Prostate-cancer (PC) is a leading cause of cancer-related deaths in men worldwide. Interleukin-(IL)-30 is a PC-progression driver, and its suppression would be strategic for fighting metastatic disease. Biocompatible Lipid-Nanoparticles (NPs) were loaded with CRISPR/Cas9gRNA to delete human(h)IL30-gene and functionalized with anti-PSCA-Abs (Cas9hIL30-PSCA-NPs). Efficiency of the NPs in targeting IL30 and metastatic potential of PC cells was examined in vivo, in xenograft models of lung metastasis, and in vitro, by using 2-Organ-on-Chip (2-OC), containing 3D-spheroids of IL30+PC-Endothelial-Cell(EC) co-cultures in circuit with either Lung-mimicking-spheroids, or Bone-marrow(BM)-niche-mimicking-scaffolds. Cas9hIL30-PSCA-NPs demonstrated circulation stability, genome editing efficiency, without off-target effects and organ toxicity. Intravenous injection of three-doses/13-days, or five-doses/20-days, of NPs in mice bearing circulating PC cells and micro-emboli substantially hindered lung metastasization. Cas9hIL30-PSCA-NPs inhibited PC cell proliferation and expression of IL30 and metastasis-drivers, such as CXCR2, CXCR4, IGF1, L1CAM, METAP2, MMP2 and TNFSF10, whereas CDH1 was up-regulated. PC-Lung and PC-BM 2-OCs revealed that Cas9hIL30-PSCA-NPs suppressed PC cell release of CXCL2/GROβ, which in vivo was associated with intra-metastatic myeloid cell infiltrates, and of DKK1, OPG and IL6, which in vitro boosted endothelial-network formation and cancer cell migration. Development of a patient-tailored nanoplatform for selective CRISPR-mediated IL30 gene deletion is a clinically valuable tool against PC progression.
Molecular therapy : the journal of the American Society of Gene Therapy. 2024 Sep 06 [Epub ahead of print]
Cristiano Fieni, Stefania Livia Ciummo, Carlo Sorrentino, Simona Marchetti, Simone Vespa, Paola Lanuti, Lavinia Vittoria Lotti, Emma Di Carlo
Department of Medicine and Sciences of Aging, "G. d'Annunzio" University" of Chieti-Pescara, 66100, Chieti, Italy; Anatomic Pathology and Immuno-Oncology Unit, Center for Advanced Studies and Technology (CAST), "G. d'Annunzio" University of Chieti-Pescara, 66100, Chieti, Italy., Department of Medicine and Sciences of Aging, "G. d'Annunzio" University" of Chieti-Pescara, 66100, Chieti, Italy., Department of Experimental Medicine, "La Sapienza" University of Rome, 00161, Rome, Italy., Department of Medicine and Sciences of Aging, "G. d'Annunzio" University" of Chieti-Pescara, 66100, Chieti, Italy; Anatomic Pathology and Immuno-Oncology Unit, Center for Advanced Studies and Technology (CAST), "G. d'Annunzio" University of Chieti-Pescara, 66100, Chieti, Italy. Electronic address: .