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Herein, a bile acid-inspired triple padlock oral gene delivery platform is developed, facilitating the protection of the therapeutic gene from gastrointestinal degradation, selective intestinal accumulation through a bile acid-specific transporter, and transportation of pDNA NPs through the enterohepatic recycling system. This nonviral oral gene delivery nanoparticle exhibits excellent gene expression kinetics in in vitro, in vivo, and ex vivo studies. A single oral dose leads to maintaining normoglycemia for up to 7 days in three different diabetes mouse models and 14 days in diabetic monkeys. Also, the optimized dosage form can reduce nonfast blood glucose levels and hemoglobin A1C within a normal range from the last stage diabetes conditions with a reduction of weight gain from changes of food uptake behavior after treatment once weekly for 20 weeks. Taken together, the current findings could improve the current painful treatment experience of diabetics and thus improve their quality of life.

Intermittent subcutaneous (S.C.) injection of teriparatide [PTH (1-34)] is one of the effective therapies to cure osteoporosis. However, a long-term repeated administration of teriparatide by S.C. to the patients is highly challenging. Herein, a triple padlock nanocarrier prepared by a taurocholic acid-conjugated chondroitin sulfate A (TCSA) is designed to develop an oral dosage form of recombinant human teriparatide (rhPTH). Oral administration of TCSA/rhPTH to the bilateral ovariectomized (OVX) rats resulted in the recovery of the bone marrow density and healthy serum bone parameters from the severe osteoporotic conditions. Also, it enhanced new bone formation in the osteoporotic tibias. This triple padlock oral delivery platform overcame the current barriers associated with teriparatide administration and exhibited a promising therapeutic effect against osteoporosis.

Type 2 diabetes mellitus (T2DM) is a chronic and progressive hyperglycemic condition. Glucagon-like peptide-1 (GLP1) is an incretin secreted from pancreatic β-cells and helps to produce insulin to balance the blood glucose level without the risk of hypoglycemia. However, the therapeutic application of GLP1 is limited by its intrinsic short half-life and rapid metabolic clearance in the body. To enhance the antidiabetic effect of GLP1, we designed a human cysteine-modified IgG1-Fc antibody-mediated oral gene delivery vehicle, which helps to produce GLP1 sustainably in the target site with the help of increased half-life of the Fc-conjugated nanocarrier, protects GLP1 from acidic and enzymatic degradation in the gastrointestinal (GI) tract, uptakes and transports the GLP1 formulation through the neonatal Fc receptor (FcRn), and helps to release the GLP1 gene in the intestine. Our formulation could reduce the blood glucose from about an average of 320 mg/dL (hyperglycemic) to 150 mg/dL (normal blood glucose concentration) in diabetic mice, which is about 50% reduction of the total blood glucose concentration. GLP1 (500 μg) complexed with the IgG1-Fc carrier was proven to be the optimal dose for a complete reduction of hyperglycemic conditions in diabetic mice. A significant amount of insulin production and the presence of GLP1 peptide were observed in the pancreatic islets of oral GLP1 formulation-treated diabetic mice in immunohistochemistry analysis compared to nontreated diabetic mice. The orally given formulation was completely nontoxic according to the histopathology analysis of mice organ tissues, and no mice death was observed. Our antibody-mediated oral gene delivery system is a promising tool for various oral therapeutic gene delivery applications to treat diseases like diabetes.

Pharmacological-based treatment approaches have been used over time to prevent postlaparotomy adhesion. However, the rapid elimination of therapeutics from the peritoneum, and their unwanted side effects, easy flow from the wound site by gravity, and low therapeutic efficacy increase the urgent need for the next generation of antiadhesion agents. This article represents the development of biocompatible and biodegradable antiadhesion agents that consist of carboxymethyl cellulose (CMC) and pullulan with three different types of physical characteristics such as the solution type (ST), film type (FT), and thermosensitive type (TST). These antiadhesion agents that contain no drugs exhibit excellent physical characteristics and superior stability over 30 days in the operative sites without any toxicity and side effects that make the compositions strong candidates as novel antiadhesion agents. Also, the proposed samples reveal superior antiadhesion and tissue regeneration properties in Sprague-Dawley (SD) rats after surgery over Medicurtain. Medicurtain effectively prevented postlaparotomy adhesion in ∼42% of experimental animals, whereas ST 2.25-10, ST 2.5-5, ST 2.5-10, FT 20, and TST 1.5 were effective in 100% of animals. Thus, we believe these antiadhesion agents could be promising to reduce adhesion-related complications during and post-surgical operations and deserve consideration for further study for clinical purposes.

Intermittent subcutaneous (S.C.) injection of teriparatide [PTH (1-34)] is one of the effective therapies to cure osteoporosis. However, a long-term repeated administration of teriparatide by S.C. to the patients is highly challenging. Herein, a triple padlock nanocarrier prepared by a taurocholic acid-conjugated chondroitin sulfate A (TCSA) is designed to develop an oral dosage form of recombinant human teriparatide (rhPTH). Oral administration of TCSA/rhPTH to the bilateral ovariectomized (OVX) rats resulted in the recovery of the bone marrow density and healthy serum bone parameters from the severe osteoporotic conditions. Also, it enhanced new bone formation in the osteoporotic tibias. This triple padlock oral delivery platform overcame the current barriers associated with teriparatide administration and exhibited a promising therapeutic effect against osteoporosis.

Oral siRNA delivery is an ideal way to translate siRNA therapeutic effects in the clinic due to its ability to be administered in convenient and multiple dosages. However, an effective oral delivery system requires overcoming both a hostile gastrointestinal (GI) environment and non-specific targeting. Here, an HTsRP-NC system is a new oral siRNA delivery system consisting of a siRNA/protamine (sRP) nano-complex protected by a multi-functional hyaluronic acid-taurocholic acid (HA-TCA) conjugate. The HTsRP-NC promotes cell penetration and enhances endosomal escape in cancer cells. Moreover, protection of the sRP by HA-TCA from the hostile GI environment helps the AKT siRNA complex to reach the liver through the utilization of a TCA-mediated enterohepatic bile acid recycling system. AKT siRNA was released by 90% in presence of hyaluronidase in the tumor cells which indicate the potential use of HTsRP-NCs for siRNA delivery to treat tumor. After HA receptor (CD44)-mediated cellular uptake of the HTsRP-NC by the liver cancer cells, functional expression of AKT siRNA leads to the suppression of metastatic liver cancer growth in a colorectal liver metastasis (CLM) murine model. Tumor nodules were reduced by more than 1 mm size compared to control group and tumor cells were suppressed by 50% after HTsRP-NCs treatment with AKT siRNA. Overall, oral administration of the HTsRP-NC supports its potential in therapeutic applications for the effective treatment of CLM.

Most currently available commercial vaccines are delivered by systemic injection. However, needle-free oral vaccine delivery is currently of great interest for several reasons, including the ability to elicit mucosal immune responses, ease of administration, and the relatively improved safety. This review summarizes the biological basis, various physiological and immunological barriers, current delivery systems with delivery criteria, and suggestions for strategies to enhance the delivery of oral vaccines. In oral vaccine delivery, basic requirements are the protection of antigens from the GI environment, targeting of M cells and activation of the innate immune response. Approaches to address these requirements aim to provide new vaccines and delivery systems that mimic the pathogen’s properties, which are capable of eliciting a protective mucosal immune response and a systemic immune response and that make an impact on current oral vaccine development.

Bioinspired materials have received substantial attention across biomedical, biological, and drug delivery research because of their high biocompatibility and lower toxicity compared with synthetic materials. Bile acids, well-established biomimetic biomolecules, have been reported with respect to their potential applications as carriers of drugs or imaging contrast agents and, most importantly, as oral absorption enhancers. This review introduced the potential mechanisms involved in the oral absorption of bile acids and their derivatives and further focused on the intelligent applications of bile acids or modified bile acids that respond to biological cues as potential oral absorption enhancers for peptides and macromolecular drugs. Our investigations via the modifications of bile acids with various linkers have demonstrated their effects on the degree of oral absorption. Furthermore, we summarized the reports regarding the development of bile acid formulations for the oral delivery of optical imaging contrast agents for GI tract imaging, as well as anticancer drug delivery. Our opinions regarding the utilization of bile acids for biological and biomedical applications provide clear and concise guidance to investigators with respect to the merits and demerits of bile acid use and the selection of appropriate bile acids based on the requirements for improved biomedical applications.

Thermogenic program (also known as browning) is a promising and attractive anti-obesity approach. Islet amyloid polypeptide (IAPP) and irisin have emerged as potential browning hormones that hold high potential to treat obesity. Here, we have constructed a dual browning gene system containing both IAPP and irisin (derived from fibronectin type III domain containing 5; FNDC5) combined with 2A and furin self-cleavage sites. Intraperitoneal administration of the construct complexed with a linear polyethylenimine into diet-induced obese mice demonstrated the elevation of anti-obesogenic effects characterized as the decreased body weight, adiposity, and levels of glucose and insulin. In addition, the construct delivery increased energy expenditure and the expression of core molecular determinants associated with browning. The additional advantages of the dual browning gene construct delivery compared to both single gene construct delivery and dual peptide delivery can be emphasized on efficacy and practicability. Hence, we have concluded that dual browning gene delivery makes it therapeutically attractive for diet-induced obesity treatment.

The combinatorial peptidergic therapy of islet amyloid polypeptide (IAPP) and leptin (LEP) analogues was once an optimistic option in treating obese animals and patients. However, the need for frequent administrations and its negative side effects prevent it from being a viable choice. Here, we developed a combinatorial gene therapy of IAPP and LEP, where two genes are inserted into a single plasmid with self-cleaving furin and 2A sites to treat diet-induced obese (DIO) mice. The developed plasmid DNA (pDNA) individually produced both IAPP and LEP peptides in vitro and in vivo. The pDNA was delivered with a non-viral polymeric carrier, and its once-a-week administrations demonstrated a synergistic loss of body weight and significant reductions of fat mass, blood glucose, and lipid levels in DIO mice. The results suggest that the combinatorial gene therapy would have higher potential than the peptidergic approach for future translation due to its improved practicability.

Preparation of hIgG1-Fc-9Arg. A. C-terminal 9Arg extension of human IgG1-Fc by PCR. Lane 1. hIgG1-Fc-9Arg PCR (annealing Tm:55, 850 bp), Mw. DNA ladder, lane 2. hIgG1-Fc-9Arg PCR (annealing Tm:52, 850 bp). Template for PCR was an Avastin heavy chain. B. DNA sequence analysis of hIgG1-Fc-9Arg expression vector. C. SDS-PAGE with reduced hIgG1-Fc-9Arg. Mw. protein ladder (10–245 kDa), lane1. 0.1 μg, lane 2. one μg, land 3. ten μg and PAGE with non-reduced hIgG1-Fc-9Arg (lane 1) and Mw (protein ladder (10–245 kDa)). (PPTX 314 kb)

Anti-diabetic effect and tissue toxicity of the complex. A Anti-diabetic effect of orally administered hIgG1-Fc-9Arg/pGLP-1 complex (20/1) and H&E histology analysis in balb/c mice (n = 3, male, 5–7 weeks) as a normal mouse and lepdb/db mice (n = 4, male, seven weeks) as a db/db mouse. (a) Serum insulin concentration, (b) serum GLP-1 concentration, (c) body weight, (d) blood glucose level at day 44 after continuous oral administration of the complex and its comparison of with the blood glucose level before administration of the complex (day 0). B H&E histology analysis of jejunum tissue from the small intestine after oral administration of the complex. (PPTX 1853 kb)

Complexation of hIgG1-Fc-9Arg with pAcGFP-N1. (PPTX 36 kb)

The strategy of the intestinal receptor-mediated delivery of therapeutic gene. (PPTX 547 kb)

Endosomal trafficking of hIgG1-Fc-9Arg complex A. Cellular uptake and endosomal escape of FITC-hIgG1-Fc-9Arg in Caco-2 cells. B. Determination of bobo-3-pDNA complexed with hIgG1-Fc-9Arg on 50/1 weight ratio for tracking complexes mediated FcRn and evaluation of presence pDNA in Caco-2 cells. Lipofectamine used as positive control. Scale bar is 50Â Îźm. (PPTX 1263 kb)

Purpose The safe and functional delivery of progesterone through the vaginal route remains an unmet clinical need. The purpose of this work is to prepare a new progesterone (P4) gel for vaginal application using a thermosensitive mucoadhesive polymer, glycol chitin (GC). Method Thermogelling, mucoadhesive, mechanical, and viscoelastic properties of GC and the new formulation were evaluated using rheometry. In vitro release profile and the bioactivity of P4 were determined using vaginal fluid simulant (VFS) pH 4.2, and PR-reporter gene assay, respectively. In vitro safety of the formulations was tested using (VK2/E6E7) vaginal epithelial cell line and Lactobacillus Crispatus. Finally, in vivo safety and the efficacy of this formulation were evaluated using an endometrial hypoplasia mouse model. Results Results shows the aqueous solution of 5%; (w/v) GC loaded with 0.1%; (w/v) P4 prepared in pH 4.2, (GC-P4), forms a thermosensitive mucoadhesive hydrogel and can maintain stable physical properties at 37°C. GC-P4 gel release 50% of P4 in 4 h after exposure to VFS, and no significant decrease in % viability of VK2/E6E7 or Lactobacillus was found after exposure to 5% GC or GC-P4. GC-P4 does not exhibit obvious toxicities to vaginal tissue in vivo even after repeated application. Efficacy studies indicated that GC-P4 was capable of preventing the progression of simple endometrial hyperplasia (SEH) to complex atypical endometrial hyperplasia (CAEH) in vivo. Conclusions Results indicates that GC-P4 retains many characteristics for an effective vaginal delivery system for P4. Therefore we believe that GC-P4 formulation is a promising alternative to current vaginal P4 formulation. Electronic supplementary material The online version of this article (doi:10.1007/s11095-014-1616-8) contains supplementary material, which is available to authorized users.

Despite the development of a myriad of anticancer drugs that appeared promising in preclinical ovarian cancer animal models, they failed to predict efficacy in clinical testing. To improve the accuracy of preclinical testing of efficacy and toxicity, including pharmacokinetic and pharmacodynamic evaluations, a novel animal model was developed and characterized. In this study, murine ID8 (epithelial ovarian cancer [EOC]) cells as injected cell suspensions (ICS) and as intact cultured monolayer cell sheets (CS) were injected or surgically grafted, respectively, into the left ovarian bursa of 6-8 week-old, female C57BL/6 black mice and evaluated at 8 and 12 weeks after engraftment. Tumor volumes at 8 weeks were as follows: 30.712 ± 18.800 mm(3) versus 55.837 ± 10.711 mm(3) for ICS and CS, respectively, p = 0.0990 (n = 5). At 12 weeks, tumor volumes were 128.129 ± 44.018 mm(3) versus 283.953 ± 71.676 mm(3) for ICS and CS, respectively, p = 0.0112 (n = 5). The ovarian weights at 8 and 12 weeks were 0.02138 ± 0.01038 g versus 0.04954 ± 0.00667 g for ICS and CS, respectively (8 weeks), p = 0.00602 (n = 5); and 0.10594 ± 0.03043 g versus 0.39264 ± 0.09271 g for ICS and CS, respectively (12 weeks), p = 0.0008 (n = 5). These results confirm a significant accelerated tumorigenesis in CS-derived tumors compared with ICS-derived tumors when measured by tumor volume/time and ovarian weight/time. Furthermore, the CS-derived tumors closely replicated the metastatic spread found in human EOC and histopathological identity with the primary tumor of origin.

Supplemental digital content is available in the text., Objectives AL3818 (anlotinib) is a receptor tyrosine kinase inhibitor targeting vascular endothelial growth factor receptors (VEGFR1, VEGFR2/KDR, and VEGFR3), stem cell factor receptor (C-kit), platelet-derived growth factor (PDGFβ), and fibroblast growth factor receptors (FGFR1, FGFR2, and FGFR3). This study evaluates the efficacy of AL3818 studying tumor regression in an orthotopic murine endometrial cancer model. Methods We tested the cytotoxicity of AL3818 on a panel of 7 human endometrial cancer cell lines expressing either wild-type or mutant FGFR2 and also assessed the in vivo antitumor efficacy in a murine, orthotopic AN3CA endometrial cancer model. AL3818 was administered daily per os either alone or in combination with carboplatin and paclitaxel, which represent the current standard of adjuvant care for endometrial cancer. Results AL3818 significantly reduces AN3CA cell number in vitro, characterized by high expression of a mutated FGFR2 protein. Daily oral administration of AL3818 (5 mg/kg) resulted in a complete response in 55% of animals treated and in a reduced tumor volume, as well as decreased tumor weights of AN3CA tumors by 94% and 96%, respectively, following a 29-day treatment cycle. Whereas carboplatin and paclitaxel failed to alter tumor growth, the combination with AL3818 did not seem to exhibit a superior effect when compared with AL3818 treatment alone. Conclusions AL3818 shows superior efficacy for the treatment of endometrial cancer irresponsive to conventional carboplatin and paclitaxel combination and warrants further investigation.

Convenient multiple dosing makes oral administration an ideal route for delivery of therapeutic siRNA. However, hostile GI environments and nonspecific biological trafficking prevent achieving appropriate bioavailability of siRNA. Here, an orally administered AuNP-siRNA-glycol chitosan-taurocholic acid nanoparticle (AR-GT NPs) was developed to selectively deliver Akt2 siRNA and treat colorectal liver metastases (CLM). AR-GT NPs are dual padlocked nonviral vectors in which the initially formed AuNP-siRNA (AR) conjugates are further encompassed by bifunctional glycol chitosan-taurocholic acid (GT) conjugates. Covering the surface of AR with GT protected the Akt2 siRNA from GI degradation, facilitated active transport through enterocytes, and enhanced selective accumulation in CLM. Our studies in CLM animal models resulted in the reduction in Akt2 production, followed by initiation of apoptosis in cancer cells after oral administration of Akt2 siRNA-loaded AR-GT. This therapeutic siRNA delivery system may be a promising approach in treating liver-associated diseases.

We synthesized a new cationic AB2 miktoarm block copolymer consisting of one poly (ethylene glycol) (PEG) block and two cationic poly (l-lysine) (PLL) blocks, wherein the PLL blocks were conjugated to the PEG blocks with or without a bioreducible linker (disulfide bonds). Bioreducible and non-bioreducible miktoarm block copolymers (mPEG-(ss-PLL)2 and mPEG-PLL2) were prepared for efficient gene delivery as a non-viral gene delivery approach. Both cationic copolymers (bioreducible and nonbioreducible) efficiently formed the nanopolyplexes with plasmid DNA (pDNA) through electrostatic interaction at different weight ratio of polymer and pDNA. Gene condensation ability of the polymers and release of the DNA under reduction condition were measured by gel electrophoresis. Dynamic light scattering (DLS) and field-emission transmission electron microscopy (FE-TEM) were used to measure the average hydrodynamic diameter and morphology of the nanoparticles, respectively. The bioreducible nanopolyplexes showed lower cytotoxicity and higher gene expression than the non-reducible nanopolyplexes in cancer cells.

The number of people suffering from insulin-independent type 2 diabetes mellitus (T2DM) is ever increasing on a yearly basis. Current anti-diabetic medications often result in adverse weight gain and hypoglycemic episodes. Hypoglycemia can be avoided with glucagon-like peptide (GLP)-1 receptor agonists, which are expensive and require daily injections that may result immune activation. This study demonstrates the use of non-viral vector based oral delivery of GLP-1 gene through enterohepatic recycling pathways of bile acids. Oral administration of the plasmid DNA (pDNA) encoding GLP-1 decreased diabetic glucose levels to the normoglycemic range with significant weight reduction in a high-fat diet (HFD) induced diabetic mouse model and a genetically engineered T2DM rat model. This novel oral GLP1 delivery system is an attractive alternative to treat late-stage T2DM conditions that require repeated insulin injection and can potentially minimize the occurrence of hypoglycemic anomalies.

A high-throughput, microfluidic flow cell array (MFCA) system has been modified to enable drug screening against small-volume cell-, and tissue cultures. The MFCA is composed of a 3D channel network that simultaneously flows fluids through forty-eight 830 μm by 500 μm flow cells, which physically divide and fluidically seal an existing culture into multiple compartments when docked onto the surface of a cell or tissue culture dish. The modified system provides temperature (37 °C) and CO2/pH level controls, while continuously flowing solutions (media or other liquid such as drug suspensions) over the cells/tissues. These assays were enhanced and validated using inverted microscopy and fluorescent staining techniques which also allow real time viability and toxicity assessments. This work presents the results of this new generation in vitro drug testing assay performed using this modified MFCA system. This setup allows the testing of 48 drug combinations on 48 different cell-, tissue specimen at once under flow conditions. All 48 flow cells were utilized to test 5 different concentrations of cisplatin (CDDP). CDDP solutions in various concentrations were continually flowed over cultured human ovarian cancer cells for 48 h. Viability assessments were performed using red-orange calcein and SYTOX ® Green nucleic acid stains. Cells were imaged at the beginning and end of the experiment (48 h). In order to compare and validate MFCAs suitability as drug screening assay, MTT assays were performed on cells. We found that both, MTT and MFCA assays generated dose-response curves with similar profiles. Innovative advantages of the MFCA system include the ability of handling smaller amounts of solutions compared to conventional and current state of the art drug screening and cell viability/toxicity methods. It also provides the ability to continually deliver fresh solution to the cell samples, while eliminating wastes that are produced. Based on our here reported findings MFCA may have a strong potential of providing a more physiological model than current state of the art static MTT assays.

A leading cause of death and suffering in patients with abdominal or pelvic malignancies is progression of peritoneal surface disease. Changes in the use of chemotherapy have shown significant survival benefits for intraperitoneal or combined intraperitoneal and intravenous treatment following optimal surgical cytoreduction. However, broader clinical use of intraperitoneal therapy has not reached its full potential due to limited efficacy, accessibility and nonspecific toxicity. To overcome these problems, we developed a mucoadhesive hybrid gel (HG) for a local, intraperitoneal drug delivery. In vivo studies confirmed reliable adherence and residence of the gel to the peritoneal sidewall for at least 72 h exhibiting no signs of tissue toxicity. Functionally active CDDP was released from HG within 2h and was equal to free CDDP in vitro. Moreover, intraperitoneal application of HG-CDDP significantly enhanced CDDP accumulation in the genomic DNA of peritoneal tissues compared to the same CDDP dose administered intravenously. These findings indicate the potential application of this hybrid gel as a mucoadhesive drug carrier amendable to use for intraperitoneal drug delivery and possible expansion for use on other mucosal surfaces of the female reproductive tract.

In this study, a new thermo-sensitive polymer, glycol chitin, was synthesized by controlled N-acetylation of glycol chitosan and evaluated as a thermogelling system. The physico-chemical properties of glycol chitins with different degrees of acetylation (DA) were investigated in terms of degradation, cytotoxicity, rheological properties, and in vitro and in vivo gel formation. Aqueous solutions of glycol chitins were flowable freely at room temperature but quickly became a durable gel at body temperature. Thermo-reversible sol-gel transition properties were observed with fast gelation kinetics. Glycol chitins with higher DA showed faster degradation in the presence of lysozyme. They exhibited no significant biological toxicity against human cell lines. An anti-cancer drug, doxorubicin, could be incorporated into the hydrogel by a simple mixing process and released in a sustained pattern over 13 days. Our findings suggest that glycol chitins could be useful as a new thermogelling biomaterial for drug delivery and injectable tissue engineering.

Multidrug resistance (MDR) is a barrier to successful cancer chemotherapy. Although MDR is associated with overexpression of ATP-binding cassette (ABC) membrane transporters, mechanisms behind their up-regulation are not entirely understood. The cleaved form of the Notch1 protein, intracellular Notch1 (N1 IC ), is involved in transcriptional regulation of genes. To test whether Notch1 is involved in the expression of multidrug resistance-associated protein 1 (ABCC1/MRP1; herein referred to as ABCC1), we measured N1 IC and presenilin 1 (PSEN1), the catalytic subunit of γ-secretase required for Notch activation. We observed higher levels of N1 IC and PSEN1 proteins as well as higher activity of N1 IC in ABCC1-expressing MDR MCF7/VP cells compared with parental MCF7/WT cells. Reducing N1 IC levels in MCF7/VP cells with either a γ-secretase inhibitor or shRNA led to reduction of ABCC1. By contrast, ectopic expression of N1 IC in MCF7/WT cells led to increased expression of ABCC1 and associated drug resistance, consistent with expression of this transporter. Inhibition of ABCC1 reversed drug resistance of N1 IC -overexpressing stable cells. Using an ABCC1 promoter construct, we observed both its reduced transcriptional activity after blocking the generation of N1 IC and its increased transcriptional activity in stable cells overexpressing N1 IC . ChIP and gel-shift assays revealed an interaction between a specific promoter region of ABCC1 and the N1 IC -activated transcription factor CBF1, suggesting that the regulation of ABCC1 expression by Notch1 is mediated by CBF1. Indeed, deletion or site-directed mutagenesis of these CBF1 binding sites within the ABCC1 promoter region attenuated promoter-reporter activity. Overall, our results reveal a unique regulatory mechanism of ABCC1 expression.

Liver metastasis (CLM) is the lethal secondary hepatic cancer with 5 - 20 months' average survival rate and mostly originated from colorectal cancer (CRC). Surgery and systemic chemotherapy are the currently best options to treat patients. However, over 60% of recurrence rate and severe side effects such as sinusoidal obstruction syndrome and nodular regenerative hyperplasia from the systemic chemotherapy significantly impact on patient's recovery from the treatment. To overcome these side effects and also achieve the synergistic therapeutic effect, we developed a dual padlock system (AR-GT) for oral siRNA delivery to regulate PI3K/Akt signaling pathway, which is highly associated with the progression of CRC to CLM. AR-GT is an about 130 nm nanoparticle consisted of a gold nanoparticle core anchoring siAkt RNA (AR) and a single outer layer of glycol chitosan-taurocholic acid (GT) conjugate. GT layer plays several roles in oral siRNA delivery such as the protection of siRNA in the gastrointestinal (GI) environment, facilitation of ileac uptake of AR-GT in the small intestine and transportation of AR-GT through enterohepatic circulation into the liver. In vitro characterization of AR-GT demonstrated the protection of siRNA under simulated GI pHs for 5 days, facilitation of the trans-epithelial transport in the intestinal Caco2 and ASBT positive cells, and reduction of Akt protein by successful release of siAKT from the AR-GT. Following oral delivery of AR-GT to CLM mouse model, we observed high biodistribution of AR-GT in the ileum, liver, and kidney through intestinal uptake, enterohepatic transportation, and renal excretion. Moreover, CLM mouse after oral delivery of AR-GT (100 μg/kg) manifested about 58 % reduction of the numbers of tumor nodules in the liver and near 100% survival benefit compared to control mouse fed with either saline or AR-Chitosan nanoparticles. Further analysis of liver samples obtained from CLM mouse after oral delivery of AR-GT indicated about 38% reduction of pAkt expression and the enhancement of apoptotic protein expressions such as 2 times or 1.5 times increase of caspase 9 and bax. Theses observation suggests survival benefit and tumor nodule reduction in CLM mouse root on successful AR-GT mediated oral siAkt RNA delivery. we conclude AR-GT oral siRNA delivery system is beneficial in the CLM treatment and further suggest its application to target other cancers. Citation Format: Sung-Hun Kang, Vishnu Revuri, Sungpil Cho, In-Kyu Park, Kwang Jae Cho, Woo Kyun Bae, Yong-Kyu Lee. Oral siRNA delivery for colorectal liver metastases cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5930.

We describe here a new method to estimate the oral drug permeability from the small intestine using surface plasmon resonance (SPR) technology. The interaction between drugs and brush border membrane (BBM) surfaces immobilized on biosensor chip were evaluated by measuring the SPR response signal.BBM vesicles, isolated from Sprague-Dawley (SD) rats, were immobilized onto the L1 chip composed of dextran derivatives with modified lipophilic residues. A SPR (BIAcore 3000) was used with L1 chip, and it was carried out in a running buffer, HEPES-buffered saline containing 0.1% DMSO. Fourteen drugs for the SPR experiments were flowed over the BBM immobilized L1 chip, and the response levels according to the BBM surfaces were evaluated directly in a continuous flow system.The immobilized BBM surface on L1 chip was very stable, and it was regenerated by injecting a new BBM vesicle solution. It was evident that drug binding events, using BBM surfaces, directly provides information that predicts the Fa value in human for transcellularly absorbed drugs. The throughput to assay each drug at a single concentration is 100 drugs for 24 h.The interaction between drugs and small intestinal surfaces was successfully assayed using SPR technology, and this SPR analysis exhibited advantages: lack of labeling requirements, the real-time acquirement of various results, and the repeated use for various drugs.

To prepare the surface generated by small intestinal brush border membrane vesicles (BBMVs) for the surface plasmon resonance (SPR) analysis, which allows the real-time measurement of binding events occurring on the intestinal membrane.BBMVs were isolated from Sprague-Dawley rats, suspended in HEPES-buffered saline, and flowed over the surface of a SPR sensor chip composed of dextran derivatives modified with lipophilic residues. The surface coverage was determined from binding of bovine serum albumin to BBMV-immobilized sensor chip. The performance of BBMVs immobilized was evaluated by their interaction with otilonium bromide and bile salts.The stable BBMV surface was achieved when BBMV suspension was flowed over the sensor chip for 8 h at a rate of 2 microl/min. The flow of otilonium bromide resulted in an increased SPR signal because of its binding to calcium channel, which is known to be distributed over the gastrointestinal tact. When bile salts were flowed over ileal and duodenal BBMV surfaces, respectively, a slightly higher SPR signal was observed in the ileal BBMV surface, indicating the specific interaction of bile salts with bile acid transporters.BBMV surfaces may be useful for the estimation of binding events on the intestinal membrane by SPR analysis, especially for the drugs that are orally administrated.

Endometrial cancer (EC) is the most common uterine malignancy in industrialized countries and is the most frequent gynecologic cancer in the United States with an estimated 60,050 new cases in 2016. Traditionally observed in peri-, postmenopausal women, the incidence rate of EC has steadily increased among younger women of reproductive age over the past two decades partially owing to a more sedentary lifestyle and rampant obesity epidemic. Surgery in combination with either radio- and or chemotherapy is commonly curative for the early stage of the disease. However, patients with recurrent or metastatic disease, who only respond poorly to cytotoxic chemotherapy are in desperate need of therapeutic alternatives as are patients of reproductive age. Thus, innovative, safe and more effective therapies are mandated. Several studies have demonstrated the increased expression of various tyrosine kinase receptors involved in the development of EC. We hypothesized that a targeted therapeutic approach using a pan-tyrosine kinase inhibitor would prevent the proliferation and progression of EC. Anlotinib (AL3818) is a multi-targeted receptor tyrosine kinase inhibitor targeting vascular endothelial growth factor receptors 1 to 3 (VEGFR1-3), stem cell factor receptor (C-kit), platelet-derived growth factor (PDGFβ), and fibroblast growth factor receptors 1 to 3 (FGFR1-3). We tested a panel of seven human endometrial cancer cell lines (AN3Ca, Ishikawa, HEC1A, HEC1B, KLE, MFE280 and MFE296) to assess the cytotoxicity of anlotinib in vitro. A lower IC50 value was observed in AN3Ca cells (25 nM), a cell line characterized by a high level of expression of an FGFR2 mutant protein, while cell lines expressing FGFR2 wild-type are less sensitive to anlotinib (HEC1A: 33 μM, or HEC1B:36 μM). Somatic mutations of FGFR2 have been observed in 12% of EC, indicating the potential for this drug in a subset of EC patients. In summary, these results suggest improved efficacy of anlotinib for the treatment of ECs expressing an increased level of FGFR2 mutant proteins. The safety and efficacy of anlotinib are currently being evaluated in an orthotopic AN3Ca mouse model of EC. Experimental groups include a tumor bearing control not subjected to any treatment, anlotinib alone, carboplatin/paclitaxel, a combination of anlotinib and carboplatin/paclitaxel. Primary study endpoints include tumor growth retardation, the delayed onset of local metastases as well as reduced toxicity as secondary endpoint. Citation Format: Sebastien Taurin, Chieh-Hsiang Yang, Maria Reyes, Sungpil Cho, Elke A. Jarboe, Theresa L. Werner, Demetrius M. Coombs, Paul Chen, Margit M. Janát-Amsbury. Treatment of endometrial cancer cells with a new small tyrosine kinase inhibitor targeting mutated fibroblast growth factor receptor-2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3244. doi:10.1158/1538-7445.AM2017-3244

To develop and characterize the pre-clinical suitability of a syngeneic mouse epithelial ovarian cancer model in immunocompetent hosts.ID8 mouse ovarian surface epithelium cells were implanted into the left ovarian bursa of C57BL/6 mice. Using conventional as well as ultrasound-based techniques and histopathological analysis, the tumor weights, volumes, metastases, ascites and vascularity were observed over a period of 16 weeks.Ovarian weights and volume increased 12- and 7-fold, respectively. Ultrasound measurements of ovarian ID8 tumors correlated with the actual size obtained following surgical excision. Ascites and metastasis were first observed at 12 weeks post-orthotopic implantation. Histopathological analysis indicated similarities between orthotopically-generated ovarian tumors and human ovarian tumors. However, there was less evidence of angiogenesis in this animal model.The development of this mouse model closely replicates characteristics seen in human ovarian cancer with feasibility of using ultrasound to assess tumor formation, progression and vascularization.

Cancer stem cells (CSCs) are a small subset of tumor cells capable of self-renewal, tumor initiation, and growth. Isolation, expansion, and differentiation of CSCs in the proper stem niches remain a topic of great debate. To address some of the challenges, we developed bioengineered-induced cancer like stem cells (iCLSCs) through in vitro oncogenic reprograming of C57BL/6 mouse embryonic stem cells (mESCs). We constructed retroviral plasmids through integration of well-defined oncogenic elements such as SV40LTg and HrasV12 into a mouse stem virus long terminal repeat (MSCV-LTR) plasmid fused with either a GFP or RFP gene. We successfully acquired these bioengineered-iCLSCs through retroviral introduction alongside with FACS sorting of positively infected mESCs. We then cultured the bioengineered-iCLSCs on mouse embryonic fibroblast (mEF) feeder cells. Bioengineered-iCLSCs expressed the introduced oncogenic genes and showed enhanced proliferation similar to cancer cells. In addition, we ascertained the unhampered stem properties of the bioengineered-iCLSCs by examining the maintenance of alkaline phosphatase activity as well as the expression of mouse stem cell marker SSEA-1. Furthermore, the orthotopic inoculation of these bioengineered-iCLSCs into either the mammalian fat pad or the ovarian bursa of female C57BL/6 mice lead to the formation of teratomas in supportive niches. Interestingly, the teratomas formed in the ovarian bursa exhibited malignant and immature features, whereas the iCLSCs introduced to the mammary fat pad were not able to generate teratomas with similar features. These observations call for additional research to identify the dependency on a supportive niche and their role in metastasis based on a tumor-favorable microenvironment. Our study helped to establish the use of in vitro oncogenic reprograming to generate bioengineered-iCLSCs and suggests the potential application for the initiation of organ-specific, malignant tumor formation in an orthotopic small animal cancer model. Citation Format: Sungpil Cho, Hongsuk Park, Elke A. Jarboe, C Matthew Peterson, You Han Bae, Margit Matthew Janat-Amsbury. Bioengineered-induced cancer-like stem cells: characterization and applications. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2502.

Attempted surgical removal of advanced stage peritoneal cancers is mostly insufficient and often fails in areas affected by disseminated disease. But the successful elimination of peritoneal surface spread is known to have a significant impact on patient survival. However, standard therapies such as surgical cytoreduction and systemic chemotherapy combined with radiation have only shown limited-efficacy, accessibility, and nonspecific toxicity as well as frequent development of multidrug resistance (MDR). To overcome those limitations, we previously reported about the development of a mucoadhesive, chitosan-hybrid gel (CS(BCDDP)) embedded with cisplatin (CDDP) containing alginate beads for intraperitoneal administration (# 1948, AACR 2012). Here we report additional results illustrating CDDP release from CS(BCDDP) at different pH in vitro, CDDP accumulation to genomic DNA (gDNA) isolated from tissues as well as toxicity data in vivo. To determine the amount of CDDP released from CS (BCDDP) in vitro, B50μgCDDP, CDDP-chitosan (CS50μgCDDP) and CS (B50μgCDDP) pellets (8 mm ø, 1.5 mm thickness) were prepared and placed into a 6 well plate containing 2 ml/well PBS (pH 6, 7, 7.4 and 8). Multiple samples were collected during the 24 h incubation (37°C, 100rpm) period. The majority (75%) of CDDP was successfully released from CS(BCDDP) within the first 2 h at pH 7.4 showing the rate of drug release to be inversely correlated with pH yielding a more rapid release under acidic pH conditions. To assess CDDP accumulation to gDNA, inductively coupled plasma mass spectrometry (ICP-MS) was performed with tissue samples obtained from the left peritoneal sidewall of female nu/nu mice 24 hours after treatment with CS50μgCDDP, CS(B50μgCDDP), intravenously (i.v.) CDDP(IV50μgCDDP) and intraperitoneal (i.p) CDDP (IP50μgCDDP), respectively. The results demonstrated a greater than three-time enhancement of CDDP-accumulation to the gDNA from CS(B50μgCDDP) when compared to CS50μgCDDP and IV50μgCDDP administration. In addition, CS(B50μgCDDP) showed similar levels of CDDP adduction compared to direct IP50μgCDDP administration. Moreover, assessment of CDDP accumulation in kidneys and blood 24 h following treatment with either IP50μgCDDP or CS(B50μgCDDP), demonstrated significantly decreased kidney toxicity from CS(B50μgCDDP) when compared to IP50μgCDDP. Our results, therefore strongly suggest that administration of this hybrid gel directly to mucosal surfaces such as the peritoneal cavity is feasible making it an advantageous, safe and non-toxic intraperitoneal drug delivery system for the treatment of disseminated peritoneal cancers such as advanced or recurrent ovarian cancer. Currently ongoing experiments are evaluating the efficacy and safety of our hybrid gel/CDDP in an orthotopic peritoneal cancer animal model. Citation Format: Sungpil Cho, Yongen Sun, Elke A. Jarboe, Andrew P. Soisson, Mark K. Dodson, David K. Gaffney, C.Mattew Peterson, Margit M. Janat-Amsbury. Tissue accumulation and toxicity of platinum released from a novel chitosan hybrid gel for intraperitoneal drug delivery. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5402. doi:10.1158/1538-7445.AM2014-5402

Ovarian cancer is the second most frequent invasive malignancy of the female genital tract and the most common cause of death among women with gynecologic malignancies. Although epithelial ovarian cancer (EOC) accounts for over 90% of all ovarian malignancies, the lack of proper animal models that mimic the development and consequences of human EOC limits the ability of current preclinical ovarian cancer models to predict treatment response. The orthotopic development of syngeneic EOC within its relevant tumor microenvironment is an experimental ideal. Here, we demonstrate the development and characterization of a syngeneic EOC mouse model in immunecompetent C57BL/6 mice by orthotopic implantation of ID 8 mouse ovarian cancer cells. Using conventional and ultrasound-based techniques to compare tumor weight, volume and vascularity we followed the development of primary tumors, metastases and ascites over 16 weeks. ID 8 cells were implanted directly underneath the left ovarian bursa. Normal saline was injected to the contralateral ovary as control. Ovarian tumors grew consistently throughout the study period; ovarian weight and volume increased twelve and seven fold, respectively, compared to the contralateral, non-cancerous ovary. Ultrasound measurements of primary tumors and surrounding tumor tissue correlated with the actual size after surgical tumor harvest. Abdominal ascites were first observed at 12 weeks post orthotopic ID8 implantation with volume changes correlating with changes in abdominal circumference. Metastatic lesions were identified by ultrasound at 12 weeks after orthotopic ID8 implantation; sites included peritoneum, liver, and intestine. Histopathological analysis of tumors and metastases indicated similarities between orthotopic ID8 ovarian tumors and human ovarian tumors, except a significantly lower formation of angiogenic vasculature within the ID8 tumors. This study confirms the successful development of a mouse model that closely replicates characteristics seen in human ovarian cancer patients. It shows the feasibility of using ultrasound to assess tumor formation, progression and vascularization. Furthermore, we demonstrate potential shortcomings, which could significantly limit the use of current animal models predicting therapeutic efficacy of novel agents, especially anti-angiogenic therapeutics, impacting the translation of preclinical information to actual clinical outcomes. Citation Format: Sungpil Cho, Yongen Sun, Andrew P. Soisson, Mark K. Dodson, C. Matthew Peterson, Elke A. Jarboe, Anne M. Kennedy, Margit M. Janat-Amsbury. Characterization and evaluation of a mouse ovarian cancer model for its preclinical suitability. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 325. doi:10.1158/1538-7445.AM2013-325

Development of multidrug resistance is a major impediment to the successful treatment of cancer. Although many cellular mechanisms are involved in multidrug resistant cancers, overexpression of ATP-binding cassette (ABC) membrane transporters represents one of the main determinants of the multidrug resistance phenotype. We have previously demonstrated a positive correlation of expression of intracellular Notch1 (NIC) and the ABCC1 transporter; we found that NIC contributes to multidrug resistance by upregulation of ABCC1 (S Cho, M Lu et al, PNAS, in press, 2011). In order to determine whether there is a functional relationship between the two proteins, we performed drug efflux experiments. We treated drug-sensitive MCF-7 and multidrug resistant (MDR) MCF-7/VP breast cancer cells with the anticancer drug doxorubicin and measured the amount of drug efflux by flow cytometry over time. The MDR MCF-7/VP cells, which have high levels of activated Notch1 and ABCC1 proteins, exhibited increased doxorubicin efflux, compared to the drug-sensitive MCF-7 cells. Since γ-secretase (GS) is required for the processing of Notch into its active form, NIC, we blocked GS activity using a GS inhibitor, DAPT, in MCF-7/VP cells and assayed drug efflux from them. We found that inhibition of Notch signaling by DAPT blocks ABCC1 mediated drug efflux. To better understand the connection between Notch, ABCC1, and drug efflux, we compared drug efflux in MCF-7 cells stably overexpressing NIC (ICN1-c4, which also overexpress ABCC1), and the “empty vector” control MCF-7 cells (pcDNA3). As expected, we saw increased drug efflux in the ICN1-c4 cells compared to MCF7/pcDNA3 cells. To establish whether the observed drug efflux is ABCC1-specific, we treated ICN1-c4 cells with either the ABCC1-specific inhibitors reversan or probenecid, or with an ABCC1 siRNA to knockdown ABCC1 expression, and then examined these cells for changes in drug efflux. We found that inhibition of ABCC1, by reducing ABCC1 activity or levels in the presence of activated Notch1, blocks drug efflux from ICN1-c4 cells. In addition, we also found that the ABCC1 inhibitor reversan sensitizes ICN1-c4 cells to doxorubicin, and knockdown of ABCC1 reverses the resistance of ICN1-c4 cells to doxorubicin. These results strongly support the idea that Notch1 exerts its effect on ABCC1-mediated drug efflux to a large extent through its induction of ABCC1. Overall, our results reveal a novel functional relationship between expression of Notch and ABCC1. (Supported in part by CA40570 [to WTB] and in part by UIC.) Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 772. doi:1538-7445.AM2012-772

Gynecological malignancies such as cervical, endometrial and ovarian cancer still are the leading causes of death for women worldwide. However, standard therapies such as surgical cytoreduction and systemic chemotherapy combined with radiation have shown limited-efficacy by accessibility as well as nonspecific toxicity. To overcome those limitations, we previously reported about the development of a muco-adhesive, chitosan-hybrid gel (CS(BCDDP)) embedded with alginate beads containing cisplatin (CDDP) for a local, intraperitoneal application (# 3232, AACR 2011). Here we report the results of CDDP release from the (CS(BCDDP)), its cellular activity in vitro and CDDP accumulation to genomic DNA (gDNA) of tissues in vivo. To determine the amount of CDDP released from (CS(BCDDP)) in vitro, we used a colorimetric SnCl2 assay following the incubation in 0.9% NaCl solution for 24 hrs. (CS(BCDDP)) demonstrated successful release of 90% CDDP within 2 hrs and consistent CDDP-release could be maintained for up to 24 hrs. To test activity of CDDP released from (CS(BCDDP)), clonogenic assay was performed with Hela (human cervical adenocarcinoma) cells. Cells treated with (CS(BCDDP)) demonstrated a significantly low number of colonies, generating 9 (± 6) colonies per 500 cells seeded compared with 233 (± 10) colonies per 500 cells treated with an empty, no drug containing control gel (CS). To further assess CDDP accumulation to gDNA of tissues in vivo, inductively coupled plasma mass spectrometry (ICP-MS) was performed with tissue samples obtained from the left peritoneal sidewall of female nude mice following treatment with a control gel mixed with CDDP (CSCDDP), (CS(BCDDP)), and intravenously (i.v.) CDDP(CDDPIV). The results demonstrated a greater than three-time enhancement of CDDP-accumulation to the gDNA from (CS(BCDDP)) when compared to CDDPIV. Moreover, (CS(BCDDP)) also demonstrated higher accumulation of CDDP to gDNA when compared with CSCDDP. Relative values were 3.2 (±0.9) for (CS(BCDDP)) and 1.3 (± 0.4) for CSCDDP compared to values obtained from CDDPIV. Our in vitro and in vivo results, strongly suggest the feasibility of applying this hybrid gel locally to mucosal surfaces of the female reproductive tract and its advantage for intraperitoneal drug accumulation. Currently ongoing experiments are evaluating the efficacy and safety of our hybrid gel/CDDP in orthotopic endometrial, ovarian and cervical cancer animal models. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1948. doi:1538-7445.AM2012-1948

Cervical cancer is one of the leading gynecological malignancies affecting women worldwide. Although surgical cytoreduction and systemic chemotherapy combined with radiation are standard therapies, accessibility to such treatment as well as nonspecific toxicity limit its efficacy. We hypothesize that the site-specific local application of anticancer drugs utilizing innovative delivery technologies will improve efficacy, reduce toxicity and provide cost-effective, accessible therapeutic options. To test this hypothesis, we developed a mucoadhesive, polymeric hybrid chitosan gel embedding drug containing alginate beads. An optimized gel was achieved by mixing 1% chitosan (w/v) with glycerol dissolved in acetic acid (AA) solution at a volume ratio of 1:2 or 1:3 (2% AA:glycerol). Alginate beads were formed by dropping alginate solution (2%, w/v) into various concentrations of CaCl2 solution. In testing swelling properties, dried alginate beads transforming into wet ones were capable of releasing the drug load. Beads obtained from 1% CaCl2 solution demonstrated optimal and stable swelling properties that were 40 times greater than other formulations. Such beads, loaded with cisplatin (CDDP) dissolved in 0.9% NaCl were prepared. The loaded CDDP concentration was confirmed by colorimetric assay with Sn2Cl2 after re-swelling of the dried beads. Drug loaded-beads contained an average of 0.2mg of CDDP per 10 mg of dried beads. There was a linear correlation (R2 = 0.92) between the swelling ratio of the alginate beads and the amount of CDDP released, allowing the ability to estimate CDDP concentration based on the swelling ratio. Drug-loaded dried alginate beads were placed into a chitosan solution resulting in a hybrid hydrogel. To test the feasibility of local application of the gel on a mucosal surface, we investigated the swelling and mucoadhesive properties on a 4cm2 section of mouse peritoneal sidewall. We confirmed the complete swelling of beads within the hybrid gel suggested sufficient release of CDDP from the beads onto a biological surface. Mucoadhesive properties and toxicity were evaluated in vivo by surgically placing a 1cm2 sized gel pellet into the left peritoneal sidewall of immune competent female mice. The hybrid gel remained in place for 72 hours exhibiting no signs of local inflammation, tissue damage or systemic toxicity. Our results strongly suggest the feasibility of application of this hybrid gel to various mucosal surfaces. Ongoing experiments will evaluate the efficacy and safety of our hybrid gel/CDDP as a radiation enhancer in the treatment of cervical cancer and for the treatment of cervical dysplasia. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3232. doi:10.1158/1538-7445.AM2011-3232