Ginsenoside Rh2

Interferon gamma (IFNγ) can amplify immune cell-mediated anti-tumor immunity, as well as directly kill cancer cells. Ginsenoside Rh2 (Rh2), a bioactive compound in traditional Chinese medicine, exhibits anti-cancer effects such as inhibiting proliferation and metastasis. Our earlier research found that Rh2 combined with IFNγ enhanced CXCL10 secretion in cancer cells. Here, we explored whether Rh2 and IFNγ exerted more potent anti-cancer activity in vitro and in vivo, along with its mechanisms and clinical value. Our data showed that Rh2 in combination with IFNγ resulted in a remarkably increased cytotoxicity in colorectal cancer cells including HT29, LoVo and T84 cell lines. Consistently, intratumoral injection with Rh2 plus IFNγ further restricted the HT29 tumor growth in vivo, and importantly, it was demonstrated to be safe for mice. Meanwhile, the combo treatment activated the stimulator of interferon genes (STING) pathway in cancer cells, promoting the transcription of downstream type I interferon. RNA sequencing revealed a dramatically transcriptional alteration in cancer cells with combo treatment and indicated that Rh2 further augmented the activation of interferon signaling pathway, compared with the IFNγ alone. Inhibition of janus kinase (JAK) by ruxolitinib could significantly rescue the cell death-triggered by the combo treatment. Then, a gene set named Rh2+IFNγ signature genes (RISG) was defined, which contained top 20 significantly upregulated genes from the combo treatment. Patients who exhibited a favorable response to the immunotherapy had a higher expression of RISG in tumor compared with those who did not respond. And the high expression of RISG was correlated with better clinical outcome in patients with colorectal cancer (CRC) and skin cutaneous melanoma (SKCM). Herein, the combination of Rh2 with IFNγ served as a promising strategy for cancer treatment, and its-derived RISG gene set also exhibited potential value in predicting clinical outcome. Schematic diagram of the anti-cancer effect of Rh2 combined with IFNγ. The schematic diagram illustrated that ginsenoside Rh2 in combination with IFNγ robustly activated the interferon signals in cancer cells, ultimately leading a significant cell death of cancer cells. ISGs, interferon-stimulated genes. Created with BioRender.com.

Macrophage-hitchhiking drug delivery across the blood-brain barrier (BBB) has garnered considerable attention for its potential in glioblastoma (GBM) treatment. However, challenges such as the complexity of in vitro synthesis and the risk of macrophage polarization toward protumor phenotypes hinder its practical application. We developed a receptor ligand-free nanoshuttle that can hitchhike on macrophages in vivo for efficient BBB penetration and targeted drug delivery while inducing macrophage polarization toward antitumor phenotypes for GBM therapy. The nanoshuttle was constructed through surface engineering of a Fe-containing metal-organic framework (MOF) to optimize its hydrophilicity and was loaded with ginsenoside Rh2 (Rh2). Upon intravenous injection, the nanoshuttles specifically adsorbed proteins from the complement system, forming a protein corona that enhanced macrophage endocytosis and facilitated the in vivo synthesis of the macrophage-hitchhiking nanoshuttles. Exploiting the chemotaxis of macrophages toward GBM tissue, the macrophage-hitchhiking nanoshuttles efficiently penetrated the BBB and target GBM tissue. The intrinsic iron species of the nanoshuttle regulated iron metabolism in the tumor microenvironment, directing macrophage polarization toward an antitumor M1 phenotype for immunotherapy. Combined with the ^•OH generating capacity of Fe-MOF and the tumor-killing effects of Rh2, these nanoshuttles effectively suppressed tumor growth and improved survival in a GBM mouse model.