Mutation-specific non-canonical pathway of PTEN as a distinct therapeutic target for glioblastoma.

PTEN is one of the most frequently altered tumor suppressor genes in malignant tumors. The dominant-negative effect of PTEN alteration suggests that the aberrant function of PTEN mutation might be more disastrous than deletion, the most frequent genomic event in glioblastoma (GBM). This study aimed to understand the functional properties of various PTEN missense mutations and to investigate their clinical relevance. The genomic landscape of PTEN alteration was analyzed using the Samsung Medical Center GBM cohort and validated via The Cancer Genome Atlas dataset. Several hotspot mutations were identified, and their subcellular distributions and phenotypes were evaluated. We established a library of cancer cell lines that overexpress these mutant proteins using the U87MG and patient-derived cell models lacking functional PTEN. PTEN mutations were categorized into two major subsets: missense mutations in the phosphatase domain and truncal mutations in the C2 domain. We determined the subcellular compartmentalization of four mutant proteins (H93Y, C124S, R130Q, and R173C) from the former group and found that they had distinct localizations; those associated with invasive phenotypes ('edge mutations') localized to the cell periphery, while the R173C mutant localized to the nucleus. Invasive phenotypes derived from edge substitutions were unaffected by an anti-PI3K/Akt agent but were disrupted by microtubule inhibitors. PTEN mutations exhibit distinct functional properties regarding their subcellular localization. Further, some missense mutations ('edge mutations') in the phosphatase domain caused enhanced invasiveness associated with dysfunctional cytoskeletal assembly, thus suggesting it to be a potent therapeutic target.

Topical application of autophagy-activating peptide improved skin barrier function and reduced acne symptoms in acne-prone skin.

BACKGROUND: Recent studies about the important roles of autophagy signaling in sebaceous lipogenesis and epidermal differentiation suggest potential benefits of autophagy activation in acne. AIMS: To investigate the effects of an autophagy activator on acne-prone skin. METHODS: Autophagy signaling in human immortalized SZ95 sebocytes, normal human epidermal keratinocytes, and 3D reconstituted skin was examined. Effects of an autophagy-activating peptide on sebaceous lipogenesis were measured by fluorescence microscopic analysis. The clinical efficacy in acne-prone skin was evaluated through an eight-week, double-blind, randomized, vehicle-controlled study. Changes in skin surface lipid compositions were further analyzed. RESULTS: In cultured sebocytes and keratinocytes, the investigated autophagy-activating peptide increased LC3-II expression, indicating a stimulation of autophagy signaling. Testosterone and linoleic acid treatment induced lipogenesis in cultured sebocytes and is further inhibited by the autophagy activator peptide treatment. Increased expression of differentiation marker proteins in cultured keratinocytes was also observed by autophagy-activating peptide. In clinical study, reduction of closed comedones and the amount of skin surface lipids as well as of trans-epidermal water loss (TEWL) were observed in acne-prone skin after autophagy-activating peptide application. In addition, reduction of squalene and increase in cholesterol were observed after an 8-week application. CONCLUSIONS: Topical application of an autophagy activator downregulated sebaceous lipogenesis and improved the skin barrier function. Considering the important roles of sebum and skin barrier function in acne pathogenesis, autophagy activation might represent a new therapeutic option in early forms of acne.

Sphere-Forming Culture for Expanding Genetically Distinct Patient-Derived Glioma Stem Cells by Cellular Growth Rate Screening.

Diffusely infiltrating gliomas (DIGs) are difficult to completely resect and are associated with a high rate of tumor relapse and progression from low- to high-grade glioma. In particular, optimized short-term culture-enriching patient-derived glioma stem cells (GSCs) are essential for customizing the therapeutic strategy based on clinically feasible in vitro drug screening for a wide range of DIGs, owing to the high inter-tumoral heterogeneity. Herein, we constructed a novel high-throughput culture condition screening platform called 'GFSCAN', which evaluated the cellular growth rates of GSCs for each DIG sample in 132 serum-free combinations, using 13 previously reported growth factors closely associated with glioma aggressiveness. In total, 72 patient-derived GSCs with available genomic profiles were tested in GFSCAN to explore the association between cellular growth rates in specific growth factor combinations and genomic/molecular backgrounds, including isocitrate dehydrogenase 1 (IDH1) mutation, chromosome arm 1p and 19q co-deletion, ATRX chromatin remodeler alteration, and transcriptional subtype. GSCs were clustered according to the dependency on epidermal growth factor and basic fibroblast growth factor (E&F), and isocitrate dehydrogenase 1 (IDH1) wild-type GSCs showed higher E&F dependencies than IDH1 mutant GSCs. More importantly, we elucidated optimal combinations for IDH1 mutant glioblastoma and lower grade glioma GSCs with low dependencies on E&F, which could be an aid in clinical decision-making for these DIGs. Thus, we demonstrated the utility of GFSCAN in personalizing in vitro cultivation to nominate personalized therapeutic options, in a clinically relevant time frame, for individual DIG patients, where standard clinical options have been exhausted.

Anti-Wrinkle Benefits of Peptides Complex Stimulating Skin Basement Membrane Proteins Expression.

The dermal-epidermal junction (DEJ) provides a physical and biological interface between the epidermis and the dermis. In addition to providing a structural integrity, the DEJ also acts as a passageway for molecular transport. Based on the recently reported importance of the DEJ in skin aging, novel peptide derivatives have been tested for their effects on basement membrane (BM) protein expressions in cultured human epidermal keratinocytes. As a result, protein expressions of collagen XVII, laminin and nidogen were stimulated by the test peptide and peptides complex. Further ex vivo evaluation using excised human skin, confirmed that the topical application of the peptides complex significantly increased dermal collagen expression, as well as expressions of collagen XVII and laminin. Interestingly, while the origin of the laminin protein is epidermal keratinocytes, the immunohistochemical staining of skin showed that laminin was only detected in the uppermost layer of the dermis, which suggests a tight assembly of laminin protein onto the dermal side of the DEJ. These results suggest that a peptide complex could improve the structural properties of the DEJ through its ability to stimulate BM proteins. In order to evaluate the anti-wrinkle benefits of the peptide complex in vivo, a clinical study was performed on 22 healthy Asian female volunteers older than 40 years. As a result, significant improvements in skin wrinkles for all of the five sites were observed after two weeks, as assessed by skin topographic measurements. Collectively, these results demonstrate the anti-aging efficacy of the peptides complex.

PIP4K2A as a negative regulator of PI3K in PTEN-deficient glioblastoma.

Glioblastoma (GBM) is the most malignant brain tumor with profound genomic alterations. Tumor suppressor genes regulate multiple signaling networks that restrict cellular proliferation and present barriers to malignant transformation. While bona fide tumor suppressors such as PTEN and TP53 often undergo inactivation due to mutations, there are several genes for which genomic deletion is the primary route for tumor progression. To functionally identify putative tumor suppressors in GBM, we employed in vivo RNAi screening using patient-derived xenograft models. Here, we identified PIP4K2A, whose functional role and clinical relevance remain unexplored in GBM. We discovered that PIP4K2A negatively regulates phosphoinositide 3-kinase (PI3K) signaling via p85/p110 component degradation in PTEN-deficient GBMs and specifically targets p85 for proteasome-mediated degradation. Overexpression of PIP4K2A suppressed cellular and clonogenic growth in vitro and impeded tumor growth in vivo. Our results unravel a novel tumor-suppressive role of PIP4K2A for the first time and support the feasibility of combining oncogenomics with in vivo RNAi screen.

FoxM1 Promotes Stemness and Radio-Resistance of Glioblastoma by Regulating the Master Stem Cell Regulator Sox2.

Glioblastoma (GBM) is the most aggressive and most lethal brain tumor. As current standard therapy consisting of surgery and chemo-irradiation provides limited benefit for GBM patients, novel therapeutic options are urgently required. Forkhead box M1 (FoxM1) transcription factor is an oncogenic regulator that promotes the proliferation, survival, and treatment resistance of various human cancers. The roles of FoxM1 in GBM remain incompletely understood, due in part to pleotropic nature of the FoxM1 pathway. Here, we show the roles of FoxM1 in GBM stem cell maintenance and radioresistance. ShRNA-mediated FoxM1 inhibition significantly impeded clonogenic growth and survival of patient-derived primary GBM cells with marked downregulation of Sox2, a master regulator of stem cell phenotype. Ectopic expression of Sox2 partially rescued FoxM1 inhibition-mediated effects. Conversely, FoxM1 overexpression upregulated Sox2 expression and promoted clonogenic growth of GBM cells. These data, with a direct binding of FoxM1 in the Sox2 promoter region in GBM cells, suggest that FoxM1 regulates stemness of primary GBM cells via Sox2. We also found significant increases in FoxM1 and Sox2 expression in GBM cells after irradiation both in vitro and in vivo orthotopic tumor models. Notably, genetic or a small-molecule FoxM1 inhibitor-mediated FoxM1 targeting significantly sensitized GBM cells to irradiation, accompanying with Sox2 downregulation. Finally, FoxM1 inhibition combined with irradiation in a patient GBM-derived orthotopic model significantly impeded tumor growth and prolonged the survival of tumor bearing mice. Taken together, these results indicate that the FoxM1-Sox2 signaling axis promotes clonogenic growth and radiation resistance of GBM, and suggest that FoxM1 targeting combined with irradiation is a potentially effective therapeutic approach for GBM.

Effects of Indoor Rowing Exercise on the Body Composition and the Scoliosis of Visually Impaired People: A Preliminary Study.

OBJECTIVE: To evaluate the effects of rowing exercise on body composition, laboratory data, fitness and scoliosis in visually impaired people. The majority of visually impaired people do not participate in active sports due to efficiency and safety issues. Rowing is a safe whole-body exercise with aerobic and anaerobic components. METHODS: Twenty subjects were recruited from among those admitted to a facility for visually impaired people (16 men and 4 women). Laboratory data, body composition, physical fitness, Cobb's angle, and fall index were checked before and after 6 weeks (5 days a week) of indoor rowing using Concept2 Model E. RESULTS: After the training, fat mass and total body fat percent decreased significantly. In the fitness test, back strength and trunk flexion score increased significantly. Laboratory data showed significant increases in serum protein and albumin and decreases in low-density lipoprotein (LDL) cholesterol. There were 9 subjects with scoliosis and after the training Cobb's angle decreased by 1.11°±1.55°, though this was not statistically significant. CONCLUSION: Visually impaired people frequently have abnormal body composition, low physical fitness, and scoliosis. A rowing exercise program can be helpful, with a positive effect on body composition and physical fitness; however, with respect to scoliosis, we need an earlier intervention program in visually impaired people.

Selectively impaired fasciculus gracilis in cervical spinal cord injury.

Selective lesions of the fasciculus gracilis have been reported only in cases of nontraumatic spinal cord disease. We present the case of a 54-year-old man who developed persistent hypesthesia and abnormal vibratory sensation below the T6 segmental level after injuring his cervical spine after a fall. Cervical magnetic resonance imaging (MRI) revealed ossification of the posterior longitudinal ligament, spinal stenosis, and a C3-4 spinal cord injury. A thoracic MRI did not reveal a spinal cord lesion. Lower thoracic dermatomal somatosensory evoked potentials showed delayed latency. The findings in this case indicate selective injury to the fasciculus gracilis at the level of the cervical spinal cord.

Maternal Psychosocial Factors that Affect Breastfeeding Adaptation and Immune Substances in Human Milk.

PURPOSE: This study was to identify relationships of maternal psychosocial factors including mother's mood state, childcare stress, social support and sleep satisfaction with breastfeeding adaptation and immune substances in breast milk, especially secretory immunoglobulin A (sIgA) and transforming growth factor-beta 2 (TGF-beta2). METHODS: Data were collected from 84 mothers who delivered full-term infants by natural childbirth. Structured questionnaires and breast milk were collected at 2~4 days and 6 weeks postpartum. Data were analyzed using descriptive statistics, Pearson's correlation, multiple linear regression, and generalized estimating equation (GEE). RESULTS: Scores for the breastfeeding adaptation scale were significantly related with child care stress, mood state and social support. Mother's anger was positively correlated with the level of sIgA in colostrum (p<.01). Immune substances of breastmilk was significantly influenced by time for milk collection (p<.001) and the type of breastfeeding (sIgA, p<.001, TGF-beta2, p=.003). Regression analysis showed that breastfeeding adaptation could be explained 59.1% by the type of breastfeeding, childcare stress, the Profile of Mood States, emotional support and sleep quality (F=16.67, p<.001). CONCLUSION: The findings from this study provide important concepts of breastfeeding adaptation program and explanation of psychosocial factors by immune substances in breast milk. Future research, specially, bio-maker research on breast milk should focus on the ways to improve breastfeeding adaptation.

Effects of the off-loading brace on the activation of femoral muscles -a preliminary study-.

OBJECTIVE: To provide the off-loading knee brace was designed relief for the pain associated with osteoarthritis by reduce loads on the degenerative compartment of the knee. This study examined the effects of the off-loading knee brace on activation of femoral muscles during squatting, slow and fast walking exercise in healthy young individuals. METHOD: Ten healthy male subjects without a history of knee pain were recruited. Each subject was asked to do squatting, slow and fast walking exercises with a brace secured to the dominant leg. The same exercises were repeated without the brace. Surface electromyographic (sEMG) data was collected from the vastus medialis oblique (VMO), vastus lateralis (VL) and biceps femoris (BF) muscles from the dominant side of the leg. All dynamic root mean squre (RMS) values of sEMG were standardized to static RMS values of the maximal isometric contraction and expressed as a percentage of maximal activity. RESULTS: We found that VMO activity was significantly decreased with application of the off-loading knee brace during squatting and fast walking exercise. However there were no significant differences in VMO activity with application of the off-loading knee brace during slow walking exercise. CONCLUSION: These results suggest that the external moment of the brace which effectively stabilized the patella in the movement in which the knee joints become relatively unstable. The brace could be useful in the short term, but for long-term use, weakening of the VMO is predicted. Therefore the program of selective muscular strength strengthening for the VMO should be emphasized.

[Analysis of willingness-to-quit cigarette price among Korean male adults].

OBJECTIVES: The purpose of this study was to estimate the willingness to quit cigarette price among Korean male adults, and to examine he factors affecting the willingness to quit cigarette price. METHODS: The data was collected by a random digit dial telephone survey. 702 samples were analyzed by using t-tests, ANOVA and OLS regression analysis. To estimate the willingness to quit cigarette price, smokers were asked dichotomous questions with open-ended follow-up and the starting point of the price was randomized by one of 5 bid prices elicited from a pilot study. RESULTS: The mean of the willingness to quit cigarette price was 4,287 Won per package, which was about 2,000 Won higher than the mean of the actual price the smokers now paid. About 41% of respondents were willing to quit smoking if the price of cigarette would be increased by 3,000 Won, and if the price would be increased by 20,000 Won, all respondents were willing to quit smoking. The factors associated with the willingness to quit cigarette price were the place of residence, the amount of smoking and the degree of exposure to smoking through the mass media. CONCLUSIONS: The results showed that to get people to quit smoking, increasing the cigarette price would obviously be effective and much higher prices have a greater effect. Furthermore, to enlarge the effect of increased cigarette prices, providing more cessation programs to small towns, reducing the amount of smoking and decreasing or prohibiting advertisements of cigarettes and smoking in the mass media will be efficient.

[The effect of cigarette price on smoking behavior in Korea].

OBJECTIVES: To determine the impact of cigarette prices on the decision to initiate and quit smoking by taking into account the interdependence of smoking and other behavioral risk factors. METHODS: The study population consisted of 3,000 male Koreans aged > or =20. A survey by telephone interview was undertaken to collect information on cigarette price, smoking and other behavioral risk factors. A two-part model was used to examine separately the effect of price on the decision to be a smoker, and on the amount of cigarettes smoked. RESULTS: The overall price elasticity of cigarettes was estimated at -0.66, with a price elasticity of -0.02 for smoking participation and -0.64 for the amount of cigarettes consumed by smokers. The inclusion of other behavioral risk factors reduced the estimated price elasticity for smoking participation substantially, but had no effect on the conditional price elasticity for the quantity of cigarettes smoked. CONCLUSIONS: From the public health and financial perspectives, an increase in cigarette price would significantly reduce smoking prevalence as well as cigarette consumption by smokers in Korea.