Rapamycin (Rap), a small-molecule inhibitor of mTOR, is an immunosuppressant, and several Rap analogues are cancer chemotherapeutics. Further pharmacologic development will be significantly facilitated if in vivo reporter models are available to enable monitoring of molecular-specific pharmacodynamic actions of Rap and its analogues. Herein we present the use of a Gal4→Fluc reporter mouse for the study of Rapinduced mTOR/FKBP12 protein-protein interactions in vivo with the use of a mouse two-hybrid transactivation strategy, a derivative of the yeast two-hybrid system applied to live mice. Upon treatment with Rap, a bipartite transactivator was reconstituted, and transcription of a genomic firefly luciferase reporter was activated in a concentration-dependent (K d = 2.3 nmol/L) and FK506-competitive (K i = 17.1 nmol/L) manner in cellulo, as well as in a temporal and specific manner in vivo. In particular, after a single dose of Rap (4.5 mg/ kg, i.p.), peak Rap-induced protein-protein interactions were observed in the liver at 24 hours post treatment, with photon flux signals 600-fold over baseline, which correlated temporally with suppression of p70S6 kinase activity, a downstream effector of mTOR. The Gal4→Fluc reporter mouse provides an intact physiologic system to interrogate protein-protein interactions and molecular-specific pharmacodynamics during drug discovery and lead characterization. Imaging protein interactions and functional proteomics in whole animals in vivo may serve as a basic tool for screening and mechanism-based analysis of small molecules targeting specific protein-protein interactions in human diseases. Mol Cancer Ther; 9(10); 2752-60. ©2010 AACR.

The use of radioactive colloidal phosphorus 32 (32P) in the treatment of epithelial ovarian cancer continues to be controversial. One institution's experience with the use of 32P in 30 patients with epithelial ovarian cancer was reviewed retrospectively. One hundred percent of attempts at placement of 32P intra-abdominally were ultimately successful. The complication rate was 11%. Mean clinical (asymptomatic) disease-free survival in patients with stage III ovarian cancer was as follows: 26 months based on absence of disease at reassessment surgery; 26 months based on microscopic residuum; and 30 months based on minimal (< 5 mm) residuum. Mean disease-free survival in patients with early-stage (stages IC through IIC) ovarian cancer was 66 months.

Background: The diversity of cancer genome significantly influences drug response and impairs the discovery of biomarker for patient stratification. As important preclinical models, patient derived cell lines are widely used in drug discovery and biomarker screening, yet complicated by intratumoral heterogeneity. Methods: We created a One Patient Panel (OPP), that is, a set of cell lines (26 lines in this case) derived from tumor tissue from one hepatocellular carcinoma (HCC) patient, and systematically integrated the OPP by transcriptional profiling, chromosomal copy number variations (CNVs) and whole-exome sequencing. The pharmacological profiles of more than 70 targeted therapeutic agents were also examined in OPP. Results: CNVs data showed that 10 chromosomes contained no significant qualitative differences, while 13 chromosomes had obvious cytogenetical and cell-line specific alterations, among the 26 cell lines. Likewise, transcriptional profiling and drug response analysis indicated the presence of clinically relevant intratumoral heterogeneity in OPP. For examples, MEK pathway was highly active in 6 (23.1%) lines and completely inactive in 3 (11.5%) lines, mTOR pathway was active in about 50% of lines, and Sonic Hedgehog pathway was active in above 30% of lines. Yet, based on the drug screening data, 80% of drugs had similar response patterns among those lines in OPP, suggesting a close genetic background among them. Interestingly, FGFR and MEK inhibitors have dramatically distinct drug response patterns among those cell lines. In response to FGFR inhibitors, there were four sensitive lines with the IC50 lower than 250 nM and five resistant lines with IC50 higher than 5 μM. For MEK inhibitors, the IC50s of three sensitive lines were lower than 50 nM, while the resistant IC50s were higher than 1 μM. Thus, a maximum of 30-folds difference in sensitivity to targeted agents was observed among cell lines in OPP. Moreover, there were no overlaps between the two groups of sensitive lines to FGFR or MEK inhibitors. Despite response variations, different inhibitors against the same molecular target, such as BGJ398 (Novartis) and AZD4547 (Astrazeneca), both against FGFR, showed comparable sensitivity in the same cell lines, indicating those inhibitors might have same biomarker(s). Conclusions: Taken together, the OPP recapitulates the intratumoral genetic, molecular and pharmacological heterogeneity in HCC. Importantly ,the OPP provides a powerful platform for optimizing drug selection and biomarker discovery, which may speed up the emergence of precision therapeutic regimens. Citation Format: Qiang Gao, William Niu, Andrew Tse, Jeffrey Lin, Jian Zhou, Simon Shung, Jia Fan. Intratumoral heterogeneity and precise personalized therapy revealed by one patient panel in hepatocellular carcinoma. [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 LB-223. doi:10.1158/1538-7445.AM2014-LB-223

Background: Data on the patient outcomes for newer β-lactam–β-lactamase inhibitor (BLBI) drugs compared to carbapenem-containing combination antibiotics for multidrug-resistant (MDR)–Pseudomonas aeruginosa infections are limited. Methods: This retrospective, case–control observational study was based on chart review of the patients managed at the University of Kentucky. Results: In total, 143 patients with MDRO Pseudomonas aeruginosa infections were identified and divided into 2 groups: 1 group received newer BLBI combinations with or without aminoglycosides or polymyxins, for at least 72 hours, and the control group received carbapenem containing combination antibiotics or other antibiotics. Baseline characteristics and patient outcomes are shown in Table 1. Discussion: The newer BLBI combinations group consisted of 60.8% MDR Pseudomonas bacteremia, whereas the control group had 68.4% of MDR Pseudomonas respiratory cultures. Overall, the use of newer BLBI combinations such as ceftazidime/avibactam, ceftolozane/tazobactam, and meropenem/vaborbactam was associated with lower rates of acute kidney injury (AKI), shorter LOS, and lower mortality rates compared to the control group, and these differences were statistically significant. Because the 2 populations of patient differed significantly based on the site of infection (sepsis vs pneumonia), the data were reanalyzed to evaluate the impact of therapy on the occurrence of AKI, LOS, and mortality based on the site of infection. Only those patients with sepsis who received the newer combination drugs had significantly better rates of AKI, lower LOS, and had lower rates of mortality. The 2 treatment arms were not statistically different when comparing patients with pneumonia. Additionally, the use of these new combination therapies did not make a difference regarding readmission rates or duration of bacteremia for the patients included in the study.Funding: NoDisclosures: None