E.B. Lee

Efficient Gene Delivery with Osmotically Active and Hyperbranched Poly(ester amine)s

Degradable and hyperbranched poly (ester amine)s (PEAs) were successfully synthesized by Michael addition reaction between hydrophilic glycerol triacrylate (GTA) and low-molecular-weight polyethylenimine (LMW-PEI) and evaluated as nonviral gene carriers. PEAs effectively condensed DNA with particle sizes below 200 nm and suitable surface charges (15-45 mV), suitable for intracellular delivery. PEAs degraded in a controlled fashion showing half-lives of more than 12 days and were essentially nontoxic in three different cell lines. Elevated transfection levels by luciferase assay revealed the superiority of PEAs over PEI 25K and Lipofectamine. PEAs synthesized using 1:4 mol ratio of GTA to PEI [GTA/PEI-1.2(1:4)] showed highest transfection efficiency in HepG2 cells. PEAs showed significant gene expression in vitro as well as in vivo through aerosol administration. Reduction in packed cell volume (PCV) of cells when treated with polyplexes supported the hyperosmotic effect of PEAs. Effect of bafilomycin A1 on transfection efficiency of PEAs on 293T cells indicated its endosomal buffering capacity. High transfection efficiency was attributed to the synergism from hyperosmotic glycerol backbone in the PEAs and endosomal buffering capacity of PEI amine groups. Therefore, this convergence of osmotically active biodegradable PEAs suggests their potential as a safe and efficient gene delivery vector.

Inhaled Fluorescent Magnetic Nanoparticles Induced Extramedullary Hematopoiesis in the Spleen of Mice

Inhaled Fluorescent Magnetic Nanoparticles Induced Extramedullary Hematopoiesis in the Spleen of Mice: Jung‐Taek Kwon, et al. Laboratory of Toxicology, College of Veterinary Medicine, Seoul National University, Korea

miRNA expression analysis in cortical dysplasia: Regulation of mTOR and LIS1 pathway

Cortical dysplasia (CD) is a common cause of epilepsy in children and is characterized by focal regions of malformed cerebral cortex. The pathogenesis and epileptogenesis of CD have not been fully elucidated, and in particular, the potential role of epigenetics has not been examined. miRNA microarray was performed on surgical specimens from CD (n=8) and normal control (n=2) children. A total of 10 differentially expressed miRNAs (DEmiRs) that were up-regulated in CD were identified including hsa-miR-21 and hsa-miR-155. The microarray results were validated using quantitative real-time PCR. After searching for the putative target genes of the DEmiRs, their biological significance was further evaluated by exploring the pathways in which the genes were enriched. The mammalian target of rapamycin (mTOR) signaling pathway was the most significantly associated, and the pathway of lissencephaly gene in neuronal migration and development was also noted. This study suggests a possible role for miRNAs in the pathogenesis of CD, especially in relation to the mTOR signaling pathway. Future studies on the epigenetic mechanisms underlying CD pathogenesis and epileptogenesis are needed.

Repeated Aerosol Delivery of Carboxyl-terminal Modulator Protein Suppresses Tumor in the Lungs of K-rasLA1 Mice

RATIONALEnDifficulties in achieving long-term survival of patients with lung cancer treated with conventional therapies suggest that novel approaches are required. Recent advances in aerosol-mediated gene delivery have provided the possibility of an alternative for the safe and effective treatment of lung cancer.nnnOBJECTIVESnTo investigate the repeated effect of carboxyl-terminal modulator protein (CTMP) on multistage lung tumorigenesis. In this study, we addressed this question by studying the effects of lentivirus-based CTMP in the lungs of 9- and 13-week-old K-ras(LA1) mice, a model of lung cancer.nnnMETHODSnAn aerosol of lentivirus-based CTMP was delivered into 9- and 13-week-old K-ras(LA1) mice, a model of lung cancer, through a nose-only inhalation system twice a week for 4 weeks. The effects of CTMP on lung cancer progression and Akt-related signals were evaluated.nnnMEASUREMENTS AND MAIN RESULTSnLong-term repeated delivery of CTMP effectively reduced tumor progression in the lungs at different stages of development. Lentiviral-CTMP inhibited protein synthesis and cell cycle and altered Akt signaling pathway in the lungs of 9-week-old K-ras(LA1) mice, and increased apoptosis was observed in the lungs of 13-week-old K-ras(LA1) mice.nnnCONCLUSIONSnLong-term repeated viral delivery of CTMP may provide a useful tool for designing lung tumor treatment.

THU0179 Tofacitinib, an Oral Janus Kinase Inhibitor, for the Treatment of Rheumatoid Arthritis: Safety and Efficacy in Open-label, Long-term Extension up to 6 Years

Background Tofacitinib is an oral Janus kinase inhibitor for the treatment of rheumatoid arthritis (RA). Objectives To report tofacitinib safety, tolerability, and durability of response up to 72 months (mo) in long-term extension (LTE) studies. Methods Data were from 2, open-label studies: A3921024 (NCT00413699 [ongoing; database unlocked as of April 2014 data cut-off]) and A3921041 (NCT00661661). Patients (pts) had RA and participated in randomised Phases (P)1/2/3 tofacitinib studies. Treatment was initiated with tofacitinib 5 or 10 mg BID as monotherapy or with background DMARDs; data for both doses ± background DMARDs were pooled. Primary endpoints: AEs and laboratory safety. Confirmed data are reported for decreased haemoglobin (HgB), neutrophil, and lymphocyte counts, and increases >50% from baseline (BL) in creatinine. Secondary endpoints: ACR responses, DAS28-4(ESR), and HAQ-DI. Safety data were included over 84 mo and efficacy up to Mo 72 (n≤29 pts, post-Mo 72). Results 4858 pts were treated (mean [max] duration: 918 [2535] days). BL data were from index studies for 91% of pts. Total tofacitinib exposure was 12 359 pt-years (py). In total, 1747 pts (36.0%) discontinued (AEs: 882 [18.2%]; insufficient clinical response: 133 [2.7%]). Most common classes of AEs: infections and infestations (63.4%), musculoskeletal/connective tissue disorders (33.9%), and GI disorders (29.9%). Most frequently reported AEs: nasopharyngitis (16.3%), upper respiratory tract infection (14.5%), and urinary tract infection (10.3%). SAEs occurred in 23.0% of pts (incidence rate [IR] 9.9/100 py [95% confidence interval [CI]; 9.4, 10.5]) and serious infections in 7.2% (IR 2.9/100 py [95% CI; 2.6, 3.2]). Malignancies (excluding NMSC) were reported in 2.5% of pts (IR 1.0/100 py [95% CI; 0.8, 1.2]). IRs for SAEs, serious infections, and malignancies up to Mo 84 did not increase vs previously reported data (Mo 72).1 Decreased Hgb (>2g/dL change from BL or Hgb <8 g/dL) occurred in 6.1% of pts and increased aminotransferases (>3× ULN) in 1.6% (ALT) and <1.0% (AST) of pts. Moderate to severe neutropenia (absolute neutrophil count [ANC] 0.5–1.5×103/mm3) was reported in 1.3% of pts. No pts had ANC <0.5×103/mm3. Absolute lymphocyte counts <0.5×103/mm3 were reported in 1.1% of pts. Increases >50% from BL in creatinine occurred in 3.1% of pts. ACR20, ACR50 and ACR70 response rates for tofacitinib were sustained to Mo 72 (80.8%, 61.5% and 35.9%). Mean DAS28-4(ESR) was 6.29 at BL, 3.74 at LTE Mo 1 and 3.32 at Mo 72. Mean HAQ-DI score was 1.42 at BL, 0.81 at LTE Mo 1 and 0.77 at Mo 72. Conclusions A consistent safety profile and sustained efficacy up to 72 mo was observed in pts with RA receiving tofacitinib 5 or 10 mg BID in LTE studies. References Wollenhaupt J et al. J Rheumatol 2014; 41: 837-852 Acknowledgements Previously presented (Wollenhaupt J et al. Arthritis Rheum 2014; 66 (11): S375 abs 849) and reproduced with permission from Arthritis and Rheumatism. All aspects of this study were funded by Pfizer Inc. Editorial support was provided by Claire Cridland of Complete Medical Communications and funded by Pfizer Inc. Disclosure of Interest J. Wollenhaupt Consultant for: Pfizer Inc, Speakers bureau: Pfizer Inc (speaker fees), J. Silverfield Grant/research support from: Pfizer Inc, Speakers bureau: Pfizer Inc (speaker fees), E. B. Lee Consultant for: Pfizer Inc, S. P. Wood Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, K. Terry Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, H. Nakamura Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, K. Kwok Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, A. Anisfeld Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, C. Nduaka Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, L. Wang Shareholder of: Pfizer Inc, Employee of: Pfizer Inc

Suppression of A549 lung cancer cell migration by precursor let-7g microRNA.

Let-7g miRNAs, short non-coding RNAs approximately 21 nucleotides long, repress protein translation by binding to the 3UTR of target mRNAs. Aberrant expression of let-7g is associated with the poor prognosis of lung cancer patients. Compared to normal lung cells, let-7g expression is absent in non-small cell lung cancer (NSCLC) cells. Furthermore, K-Ras and HMGA2 are well known as targets of let-7g. In this study, we evaluated the potential role of precursor (pre)-let-7g in lung cancer cell metastasis, focusing on the two targets of let-7g, HMGA2 and K-Ras. We found that pre-let-7g inhibited the migration of A549 lung cancer cells through HMGA2-mediated E2F1 down-regulation. Thus, our results suggest that pre-let-7g could be used as a suitable target for the suppression of lung cancer cell migration.

High dietary inorganic phosphate enhances cap-dependent protein translation, cell-cycle progression, and angiogenesis in the livers of young mice

Inorganic phosphate (P(i)) plays a key role in diverse physiological functions. Recent studies have indicated that P(i) affects Akt signaling through the sodium-dependent phosphate cotransporter. Akt signaling, in turn, plays an important role in liver development; however, the effects of high dietary P(i) on the liver have not been investigated. Here, we examined the effects of high dietary phosphate on the liver in developing mice. We found that high dietary P(i) increased liver mass through enhancing Akt-related cap-dependent protein translation, cell cycle progression, and angiogenesis. Thus careful regulation of P(i) consumption may be important in maintaining normal development of the liver.

THU0131 Tofacitinib (CP-690,550), an oral janus kinase inhibitor: Analysis of malignancies across the rheumatoid arthritis clinical programme

Background Tofacitinib (CP-690,550) is a novel, oral Janus kinase inhibitor that is being investigated as a targeted immunomodulator and disease-modifying therapy for rheumatoid arthritis (RA). Objectives To evaluate the malignancies that occurred in the tofacitinib RA programme from the Phase (P) 2, 3, and long-term extension (LTE) studies up to 29 March 2011. Methods Data were pooled from 6 randomized P2, 5 randomized P3 studies and 2 open-label LTE studies. Patients (pts) in P3 and LTE studies were treated with tofacitinib 5 or 10 mg twice daily. Analyses included malignancy data from 1608 pts in P2, 3315 pts in P3, and 3227 pts in LTE studies (LTE pts rolled over from the P2 and P3 studies). Results A total of 4789 patients (5651 pt-yr) received tofacitinib in the P2, P3 and LTE studies. Fifty pts receiving tofacitinib (all doses) reported malignancies (excluding non-melanoma skin cancer [NMSC]); the most common were lung (12 cases) and breast cancer (9 cases). There were 3 lymphoma cases. The overall incidence rate (IR, events per 100 pt-yr) for all malignancies (excluding NMSC) was 0.89 (95% confidence interval [CI]: 0.67,1.17). The IRs (95% CI) of all malignancies (excluding NMSC) broken down into 0-6, 6-12, 12-18, 18-24 and >24 months based on exposure to study drug were 0.75 (0.46,1.23), 0.73 (0.41,1.28), 0.97 (0.48,1.94), 1.28 (0.53,3.08), and 1.37 (0.71,2.63), respectively. The number of cases in each time interval was small, with resultant wide CIs. The standardised incidence ratio (SIR) (95% CI) (as compared with the Surveillance Epidemiology and End Result database covering the general population) for all malignancies (excluding NMSC), lung, breast cancer and lymphomas in the tofacitinib group were 1.11 (0.82-1.47), 2.16 (1.12,3.77), 0.82 (0.38,1.56) and 1.74 (0.36,5.10), respectively. Twenty-one pts experienced NMSCs, for an IR of 0.37 (95% CI: 0.24,0.57). By comparison, the IR of NMSC in patients treated with anti-TNF was 0.47 (0.37-0.59) in a meta-analysis of randomized controlled trials and ranged from 0.23 to 0.35 in a meta-analysis of registries.1,2 Conclusions The malignancies that occurred in the tofacitinib RA programme are consistent with the type and distribution of malignancies expected for patients with moderate to severe RA. The IRs for all malignancies (excluding NMSC), lung cancer, breast cancer and lymphomas are consistent with published estimates in RA patients treated with biologic and non-biologic DMARDs.3-6 Longer follow-up is necessary to further evaluate the potential risk of malignancies in the CP RA programme. References Askling J, et al. Pharmacoepidemiol Drug Saf 2011;20:119-30. Mariette X, et al. Ann Rheum Dis 2011;70:1895-904. Carmona L, et al. Semin Arthritis Rheum 2011;41:71-80. Pallavicini FB, et al. Autoimmun Rev 2010;9:175-80. Simon TA, et al. Ann Rheum Dis 2009;68:1819-26. Wolfe F, Michaud K. Arthritis Rheum 2007;56:2886-95. Disclosure of Interest X. Mariette Consultant for: Pfizer Inc, J. Curtis Grant/Research support from: Pfizer Inc, Consultant for: Pfizer Inc, E. Lee Consultant for: Pfizer Inc, R. Riese Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, I. Kaplan Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, R. Chew Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, J. Geier Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, L. Wang Shareholder of: Pfizer Inc, Employee of: Pfizer Inc, J. Bradley Shareholder of: Pfizer Inc, Employee of: Pfizer Inc

Costs of illness and quality of life in patients with systemic lupus erythematosus in South Korea

Objective To assess the costs of illness, health-related quality of life (HRQOL) and their associated factors in patients with systemic lupus erythematosus (SLE) in South Korea. Method Two hundred and one patients with SLE were enrolled at the Rheumatology clinic of Seoul National University Hospital. Direct, indirect and total costs and HRQOL were measured using hospital electronic data and face-to-face interview. Socio-demographic and clinical factors associated with cost of illness and HRQOL were analyzed using multiple regression and multivariate logistic regression. Results The average total cost of illness was estimated to be KRW 9.82 million (US

Low dietary inorganic phosphate affects the lung growth of developing mice

8993) per year, of which 41.6% was accounted for by direct costs and 58.4% by indirect costs. In multivariate regression, patients with renal involvement and those with depression incurred an average increment in annual total costs of 37.6% (pu2009=u20090.050) and 49.1% (pu2009=u20090.024), respectively, and an average increment in annual direct costs of 26.4% (pu2009=u20090.050) and 43.3% (pu2009=u20090.002), respectively, compared with patients without renal involvement and depression, respectively. In addition, disease damage was positively associated with an average increment in annual total and direct costs (55.3%, pu2009=u20090.006; 33.3%, pu2009=u20090.013, respectively), and the occurrence of indirect costs (OR 2.21, 1.09–4.88). There was no significant difference in HRQOL between patients with and without renal involvement (0.655 vs. 0.693, pu2009=u20090.203) Conclusion Renal involvement, depression, and disease damage were major factors associated with higher total and medical costs for patients with SLE in South Korea. Effective treatment of renal disorders and depression may reduce the high economic burden of SLE.