Identification of MYC as an antinecroptotic protein that stifles RIPK1-RIPK3 complex formation

The underlying mechanism of necroptosis in relation to cancer is still unclear. Here, MYC, a potent oncogene, is an antinecroptotic factor that directly suppresses the formation of the RIPK1-RIPK3 complex. Gene set enrichment analyses reveal that the MYC pathway is the most prominently down-regulated signaling pathway during necroptosis. Depletion or deletion of MYC promotes the RIPK1-RIPK3 interaction, thereby stabilizing the RIPK1 and RIPK3 proteins and facilitating necroptosis. Interestingly, MYC binds to RIPK3 in the cytoplasm and inhibits the interaction between RIPK1 and RIPK3 in vitro. Furthermore, MYC-nick, a truncated form that is mainly localized in the cytoplasm, prevented TNF-induced necroptosis. Finally, down-regulation of MYC enhances necroptosis in leukemia cells and suppresses tumor growth in a xenograft model upon treatment with birinapant and emricasan. MYC-mediated suppression of necroptosis is a mechanism of necroptosis resistance in cancer, and approaches targeting MYC to induce necroptosis represent an attractive therapeutic strategy for cancer

Structure-Activity Relationship Analysis of YM155 for Inducing Selective Cell Death of Human Pluripotent Stem Cells

Despite great potential for regenerative medicine, the high tumorigenic potential of human pluripotent stem cells (hPSCs) to form undesirable teratoma is an important technical hurdle preventing safe cell therapy. Various small molecules that induce the complete elimination of undifferentiated hPSCs, referred to as “stemotoxics,” have been developed to facilitate tumor-free cell therapy, including the Survivin inhibitor YM155. In the present work, based on the chemical structure of YM155, total 26 analogs were synthesized and tested for stemotoxic activity toward human embryonic stem cells (hESCs) and induced PSCs (iPSCs). We found that a hydrogen bond acceptor in the pyrazine ring of YM155 derivatives is critical for stemotoxic activity, which is completely lost in hESCs lacking SLC35F2, which encodes a solute carrier protein. These results suggest that hydrogen bonding interactions between the nitrogens of the pyrazine ring and the SLC35F2 protein are critical for entry of YM155 into hPSCs, and hence stemotoxic activity

Drug Repositioning for Cancer Therapy Based on Large-Scale Drug-Induced Transcriptional Signatures.

An in silico chemical genomics approach is developed to predict drug repositioning (DR) candidates for three types of cancer: glioblastoma, lung cancer, and breast cancer. It is based on a recent large-scale dataset of ~20,000 drug-induced expression profiles in multiple cancer cell lines, which provides i) a global impact of transcriptional perturbation of both known targets and unknown off-targets, and ii) rich information on drug's mode-of-action. First, the drug-induced expression profile is shown more effective than other information, such as the drug structure or known target, using multiple HTS datasets as unbiased benchmarks. Particularly, the utility of our method was robustly demonstrated in identifying novel DR candidates. Second, we predicted 14 high-scoring DR candidates solely based on expression signatures. Eight of the fourteen drugs showed significant anti-proliferative activity against glioblastoma; i.e., ivermectin, trifluridine, astemizole, amlodipine, maprotiline, apomorphine, mometasone, and nortriptyline. Our DR score strongly correlated with that of cell-based experimental results; the top seven DR candidates were positive, corresponding to an approximately 20-fold enrichment compared with conventional HTS. Despite diverse original indications and known targets, the perturbed pathways of active DR candidates show five distinct patterns that form tight clusters together with one or more known cancer drugs, suggesting common transcriptome-level mechanisms of anti-proliferative activity

A Novel Human scFv Library with Non-Combinatorial Synthetic CDR Diversity.

The present work describes the construction and validation of a human scFv library with a novel design approach to synthetic complementarity determining region (CDR) diversification. The advantage of synthetic antibody libraries includes the possibility of exerting fine control over factors like framework sequences, amino acid and codon usage, and CDR diversity. However, random combinatorial synthesis of oligonucleotides for CDR sequence diversity also produces many clones with unnatural sequences and/or undesirable modification motifs. To alleviate these issues, we designed and constructed a novel semi-synthetic human scFv library with non-combinatorial, pre-designed CDR diversity and a single native human framework each for heavy, kappa, and lambda chain variable domains. Next-generation sequencing analysis indicated that the library consists of antibody clones with highly nature-like CDR sequences and the occurrence of the post-translational modification motifs is minimized. Multiple unique clones with nanomolar affinity could be isolated from the library against a number of target antigens, validating the library design strategy. The results demonstrate that it is possible to construct a functional antibody library using low, non-combinatorial synthetic CDR diversity, and provides a new strategy for the design of antibody libraries suitable for demanding applications

The list of active DR candidates.

<p>The list of active DR candidates.</p

Performance evaluations using the public anti-cancer HTS dataset as a benchmark.

<p>The seven classifiers (S, T, E, ST, SE, TE, and STE) were evaluated based on the AUCs of the ROC curve for glioblastoma, lung cancer, and breast cancer. Only compounds in the core set were evaluated. The AUC values were calculated by averaging 100 rounds of 3-fold cross validation. (A) Typical examples of performance evaluation using the HTS data set for glioblastoma (AID45), lung cancer (AID5), and breast cancer (AID97). The AUCs were independently calculated using two distinct sets of hit compounds as a benchmark (or positives)—i) the hit compounds of known anti-cancer activity (red lines) and ii) the novel hits (green lines). The distribution of AUCs using (B) the compounds of known anti-cancer activity as a benchmark, and (C) the novel hits as a benchmark.</p

Pathway enrichment pattern of the eight active DR candidates for glioblastoma.

<p>The p-values and their adjusted q-values were calculated by hypergeometric test and the Benjamini-Hochberg method, respectively.</p

Prognostic Value of Functional Assessment of Cancer Therapy-General (FACT-G) in Advanced Non-Small-Cell Lung Cancer Treated with Korean Medicine

Objectives. The impact of health-related quality of life (HRQoL) on survival has been investigated in patients with various cancers. Here, we evaluated the prognostic value of HRQoL using the Functional Assessment of Cancer Therapy-General (FACT-G) in advanced non-small-cell lung cancer (NSCLC) patients treated with Korean medicine. Methods. A retrospective review of medical records and FACT-G scores of patients with advanced NSCLC who received treatment with Korean medicine was conducted. The reliability of the FACT-G was determined using Cronbach’s alpha and calculating floor-and-ceiling effects. Correlations between FACT-G scores were estimated using Pearson’s correlation analysis. Overall survival was calculated using the Kaplan–Meier method, and the prognostic impact of FACT-G scores and patients’ characteristics was evaluated with Cox proportional hazards regression. Results. Of the 165 enrolled patients, 115 (70%) had extrathoracic metastasis and 139 (84%) had undergone prior anticancer treatment. The median overall survival was 10.1 months. The mean FACT-G score was 65.0, and Cronbach’s alpha for the FACT-G was 0.917. Age ≥65 years, male sex, smoking history, squamous-cell carcinoma, Eastern Cooperative Oncology Group Performance Status (ECOG-PS) ≥2, and presence of extrathoracic metastasis were associated with an increased risk of mortality. High FACT-G total scores, physical well-being (PWB), emotional well-being, and functional well-being were associated with prolonged survival. After adjusting for age, sex, smoking history, ECOG-PS, histological type, and presence of extrathoracic metastasis, a high FACT-G total score (hazard ratio (HR): 0.99, p=0.032) and high PWB score (HR: 0.94, p<0.001) were associated with prolonged survival as independent prognostic factors in patients with advanced NSCLC. Conclusion. The FACT-G total score and PWB score as HRQoL measurements were significant prognostic factors for survival in advanced NSCLC patients treated with Korean medicine. This finding implies that the FACT-G can be used in clinical practice as a predictor of survival in patients with advanced NSCLC

Significance of serum ferritin as a prognostic factor in advanced hepatobiliary cancer patients treated with Korean medicine: a retrospective cohort study

Abstract Background Advanced hepatobiliary cancers are highly lethal cancers that require precise prediction in clinical practice. Serum ferritin level increases in malignancy and high serum ferritin level is associated with poor survival in various cancers. This study aimed to identify whether serum ferritin could independently predict the overall survival (OS) of patients with advanced hepatobiliary cancers. Methods The retrospective cohort study was performed by reviewing medical records of patients with advanced hepatobiliary cancers from June 2006 to September 2016. The demographic and clinicopathological characteristics as well as the biochemical markers were evaluated at the initiation of Korean medicine (KM) treatment. The OS was calculated using Kaplan-Meier estimates. The Cox proportional hazard model was used to identify the independent prognostic significance of serum ferritin for survival. Results The median OS of all subjects was 5.1 months (range, 0.5–114.9 months). The median OS of group with low ferritin levels and that with high ferritin levels was 7.5 months (range, 0.7–114.9 months) and 2.8 months (range, 0.5–22.8 months), respectively (P < 0.001). The results of the univariate analysis showed that the Eastern Cooperative Oncology Group Performance Status (ECOG-PS) (P = 0.002), tumor type (P = 0.001), prior treatment (P = 0.023), serum ferritin (P < 0.001), hemoglobin (P = 0.002), total bilirubin (P = 0.002), gamma-glutamyl transpeptidase (P = 0.007), albumin (P = 0.013), white blood cell (P = 0.002), and C-reactive protein (CRP) (P < 0.001) were significant factors for the patients’ survival outcome. On multivariate analysis controlling confounding factors, ferritin (P = 0.041), CRP (P = 0.010), ECOG-PS (P = 0.010), and tumor type (P = 0.018) were identified as independent prognostic factors for survival. Conclusions These results indicate that serum ferritin is a valid clinical biochemical marker to predict survival of patients with advanced hepatobiliary cancers