Genetic blockade of insulin-like growth factor-1 receptor via recombinant adenovirus in lung cancer can be enhanced by the histone deacetylase inhibitor, vorinostat.

BACKGROUND: Many approaches have been suggested as anti-tumor therapy for targeting insulin-like growth factor 1 receptor (IGF-1R), such as monoclonal antibodies and tyrosine kinase inhibitor. We introduced recombinant adenoviruses expressing antisense, dominant negative or short hairpin RNA to IGF-1R. Moreover, we demonstrated that histone deacetylase inhibitor (vorinostat) can increase the transduction efficiency of adenoviruses by increasing CAR-induced transduction and by enhancing the transcription of the adenoviral transgene. In the present study, we showed that the combination of ad-sh (short hairpin) IGF-1R with vorinostat leads to a synergistic enhancement of IGF-1R blockade. METHODS: We measured the change in IGF-1R upon cotreatment with vorinostat and ad-shIGF-1R. Changes in transduction efficiency of ad-shIGF-1R were measured by fluorescent microscopy. Changes in apoptotic proportion and cell survival after the cotreatment were measured by the sub-G1 assay and cell counts. The effect of nuclear factor (NF)-κB activation was also measured by NF-κB p65 activation enzyme-linked immunosorbent assay. Drug interactions were analyzed upon cotreatment with ad-shIGF-1R, vorinostat and cisplatin. RESULTS: Combined treatment of ad-shIGF-1R and vorinostat synergistically suppressed the IGF-1R expression in lung cancer cell lines and also increased the transduction efficiency of ad-shIGF-1R. Ad-shIGF-1R and vorinostat cotreatment increased apoptotic cell death and synergistically suppressed cell growth compared to ad-shIGF-1R or vorinostat treatment alone. Vorinostat suppressed NF-κB activation, which was activated by ad-shIGF-1R. Moreover, triple combination of ad-shIGF-1R, vorinostat and cisplatin demonstrated synergistic cytotoxicity on lung cancer cells. CONCLUSIONS: Vorinostat enhanced the blocking capability of ad-shIGF-1R. The combined treatment of vorinostat and ad-sh-IGF-1R appears to have promising potential as a new therapeutic approach for lung cancer.

The association of down-regulated toll-like receptor 4 expression with airflow limitation and emphysema in smokers.

BACKGROUND: An association between innate immunity including Toll-like receptors (TLRs) and COPD is reported recently; TLR4 deficiency in lung can cause emphysema in animals, which is not evident in humans. We analyzed the association of TLR4 expression, airflow limitation and emphysema in smokers. METHODS: We enrolled patients of ≥40years old with smoking histories of ≥10 pack-years and who had undergone lung resection. We measured TLR4 expression in lung lysates. The severity of emphysema was evaluated on computed tomography. TLR4 expression was also evaluated immunohistochemically. RESULTS: In total, 53 patients were enrolled. Forced expiratory volume in one second per forced vital capacity (FEV1/FVC) increased (P=0.03) and emphysema score decreased (P=0.01) as TLR4 expression increased. These were still significant, in multiple regression analysis including sex, age, tuberculosis history, smoking history and inhaled corticosteroid (ICS) usage. We also classified patients as high, intermediate, and low expressers according to TLR4 expression. Although no differences in age, gender, tuberculosis, or smoking history were observed among the groups, emphysema severity increased significantly (P = 0.02) and FEV1/FVC decreased significantly (P = 0.006) in TLR4 low expresser. The difference in TLR4 expression based on immunohistochemistry was most prominent in bronchial and alveolar epithelial cells. CONCLUSION: Down-regulated TLR4 expression in lung was associated with emphysema and airflow limitation in smokers.

Combination therapy of conditionally replicating adenovirus and histone deacetylase inhibitors.

Combination therapy of adenoviral gene therapy and a histone deacetylase (HDAC) inhibitor is important due to the enhancing effect of HDAC inhibitors on adenoviral transduction and transcription. However, contradictory results have been reported on the effect of combination of CRAd (conditionally replicating adenovirus) and HDAC inhibitors. This study was designed to investigate the interaction of CRAd and HDAC inhibitors and determine the ideal way to combine the two agents. Combination of HDAC inhibitors (SK7041, SBHA and vorinostat) at pre- and post-transductional periods with CRAd enhanced the transduction of CRAd and expression of luciferase expression from Δ24-luc in vitro. However, suppression of luciferase expression from Δ24-luc injected tumor mass was observed by in vivo tumor bioluminescence imaging and drug interaction analysis also showed an antagonistic interaction that was probably related with the inhibitory effect of the HDAC inhibitor on adenoviral replication. Suppression of p21 induction by p21 siRNA reversed the suppressive effect of vorinostat on the replication of CRAd, but still failed to reverse the antagonistic interaction. Addition of vorinostat at the pre-transductional period revealed an improvement in the transduction efficiency of CRAd and also induced a synergistic interaction between CRAd and vorinostat, which was possibly related with prevention of the suppressive effect of vorinostat on adenoviral replication. In conclusion, the addition of HDAC inhibitor before CRAd injection showed synergistic antitumor effects, which warrants further investigation on the sequence of HDAC inhibitor and CRAd treatment in an animal tumor model.

Combination gene therapy of lung cancer with conditionally replicating adenovirus and adenovirus-herpes simplex virus thymidine kinase.

A major obstacle to the success of gene therapy strategies that directly target cancer cells is the low gene transfer rate. To address this problem, we had previously proposed a combination adenoviral gene therapy containing a conditionally replicating adenovirus (CRAD) expressing mutant E1 (Delta24RGD), and a replication-defective E1-deleted adenovirus to enhance the efficiency of gene transfer. Suicide/pro-drug gene therapy has an important additional benefit to the therapy of cancer. This relates to the transfer and expression of non-mammalian genes encoding enzymes that convert non-toxic pro-drugs into cellular toxins. We investigated the interaction between CRAD (Delta24RGD) and a replication-defective E1-deleted adenovirus (ad-HSTK) containing a suicide gene (HSTK: herpes simplex virus thymidine kinase gene) with respect to therapeutic gene production and tumor cell killing efficacy. Combined transduction of CRAD and ad-HSTK increased the transduction efficiency of HSTK and increased its sensitivity to ganciclovir (GCV) more efficiently than ad-HSTK alone. Transfer of medium of CRAD and ad-HSTK co-transduced cells induced the transfer of HSTK (media transferable bystander effect), and enhanced its sensitivity to GCV. In an animal tumor model, combined intratumoral injection of CRAD and ad-HSTK followed by GCV administration induced prolonged expression of HSTK and stronger growth suppression of established lung cancer xenografts than single injections. These data demonstrate that the selective replication of ad-HSTK due to the presence of mutant E1, produced by a Delta24RGD and HSTK/GCV suicide gene system, resulted in a striking improvement in anti-tumor effects in vitro and in vivo.

[King Jung-jo's medical philosophy].

King Jungjo who introduced the advent of cultural renaissance of Chosun Dynasty as little been known about his work in medicine. With a wide knowledge in medicine, he was the only one among the kings who wrote a book on medicine, called "SueMinMyoJeon". In this paper, his perspective on medicine will be looked into based on "The Annals of the Chosun Dynasty", "Seungjeongwon Ilgi", "Hong Je jun Se", "KukGoBoGam", "Ildkrok", "JeJungShinPyun", "SueMinMyoJeon" etc. King Jungo valued empiricism in the field of medicine. He deepened understandings in medicine while taking care of King Youngjo, the late king. And it led him to author "SueMinMyoJeon" himself, and further ordered the publications of "JeJungShinPyun" "MaGuaHeoiTong". These two books were conducted to include empirical cases of folklore remedy. King Jungjo's medical philosophy can be epitomized in filial piety and realization of people-serving politics, which are the essentials of Confucianism. His filial piety towards the late king, Youngjo and his mother is shown in his devotion when taking care of them. Especially the way he examined the differentiation of diseases and corresponding treatments is well described in "The Annals of the Chosun Dynasty". "JeJungShinPyun" was also published and it came handy for folk villagers in times of medical needs. Later this book influenced "BangYakHaepPyun" by Hwang Do Yeon. King Jungjo emphasized pragmatism in spreading medical knowledges, thus removing the theoretical contents that are related to Taoism, especially the ones on alchemy from "DongEuiBoGam", when publishing "SueMinMyoJeon". Even the excerpts from "SoMun" were taken out, if not practical. King Jungjo, however, discussed the importance of healthy regimen and mentioned himself practicing it from the book "IlDeukLok", which seems to be the only book that derailed from the pragmatistic track. King Jungjo put emphasis on consistency between diagnosis and treatment. In diagnosing, Meridian pulse was taken important as a means of finding the origin of disease, while deploring how doctors then neglected to study.

Blockade of NF-κB activation by IκBα gene therapy enhances radiation sensitivity and abolishes acquired resistance to radiation.

Radiation is one of the main treatment modalities in lung cancer, but low sensitivity and acquired resistance of lung cancer cells to radiation frequently result in treatment failure. The activation of nuclear factor (NF)-κB is reportedly the main mechanism by which cancer cells exhibit resistance to external stresses, such as chemotherapy or radiation therapy. In this study, we blocked the activation of NF-κB by adenovirus-expressing IκBα-SR and investigated the effect this had on radiation sensitivity. Transduction with ad-IκBα effectively blocked the activation of NF-κB by radiation in all the cancer cell lines tested, except for NCI H460. Clonogenic assay after radiation demonstrated that ad-IκBα transduction enhanced the sensitivity to radiation of the lung cancer cell lines and HeLa cells, except for NCI H460. The radiosensitizing effect of IκBα was more potent in lung cancer cell lines with radioresistance (SKMESres) (sensitizer enhancement ratio 1:61). From these findings, NF-κB blockade by ad-IκBα enhanced the radiation sensitivity and also abolished the acquired radiation resistance of lung cancer cell lines. This study provides a therapeutic rationale for combining an NF-κB-blocking strategy with radiation to both increase sensitivity and overcome acquired resistance to radiation.

The development of the friction coefficient inspection equipment for skin using a load cell.

The skin is an indispensible organ for human because it contributes to the metabolism using its own biochemical functions as well as it protects the human body from the exterior stimuli. Recently, the friction coefficient have been used as the decision index of the progress for the bacterial aliments in the field of the skin physiology and the importance of friction coefficient have been increased in the skin care market because of the needs of the well being times. In addition, the usage of friction coefficient is known to have the big discrimination ability in classification of human constitutions, which is utilized in the alternative medicine. In this study, we designed a system which used the multi axes load cell and hemi-circular probe and tried to measure the friction coefficient of hand skins repeatedly. Using this system, the relative repeatability error for the measurement of the friction coefficient was below 4 %. The coefficient is not concerned in curvatures of tips. Using this system, we will try to establish the standard for classification of constitutions.