Mouse Colon Cancer Research Articles

Investigation of artificial intelligence integrated fluorescence endoscopy image analysis with indocyanine green for interpretation of precancerous lesions in colon cancer.

Indocyanine green (ICG) has been used in clinical practice for more than 40 years and its safety and preferential accumulation in tumors has been reported for various tumor types, including colon cancer. However, reports on clinical assessments of ICG-based molecular endoscopy imaging for precancerous lesions are scarce. We determined visualization ability of ICG fluorescence endoscopy in colitis-associated colon cancer using 30 lesions from an azoxymethane/dextran sulfate sodium (AOM/DSS) mouse model and 16 colon cancer patient tissue-samples. With a total of 60 images (optical, fluorescence) obtained during endoscopy observation of mouse colon cancer, we used deep learning network to predict four classes (Normal, Dysplasia, Adenoma, and Carcinoma) of colorectal cancer development. ICG could detect 100% of carcinoma, 90% of adenoma, and 57% of dysplasia, with little background signal at 30 min after injection via real-time fluorescence endoscopy. Correlation analysis with immunohistochemistry revealed a positive correlation of ICG with inducible nitric oxide synthase (iNOS; r > 0.5). Increased expression of iNOS resulted in increased levels of cellular nitric oxide in cancer cells compared to that in normal cells, which was related to the inhibition of drug efflux via the ABCB1 transporter down-regulation resulting in delayed retention of intracellular ICG. With artificial intelligence training, the accuracy of image classification into four classes using data sets, such as fluorescence, optical, and fluorescence/optical images was assessed. Fluorescence images obtained the highest accuracy (AUC of 0.8125) than optical and fluorescence/optical images (AUC of 0.75 and 0.6667, respectively). These findings highlight the clinical feasibility of ICG as a detector of precancerous lesions in real-time fluorescence endoscopy with artificial intelligence training and suggest that the mechanism of ICG retention in cancer cells is related to intracellular nitric oxide concentration.

Oleanolic Acid Glycosides from Scabiosa caucasica and Scabiosa ochroleuca: Structural Analysis and Cytotoxicity.

In the field of research on medicinal plants from the Armenian flora, the phytochemical study of two Scabiosa L. species, S. caucasica M. Bieb. and S. ochroleuca L. (Caprifoliaceae), has led to the isolation of five previously undescribed oleanolic acid glycosides from an aqueous-ethanolic extract of the roots: 3-O-α-L-rhamnopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-β-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester, 3-O-β-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-β-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester, 3-O-β-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-β-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid, 3-O-β-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D-xylopyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-β-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester, 3-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-β-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester. Their full structural elucidation required extensive 1D and 2D NMR experiments, as well as mass spectrometry analysis. For the biological activity of the bidesmosidic saponins and the monodesmosidic saponin, their cytotoxicity on a mouse colon cancer cell line (MC-38) was evaluated.

Anticancer Effect of Cold Atmospheric Plasma in Syngeneic Mouse Models of Melanoma and Colon Cancer.

Cold atmospheric plasma (CAP) may have applications in treating various types of malignant tumors. This study assessed the anticancer effects of CAP using melanoma and colon cancer cell lines. CAP treatment significantly reduced the in vitro viability of melanoma and colon cancer cell lines and had a negligible effect on the viability of normal human melanocytes. Additionally, CAP and epidermal growth factor receptor (EGFR) inhibitor had an additive anticancer effect in a CAP-resistant melanoma cell line. Reactive oxygen and nitrogen species known to be generated by CAP enhanced the anticancer effects of CAP and EGFR inhibitors. The in vivo anticancer activities of CAP were evaluated by testing its effects against syngeneic tumors induced in mice by melanoma and colon cancer cells. CAP treatment reduced tumor volume and weight in both cancer models, with the extent of tumor reduction dependent on the duration and number of CAP treatments. Histologic examination also revealed the tumoricidal effects of CAP in both tumor models. In conclusion, CAP inhibits the growth of mouse melanoma and colon cancer cell lines in vitro and shows tumoricidal effects against mouse models of melanoma and colon cancer in vivo.

Ultra-high-performance liquid chromatography-tandem mass spectrometry-based metabolomics unveils the metabolic alterations in colon cancer mice during CT-guided radiofrequency ablation.

Colon cancer (CC) is a malignancy of the digestive tract, and computed tomography (CT)-guided radiofrequency ablation (RFA) has been extensively adopted in cancer treatment. We aimed to explore the changes in fecal metabolism after CT-guided RFA in CC mice. The orthotopic CC mice received CT-guided RFA upon modeling. Subsequently, we quantified tumor volumes and weights to assess treatment efficacy. Next, because metabolomics is useful for evaluating therapeutic validity, feces were collected for metabolomics analysis. CT-guided RFA inhibited tumor growth effectively. Additionally, metabolomics results showed that the contents of bile acids and fatty acids were downregulated in CC mouse feces. Moreover, the levels of amino acids and carbohydrates were decreased while the levels of fatty acids, organic acids, phenols, pyridines and short-chain fatty acids were elevated in feces after CC mice received CT-guided RFA. Pathway enrichment analysis revealed that those differential metabolites were closely related to fatty acids degradation and synthesis. CT-guided RFA possesses a strong ability to suppress CC development in mice, accompanied by a significant increase of fatty acid content in feces. This study proposes a novel approach and target for CC treatment, which provides hope for CC patients and establishes a solid basis for future in-depth studies.

TNFR2-targeting peptide for cancer immunotherapy

Abstract There is compelling evidence that tumor necrosis factor receptor type II (TNFR2) is preferentially expressed by highly immunosuppressive CD4+Foxp3+ regulatory T cells (Tregs), especially those present in tumor-infiltrating lymphocytes (TILs). These findings led us and others to propose and experimentally proved that targeting TNFR2 with antibodies could induce anti-tumor immune responses. In this study, we identified a TNFR2-targeting peptide P20 through phage display. P20 had in vitro activity to block TNF-TNFR2 interaction and inhibit TNFR2-mediated activation/expansion of Treg cells. The in vivo effect of P20 on tumor growth was examined with mouse tumor models including CT26 colon cancer and OVA-EG7 lymphoma. The results showed that the treatment with P20 (i.p.) resulted in a marked inhibition of tumor growth, accompanied by a reduced number of Tregs and an increased number of IFN-γ+ CD8+ cytotoxic T lymphocytes in the tumor microenvironment. Furthermore, P20 could markedly enhance the efficacy of anti-PD-L1 in the inhibition of tumor growth in a syngeneic MC38 mouse colon cancer model. Thus, P20 may be a useful anti-tumor immunotherapeutic agent and merits further investigation. (Funded by FDCT 0099/2021/A2, UM MYRG2022-00260-ICMS, UM CPG2023-00025-ICMS) There is compelling evidence that tumor necrosis factor receptor type II (TNFR2) is preferentially expressed by highly immunosuppressive CD4+Foxp3+ regulatory T cells (Tregs), especially those present in tumor-infiltrating lymphocytes (TILs). These findings led us and others to propose and experimentally proved that targeting TNFR2 with antibodies could induce anti-tumor immune responses. In this study, we identified a TNFR2-targeting peptide P20 through phage display. P20 had in vitro activity to block TNF-TNFR2 interaction and inhibit TNFR2-mediated activation/expansion of Treg cells. The in vivo effect of P20 on tumor growth was examined with mouse tumor models including CT26 colon cancer and OVA-EG7 lymphoma. The results showed that the treatment with P20 (i.p.) resulted in a marked inhibition of tumor growth, accompanied by a reduced number of Tregs and an increased number of IFN-γ+ CD8+ cytotoxic T lymphocytes in the tumor microenvironment. Furthermore, P20 could markedly enhance the efficacy of anti-PD-L1 in the inhibition of tumor growth in a syngeneic MC38 mouse colon cancer model. Thus, P20 may be a useful anti-tumor immunotherapeutic agent and merits further investigation

Targeting inhibition of prognosis-related lipid metabolism genes including CYP19A1 enhances immunotherapeutic response in colon cancer

BackgroundLipid metabolic reprogramming in colon cancer shows a potential impact on tumor immune microenvironment and is associated with response to immunotherapy. Therefore, this study aimed to develop a lipid metabolism-related prognostic risk score (LMrisk) to provide new biomarkers and combination therapy strategies for colon cancer immunotherapy.MethodsDifferentially expressed lipid metabolism-related genes (LMGs) including cytochrome P450 (CYP) 19A1 were screened to construct LMrisk in TCGA colon cancer cohort. The LMrisk was then validated in three GEO datasets. The differences of immune cell infiltration and immunotherapy response between LMrisk subgroups were investigated via bioinformatic analysis. These results were comfirmed by in vitro coculture of colon cancer cells with peripheral blood mononuclear cells, human colon cancer tissue microarray analysis, multiplex immunofluorescence staining and mouse xenograft models of colon cancer.ResultsSix LMGs including CYP19A1, ALOXE3, FABP4, LRP2, SLCO1A2 and PPARGC1A were selected to establish the LMrisk. The LMrisk was positively correlated with the abundance of macrophages, carcinoma-associated fibroblasts (CAFs), endothelial cells and the levels of biomarkers for immunotherapeutic response including programmed cell death ligand 1 (PD-L1) expression, tumor mutation burden and microsatellite instability, but negatively correlated with CD8+ T cell infiltration levels. CYP19A1 protein expression was an independent prognostic factor, and positively correlated with PD-L1 expression in human colon cancer tissues. Multiplex immunofluorescence analyses revealed that CYP19A1 protein expression was negatively correlated with CD8+ T cell infiltration, but positively correlated with the levels of tumor-associated macrophages, CAFs and endothelial cells. Importantly, CYP19A1 inhibition downregulated PD-L1, IL-6 and TGF-β levels through GPR30-AKT signaling, thereby enhancing CD8+ T cell-mediated antitumor immune response in vitro co-culture studies. CYP19A1 inhibition by letrozole or siRNA strengthened the anti-tumor immune response of CD8+ T cells, induced normalization of tumor blood vessels, and enhanced the efficacy of anti-PD-1 therapy in orthotopic and subcutaneous mouse colon cancer models.ConclusionA risk model based on lipid metabolism-related genes may predict prognosis and immunotherapeutic response in colon cancer. CYP19A1-catalyzed estrogen biosynthesis promotes vascular abnormality and inhibits CD8+ T cell function through the upregulation of PD-L1, IL-6 and TGF-β via GPR30-AKT signaling. CYP19A1 inhibition combined with PD-1 blockade represents a promising therapeutic strategy for colon cancer immunotherapy.

The disinfectant residues promote the leaching of water contaminants from plastic pipe particles

Disinfection treatment is an indispensable water purification process, but it can leave trace concentrations of disinfectant in the purified water. Disinfectants oxidation can age plastic pipes and release hazardous microplastics and chemicals into drinking water. Lengths of commercially-available unplasticized polyvinyl chloride and polypropylene random copolymer water pipe were ground into particles and exposed to micro-molar concentrations of ClO2, NaClO, trichloroisocyanuric acid, or O3 for up to 75 days. The disinfectants aged the plastic and changed its surface morphology and functional groups. Meanwhile, disinfectants could significantly promote the release of organic matter from plastic pipes into the water. ClO2 generated the highest concentrations of organic matter in the leachates from both plastics. Plasticizers, antioxidants and low molecular weight organic matter were detected in all of the leachates. Leachate samples inhibited the proliferation of CT26 mouse colon cancer and induced oxidative stress in the cells. Even trace concentrations of residual disinfectant can constitute a drinking water risk.

Abstract 483: Hederacolchiside A1 inhibits autophagy via CTSC inhibition and inhibits growth in colon cancer

Abstract Background: Autophagy is a normal regulatory process that degrades non-functional or unnecessary cellular components. The degradation of components provides materials for the synthesis of cellular components and, during tumor development, may provide energy. Therefore, autophagy regulation may have potential therapeutic uses in oncology. The triterpenoid saponin hederacolchiside A1 (HA1) has potent antitumor effects; however, its role in autophagy is still unknown. Methods: In the present study, we used human colon cancer cells (SW480 and HT29) and mouse colon cancer cells (CT26) for the main experiments, and several additional cancer cell lines for other experiments. To assess the autophagy dysregulation caused by HA1, we evaluated the protein expressions of two autophagy markers (LC3 and SQSTM1) using Western blotting and the changes in organelles using LC3 puncta assay and transmission electron microscopy. We compared the effects of HA1 and other autophagy inhibitors. Additionally, we compared the HA1-induced decrease in cathepsin C (CTSC) expression in tumors and non-tumors using publicly available data. The effect of HA1 on tumor growth was evaluated in vitro using a three-dimensional spheroid model and patient-derived colon organoids, and in vivo based on tumor growth in a subcutaneous injection mouse model. Results: HA1 treatment resulted in marked vacuolization of colon and other cancer cells. Additionally, autophagy was induced by the accumulation of LC3-II and SQSTM1. These findings were supported by increased LC3 puncta and abnormal organelle morphology. Compared to treatment with other autophagy inhibitors, HA1 treatment resulted in the formation of distinct vacuoles and increased LC3 and SQSTM1, similar to chloroquine treatment. The publicly available data showed that the lysosomal protease CTSC was increased in several cancers, including colon cancer, compared to the normal tissues. Furthermore, HA1 decreased the CTSC expression and enzymatic activity independently of the proteasome degradation, resulting in autophagy dysregulation and, subsequently, autophagy flux inhibition. HA1 inhibited cell growth and induced cell cycle arrest in the two-dimensional culture, three-dimensional spheroids, patient-derived colon cancer organoids, and the in vivo experiments. Conclusion: HA1 inhibits cell growth and induces autophagy dysregulation. Therefore, HA1 may be a potential therapeutic agent in colon cancer. Citation Format: Solbi Kim, Eunji Kim, Minju Han, Youn Seo Oh, Sora Kang, Heung Jin Jeon, Hyo Jin Lee. Hederacolchiside A1 inhibits autophagy via CTSC inhibition and inhibits growth in colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 483.

Abstract 3260: Potentiation of Type I interferon signaling leads to in vivo efficacy achieved with combination of chemotherapy and the PARP7 inhibitor RBN-2397

Abstract There are many pathways that activate the Type I interferon (IFN) response to combat tumor progression including the cytosolic nucleic acid sensing pathways. PARP7 is a monoART enzyme that was reported to negatively regulate the Type I IFN response by interacting with TBK1 during viral infection. We and others have demonstrated that PARP7 is a novel negative regulator of cytosolic nucleic acid sensing in tumor cells. RBN-2397 is the first potent and selective small molecule inhibitor of PARP7 catalytic function. We have shown that administration of RBN-2397 to CT26 tumor-bearing mice significantly decreased tumor growth and led to complete and durable regressions in a subset of mice. The antitumor effects correlated with activation of p-STAT1 and increased expression of ISGs including CXCL10, MX1, and CCL5 in tumors. A growing body of work has shown that DNA damaging agents such as chemotherapies activate the Type I IFN response by generating accumulation of nucleic acids, including DNA, in the cytosol or tumor microenvironment. These aberrant cytosolic nucleic acids are recognized by the innate immune response via cGAS leading to activation of the STING/TBK pathway and subsequent production of IFN-β and expression of ISGs (interferon induced genes). Given that PARP7 suppresses the Type I IFN response, we hypothesized that the immune-related mechanism observed with chemotherapy would be inhibited in tumors with high PARP7 expression. Combining chemo with RBN-2397 could potentiate Type I IFN response activation in PARP7 expressing tumors and enhance immune mediated anti-tumor responses. We tested the ability of platinum (cisplatin, oxaliplatin, and carboplatin) and taxane (docetaxel and paclitaxel) chemotherapy agents to activate the Type I IFN response in the CT26 mouse colon cancer cell line in vitro by gene expression analysis of several ISGs (CXCL10, CCL5, MX1, IFIT1, and IFN-β). Single agent chemo led to modest increases in the expression of ISGs. The combination of cisplatin or oxaliplatin with RBN-2397 significantly potentiated the effect of RBN-2397 and led to a synergistic increase of the ISGs in CT26 cells after 24 hours of treatment. Similar effects were obtained with the combination of docetaxel or paclitaxel with RBN-2397. To evaluate if the combinatorial effect observed in vitro translated into increased efficacy in vivo, we first identified doses of single agent cisplatin or docetaxel that increased expression of ISGs and enhanced cytosolic-DNA in CT26 tumors. Next, we verified the tolerability of validated doses in combination with RBN-2397. Finally, we performed an efficacy study in CT26 tumor bearing mice and observed that the combination of cisplatin and RBN-2397 increased tumor growth inhibition and survival relative to single agent activity. This study provides rationale for exploration of DNA damaging agents in combination with RBN-2397. Citation Format: Kaiko Kunii, Jeff J. Song, Danielle Zimmerman, Chang Liu, Kristy G. Kuplast-Barr, Jonathan J. Novak, Sunaina P. Nayak, Joseph M. Gozgit, Melissa M. Vasbinder, Kristen McEachern, Kevin W. Kuntz, Heike Keilhack, Jennifer R. Molina. Potentiation of Type I interferon signaling leads to in vivo efficacy achieved with combination of chemotherapy and the PARP7 inhibitor RBN-2397 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3260.

Abstract 1546: ADCC enhanced anti-CD25 mAb (TST010) demonstrated antitumor activity via depleting Treg cells and increasing CD8+T/Treg ratio in preclinical tumor models

Abstract Regulatory T (Treg) cells are a distinctive lineage of CD4+ T cells as characterized by expression of FoxP3 and CD25 markers. Treg cells are involved in tumor development and progression by inhibiting antitumor immunity. Accumulation of Treg cells in the tumor microenvironment (TME) is often associated with an adverse prognosis in multiple cancers and also has implication for predicting response to immunotherapy. Treg cell depletion strategy has been explored in preclinical and clinical settings to induce effective antitumor immune responses in TME. CD25, a high-affinity binding subunit alpha of the IL-2 receptor, is constitutively and highly expressed on Treg cells but also transiently upregulated on effector T cells. The high level of CD25 expression on Treg cells could deprive of IL-2 for effector T cells and inhibit their proliferation. Thus, an effective approach to selectively suppress Treg cell function is to deplete Treg cells while avoid blocking IL-2 binding to CD25 for allowing T effect cell proliferation and activation. Using our immune tolerance breaking antibody platform, we generated a humanized IgG1 subtype anti-CD25 monoclonal antibody (TST010). TST010 bound to human CD25 with high affinity and selectivity but did not block IL-2 binding to CD25. No blocking activity to IL-2 signaling on human PBMC was further confirmed by the phosphorylation of Stat5 using FACS analysis. By reducing fucosylation during cell culture process, TST010 mAb gained the enhanced ADCC activity. In an ADCC reporter assay, TST010 showed its sub-nanomolar ADCC activity against CD25-overexpression SU-DHL-1 cell line and human induced Treg cells, but much less to human activated CD8+ T effect cells due to lower CD25 expression. Similarly, in a NK cytotoxicity assay, TST010 selectively killed human induced Treg cells in contrast to activated CD8+ T cells. In vivo antitumor activity of TST010 was evaluated in the syngeneic mouse tumor models. The mouse colon cancer MC38 model was developed on the human CD25 knock-in mice. In the MC38 tumor model, TST010 markedly reduced the Treg (FoxP3+CD4+) population in both peripheral blood and TME. Importantly, the CD8+T/Treg ratio in tumor was increased starting at 2 weeks post TST010 treatment, and it resulted in a significant single agent tumor growth inhibition at a 10 mg/kg dose. Furthermore, in the LLC tumor model which does not respond well to checkpoint inhibitors, the anti-CD25 mAb showed a good combination effect with a PD-L1 inhibitor. In summary, we have discovered a novel therapeutic anti-CD25 mAb that can deplete Treg cells without blocking IL-2 signaling. As demonstrated in our preclinical tumor models, it has a good potential to induce effective antitumor immune responses in TME and tumor growth inhibition especially in combination with PD-1/PD-L1 treatment. Citation Format: Di Sun, Huanhuan Guo, Hongjun Li, Xinlai Yao, Yiqing Xu, Shenjie Zhang, Shijie Zhou, Xiaoli Zi, Lisa Zheng, Yi Gu, Xueming Qian. ADCC enhanced anti-CD25 mAb (TST010) demonstrated antitumor activity via depleting Treg cells and increasing CD8+T/Treg ratio in preclinical tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1546.

Focused ultrasound restrains the growth of orthotopic colon cancer via promoting pyroptosis.

Focused ultrasound (FUS) is a non-invasive tumor therapy technology emerging in recent years, which can treat various solid tumors. However, it is unclear whether FUS can affect the pyroptosis of colon cancer (CC) cells. Here, we analyzed the effect of FUS on pyroptosis in the orthotopic CC model. After an orthotopic CC mouse model was constructed by injecting CT26-Luc cells, BABL/C mice were allocated to the normal, tumor, FUS, and FUS + BAY11-7082 (pyroptosis inhibitor) groups. We monitored the tumor status of the mice through in vivo fluorescence image analysis. The histopathological injury of the intestinal tissue and the expression of IL-1β, IL-18, caspase-recruitment domain (ASC), cleaved caspase-1, gasdermin D (GSDMD), and NLRP3 of the CC tumors were examined utilizing hematoxylin and eosin staining, immunohistochemical assay, and Western blot. FUS restrained the fluorescence intensity of the tumors in orthotopic CC mice, while FUS-mediated suppression of the bioluminescent signal of the tumors was alleviated by BAY11-7082. FUS was found to relieve the injury of the intestinal tissues in CC mice as revealed by morphology. Furthermore, the expressions of IL-1β, IL-18, GSDMD, ASC, cleaved caspase-1, and NLRP3 of the CC tumors in the FUS group were higher than those in the tumor group, while BAY11-7082 addition partly reversed the FUS's effects on orthotopic CC model mice. Our results pointed out that FUS presented anti-tumor activity in experimental CC, and its mechanism was correlated with the promotion of pyroptosis.

In Situ Transformable Nanoplatforms with Supramolecular Cross-Linking Triggered Complementary Function for Enhanced Cancer Photodynamic Therapy.

In vivo cross-linking of nanoparticles is widely used to increase accumulation of therapeutic agents at tumor site for enhanced therapy. However, the components in nanoplatforms usually only play for one role and are independent of each other, unable to amplify their biofunctions. Herein, a complementary functioning tumor microenvironment triggered, supramolecular coordination-induced nanoparticle cross-linking strategy is constructed for enhanced photodynamic therapy. Manganese oxide (MnOx ) and polyhydroxy photosensitizer hypericin (Hyp) are coated and loaded onto lanthanide-doped upconversion nanoparticles (UCNPs) to form transformable UCNP@MnOx -Hyp. In CT26mouse colon cancer cells and xenograft tumors, UCNP@MnOx -Hyp is reduced by glutathione and H2 O2 , releasing Mn2+ and Hyp for in situ cross-linking to transform to UCNP@Mn2+ -Hyp. Compared to the simple photosensitizer-loaded UCNP@PEI-Hyp, the Mn2+ -Hyp coordination redshifts absorbance of Hyp and improves the energy transfer efficiency from UCNPs to Hyp (5.6-fold). In turn, the supramolecular coordination-induced UCNPs cross-linking exhibits enhanced luminescence recovery and increased intracellular accumulation of both UCNPs and Hyp, thus enhancing the photodynamic therapy efficacy both at cellular level (2.1-fold) and in vivo, realizing the function amplification of each component after responsive transformation and offering a new avenue for enhanced cancer therapy.

Mesenchymal stem cells can prevent or promote the progression of colon cancer based on their timing of administration

BackgroundMesenchymal stem cell (MSC) therapy has been shown to have some therapeutic effects in rodent models and patients with IBD; however, its role in colon tumor models is controversial. In this study, the potential role and mechanisms of bone marrow-derived MSCs (BM-MSCs) in colitis-associated colon cancer (CAC) were investigated.MethodsThe CAC mouse model was established with azoxymethane (AOM) and dextran sulfate sodium (DSS). The mice were administered an intraperitoneal injection of MSCs once weekly for different periods. The progression of CAC and the cytokine expression in tissues was assessed. Immunofluorescence staining was used to detect MSCs localization. Levels of immune cells in the spleen and lamina propria of the colon were detected using flow cytometry. A co-culture of MSCs and naïve T cells was performed to determine the effect of MSCs on naïve T cell differentiation.ResultsEarly administration of MSCs inhibited the occurrence of CAC, while late administration promoted the progression of CAC. The inhibitory effect of early injection in mice was characterized by the expression of inflammatory cytokines in colon tissue was decreased, and induction of T regulatory cells (Tregs) infiltration via TGF-β. The promotive effect of late injection was characterized by a shift of T helper (Th) 1/Th2 immune balance toward a Th2 phenotype through IL-4 secretion. IL-12 can reverse this shift to Th2 accumulation in mice.ConclusionMSCs can curb the progression of colon cancer by inducing Treg accumulation via TGF-β at the early stage of inflammatory transformation but promote the progression of colon cancer by inducing a shift in Th1/Th2 immune balance to Th2 through IL-4 secretion at the late stage. And the immune balance of Th1/Th2 influenced by MSCs could be reversed by IL-12.

A Cascade-Targeted Enzyme-Instructed Peptide Self-Assembly Strategy for Cancer Immunotherapy through Boosting Immunogenic Cell Death.

Immunogenic cell death (ICD) approaches by encumbering mitochondrial functions provide great promise for the treatment of malignant tumors, but these kinds of ICD strategies are still in their infancy. Here, one multifunctional drug-loaded, cascade-targeted, and enzyme-instructed self-assembling peptide nanomedicine (Comp. 4) for ICD-based cancer therapy is constructed. Comp. 4 consists of 1) lonidamine (LND) that specifically interferes with mitochondrial functions; 2) a programmed death ligand 1 (PD-L1) binding peptide sequence (NTYYEDQG) and a mitochondria-specific motif (triphenylphosphonium, TPP) that can sequentially control the cell membrane and mitochondria targeting capacities, respectively; and 3) a -GD FD FpD Y- assembly core to in situ organize peptide assemblies responsive to alkaline phosphatase (ALP). Comp. 4 demonstrates noticeable structural and morphological transformations in the presence of ALP and produces peptide assemblies in mouse colon cancer cells (CT26) with high expressions of both ALP and PD-L1. Moreover, the presence of PD-L1- and mitochondria-specific motifs can assist Comp. 4 for effective endocytosis and endosomal escape, forming peptide assemblies and delivering LND into mitochondria. Consequently, Comp. 4 shows superior capacities to in vivo induce abundant mitochondrial oxidative stress, provoke robust ICD responses, and produce an immunogenic tumor microenvironment, successfully inhibiting CT26 tumor growth by eliciting a systemic ICD-based antitumor immunity.

CLASH of the Titans: How CAR-T Cells Can Triumph Over Tumors.

CLASH of the Titans: How CAR-T Cells Can Triumph Over Tumors.

Inhibition of mouse colon cancer growth following immunotherapy with a fraction of hydatid cyst fluid

BackgroundHydatid cyst is the larval stage of the tape worm Echinococcus granulosus which is located in human and livestock viscera. There are some scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. In this study, the effects of a fraction of hydatid cyst fluid on colon cancer tumor in BALB/c mice were investigated. Materials and methodsIn this experimental work six groups of mice were challenged with mouse colon cancer cells. 5 days later when the sign of tumor growth in mice was seen, group 1–4 were injected with hydatid cyst fluid, the 78 kDa fraction, live protoscolices and BCG respectively. Group five was injected with alum alone and the sixth group left intact without any injection. The size of the tumor was measured and compared in all groups. Then blood samples of mice were evaluated for serum cytokine levels. ResultIn mice injected with hydatid cyst antigens especially a fraction of hydatid cyst fluid, tumor size was smaller than the that of control groups and the difference of tumor size in cases and control groups was statistically significant. ConclusionThe results of this study showed that injection of mice with a fraction of hydatid cyst fluid significantly inhibits the growth of mouse colon cancer and this inhibition may be related to effect of immune response to these antigens.

In vitro toxicity of naringin and berberine alone, and encapsulated within PMMA nanoparticles

Phytochemical compounds, such as naringin and berberine, have been used for many years due to their antioxidant activities, and consequently, beneficial health effects. In this study, it was aimed to evaluate the antioxidant properties of naringin, berberine and poly(methylmethacrylate) (PMMA) nanoparticles (NPs) encapsulated with naringin or berberine and their possible cytotoxic, genotoxic, and apoptotic effects on mouse fibroblast (NIH/3 T3) and colon cancer (Caco-2) cells. According to the results of the study, it was found that the 2,2-diphenyl-1-picrylhydrazyl (DPPH) inhibition antioxidant activity of naringin, berberine, and naringin or berberine encapsulated PMMA NPs, was significantly increased at higher tested concentrations due to the antioxidant effects of naringin, berberine and naringin or berberine encapsulated PMMA NPs. As a result of the cytotoxicity assay, after 24-, 48- and 72-h of exposure, all of the studied compounds caused cytotoxic effects in both cell lines. Genotoxic effects of studied compounds were not registered at lower tested concentrations. Based on these data, polymeric nanoparticles encapsulated with naringin or berberine may contribute to new treatment approaches for cancer, but further in vivo and in vitro research is required.

Lactoferrin suppresses the progression of colon cancer under hyperglycemia by targeting WTAP/m6A/NT5DC3/HKDC1 axis

BackgroundAlthough the relationship between type 2 diabetes (T2D) and the increased risk of colorectal carcinogenesis is widely defined in clinical studies, the therapeutic methods and molecular mechanism of T2D-induced colon cancer and how does hyperglycemia affect the progression is still unknown. Here, we studied the function of lactoferrin (LF) in suppressing the progression of colon cancer in T2D mice, and uncovered the related molecular mechanisms in DNA 5mC and RNA m6A levels.MethodsWe examined the effects of LF (50% iron saturation) on the migration and invasion of colon tumor cells under high concentration of glucose. Then, transcriptomics and DNA methylation profilings of colon tumor cells was co-analyzed to screen out the special gene (NT5DC3), and the expression level of NT5DC3 in 75 clinical blood samples was detected by q-PCR and western blot, to investigate whether NT5DC3 was a biomarker to distinguish T2D patients and T2D-induced colon cancer patients from healthy volunteers. Futhermore, in T2D mouse with xenografted colon tumor models, the inhibitory effects of LF and NT5DC3 protein on colon tumors were investigated. In addition, epigenetic alterations were measured to examine the 5mC/m6A modification sites of NT5DC3 regulated by LF. Utilizing siRNA fragments of eight m6A-related genes, the special gene (WTAP) regulating m6A of NT5DC was proved, and the effect of LF on WTAP/NT5DC3/HKDC1 axis was finally evaluated.ResultsA special gene NT5DC3 was screened out through co-analysis of transcriptomics and DNA methylation profiling, and HKDC1 might be a downstream sensor of NT5DC3. Mechanistically, LF-dependent cellular DNA 5mC and RNA m6A profiling remodeling transcriptionally regulate NT5DC3 expression. WTAP plays a key role in regulating NT5DC3 m6A modification and subsequently controls NT5DC3 downstream target HKDC1 expression. Moreover, co-treatment of lactoferrin and NT5DC3 protein restrains the growth of colon tumors by altering the aberrant epigenetic markers. Strikingly, clinical blood samples analysis demonstrates NT5DC3 protein expression is required to direct the distinction of T2D or T2D-induced colon cancer with healthy humans.ConclusionsTogether, this study reveals that lactoferrin acts as a major factor to repress the progression of colon cancer under hyperglycemia, thus, significantly expanding the landscape of natural dietary mediated tumor suppression.Graphical

Anticancer potential of fermented milk with autochthonous lactic acid bacteria.

The purpose of this study was to investigate the effect of fermented milk supernatants of autochthonous lactic acid bacteria, including Lactobacillus helveticus KMCH1 (ON561781), Lactococcus lactis KMCM3 (ON561782), and Lactiplantibacillus plantarum KMJC4 (ON615217), on human colon cancer (HT-29) and normal mouse fibroblast (L929) cells in vitro. Proteolytic activity, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide test, evaluation of apoptosis induction, and cell cycle arrest by flow cytometry were the assays performed in this study. The measurement of proteolytic activity of three types of fermented milk supernatant using an orthophthalaldehyde reagent showed that the fermented milk supernatant of L. helveticus KMCH1 included the highest proteolysis. Three types of fermented milk supernatant showed anticancer effects on HT-29 cell in a time- and concentration-based manner (at a concentration of 16 mg ml-1 for 72 h of incubation), while the effect of three types of supernatant on inhibition of L929 cell was 3%-10%. Besides, three types of supernatant inhibited HT-29 cell proliferation by inducing apoptosis and cell cycle arrest in the S phase. Autochthonous lactic acid bacteria strains were able to produce bioactive peptides with anticancer effects in fermented milk. Inhibition of HT-29 cell proliferation was dependent on peptide concentration.

FOF1-ATP synthase molecular motor biosensor for miRNA detection of colon cancer

AimsTo establish a FOF1-ATP synthase molecular motor biosensor to accurately identify colon cancer miRNAs. Main methodsThe FOF1-ATP synthase molecular motor is extracted by fragmentation–centrifugation and connected to the colon cancer-specific miR-17 capture probe in the manner of the ε subunit-biotin-streptavidin-biotin system. Signal probes are designed for dual-signal characterization to increase detection accuracy. The FOF1-ATPase rotation rate decreases when the signaling and capture probes are combined with the target miRNA, resulting in a decrease in ATP synthesis. miR-17 concentrations are determined by changes in ATP-mediated chemiluminescence intensity and signal probe-mediated OD450nm. Key findingsThe chemiluminescence intensity and OD450nm show a good linear relationship with the miR-17 concentration in the range of 5 to 200 nmol L−1 (R2 = 0.9985, 0.9989). The colon cancer mouse model is established for the blood samples, and miR-17 in serum and RNA extracts is quantitatively determined using the constructed sensor. SignificanceThe results are consistent with colon cancer progression, and the low concentration of miR-17 detecting accuracy is comparable to the PCR assay. In conclusion, the developed method is a direct, rapid, and promising method for miRNA detection of colon cancer.