Gryllus bimaculatus De Geer hydrolysates alleviate lipid accumulation, inflammation, and endoplasmic reticulum stress in palmitic acid-treated human hepatoma G2 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Nonalcoholic fatty liver disease (NAFLD) is one of the most common hepatic diseases closely intertwined with saturated fatty acids intake. Therefore, various studies are being conducted to find natural substances to prevent either the onset or progression of NAFLD. According to traditional medicinal literature, it has been reported that Gryllus bimaculatus De Geer (GB) has systemic detoxifying activity; however, the preventive effects of GB on NAFLD have not been elucidated to date. AIM OF STUDY: To evaluate the potential of GB as a material for the mitigation of NAFLD, we investigated the effects of GB hydrolysates on the hepatic lipid accumulation, inflammation, and endoplasmic reticulum (ER) stress in human hepatoma G2 (Hep G2) cells treated with palmitic acid (PA). METHODS: Steamed and dried GB was defatted, pulverized, and then lyophilized following hydrolyzation using Neutrase® (GB-N) or Flavourzyme® (GB-F). Hep G2 cells were incubated with GB-N or GB-F at various concentrations (0, 0.25, 0.5, and 1 mg/mL) for 24 h, and then PA was treated for another 24 h. RESULTS: The GB-N and GB-F significantly prevented the PA-induced intracellular lipid accumulation in the human liver cells (p < 0.05). Moreover, the GB-N and GB-F increased the hepatic cellular viability against the PA-treatment (p < 0.05). In addition, the GB-N and GB-F significantly ameliorated the PA-inducible proinflammatory cytokines mRNA expression, such as tumor necrosis factor-α and interleukin-1ß, compared to the PA-treated hepatic cells (p < 0.05). Furthermore, the GB-N and GB-F inhibited the PA-inducible lipogenic mRNA expression, such as fatty acid synthase, sterol regulatory element-binding protein 1c, and peroxisome proliferator-activated receptor-γ (p < 0.05). Moreover, the GB-N and GB-F alleviated the ER stress-related mRNA expression, such as glucose regulatory protein 78 and X-box binding protein increased in PA-treated cells (p < 0.05). CONCLUSIONS: These results indicate that GB-N and GB-F could be used as materials to prevent the NAFLD onset or progression with alleviating hepatic lipid accumulation, inflammation, and ER stress.

Overexpression of Replication-Dependent Histone Signifies a Subset of Dedifferentiated Liposarcoma with Increased Aggressiveness.

Liposarcoma (LPS) is an adult soft tissue malignancy that arises from fat tissue, where well-differentiated (WD) and dedifferentiated (DD) forms are the most common. DDLPS represents the progression of WDLPS into a more aggressive high-grade and metastatic form. Although a few DNA copy-number amplifications are known to be specifically found in WD- or DDLPS, systematic genetic differences that signify subtype determination between WDLPS and DDLPS remain unclear. Here, we profiled the genome and transcriptome of 38 LPS tumors to uncover the genetic signatures of subtype differences. Replication-dependent histone (RD-HIST) mRNAs were highly elevated and their regulation was disrupted in a subset of DDLPS, increasing cellular histone molecule levels, as measured using RNA-seq (the averaged fold change of 53 RD-HIST genes between the DD and WD samples was 10.9) and immunohistochemistry. The change was not observed in normal tissues. Integrated whole-exome sequencing, RNA-seq, and methylation analyses revealed that the overexpressed HMGA2 (the fold change between DD and WD samples was 7.3) was responsible for the increased RD-HIST level, leading to aberrant cell proliferation. Therefore, HMGA2-mediated elevation of RD-HISTs were crucial events in determining the aggressiveness of DDLPS, which may serve as a biomarker for prognosis prediction for liposarcoma patients.

Complete regression of metastatic de-differentiated liposarcoma with engineered mesenchymal stromal cells with dTRAIL and HSV-TK.

De-differentiated liposarcoma (DDLPS) is a rare cancer with high rates of recurrence and metastasis. Currently, treatment with doxorubicin-ifosphamide, following surgical resection, is routinely performed. However, clinical treatment of these refractory cancers require further study. We investigated the treatment of mesenchymal stromal cells (MSC) transduced with dodecameric tumor necrosis factor receptor apoptosis-inducing ligand (dTRAIL) and herpes simplex virus thymidine kinase (HSV-TK) (MSC-TR/TK), as a method to approach DDLPS therapy. First, in order to assess the efficacy of this therapy, cell viability was evaluated by apoptosis analysis of a DDLPS cell line co-cultured with patient-derived cells (PDCs) and MSC-TR/TK in vitro. In vivo, we established a lung metastasis model using the DDLPS cell line and assessed the anti-tumorigenic efficiency of dTRAIL-TK by injecting MSC-TR/TK. Results confirmed that liposarcoma cells resistant to dTRAIL in PDCs, transformed by HSV-TK, induced apoptosis effectively after treatment with toxic ganciclovir (GCV). Meanwhile, we observed that treatment of GCV after injection of MSC-TR/TK effectively eliminated lung nodules in a lung metastasis model established from LPS246 cells resistant to dTRAIL. When mice were treated with GCV two days after double injection with MSC-TR/TK, the tumor suppression effect was even more pronounced.

Combination therapy with c-met inhibitor and TRAIL enhances apoptosis in dedifferentiated liposarcoma patient-derived cells.

BACKGROUND: Liposarcoma (LPS) is a tumor derived from adipose tissue, and has the highest incidence among soft tissue sarcomas. Dedifferentiated liposarcoma (DDLPS) is a malignant tumor with poor prognosis. Recurrence and metastasis rates in LPS remain high even after chemotherapy and radiotherapy following complete resection. Therefore, the development of advanced treatment strategies for LPS is required. In the present study, we investigated the effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) treatment, and of combination treatment using TRAIL and a c-Met inhibitor on cell viability and apoptosis in LPS and DDLPS cell lines of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) treatment, and of combination treatment using TRAIL and a c-Met inhibitor. METHODS: We analyzed cell viability after treatment with TRAIL and a c-Met inhibitor by measuring CCK8 and death receptor 5 (DR5) expression levels via fluorescence activated cell sorting (FACS) in both sarcoma cell lines and DDLPS patient-derived cells (PDCs). Moreover, we validated the effects of TRAIL alone and in combination with c-Met inhibitor on apoptosis in LPS cell lines and DDLPS PDCs via FACS. RESULTS: Our results revealed that combination treatment with a c-Met inhibitor and human recombinant TRAIL (rhTRAIL) suppressed cell viability and induced cell death in both sarcoma cell lines and DDLPS PDCs, which showed varying sensitivities to rhTRAIL alone. Also, we confirmed that treatment with a c-Met inhibitor upregulated DR5 levels in sarcoma cell lines and DDLPS PDCs. In both TRAIL-susceptible and TRAIL-resistant cells subjected to combination treatment, promotion of apoptosis was dependent on DR5 upregulation. CONCLUSION: From these results, our findings validated that DR5 up-regulation caused by combination therapy with a c-Met inhibitor and rhTRAIL enhanced TRAIL sensitization and promoted apoptosis. We propose the use of this approach to overcome TRAIL resistance and serve as a novel treatment strategy for clinical trials.

Establishment of a Novel PDX Mouse Model and Evaluation of the Tumor Suppression Efficacy of Bortezomib Against Liposarcoma.

The patient-derived xenograft (PDX) model has been adopted as a major tool for studying tumorigenesis and differentiation in various carcinomas. In addition, it has been used in the development of anticancer agents. PDX models have been among the most meaningful tools used to understand the role of stromal cells and vascular cells in the body, which are major factors in cancer development and the application of therapeutic agents. Also, the establishment of PDX models from liposarcoma patients is considered to be important for understanding lipomagenesis and following drugs development. For these reasons, we developed patient-derived cell (PDC) and PDX models derived from 20 liposarcoma patients. The tissues of these patients were obtained in accordance with the principles of the Samsung Medical Center's ethics policy, and cell culture and xenografting onto the mice were performed under these principles. High-throughput drug screening (HTS) was carried out using established PDCs to select candidate drugs. Among the different candidate anticancer drugs, we tested the effect of bortezomib, which was expected to inhibit MDM2 amplification. First, we confirmed that the PDCs maintained the characteristics of liposarcoma cells by assessing MDM2 amplification and CDK4 overexpression using fluorescence in situ hybridization. Analysis of short tandem repeats and an array using comparative genomic hybridization confirmed that the PDX model exhibited the same genomic profile as that of the patient. Immunohistochemistry for MDM2 and CDK4 showed that the overexpression patterns of both proteins were similar in the PDX models and the PDCs. Specifically, MDM2 amplification was observed to be significantly correlated with the successful establishment of PDX mouse models. However, CDK4 expression did not show such a correlation. Of the anticancer drugs selected through HTS, bortezomib showed a strong anticancer effect against PDC. In addition, we observed that bortezomib suppressed MDM2 expression in a dose-dependent manner. In contrast, p21 tended to elicit an increase in PDC expression. Treatment of the PDX model with bortezomib resulted in an anticancer effect similar to that seen in the PDCs. These results support that PDCs and PDX models are among the most powerful tools for the development and clinical application of anticancer drugs for the treatment of liposarcoma patients.

Therapeutic strategies for locally recurrent and metastatic de-differentiated liposarcoma with herpes simplex virus-thymidine kinase-expressing mesenchymal stromal cells.

BACKGROUND AIMS: Major challenges in de-differentiated liposarcoma (DDLPS) therapy are the high rate of sequential recurrence (>80%) and metastasis (20-30%) following surgical removal. However, well-defined therapeutic strategies for this rare malignancy are lacking and are critically needed. METHODS: We investigated a new approach to DDLPS therapy with mesenchymal stromal cells expressing herpes simplex virus-thymidine kinase (MSC-TK). In an effort to evaluate this efficacy, in vitro cytotoxicity of MSC-TK against DDLPS cells was analyzed using an apoptosis assay. For pre-clinical study, the MSC-TK-induced reduction in recurrence and metastasis was validated in a recurrent DDLPS model after the macroscopic complete resection and lung metastasis DDLPS model. RESULTS: MSC-TK induced apoptosis in DDLPS cells by bystander effects via gap junction intracellular communication (GJIC) of toxic ganciclovir (GCV). Recurrent DDLPS models following no residual tumor/microscopic tumor resection and lung metastasis DDLPS models were established, which suggested clinical relevance. MSC-TK markedly reduced locoregional recurrence rates and prolonged recurrence-free survival, thus increasing overall survival in the recurrent DDLPS model. MSC-TK followed by GCV treatment yielded a statistically significant reduction in early- and advanced-stage lung metastasis. DISCUSSION: This therapeutic strategy may serve as an alternative or additional strategy by applying MSC-TK to target residual tumors following surgical resection, thus reducing local relapse and metastasis in these patients.

Comparison of evaluation parameters in the retinal layer between diabetic cystoid macular edema and postoperative cystoid macular edema after cataract surgery based on a hierarchical approach.

In this study, several variables related to the thickness of the retinal layer were measured via optical coherence tomography (OCT), and the clinical applicability of such measurements was evaluated to differentiate between diabetic cystoid macular edema (DCME) and postoperative cystoid macular edema (PCME). To this end, a total of 120 subjects (30 healthy individuals, 60 DCME patients, and 30 PCME patients) were selected as the experimental subjects. The six risk factors included the thicknesses for the total retina (TR), the inner retina (IR), the photoreceptor outer segments (POS), the outer retina (OR), the ganglion cell (GC), and the retinal nerve fiber layer (RNFL), and these were estimated by using a hierarchical approach through observations from OCT image scans. All of the risk factors were obtained from the OCT images captured within a 6-mm diameter from the center of the macula. The results of the experiment indicated that the proposed method can reliably differentiate between DCME and PCME. Moreover, as DCME and PCME progressed, the most significant deterioration was found in the central macular region. These results suggest that the proposed method has clinical applicability for the diagnosis of DCME and PCME from OCT images.