Granulin-epithelin precursor is an oncofetal protein defining hepatic cancer stem cells

Background and Aims

Increasing evidence has suggested that hepatocellular carcinoma (HCC) might originate from a distinct subpopulation called cancer stem cells (CSCs), which are responsible for the limited efficacy of conventional therapies. We have previously demonstrated that granulin-epithelin precursor (GEP), a pluripotent growth factor, is upregulated in HCC but not in the adjacent non-tumor, and that GEP is a potential therapeutic target for HCC. Here, we characterized its expression pattern and stem cell properties in fetal and cancerous livers.

Methods

Protein expression of GEP in fetal and adult livers was examined in human and mouse models by immunohistochemical staining and flow cytometry. Liver cancer cell lines, isolated based on their GEP and/or ATP-dependent binding cassette (ABC) drug transporter ABCB5 expression, were evaluated for hepatic CSC properties in terms of colony formation, chemoresistance and tumorigenicity.

Results

We demonstrated that GEP was a hepatic oncofetal protein that expressed in the fetal livers, but not in the normal adult livers. Importantly, GEP+ fetal liver cells co-expressed the embryonic stem (ES) cell-related signaling molecules including β-catenin, Oct4, Nanog, Sox2 and DLK1, and also hepatic CSC-markers CD133, EpCAM and ABCB5. Phenotypic characterization in HCC clinical specimens and cell lines revealed that GEP+ cancer cells co-expressed these stem cell markers similarly as the GEP+ fetal liver cells. Furthermore, GEP was shown to regulate the expression of ES cell-related signaling molecules β-catenin, Oct4, Nanog, and Sox2. Isolated GEP^high cancer cells showed enhanced colony formation ability and chemoresistance when compared with the GEP_low counterparts. Co-expression of GEP and ABCB5 better defined the CSC populations with enhanced tumorigenic ability in immunocompromised mice.

Conclusions

Our findings demonstrate that GEP is a hepatic oncofetal protein regulating ES cell-related signaling molecules. Co-expression of GEP and ABCB5 further enriches a subpopulation with enhanced CSC properties. The current data provide new insight into the therapeutic strategy.     

Biological growth and spread modeled by systems of recursions. II. Biological theory

The mathematical theory developed in Part I is applied to a selection-migration model in population genetics with sex-linked locus and to the host-vector or venereal disease epidemic model. In both models, a constant c*(xi) is found for each unit vector xi. The mathematical results imply that under certain initial conditions, the frequency of the advantageous gene in the male and female gametic outputs or the epidemic will spread at a speed c*(xi) in the direction xi as time goes to infinity. Time is measured in discrete nonoverlapping generations. In most cases, we can find a formula for c*(xi).     

Identification of plasmin as the major contaminant in immunoglobulin preparations

A proteolytic enzyme could be isolated from rabbit serum by means of DEAE cellulose, Protein A-bound Sepharose and lysine-bound Sepharose chromatographies. This enzyme was found to be the major protease contaminating IgG preparations of rabbit serum. This enzyme was identified as plasmin because it displayed an apparent Mr of 90,000 on nonreduced SDS polyacrylamide gel electrophoresis, was able to directly lyse fibrin and the chromogenic substrate H-D-Val-Leu-Lys-p-nitroanilide, and was stable after heating at 56 degrees for 30 min but broke down at 80 degrees. Its Km toward the chromogenic substrate was 0.35 mM, which agreed well with the published value for plasmin.     

Is There an Optimal Level of Open-Endedness in Prebiotic Evolution?

In this paper we explore the question of whether there is an optimal set up for a putative prebiotic system leading to open-ended evolution (OEE) of the events unfolding within this system. We do so by proposing two key innovations. First, we introduce a new index that measures OEE as a function of the likelihood of events unfolding within a universe given its initial conditions. Next, we apply this index to a variant of the graded autocatalysis replication domain (GARD) model, Segre et al. (P Natl Acad Sci USA 97(8):4112-4117, 2000; Markovitch and Lancet Artif Life 18(3), 2012), and use it to study - under a unified and concise prebiotic evolutionary framework - both a variety of initial conditions of the universe and the OEE of species that evolve from them.     

Quantitative evaluation of motor functional recovery process in chronic stroke patients during robot-assisted wrist training

This study was to investigate the motor functional recovery process in chronic stroke during robot-assisted wrist training. Fifteen subjects with chronic upper extremity paresis after stroke attended a 20-session wrist tracking training using an interactive rehabilitation robot. Electromyographic (EMG) parameters, i.e., EMG activation levels of four muscles: biceps brachii (BIC), triceps brachii (TRI, lateral head), flexor carpiradialis (FCR), and extensor carpiradialis (ECR) and their co-contraction indexes (CI) were used to monitor the neuromuscular changes during the training course. The EMG activation levels of the FCR (11.1% of decrease from the initial), BIC (17.1% of decrease from the initial), and ECR (29.4% of decrease from the initial) muscles decreased significantly during the training (P < 0.05). Such decrease was associated with decreased Modified Ashworth Scores for both the wrist and elbow joints (P < 0.05). Significant decrease (P < 0.05) was also found in CIs of muscle pairs, BIC&TRI (21% of decrease from the initial), FCR&BIC (11.3% of decrease from the initial), ECR&BIC (49.3% of decrease from the initial). The decreased CIs related to the BIC muscle were mainly caused by the reduction in the BIC EMG activation level, suggesting a better isolation of the wrist movements from the elbow motions. The decreased CI of ECR& FCR in the later training sessions (P < 0.05) was due to the reduced co-contraction phase of the antagonist muscle pair in the tracking tasks. Significant improvements (P < 0.05) were also found in motor outcomes related to the shoulder/elbow and wrist/hand scores assessed by the Fugl–Meyer assessment before and after the training. According to the evolution of the EMG parameters along the training course, further motor improvements could be obtained by providing more training sessions, since the decreases of the EMG parameters did not reach a steady state before the end of the training. The results in this study provided an objective and quantitative EMG measure to describe the motor recovery process during poststroke robot-assisted wrist for the further understanding on the neuromuscular mechanism associated with the recovery.