Novel peptide targeting CXCR4 disrupt tumor-stroma crosstalk to eliminate migrating cancer stem cells

Tiwary, K.; Lahusen, A.; Inaas, S.; Beitzinger, B.; Schmid, R.; Harms, M.; Hauff, S.; Arnold, F.; Walter, K.; Alcala, S.; Hahn, S. A.; Hessmann, E.; Kleger, A.; Azoitei, N.; Seufferlein, T.; Sainz Anding, B.; Muench, J.; Linden, M.; Hermann, P. C.

doi:10.1101/2025.03.03.641126

bioRxiv · 10.1101/2025.03.03.641126

Novel peptide targeting CXCR4 disrupt tumor-stroma crosstalk to eliminate migrating cancer stem cells

Tiwary, K.·Lahusen, A.·Inaas, S.·Beitzinger, B.·Schmid, R.·Harms, M.·Hauff, S.·Arnold, F.·Walter, K.·Alcala, S.·Hahn, S. A.·Hessmann, E.·Kleger, A.·Azoitei, N.·Seufferlein, T.·Sainz Anding, B.·Muench, J.·Linden, M.·Hermann, P. C.

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Abstract

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and metastatic malignancies worldwide. Migrating cancer stem cells (miCSCs) marked by CD133+CXCR4+ expression drives metastasis but lacks effective drug targets. Here, we show that activated pancreatic stellate cells secrete the CXCR4 ligand CXCL12 to foster stemness, epithelial-to-mesenchymal transition (EMT), and chemoresistance. Protein interaction network analyses links CXCL12/CXCR4 signaling axis and the downstream transcription factor BMI1. Knockdown experiments confirmed the BMI1s role in (mi)CSCs maintenance and survival. Novel CXCR4 inhibitors, i.e., the endogenous human peptide EPI-X4 and its derivatives (e.g., JM#21) strongly inhibited the in vitro migration of miCSCs. In particular, the most potent EPI-X4 derivate JM#21 sufficiently suppressed EMT, stemness, and self-renewal of human PDAC cell lines. In addition, JM#21 sensitized cell lines towards gemcitabine and paclitaxel. Overall, our study reveals that (mi)CSCs are enhanced and maintained via a tumor-stroma crosstalk through BMI1, ultimately promoting metastases and therapeutic resistance in PDAC. Peptide targeting of the CXCL12/CXCR4/BMI1 signaling axis via JM#21 could enhance PDAC combination therapies, offering a promising strategy against this deadly cancer. SynopsisThe study identifies a tumor-stroma interaction mediated by pancreatic stellate cells (PSCs) secreting CXCL12, which binds to CXCR4 on (mi)CSCs, fostering stemness, epithelial-to-mesenchymal transition (EMT), and chemoresistance. The CXCL12/CXCR4 axis activates the downstream BMI1 transcription factor, crucial for migration and stemness maintenance. O_FIG O_LINKSMALLFIG WIDTH=173 HEIGHT=200 SRC="FIGDIR/small/641126v1_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@14e117org.highwire.dtl.DTLVardef@c5db35org.highwire.dtl.DTLVardef@1ba7d2dorg.highwire.dtl.DTLVardef@702ccf_HPS_FORMAT_FIGEXP M_FIG C_FIG O_LICXCL12 enhances (mi)CSC populations and metastatic potential through CXCR4 signaling. C_LIO_LIBMI1 is identified as a pivotal downstream effector linking CXCR4 to EMT and stemness. C_LIO_LIJM#21 effectively blocks CXCL12-induced migration, EMT, and stemness in vitro, demonstrating superior efficacy compared to other CXCR4 inhibitors. C_LIO_LIEncapsulation of JM#21 in silica nanoparticles enhances its stability and delivery, reducing chemoresistance and miCSC populations in co-culture systems. C_LIO_LICombining JM#21 with chemotherapy significantly impairs colony formation and CSC-mediated drug resistance. C_LI

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Tiwary, K., Lahusen, A., Inaas, S., Beitzinger, B., Schmid, R., Harms, M., Hauff, S., Arnold, F., Walter, K., Alcala, S., Hahn, S. A., Hessmann, E., Kleger, A., Azoitei, N., Seufferlein, T., Sainz Anding, B., Muench, J., Linden, M., Hermann, P. C.. 2025-03-11. Novel peptide targeting CXCR4 disrupt tumor-stroma crosstalk to eliminate migrating cancer stem cells. https://doi.org/10.1101/2025.03.03.641126

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