Liver fibrosis promotes immunity escape but limits the size of liver tumor in a rat orthotopic transplantation model

AbstractLiver fibrosis plays a crucial role in promoting tumor immune escape and tumor aggressiveness for liver cancer. However, an interesting phenomenon is that the tumor size of liver cancer patients with liver fibrosis is smaller than that of patients without liver fibrosis. In this study, 16 SD rats were used to establish orthotopic liver tumor transplantation models with Walker-256 cell lines, respectively on the fibrotic liver (n = 8, LF group) and normal liver (n = 8, control group). MRI (magnetic resonance imaging) was used to monitor the size of the tumors. All rats were executed at the third week after modeling, and the immunohistochemical staining was used to reflect the changes in the tumor microenvironment. The results showed that, compared to the control group, the PD-L1 (programmed cell death protein receptor-L1) expression was higher, and the neutrophil infiltration increased while the effector (CD8+) T cell infiltration decreased in the LF group. Additionally, the expression of MMP-9 (matrix metalloproteinase-9) of tumor tissue in the LF group increased. Three weeks after modeling, the size of tumors in the LF group was significantly smaller than that in the control group (382.47 ± 195.06 mm3 vs. 1736.21 ± 657.25 mm3, P < 0.001). Taken together, we concluded that liver fibrosis facilitated tumor immunity escape but limited the expansion of tumor size.

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Liver Cirrhosis Promotes Immune Escape in Hepatocellular Carcinoma via GOLM1-mediated PD-L1 Upregulation

10.21203/rs.3.rs-117712/v1 ◽

2020 ◽

Author(s):

Meng-yun Ke ◽

Zhi-jin Li ◽

Yuan-peng Ye ◽

Feng-gang Ren ◽

Shao-ying Lu ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Cirrhosis ◽

T Cell ◽

Liver Fibrosis ◽

Immune Escape ◽

Immune Checkpoint Blockade ◽

Immune Checkpoints ◽

Control Group ◽

Tumor Immune Escape ◽

Fibrotic Liver

Abstract Background:Immune checkpoint blockade is considered a breakthrough in cancer treatment. However, the low response rates and therapeutic resistance of patients with hepatocellular carcinoma (HCC) represent significant challenges in the application of this treatment. Liver cirrhosis is a key driver of tumor immune escape. However, the mechanism underlying this outcome has never been clarified. This study sought to explore the role of liver cirrhosis in regulating tumor-infiltrating lymphocytes (TILs) and inducing tumor immunosuppression. Methods:Ninety-nine fixed HCC tissue samples were used to analyze the association between liver cirrhosis and immune escape by immunohistochemistry. H22 cells with or without GOLM1 knockdown were inoculated subcutaneously into BALB/c and BALB/c-nu/nu mice. We also created hepatocyte-specific GOLM1 transgenic mice (Alb/GOLM1 mice) and induced chemical carcinogenesis. The efficacy of anti-PD-L1 therapy combined with GOLM1 inhibition was estimated in GOLM1 -overexpressing subcutaneous model of HCC.Results:In HCC patients, low FIB-4 values and high CD8+ T cell infiltration were correlated with prolonged survival. Elevated expression of immune checkpoints and attenuated antitumor immunity were observed in CCl4-induced mice liver fibrosis models and human fibrotic livers compared to control group. GOLM1 levels were increased in livers of patients with cirrhosis and mice in response to CCl4-induced liver fibrosis. CD8+ T cell infiltrations were significantly decreased and PD-L1 expression was significantly increased in tumor tissues from Alb/GOLM1 mice compared to their corresponding control WT mice. GOLM1 induced PD-L1 expression via EGFR pathway activation. EGFR inhibitors, especially together with anti-PD-L1 therapy, improved the efficacy of immunotherapy in HCC. Conclusions:These findings illustrate the importance of liver fibrosis-induced immunosuppression as a tumor-promoting mechanism. GOLM1, which is highly upregulated in the fibrotic liver, regulates tumor microenvironmental immune escape via the EGFR/PD-L1 signaling pathway. EGFR blockade may bolster the efficacy.

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Evaluation of Rabbits Liver Fibrosis Using Gd-DTPA-BMA of Dynamic Contrast-Enhanced Magnetic Resonance Imaging

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/2791142 ◽

2021 ◽

Vol 2021 ◽

pp. 1-7

Author(s):

Qian Cui ◽

FengTai He ◽

Jiawei Hu ◽

Shuo Li ◽

Dongmei Guo ◽

...

Keyword(s):

Liver Fibrosis ◽

Normal Liver ◽

Fibrosis Stage ◽

Pharmacokinetic Parameters ◽

Control Group ◽

Resonance Imaging ◽

Fibrotic Liver ◽

Dce Mri ◽

Contrast Enhanced ◽

Advanced Liver Fibrosis

Objective. To evaluate the different pharmacokinetic parameters of the DCE-MRI method on diagnosing and staging of rabbits’ liver fibrosis. Methods. We had performed DCE-MRI for rabbits that had been divided into the experiment group and the control group. Then, rabbits’ images were transferred to a work station to get three parameters such as Ktrans, Kep, and Ve, which had been measured to calculate. After data were analyzed, ROC analyses were performed to assess the diagnostic performance of Ktrans, Kep, and Ve to judge liver fibrosis. Results. The distribution of the different liver fibrosis group was as follows: F1, n = 8; F2, n = 9; F3, n = 6; F4, n = 5. No fibrosis was deemed as F0, n = 6. Kep is statistically significant P < 0.05 for F0 and mild liver fibrosis stage, and the Kep shows AUC of 0.814. Three parameters are statistically significant for F0 and advanced liver fibrosis stage (Ktrans and Kep, P < 0.01 ; Ve, P < 0.05 ), and the Ktrans shows AUC of 0.924; the Kep shows AUC of 0.909; the Ve shows AUC of 0.848; Ktrans and Kep are statistically significant for mild and advanced liver fibrosis stages (Ktrans, P < 0.01 ; Kep, P < 0.05 ), and the Ktrans shows AUC of 0.840; the Kep shows AUC of 0.765. Both Ktrans and Kep are negatively correlated with the liver fibrosis stage. Ve is positively correlated with the liver fibrosis stage. Conclusion. Ktrans is shown to be the best DCE parameter to distinguish the fibrotic liver from the normal liver and mild and advanced fibrosis. On the contrary, Kep is moderate and Ve is worst. And Kep is a good DCE parameter to differentiate mild fibrosis from the normal liver.

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High Risk of Hepatocellular Carcinoma Development in Fibrotic Liver: Role of the Hippo-YAP/TAZ Signaling Pathway

International Journal of Molecular Sciences ◽

10.3390/ijms20030581 ◽

2019 ◽

Vol 20 (3) ◽

pp. 581 ◽

Cited By ~ 7

Author(s):

Hyuk Moon ◽

Kyungjoo Cho ◽

Sunyeong Shin ◽

Do Kim ◽

Kwang-Hyub Han ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Fibrosis ◽

Liver Cancer ◽

Signaling Pathways ◽

Signaling Pathway ◽

Genetic Alterations ◽

Molecular Signaling ◽

Fibrotic Liver ◽

Molecular Signaling Pathways ◽

End Stage

Liver cancer is the fourth leading cause of cancer-related death globally, accounting for approximately 800,000 deaths annually. Hepatocellular carcinoma (HCC) is the most common type of liver cancer, making up about 80% of cases. Liver fibrosis and its end-stage disease, cirrhosis, are major risk factors for HCC. A fibrotic liver typically shows persistent hepatocyte death and compensatory regeneration, chronic inflammation, and an increase in reactive oxygen species, which collaboratively create a tumor-promoting microenvironment via inducing genetic alterations and chromosomal instability, and activating various oncogenic molecular signaling pathways. In this article, we review recent advances in fields of liver fibrosis and carcinogenesis, and consider several molecular signaling pathways that promote hepato-carcinogenesis under the microenvironment of liver fibrosis. In particular, we pay attention to emerging roles of the Hippo-YAP/TAZ signaling pathway in stromal activation, hepatic fibrosis, and liver cancer.

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Orphan nuclear receptor NR4A2 inhibits hepatic stellate cell proliferation through MAPK pathway in liver fibrosis

PeerJ ◽

10.7717/peerj.1518 ◽

2015 ◽

Vol 3 ◽

pp. e1518 ◽

Cited By ~ 10

Author(s):

Pengguo Chen ◽

Jie Li ◽

Yan Huo ◽

Jin Lu ◽

Lili Wan ◽

...

Keyword(s):

Extracellular Matrix ◽

Cell Proliferation ◽

Liver Fibrosis ◽

Nuclear Receptor ◽

Mapk Pathway ◽

Stellate Cell ◽

Smooth Muscle Actin ◽

Control Group ◽

Orphan Nuclear Receptor ◽

Fibrotic Liver

Hepatic stellate cells (HSCs) play a crucial role in liver fibrosis, which is a pathological process characterized by extracellular matrix accumulation. NR4A2 is a nuclear receptor belonging to the NR4A subfamily and vital in regulating cell growth, metabolism, inflammation and other biological functions. However, its role in HSCs is unclear. We analyzed NR4A2 expression in fibrotic liver and stimulated HSCs compared with control group and studied the influence on cell proliferation, cell cycle, cell apoptosis and MAPK pathway after NR4A2 knockdown. NR4A2 expression was examined by real-time polymerase chain reaction, Western blotting, immunohistochemistry and immunofluorescence analyses. NR4A2 expression was significantly lower in fibrotic liver tissues and PDGF BB or TGF-βstimulated HSCs compared with control group. After NR4A2 knockdownα-smooth muscle actin and Col1 expression increased. In addition, NR4A2 silencing led to the promotion of cell proliferation, increase of cell percentage in S phase and reduced phosphorylation of ERK1/2, P38 and JNK in HSCs. These results indicate that NR4A2 can inhibit HSC proliferation through MAPK pathway and decrease extracellular matrix in liver fibrogenesis. NR4A2 may be a promising therapeutic target for liver fibrosis.

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Solanine Inhibits Immune Escape Mediated by Hepatoma Treg Cells via the TGFβ/Smad Signaling Pathway

BioMed Research International ◽

10.1155/2020/9749631 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Juwei Gao ◽

Yinyin Ying ◽

Jue Wang ◽

Yiyi Cui

Keyword(s):

Flow Cytometry ◽

Liver Cancer ◽

Tumor Volume ◽

Immune Escape ◽

Tumor Model ◽

Treg Cells ◽

Inhibitory Effect ◽

Control Group ◽

Transplanted Tumor ◽

Tumor Tissues

Objective. To observe the inhibitory effect of solanine on regulatory T cells (Treg) in transplanted hepatoma mice and to study the mechanism of solanine inhibiting tumor growth. Methods. The levels of Treg cells and IL-2, IL-10, and TGFβ in the blood of patients with liver cancer were detected by flow cytometry and ELISA, respectively. A mouse hepatocellular carcinoma (HCC) graft model was established and randomly divided into four groups: control group, solanine group, TGFβ inhibitor group (SB-431542), and solanine +TGFβ inhibitor combined group. Tumor volume of each group was recorded, tumor inhibition rate was calculated, and tumor metastasis was counted. The proportion of CD4+CD25+Foxp3+ Treg in transplanted tumor tissues was detected by flow cytometry. The expression levels of Foxp3 and TGFβ in transplanted tumor tissues were detected by quantitative fluorescence PCR. Results. Compared with healthy people, Treg cells and IL-2, IL-10, and TGFβ contents in peripheral blood of liver cancer patients were increased. The results of the transplanted tumor model in mice showed that the tumor volume of the transplanted mice in the solanine group and the TGFβ inhibitor mice was reduced compared with the control group. The combined group had the smallest tumor volume. The proportion of CD4+CD25+Foxp3+ Treg in the transplanted tumor tissues of mice in the solanine treatment group was significantly lower than that in the control group. The expressions of Foxp3 and TGFβ in the transplanted tumor tissues of mice in the solanine group were significantly lower than those in the control group. Conclusion. Solanine may enhance the antitumor immune response by downregulating the proportion of CD4+CD25+ Treg and the expression of Foxp3 and TGFβ in tumor tissues.

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A Preliminary Study on the Diagnostic Efficacy of Proton Magnetic Resonance Spectroscopy at 3.0T in Rabbit With VX2 Liver Tumor

Technology in Cancer Research & Treatment ◽

10.1177/15330338211036852 ◽

2021 ◽

Vol 20 ◽

pp. 153303382110368

Author(s):

Ruikun Liao ◽

Dan Zhang ◽

Xiaojiao Li ◽

Jiang Ma ◽

Jiayi Yu ◽

...

Keyword(s):

Magnetic Resonance ◽

Liver Cancer ◽

Magnetic Resonance Spectroscopy ◽

Liver Tumor ◽

Roc Curve ◽

Normal Liver ◽

Resonance Spectroscopy ◽

Diagnostic Efficacy ◽

Cutoff Value ◽

Vx2 Tumor

Background: To investigate the diagnostic efficacy of choline (Cho) value of magnetic resonance spectroscopy (MRS) in rabbit with VX2 liver tumor via comparative and quantitative analysis with the choline compounds concentration measured by enzyme linked immunosorbent assay (ELISA). Methods: MRS was performed on normal liver and VX2 tumor. The Cho value of VX2 tumor was compared with that of normal liver. Tissues were harvested for ELISA to detect the concentrations of acetylcholine (ACh), glycophorophosphygholine (GPC) and phosphochorine (PC). The diagnostic performance of Cho value and concentrations of choline compounds were assessed by receiver operating characteristic (ROC) curve and area under ROC curve (AUC). The specificity and sensitivity were discussed by the maximum Youden’s index. Results: The concentration of ACh was obviously higher than that of GPC and PC both in VX2 tumor and normal liver ( P < 0.01). Furthermore, the concentration differences among ACh, GPC and PG were the third power of 10. Both the ACh concentration and Cho value of MRS in VX2 tumor were significantly higher than those in normal liver ( P < 0.01). The AUC of ACh in VX2 tumor was 0.883, when the cutoff value was 7259000, the sensitivity and specificity of the diagnosis of liver cancer were 94.4% and 77.8%, respectively. The AUC of Cho in VX2 tumor was 0.807, when the cutoff value was 28.35, the sensitivity and specificity of the diagnosis of liver cancer were 83.3% and 77.8%, respectively. Conclusion: The change of Cho value in MRS between liver cancer and normal liver was consistent with the changes of concentrations of choline compounds measured by ELISA, especially the change of ACh concentration. The diagnostic efficiency of Cho value and that of choline compounds concentration in liver cancer were extremely similar, with the AUC more than 0.8. We conclude that MRS may be applied as an important, non-invasive biomarker for the diagnosis of liver cancer.

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Diagnostic and prognostic potential of kallistatin in assessment of liver parenchyma changes in patients with non-alcoholic fatty liver disease and hypertension kallistatin in patients with NAFLD and hypertension

Imaging ◽

10.1556/1647.2021.00046 ◽

2021 ◽

Author(s):

Natalia M. Zhelezniakova ◽

Anastasiia O. Rozhdestvenska ◽

Olha V. Stepanova

Keyword(s):

Liver Disease ◽

Liver Fibrosis ◽

Fatty Liver ◽

Fatty Liver Disease ◽

Liver Steatosis ◽

Liver Parenchyma ◽

Control Group ◽

Fibrotic Liver ◽

Alcoholic Fatty Liver ◽

Alcoholic Fatty Liver Disease

Abstract Background and aim Non-alcoholic fatty liver disease (NAFLD) is closely linked to hypertension (HT). An important issue remains the search for non-invasive tests to NAFLD detection in the early stages of liver fibrosis. The objective of the study was to evaluate the diagnostic and prognostic value of kallistatin in assessing the liver fibrosis progression in NAFLD and HT patients. Patients and methods One hundred fifteen patients with NAFLD with and without HT were examined, the control group consisted of 20 relatively healthy volunteers. Plasma kallistatin level measurement, ultrasound steatometry and elastography were performed in all patients. Results Kallistatin level was 65.03 ng mL−1 (95% CI 61.38; 68.68), 83.42 ng mL−1 (95% CI 81.89; 84.94) and 111.70 ng mL−1 (95% CI 106.14; 113.22) in patients with NAFLD and HT, isolated NAFLD and control group, respectively. There were significant differences in the liver parenchyma condition between groups. Kallistatin levels strongly inversely correlated with the attenuation coefficient and the mean liver stiffness in NAFLD and HT (rs = −0.70) and in the isolated NAFLD patients (rs = −0.56; rs = −0.68, respectively). Kallistatin level was 71.82 ng mL−1 (95% CI 70.16; 79.51) and 58.62 ng mL−1 (95% CI 55.81; 64.45) in patients with HT stage I and HT stage II, respectively (P < 0.001). Conclusions Concomitant HT in NAFLD patients is associated with greater severity of fatty and fibrotic liver changes. The course of NAFLD is accompanied by decrease in kallistatin level. Increased degree of liver steatosis and fibrosis, inflammation activity, increased BMI and increased stage of HT lead to inhibition of kallistatin activity. Kallistatin may be considered as a biomarker for progression assessment of NAFLD with or without HT.

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Deficient necroptosis pathway as a negative prognostic factor in acute myeloid leukemia.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.11611 ◽

2017 ◽

Vol 35 (15_suppl) ◽

pp. 11611-11611

Author(s):

Silvia Lo Monaco ◽

Giovanni Marconi ◽

Maria Chiara Fontana ◽

Cristina Papayannidis ◽

Eugenio Fonzi ◽

...

Keyword(s):

Overall Survival ◽

Prognostic Factors ◽

Molecular Mechanisms ◽

Immune Escape ◽

Rank Test ◽

Control Group ◽

Fisher Exact Test ◽

Copy Number Loss ◽

Tumor Immune Escape ◽

Exact Test

11611 Background: Necroptosis is a type of necrotic cell death involving several genes transcription and activation of molecular mechanisms as death receptors, interferon, toll-like receptors, intracellular RNA and DNA sensors.The process is leading by the family of receptor-interacting protein kinase ( RIPK3, RIPK2, RIPK1) and the MLKL substrate. Losses of RIPK3 or MLKL, as well as deficiency in apoptosis, could allow tumor cells to escape the immunomediated cells death (ICD). Methods: We performed SNP Arrays (Cytoscan HD and SNP 6.0, Affymetrix) on a cohort of 300 non-M3 AML patients at diagnosis and we analyzed the Overall Survival (OS) of our patients with deficiency on necroptosis pathways. Survival was analyzed with Kaplan-Mayer method and Log-Rank test. We further analyze the relevance of different prognostic factors by the use of COX-Hazard Ratio statistical analysis. Results: We find that 18 patients presented a loss of RIPK1 or MLKL (nobody presented losses in RIPK3/RIPK2) and 13/18 patients were older than 65 years old. The Overall Survival (OS) of patients with alterations in these genes is significantly lower than control group, with a median OS of 3 vs 6 month respectively (p<.0.001). With Fisher Exact Test we further demonstrate that copy number loss of RIPK1 or MLKL are associate to loss of TP53 or FANCAgenes, complex karyotype and advanced age. COXHR model with RIPK1 or MLKL loss, BRACA1 loss, TP53 mutation, FANCA loss, secondary disease and diagnosis karyotype considered as categorical variable shows that necroptosis deficiency (HR 1.98, CI 95% 1.04-3.78), TP53mutation , and secondary AML are independent negative prognostic factors in an optimal model. Conclusions: Our study shows that losses in necroptosis pathways are an uncommon alteration in AML, prevalent in old population. Moreover, we hypothesize that the loss of genes involved in necroptosis could be a real mechanism of tumor immune-escape and could be a rational to select patients that have high probability to be resistant at chemotherapy promoting ICD mechanism. Acknowledgment: ELN,AIL,AIRC, progetto Regione-Università 2010-12, FP7 NGS-PTL project,HARMONY.

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Salivary duct carcinoma with striking neutrophil-tumor cell cannibalism

CytoJournal ◽

10.4103/1742-6413.84222 ◽

2011 ◽

Vol 8 ◽

pp. 15 ◽

Cited By ~ 7

Author(s):

Payam Arya ◽

Walid E. Khalbuss ◽

Sara E. Monaco ◽

Liron Pantanowitz

Keyword(s):

Tumor Cell ◽

Immune Escape ◽

Surgical Excision ◽

Salivary Duct Carcinoma ◽

High Grade ◽

Salivary Duct ◽

Melanoma Tumor ◽

Interesting Phenomenon ◽

Tumor Immune Escape ◽

Duct Carcinoma

Cannibalism of neutrophils by tumor cells has previously been reported in certain carcinomas, lymphoma and melanoma. Tumor cannibalism is believed to serve as a tumor-immune escape mechanism, associated with high-grade aggressive cancers with a significantly increased metastatic potential. This interesting phenomenon has not been previously documented in association with salivary gland tumors. We report, for the first time, striking neutrophil-tumor cell cannibalism associated with a high grade, aggressive and metastatic salivary duct carcinoma of the parotid gland highlighted within cytological and surgical excision pathology specimens.

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Biological and Phytochemical Screening of Fumaria indica extract on Chemically Induced Hepatocellular Carcinoma with Reference to Biochemical Parameters

International Journal of Pharmaceutical Quality Assurance ◽

10.25258/ijpqa.10.2.1 ◽

2019 ◽

Vol 10 (02) ◽

Author(s):

Irfan Aziz ◽

Birendra Shrivastava ◽

Chandana Venkateswara Rao2 ◽

Sadath Ali

Keyword(s):

Free Radicals ◽

Liver Disease ◽

Liver Cancer ◽

Early Stage ◽

Food Additives ◽

Ethanolic Extract ◽

Hepatoprotective Activity ◽

Control Group ◽

Industrial Chemicals ◽

Toxic Industrial Chemicals

Liver disease or liver cancer is the sixth most common cancer and the third leading cause of cancer mortality in the world. Hepatitis viral infection, food additives, alcohol, fungal toxins (aflatoxins), toxic industrial chemicals, air and water pollutants are the major risk factors of liver cancer. Moreover, due to high tolerance of liver, HCC is seldom detected at an early stage and once detected treatment faces a poor prognosis in most cases.Fumaria indica possesses hepatoprotective activity as evidenced by the significant and dose dependent restoring the activities of entire liver cancer marker enzymes, diminution in tumor incidence, decrease in lipid peroxidation (LPO) and increase in the level of antioxidant enzymes (GSH, CAT, SOD, GPx and GST) through scavenging of free radicals, or by enhancing the activity of antioxidant, which then detoxify free radicals. These factors protect cells from ROS damage in NDEA and CCl4-induced hepatocarcinogenesis. Histopathological observations of liver tissues too correlated with the biochemical observations. Thus, present investigation suggested that the Fumaria indica would exert a chemoprotective effect by reversing the oxidant-antioxidant imbalance during hepatocarcinogenesis induced by NDEA and CCl4. Besides Fumaria indicais very much effective in preventing NDEA-induced multistage hepatocarcinogenesis possibly through antioxidant and antigenotoxic nature, which was confirmed by various liver injury and biochemical tumour markers enzymes. The hepatoprotective activity of a Fumaria indicaof 50 % ethanolic extract was studied using rats. The animals received a single intraperitoneal injection of N-nitrosodiethylamine 200mg/kg body wt followed by subcutaneous injection of CCl4 in a dose of 3 ml/kg body wt. Fumaria indica extract dose dependently and significantly the increase in serum hepatic enzyme levels after NDEAand CCl4 treatment compared to the toxin control group. The results of this study confirmed the antioxidant and hepatoprotective activity of the Fumaria indicaextract against carbon tetrachlorideand N-nitrosodiethylamine induced hepatotoxicity in rats. In addition to this, studies on molecular aspect of hepatoprotective therapy will give mechanistic information in hepatoprotective therapy and also critical balance should be there between the animal model and clinical research. The hepatoprotective properties of Fumaria indicashould provide useful information in the possible application in hepatic liver disease.

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