Kepeng Wang

The lack of tumor-specific targeting by oncolytic viruses (OVs) restricts their therapeutic potential. Here, we report a “cap-linker” strategy to modify the vesicular stomatitis virus (VSV) glycoprotein (G protein, VSV-G ^IN ). The modified VSV (Pro-VSV ^IN ) loses its ability to infect cells until present in the tumor microenvironment, where the linker is cleaved by tumor-enriched matrix metalloproteinases (MMPs). When Pro-VSV ^IN is armed with a single-chain IL-12 (Pro-VSV ^IN -IL-12) and systemically injected into MC38 tumor-bearing mice, Pro-VSV ^IN -IL-12 eradicates established tumors while avoiding systemic toxicity, and elicits long-term anti-tumor immune memory. Systemic administration of Pro-VSV ^IN -IL-12 also markedly inhibits the growth of metastatic cancers in the lung. The efficacy of Pro-VSV ^IN -IL-12 was further improved when combined with PD-1 checkpoint blockade. Compared to WT VSV ^IN that carries IL-12, intravenously injected Pro-VSV ^IN -IL-12 leads to strong Th1 immunity, near abolishment of regulatory T cells, and potent CD8 ⁺ T cell activation and clonal expansion in tumors. This preclinical work demonstrates a new tumor-specific targeting strategy of VSV for enhanced immunotherapy safety with superior efficacy.

Supplementary Fig. S2. Endoplasmic reticulum stress and unfolded protein response in normal physiology and cancer Tumor cells frequently encounter endogenous and exogenous stresses that influence protein folding in the ER, including hypoxia, inflammation, nutrient deprivation, altered glycolysis, DNA damage, genomic instability, oncogene activation, and proteasome dysfunction, can cause excessive accumulation of misfolded or unfolded proteins within the ER lumen, a condition termed "ER stress." As a response to prolonged ER stress conditions, the ER responds to the ER stress by activating intracellular signal transduction pathways, collectively referred to as the UPR, to restore-ER homeostasis (protective effect) or activate cell death (cytotoxic effect). The UPR signaling cascade comprises three parallel pathways that employ unique signal transduction mechanisms and are initiated by activating three conserved ER-stress sensors: IRE1, ATF6, and PERK. Furthermore, in mammals and under resting conditions, the critical ER-resident chaperone, GRP78/BiP, prevents the activation and blocks the downstream signaling of UPR by binding to the N-terminal side of stress sensor proteins, IRE-1, PERK, and ATF6 and maintains these sensors in an inactive state. When ER stress is triggered, and the ER luminal load of unfolded or misfolded proteins increases, the GRP78/BiP dissociates, activating IRE1, PERK, and ATF6 signaling cascades, leading to the activation of these UPR signaling pathways. Upon activation, the UPR can regulate many cellular events of tumor cells, including autophagy, ER biogenesis, cell survival, metastasis, angiogenesis, migration and invasion, chemoresistance, glycolysis, and proliferation. ER, Endoplasmic reticulum; UPR, unfolded protein response; PREK, protein kinase R (PKR) like endoplasmic reticulum kinase; ATF, activating transcription factor; IRE1, inositol requiring enzyme 1; GRP78, 78-kDa glucose-regulated protein/immunoglobulin-binding protein; CHOP, C/EBP homologous protein; AMPK, AMP-activated protein kinase; XBP1, X-box binding protein 1; Dr5, death receptor 5; GADD34, growth arrest and DNA damage-inducible protein 34; ERO1α, ER oxidoreductin 1; GCN2, general control non-derepressible 2.

Supplementary Fig. S1. The structure of mTOR complexes and overview of the PI3K/AKT/mTOR signaling pathway. The mTOR signaling pathway is implicated in a variety of major cellular processes, from cell growth to apoptosis. mTOR nucleates at least two distinct multi-protein complexes, mTORC1 and mTORC2. mTORC1 activation occurs in the response of growth factors through PI3K/Akt and RAS/MAPK pathways. Human mTOR can assemble at least two larges, physically, and functionally distinct polyprotein complexes, mTORC1, and mTORC2. mTORC1 and mTORC2 share several common proteins, including the catalytic subunit mTOR, mLST8 and DEPTOR, and Tel2/Tti1 complex. In addition, each complex has specific proteins. The activity of mTORC1 is regulated by multiple signals, including nutrient availability such as amino acids (leucine, arginine, and glutamine), intracellular energy level (Low ATP/High AMP), and extracellular growth factors such as insulin and cytokine receptors. A receptor-mediated activation of PI3K leads to PIP3 being produced from PIP2. PIP3 then promoted the recruitment of PDK1, AKT, and mTORC2 complex, facilitating the phosphorylation and full activation of AKT at Thr308 and Ser473 via PDK1 and mTORC2, respectively. Activated AKT phosphorylates TSC2 and induces the dissociation of TSC1 and TSC2 complex, which results in the dissociation of TSC2 from lysosomes and loss of their inhibitory action on RHEB/mTORC1 axis. Upon activation, mTOC1 promotes basic cellular events, including Cap-dependent translation, fatty acid synthesis, nucleoid acid synthesis, ribosomal biogenesis, metabolic reprogramming to support rapid cancer cell growth and survival, metastasis and invasion, and angiogenesis. mTORC2 has different subcellular localization and phosphorylates and activates AGC kinase family members, including AKT at Ser473 (AKT-pSer473), SGK, and PKC in response to upstream signals such as IGF1. Similar to mTORC1, PI3K acts as a critical regulator of mTORC2 by inducing the binding of mTORC2 to the ribosome. mTOR, mammalian/mechanistic target of rapamycin; PI3K, phosphatidylinositol 3-kinase; RAPTOR, Regulatory associated protein of mTOR; PRAS40, Proline-rich AKT substrate 40ku; mLST8, Mammalian LST8, also called G protein b-subunit like protein; DEPTOR, DEP domain-containing-mTOR interacting protein; Tel2, Telomere maintenance 2; Tti1, Tel2-interacting protein 1; RICTOR, Rapamycin insensitive companion of mTOR; mSIN1, mitogen-activated protein kinase-associated protein 1; PROTOR, Protein observed with Rictor; SGK, serum glucose kinase; PKC, protein kinase C; IGF1; Insulin-like growth factor 1; AMPK, AMP-activated protein kinase; REDD1, regulated in development and DNA damage response 1; HIF-1α, hypoxia-inducible factor-1α; VEGF, vascular endothelial growth factor; PTEN, phosphatase and tensin homolog deleted on chromosome 10; NF-κB, nuclear factor-κB; ULK1, UNC-51-like kinase 1; 4E-BP1, initiation factor 4E-binding protein 1; RHEB, Ras homolog enriched in brain; TSC1 and TSC2, tuberous sclerosis proteins 1 and 2; GSK3β, glycogen synthase kinase 3β; MMP, matrix metallopeptidase; FKBP12, FK506-binding protein; PIP2, phosphatidylinositol 4,5 biphosphate; PDK1, phosphoinositol-dependent-kinase; PtdIns(3,4,5)P3, Phosphatidylinositol (3,4,5)-trisphosphate.

Cancer immunotherapy has recently achieved a breakthrough status, however, it is not effective in all cancer types. Genetically engineered oncolytic viruses (OVs) with immunomodulators are promising new therapeutic modalities for cancer. CARG-2020 is an engineered trivalent oncolytic viral construct that specifically expresses three immune modulators that inhibit IL-17RA signaling and regulate PD-L1 expression by shRNAs, along with the cytokine IL-12 which activates multiple tumoricidal pathways. Previous work showed that intratumoral (i.t.) injection of CARG-2020 exhibits robust efficacy against established colorectal cancer (CRC). In this study, we report that systemic delivery of CARG-2020 via the intravenous (i.v.) route can successfully control CRC growth. To further expand the scope of CARG-2020 as a pan-cancer candidate, we also show that CARG-2020 works in two additional mouse models of melanoma and triple-negative breast cancer. Administration of CARG-2020 resulted in increased accumulation of CD8⁺ T lymphocytes in the tumors, and depletion of these T cells results in poor tumor regression mediated by CARG-2020. Overall, our study shows a broad-spectrum efficacy of CARG-2020 in solid tumors and demonstrates the potential of CARG-2020 to be developed as a clinical candidate for the treatment of multiple human cancers that are surgically accessible.

Emerging evidence indicates a correlation between inflammation and the development and progression of cancer. Among the various inflammatory signals, interleukin-17 (IL-17) family cytokines serve as a critical link between inflammation and cancer. IL-17 is a highly versatile pro-inflammatory cytokine that plays a pivotal role in host defense, tissue repair, the pathogenesis of inflammatory diseases, and cancer progression. During the early stages of tumorigenesis, IL-17 signaling directly promotes the proliferation of tumor cells. Conversely, IL-17 has been shown to exhibit antitumor immunity in several models of grafted subcutaneous tumors. Additionally, dynamic changes in the microbiome can influence the secretion of IL-17, thereby affecting tumor development. The specific role of IL-17 is contingent upon its functional classification, spatiotemporal characteristics, and the stage of tumor development. In this review, we introduce the fundamental biology of IL-17 and the expression profile of its receptors in cancer, while also reviewing and discussing recent advancements regarding the pleiotropic effects and mechanisms of IL-17 in inflammation-related cancers. Furthermore, we supplement our discussion with insights into the mechanisms by which IL-17 impacts cancer progression through interactions with the microbiota, and we explore the implications of IL-17 in cancer therapy. This comprehensive analysis aims to enhance our understanding of IL-17 and its potential role in cancer treatment.

Supplementary Fig. S2. Endoplasmic reticulum stress and unfolded protein response in normal physiology and cancer Tumor cells frequently encounter endogenous and exogenous stresses that influence protein folding in the ER, including hypoxia, inflammation, nutrient deprivation, altered glycolysis, DNA damage, genomic instability, oncogene activation, and proteasome dysfunction, can cause excessive accumulation of misfolded or unfolded proteins within the ER lumen, a condition termed "ER stress." As a response to prolonged ER stress conditions, the ER responds to the ER stress by activating intracellular signal transduction pathways, collectively referred to as the UPR, to restore-ER homeostasis (protective effect) or activate cell death (cytotoxic effect). The UPR signaling cascade comprises three parallel pathways that employ unique signal transduction mechanisms and are initiated by activating three conserved ER-stress sensors: IRE1, ATF6, and PERK. Furthermore, in mammals and under resting conditions, the critical ER-resident chaperone, GRP78/BiP, prevents the activation and blocks the downstream signaling of UPR by binding to the N-terminal side of stress sensor proteins, IRE-1, PERK, and ATF6 and maintains these sensors in an inactive state. When ER stress is triggered, and the ER luminal load of unfolded or misfolded proteins increases, the GRP78/BiP dissociates, activating IRE1, PERK, and ATF6 signaling cascades, leading to the activation of these UPR signaling pathways. Upon activation, the UPR can regulate many cellular events of tumor cells, including autophagy, ER biogenesis, cell survival, metastasis, angiogenesis, migration and invasion, chemoresistance, glycolysis, and proliferation. ER, Endoplasmic reticulum; UPR, unfolded protein response; PREK, protein kinase R (PKR) like endoplasmic reticulum kinase; ATF, activating transcription factor; IRE1, inositol requiring enzyme 1; GRP78, 78-kDa glucose-regulated protein/immunoglobulin-binding protein; CHOP, C/EBP homologous protein; AMPK, AMP-activated protein kinase; XBP-1, X-box binding protein 1; Dr5, death receptor 5; GADD34, growth arrest and DNA damage-inducible protein 34; ERO1α, ER oxidoreductin 1; GCN2, general control non-derepressible 2.

Supplementary Fig. S2. Endoplasmic reticulum stress and unfolded protein response in normal physiology and cancer Tumor cells frequently encounter endogenous and exogenous stresses that influence protein folding in the ER, including hypoxia, inflammation, nutrient deprivation, altered glycolysis, DNA damage, genomic instability, oncogene activation, and proteasome dysfunction, can cause excessive accumulation of misfolded or unfolded proteins within the ER lumen, a condition termed "ER stress." As a response to prolonged ER stress conditions, the ER responds to the ER stress by activating intracellular signal transduction pathways, collectively referred to as the UPR, to restore-ER homeostasis (protective effect) or activate cell death (cytotoxic effect). The UPR signaling cascade comprises three parallel pathways that employ unique signal transduction mechanisms and are initiated by activating three conserved ER-stress sensors: IRE1, ATF6, and PERK. Furthermore, in mammals and under resting conditions, the critical ER-resident chaperone, GRP78/BiP, prevents the activation and blocks the downstream signaling of UPR by binding to the N-terminal side of stress sensor proteins, IRE-1, PERK, and ATF6 and maintains these sensors in an inactive state. When ER stress is triggered, and the ER luminal load of unfolded or misfolded proteins increases, the GRP78/BiP dissociates, activating IRE1, PERK, and ATF6 signaling cascades, leading to the activation of these UPR signaling pathways. Upon activation, the UPR can regulate many cellular events of tumor cells, including autophagy, ER biogenesis, cell survival, metastasis, angiogenesis, migration and invasion, chemoresistance, glycolysis, and proliferation. ER, Endoplasmic reticulum; UPR, unfolded protein response; PREK, protein kinase R (PKR) like endoplasmic reticulum kinase; ATF, activating transcription factor; IRE1, inositol requiring enzyme 1; GRP78, 78-kDa glucose-regulated protein/immunoglobulin-binding protein; CHOP, C/EBP homologous protein; AMPK, AMP-activated protein kinase; XBP-1, X-box binding protein 1; Dr5, death receptor 5; GADD34, growth arrest and DNA damage-inducible protein 34; ERO1α, ER oxidoreductin 1; GCN2, general control non-derepressible 2.