.
Tumors were enzymatically dissociated into single-cell suspensions. After removal of dead cells, viable cells (>80%) were resuspended in PBS with 0.5% BSA and processed for scRNA-seq. Single-cell libraries were generated using a droplet-based platform according to the manufacturer’s protocol, including reverse transcription, cDNA amplification, and library indexing.
Libraries were sequenced on an Illumina NovaSeq platform, yielding ~200,000 reads per cell with a median detection of ~2,000–2,500 genes. Raw reads were filtered with fastp (4), aligned to the mouse mm10 genome, and processed to generate gene-barcode matrices for downstream analysis in Seurat.
Bulk RN..., # Data from: Engineering a spatiotemporal macrophage circuit via STING phase separation to override immune suppression in pancreatic cancer

Dataset DOI: [10.5061/dryad.kkwh70shs](https://doi.org/10.5061/dryad.kkwh70shs)

## Description of the data and file structure

#### File: scRNA-Seq.zip

**Description:** This compressed folder contains raw and processed single-cell RNA sequencing data used in the study.

Six data files are included, representing two experimental groups (control and treatment), with three biological replicates per group:

PBS-1, PBS-2, PBS-3: Control group (phosphate-buffered saline–treated samples)

PMMB-1, PMMB-2, PMMB-3: Experimental group (treated samples)

In each subfolder of this dataset, you will find three essential files that together form the Gene Expression Matrix for a single sample.

Here is a description of what each file contains:

1\. ‘barcodes.tsv.gz’：A list of all unique cell barcodes identified in the sample.

Each line represents a single cell...,