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Updated: Jul 26, 2026
Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models
Published on: December 9, 2016
Single-cell and spatial transcriptomics reveal the interaction between PRRX2-driven epithelial cells and SPP1+
Jingmei Liu1,2, Zhilong Li3, Guanhua Liu3
1Department of Gastroenterology, Cancer Hospital Affiliated to Shanxi Medical University/Shanxi Province Cancer Hospital/Shanxi Hospital Affiliated to Cancer Hospital, Chinese Academy of Medical Sciences, Taiyuan, Shanxi 030013, China.
Background:
The development of gemcitabine resistance significantly diminishes treatment efficacy and worsens prognosis in patients with pancreatic ductal adenocarcinoma (PDAC). A comprehensive understanding of the molecular and cellular mechanisms driving gemcitabine resistance is thus essential for optimizing treatment strategies and improving survival outcomes in PDAC.
Methods:
An integrated analysis of bulk RNA sequencing (RNA-seq), single-cell RNA sequencing (scRNA-seq), and spatial transcriptomics was performed to elucidate the molecular mechanisms of gemcitabine resistance. Tumor-associated epithelial and myeloid cell subsets were identified, and their interactions were characterized using clustering, pseudotime analysis, and cell-to-cell communication networks. Findings were validated through molecular and cellular experiments in PDAC cell lines, as well as multicolor immunofluorescence staining in clinical cohort samples.
Results:
The findings revealed that PRRX2+ epithelial cells (EC) and SPP1+ tumor-associated macrophages (TAM) were enriched in gemcitabine-resistant samples, with robust cell-to-cell communication observed between these two populations. PRRX2+ EC cells and SPP1+ TAM cells interacted through the TGFB1/TGFBR2 axis, promoting epithelial-mesenchymal transition (EMT) and extracellular matrix (ECM) remodeling, thereby contributing to drug resistance. The transcription factor PRRX2 was identified as a central regulator of this resistance mechanism, amplifying TGF-β signaling through TGFB1 to induce EMT and activating Fibronectin 1 to remodel the ECM, thereby enhancing an immunosuppressive tumor microenvironment. Furthermore, experimental downregulation of PRRX2 in pancreatic cancer cell lines resulted in reduced tumor cell migration. High expression levels of PRRX2+EC/SPP1+TAM were associated with poor prognosis and may indicate susceptibility to Dasatinib and antibody-drug conjugates (ADCs) targeting TROP2, MUC1, and CEACAM5.
Conclusions:
The results revealed the critical role of PRRX2+EC/SPP1+TAM cell communication in mediating gemcitabine resistance in PDAC. These findings provide new insights into potential therapeutic strategies, highlighting the importance of targeting the PRRX2+EC/SPP1+TAM axis and supporting the use of multidrug regimens to overcome resistance and improve patient outcomes in PDAC.
Insights
Gemcitabine resistance in pancreatic cancer is driven by communication between PRRX2+ epithelial cells and SPP1+ macrophages. Targeting this axis offers new therapeutic strategies for pancreatic ductal adenocarcinoma (PDAC).
Area of Science:
- Oncology
- Cancer Biology
- Translational Research
Background:
- Gemcitabine resistance is a major challenge in pancreatic ductal adenocarcinoma (PDAC) treatment, negatively impacting patient prognosis.
- Understanding the molecular mechanisms of gemcitabine resistance is crucial for developing effective therapeutic strategies in PDAC.
Purpose of the Study:
- To elucidate the molecular and cellular mechanisms underlying gemcitabine resistance in pancreatic ductal adenocarcinoma (PDAC).
- To identify specific cell populations and their interactions contributing to drug resistance in PDAC.
Main Methods:
- Integrated analysis of bulk RNA sequencing (RNA-seq), single-cell RNA sequencing (scRNA-seq), and spatial transcriptomics.
- Identification and characterization of tumor-associated epithelial and myeloid cell subsets and their communication networks.
- Validation using PDAC cell line experiments and clinical cohort sample analysis.
Main Results:
- PRRX2+ epithelial cells (EC) and SPP1+ tumor-associated macrophages (TAM) were enriched in gemcitabine-resistant PDAC samples.
- PRRX2+ EC and SPP1+ TAM interact via the TGFB1/TGFBR2 axis, promoting epithelial-mesenchymal transition (EMT) and extracellular matrix (ECM) remodeling, driving drug resistance.
- PRRX2 acts as a key regulator, amplifying TGF-β signaling and activating Fibronectin 1, creating an immunosuppressive tumor microenvironment and reducing tumor cell migration.
Conclusions:
- PRRX2+EC/SPP1+TAM cell communication critically mediates gemcitabine resistance in PDAC.
- Targeting the PRRX2+EC/SPP1+TAM axis presents a promising therapeutic strategy for overcoming gemcitabine resistance.
- Multidrug regimens targeting this axis may improve patient outcomes in PDAC.
