Drug Intervention Studies Research Articles

A mixed-methods approach exploring acceptability and feasibility of trials designed to test drugs targeting prevention of post-traumatic osteoarthritis after knee injury.

To explore key stakeholder views around feasibility and acceptability of trials seeking to prevent post-traumatic osteoarthritis (PTOA) following knee injury, and provide guidance for next steps in PTOA trial design. Healthcare professionals, clinicians, and/or researchers (HCP/Rs) were surveyed, and the data were presented at a congress workshop. A second and related survey was then developed for people with joint damage caused by knee injury and/or osteoarthritis (PJDs), who were approached by a UK Charity newsletter or Oxford involvement registry. Anonymized data were collected and analyzed in Qualtrics. Survey responses (n = 19 HCP/Rs, 39 PJDs) supported studies testing pharmacological agents preventing PTOA. All HCP/Rs and 30/31 (97%) PJDs supported the development of new treatments that improved or delayed knee symptoms and damage to knee structure. PJDs thought that improving structural knee damage was more important than knee symptoms. Both groups found studies more acceptable as expected future benefit and risk of PTOA increased. All drug delivery routes were acceptable. Workshop participants (around n = 60) reflected survey views. Discussions suggested that stratifying using molecular testing for likely drug response appeared to be more acceptable than using characteristics such as sex, age, and BMI. Our findings supported PTOA drug intervention studies, including situations where there is low risk of disease, no expected benefit of treatment, and frequent treatment administration. PJDs appeared less risk-averse than HCP/Rs. This work reinforces the benefits of consensus and involvement work in the co-creation of PTOA drug trial design. Involvement of key stakeholders, such as PJDs with different risks of OA and regulatory representatives, are critical for trial design success.

A unified framework for prediction of liver steatosis dynamics in response to different diet and drug interventions

Background & AimsNon-alcoholic fatty liver disease (NAFLD) is a common metabolic disorder, characterized by the accumulation of excess fat in the liver, and is a driving factor for various severe liver diseases. These multi-factorial and multi-timescale changes are observed in different clinical studies, but these studies have not been integrated into a unified framework. In this study, we aim to present such a unified framework in the form of a dynamic mathematical model. MethodsFor model training and validation, we collected data for dietary or drug-induced interventions aimed at reducing or increasing liver fat. The model was formulated using ordinary differential equations (ODEs) and the mathematical analysis, model simulation, model formulation and the model parameter estimation were all performed in MATLAB ResultsOur mathematical model describes accumulation of fat in the liver and predicts changes in lipid fluxes induced by both dietary and drug interventions. The model is validated using data from a wide range of drug and dietary intervention studies and can predict both short-term (days) and long-term (weeks) changes in liver fat. Importantly, the model computes the contribution of each individual lipid flux to the total liver fat dynamics. Furthermore, the model can be combined with an established bodyweight model, to simulate even longer scenarios (years), also including the effects of insulin resistance and body weight. To help prepare for corresponding eHealth applications, we also present a way to visualize the simulated changes, using dynamically changing lipid droplets, seen in images of liver biopsies. ConclusionIn conclusion, we believe that the minimal model presented herein might be a useful tool for future applications, and to further integrate and understand data regarding changes in dietary and drug induced changes in ectopic TAG in the liver. With further development and validation, the minimal model could be used as a disease progression model for steatosis.

Fabrication of a 3D bioprinting model for posterior capsule opacification using GelMA and PLMA hydrogel-coated resin.

Posterior capsule opacification (PCO) remains the predominant complication following cataract surgery, significantly impairing visual function restoration. In this study, we developed a PCO model that closely mimics the anatomical structure of the crystalline lens capsule post-surgery. The model incorporated a threaded structure for accurate positioning and observation, allowing for opening and closing. Utilizing 3D printing technology, a stable external support system was created using resin material consisting of a rigid, hollow base and cover. To replicate the lens capsule structure, a thin hydrogel coating was applied to the resin scaffold. The biocompatibility and impact on cellular functionality of various hydrogel compositions were assessed through an array of staining techniques, including calcein-AM/PI staining, rhodamine staining, BODIPY-C11 staining and EdU staining in conjunction with transwell assays. Additionally, the PCO model was utilized to investigate the effects of eight drugs with anti-inflammatory and anti-proliferative properties, including 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), THZ1, sorbinil, 4-octyl itaconate (4-OI), xanthohumol, zebularine, rapamycin and caffeic acid phenethyl ester, on human lens epithelial cells (HLECs). Confocal microscopy facilitated comprehensive imaging of the PCO model. The results demonstrated that the GelMA 60 5% + PLMA 2% composite hydrogel exhibited superior biocompatibility and minimal lipid peroxidation levels among the tested hydrogels. Moreover, compared to using hydrogel as the material for 3D printing the entire model, applying surface hydrogel spin coating with parameters of 2000 rpm × 2 on the resin-based 3D printed base yielded a more uniform cell distribution and reduced apoptosis. Furthermore, rapamycin, 4-OI and AICAR demonstrated potent antiproliferative effects in the drug intervention study. Confocal microscopy imaging revealed a uniform distribution of HLECs along the anatomical structure of the crystalline lens capsule within the PCO model, showcasing robust cell viability and regular morphology. In conclusion, the PCO model provides a valuable experimental platform for studying PCO pathogenesis and exploring potential therapeutic interventions.

Abstract LB_B04: Deciphering FGFR3-TACC3 oncogenic fusions

Abstract Chromosomal rearrangements of the fibroblast growth factor receptor (FGFR) genes that give rise to fusions are one of several mechanisms by which the FGF/FGFR signaling axis can become deregulated and result in enhanced signaling in cancer. We have previously identified truncation of exon (E) 18 of FGFR2 as a potent single-driver alteration in cancer, independently of the rearrangement (RE) partner. In contrast, the same does not appear to hold true for its E18-truncated ortholog FGFR3. We mined human oncogenomic datasets from Hartwig Medical Foundation (>2,500 WGS profiles) and Foundation Medicine (>200,000 hybrid-capture panel-seq profiles) for alterations affecting FGFR3. Examination of the structural variants affecting FGFR3 uncovered that 85% of all E18-truncating FGFR3 REs involved transforming acidic coiled-coil-containing protein 3 (TACC3) as the downstream fusion partner gene, while REs with intergenic regions and other known protein-encoding genes were much less frequent. Additionally, there was a clear predominance of self-interacting domains (95%) among all FGFR3 RE partners, with the coiled-coil domain being the most recurrent, suggesting enhanced receptor dimerization and downstream signaling capacity for the majority of the FGFR3 fusions. Our findings led to the generation of a compendium of FGFR3 structural variants which we then functionally tested. In vitro testing of mouse mammary epithelial cells expressing Fgfr3ΔE18-Tacc3E7-E16 fusion variants showed that both E18-truncation and a fusion partner were required for 3D outgrowth and signaling induction. In vivo evaluation of the oncogenic capacity of Fgfr3 variants using somatic delivery of lentiviruses to the mouse mammary gland via intraductal injection and lung via intratracheal injection found that Fgfr3ΔE18-Tacc3E7-E16 fusion variants rapidly induced mammary and lung tumor formation in wild-type and Wap-Cre;Cdh1F/F, and in wild-type and Trp53F/F mice, respectively. In contrast, Fgfr3full-length(FL), Fgfr3ΔE18 and Fgfr3FL–Tacc3E7-E16 were non-tumorigenic. Noteworthy, mammary tumors driven by Fgfr3ΔE18-Tacc3E7-E16 fusions were sensitive to the FGFR inhibitor AZD4547 during a drug intervention study. In summary, our findings show that E18-truncating FGFR3 alterations are recurrent across human cancers and, as opposed to FGFR2 E18-truncating alterations, FGFR3 appears to depend both on E18-truncation and an additional fusion partner with dimerizing capacity. Notably, TACC3 is the most recurrent 3’ fusion partner across all FGFR3 E18-truncating alterations, and somatic tumor modeling has shown that Fgfr3ΔE18-Tacc3 fusions drive mouse mammary and lung tumorigenesis, and are sensitive to FGFR targeted therapy. Citation Format: Julia Yemelyanenko, Jinhyuk Bhin, Sjoerd Klarenbeek, Ji-Ying Song, Catrin Lutz, Marieke van de Ven, Shridar Ganesan, Lodewyk F. A. Wessels, Daniel Zingg, Jos Jonkers. Deciphering FGFR3-TACC3 oncogenic fusions [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr LB_B04.

Age-dependent effect of vitamin D supplementation on musculoskeletal health in chronic spinal cord injury patients: A pilot study

Objective To determine the effect of vitamin D supplementation on changes in body composition associated with musculoskeletal health status in patients with chronic SCI and vitamin D deficiency as a response to age. Design Prospective drug-intervention study. Setting Department of rehabilitation medicine, Veterans Health Service Medical Center. Participants Seventeen patients with vitamin D insufficiency/deficiency (<30 ng/mL) and chronic SCI were divided into two groups: groups A <65 years (n = 8) and B ≥65 years of age (n = 9). Interventions Both groups received 800 IU/day cholecalciferol for 12 weeks. Outcome measures We used blood samples to evaluate metabolites related to vitamin D, testosterone (T), lipid profiles, and sex hormone-binding globulin (SHBG). Bioelectrical impedance analysis (BIA) was used to evaluate body composition. Results Group A had significantly better baseline clinical characteristics for all BIA measurements. SHGB was significantly higher in Group B (P = 0.003) and albumin was significantly higher in Group A (P = 0.000). When comparing pre- to post-treatment, Group A showed a significant improvement in T (P = 0.042), total cholesterol (P = 0.035), and triglyceride (P = 0.025) levels, whereas Group B significantly increased vitamin D (P = 0.038) and protein mass (PM) (P = 0.034) levels. Conclusion This study suggested that addressing vitamin D deficiency in patients with SCI had different effects in young and older adults, with both groups showing positive changes in body composition. Particularly, the increase in PM on BIA measurements in elderly patients at high risk of sarcopenia was encouraging.

O-263 Effect of clindamycin and a live biotherapeutic containing lactobacilli on the reproductive outcomes of IVF patients with abnormal vaginal microbiota: a double-blind, placebo-controlled multicentre trial

Abstract Study question Does treatment of abnormal vaginal microbiota improve the reproductive outcomes of IVF patients? Summary answer Data is being analysed and will be presented at ESHRE 2023. What is known already An increasing number of studies reported an association between abnormal genital tract microbiota and adverse reproductive outcomes in IVF patients. One hypothesis could be that vaginal microbiota ascend to the endometrium where the microbiota hampers implantation. To the best of our knowledge, this is the first study powered to investigate causality between abnormal genital tract microbiota - in this study defined by a bacterial vaginosis like vaginal microbiota - and clinical pregnancy rate in IVF patients. Study design, size, duration Double-blind, placebo-controlled multicentre trial in IVF patients diagnosed with abnormal vaginal microbiota and subsequently randomised into three parallel groups 1:1:1. The first group received clindamycin 300 mg ×2 daily for 7 days followed by vaginal Lactobacillus crispatus until the clinical pregnancy scan. The second group received clindamycin and placebo, whereas the third group received placebo/placebo. A total of 1518 patients were screened, and 338 patients were randomised. The study duration was from 2017 to 2023. Participants/materials, setting, methods IVF patients with any cause of infertility embarking on their first, second or third IVF stimulation cycle or embryo transfer were approached for inclusion. At a minimum 12 days prior to embryo transfer, patients were screened for abnormal vaginal microbiota defined by a qPCR assay, targeting high quantitative loads of Fannyhessea vaginae and Gardnerella spp. Patients were excluded if they had intrauterine malformations like polyps, septum, and fibroma. Main results and the role of chance Data and safety monitoring board has decided not to open the randomization code yet. The final dataset is being completed. Data is being analysed and will be presented at ESHRE 2023. Limitations, reasons for caution Data is being analysed and will be presented at ESHRE 2023. Wider implications of the findings The present drug intervention study used clindamycin and a live biotherapeutic containing Lactobacillus crispatus. The results of the study could have a worldwide important clinical impact on daily IVF practice as currently there is no evidence that screening and treating IVF patients with abnormal vaginal microbiota improves the reproductive outcome. Trial registration number EudraCT 2016-002385-31

Urine titin as a novel biomarker for Duchenne muscular dystrophy

Duchenne muscular dystrophy (DMD) is the most severe form of muscular dystrophy that is caused by lack of dystrophin, a critical structural protein in skeletal muscle. DMD treatments, and quantitative biomarkers to assess the efficacy of potential treatments, are urgently needed. Previous evidence has shown that titin, a muscle cell protein, is increased in the urine of patients with DMD, suggesting its usefulness as a DMD biomarker. Here, we demonstrated that the elevated titin in urine is directly associated with the lack of dystrophin and urine titin responses to drug treatment. We performed a drug intervention study using mdx mice, a DMD mouse model. We showed that mdx mice, which lack dystrophin due to a mutation in exon 23 of the Dmd gene, have elevated urine titin. Treatment with an exon skipper that targets exon 23 rescued muscle dystrophin level and dramatically decreased urine titin in mdx mice and correlates with dystrophin expression. We also demonstrated that titin levels were significantly increased in the urine of patients with DMD. This suggests that elevated urine titin level might be a hallmark of DMD and a useful pharmacodynamic marker for therapies designed to restore dystrophin levels.

Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles.

Small extracellular vesicles (sEV) can be released from all cell types and carry protein, DNA, and RNA. Signaling molecules serve as indicators of the physiological and pathological state of a cell. However, there is no standard method for sEV isolation, which prevents downstream biomarker identification and drug intervention studies. In this article, we provide a detailed protocol for the isolation and purification of 50-200 nm sEV by a flow cell sorter. For this, a 50 µm nozzle and 80 psi sheath fluid pressure were selected to obtain a good sorting rate and stable side stream. Standard sized polystyrene microspheres were used to locate populations of 100, 200, and 300 nm particles. With additional optimization of the voltage, gain, and forward scatter (FSC) triggering threshold, the sEV signal could be separated from the background noise. These optimizations provide a panel of critical sort settings that enables one to obtain a representative population of sEV using FSC vs. side scatter (SSC) only. The flow cytometry-based isolation method not only allows for high-throughput analysis but also allows for synchronous classification or proteome analysis of sEV based on the biomarker expression, opening numerous downstream research applications.

HDAC6 inhibition attenuates renal injury by reducing IL-1β secretion and RIP kinase mediated necroptosis in acute oxalate nephropathy

BackgroundPathogenesis of acute kidney injury is driven by necro-inflammation, which is comprised of IL-1β mediated inflammation and RIP-1 mediated tubular necroptosis. HDAC6 is reported to regulate both inflammation and cell death. In the present study, we explored the role of HDAC6 in the lysosomal exocytosis of IL-1β and RIP-1 mediated necroptosis in the context of oxalate nephropathy. MethodsRaw 264.7 macrophages and NRK52E stimulated with oxalate crystals and LPS with or without HDAC6 inhibitor for in vitro experiments. Acute oxalate nephropathy was induced in C57BL/6 mice by injecting sodium oxalate (75 mg/kg). For the drug intervention study, Tubastain A (TSA) was given an hour before injection of sodium oxalate. Mice were sacrificed 24 hrs after the oxalate injection, blood and kidney were harvested. Blood samples were analyzed for BUN and IL-1β levels. Renal tissues were analyzed for histology, immunohistochemistry, RNA, and protein expression. ResultsHDAC6 and IL-1β upregulated in crystal stimulated macrophages and acute oxalate nephropathy. Pre-treatment of macrophages with TSA reduced IL-1β in supernatant without affecting the expression of pro-IL-1β and mature IL-1β in cell lysate. The effect of TSA on IL-1β secretion was influenced by tubulin acetylation. Renal epithelial cell NRK52E stimulated with crystals showed upregulation of necroptosis pathway markers and concentration-dependent cell death. TSA inhibited RIP-1, RIP3, and MLKL expression along with p-MLKL in stimulated epithelial cells. TSA treatment of oxalate nephropathy mice showed decreased inflammation and tubular cell death by regulating IL-1β and necroptosis and reduced renal injury. ConclusionThis study highlights the role of HDAC6 in regulating the tubulin-mediated secretion of IL-1β and RIP kinase mediated necroptosis in acute oxalate nephropathy.

Effect of clindamycin and a live biotherapeutic on the reproductive outcomes of IVF patients with abnormal vaginal microbiota: protocol for a double-blind, placebo-controlled multicentre trial

IntroductionRecent studies in in vitro fertilisation (IVF) patients have associated abnormal vaginal microbiota (AVM) with poor clinical pregnancy rates of 6%–9% per embryo transfer. The biological plausibility for this finding...

Characteristics of Clinical Trials relating to COVID-19 registered at ClinicalTrials.gov.

Since the beginning of the COVID-19 outbreak in China in December 2019, the epidemic has continued to spread globally. Despite continuous reports of clinical trials being launched, no studies have yet systematically summarized and analysed their characteristics. Our objective is to do this by reviewing trials registered at ClinicalTrials.gov. We searched the ClinicalTrials.gov database and retrieved all clinical trials on COVID-19 registered up to and including 3 April 2020. We summarized the characteristics of the trials, presenting the results of all trials, all intervention trials and drug intervention (including vaccines and traditional Chinese medicine) trials. We identified 306 COVID-19-related clinical trials. Seven of the studies had been withdrawn, leaving 299 active trials. Of the trials, 28.8% were planned to be conducted in Asia, 26.8% in Europe and 18.7% in North America. Most (73.0%) proposed trials expected to recruit fewer than 500 people, and only 22.1% of the studies included children (aged <18years). About two-thirds (67.2%) of the studies were funded by the own resources of medical or research institutions. Of intervention trials, 73.9% used random allocation, and 73.4% used parallel assignment. Only 36.7% of the intervention trials used blinding. In terms of drug trials, 147 trials were drug intervention studies, covering 80 conventional drugs and seven traditional Chinese medicine drugs. Antiviral drugs and antimalarial drugs were the most commonly studied drugs with 52 and 45 trials registered, respectively. Five registered clinical trials were on vaccines. A large number of COVID-19-related trials have been registered within the first 4months since the first infection was reported. These involve a large number of different drugs, the most common being antiviral drugs and antimalarial drugs. More attention should be paid to adequate blinding in future trials.

Analysis of 30 clinical trials on rare neurological diseases in children

Objective To explore the methods for generating evidence on health outcomes in children with rare diseases. Methods The data from 30 clinical trials on rare neurological diseases in children from January 2008 to December 2018 were collected and analyzed.Statistical analysis was conducted on the relationship between the study sponsor and the study center, the number of participants and the prevalence rate. Results Thirty studies involved 6 types of diseases, including 14 kinds of diseases.(1) All global multicenter studies (14 items) were initiated by pharmaceutical companies, whereas most of single-center studies (6/7 kinds, 86%) and multiple centers within one country(7/9 kinds, 78%) were initiated by investigators.There was a significant correlation between the research center and the research sponsor(P<0.001). (2) Most of the drugs studied were selected based on previous clinical trials (9/30 items) and animal experiments (9/30 items). (3) The median number of participants included 39 cases (10-215 cases), and 60%(18/30 items) of the studies was fewer than 50 cases.(4) Study design: 53%(16/30 items) of studies were randomized controlled studies, 33%(10/30 items) studies were open-label single-arm studies, and 14% (4/30 items) were randomized cross-over trials.Seventy-five percent(12/16 items)of randomized controlled studies were initiated by pharmaceutical companies, 50%(5/10 items) open-label single-arm studies and all randomized cross-over trials were initiated by investigators.There was a statistical correlation between the study sponsors and the study design method (χ2=7.602, P=0.022). (5) Outcome index: Scale score was used as the primary outcome in half of studies.Other studies used symptom improvement or pathological changes. Conclusions Clinical trials in rare diseases enrolled fewer participants than that in non-rare diseases, and the study design method was relatively simple.Therefore, it is necessary to further improve the level of evidence of clinical research of rare diseases through global and multi-center recruitment, initiation of pharmaceutical companies and improving the study design method. Key words: Child; Rare diseases; Drug intervention study; Research design method

A standardised framework to identify optimal animal models for efficacy assessment in drug development.

IntroductionPoor translation of efficacy data derived from animal models can lead to clinical trials unlikely to benefit patients–or even put them at risk–and is a potential contributor to costly and unnecessary attrition in drug development.ObjectivesTo develop a tool to assess, validate and compare the clinical translatability of animal models used for the preliminary assessment of efficacy.Design and resultsWe performed a scoping review to identify the key aspects used to validate animal models. Eight domains (Epidemiology, Symptomatology and Natural History–SNH, Genetic, Biochemistry, Aetiology, Histology, Pharmacology and Endpoints) were identified. We drafted questions to evaluate the different facets of human disease simulation. We designed the Framework to Identify Models of Disease (FIMD) to include standardised instructions, a weighting and scoring system to compare models as well as factors to help interpret model similarity and evidence uncertainty. We also added a reporting quality and risk of bias assessment of drug intervention studies in the Pharmacological Validation domain. A web-based survey was conducted with experts from different stakeholders to gather input on the framework. We conducted a pilot study of the validation in two models for Type 2 Diabetes (T2D)–the ZDF rat and db/db mouse. Finally, we present a full validation and comparison of two animal models for Duchenne Muscular Dystrophy (DMD): the mdx mouse and GRMD dog. We show that there are significant differences between the mdx mouse and the GRMD dog, the latter mimicking the human epidemiological, SNH, and histological aspects to a greater extent than the mouse despite the overall lack of published data.ConclusionsFIMD facilitates drug development by serving as the basis to select the most relevant model that can provide meaningful data and is more likely to generate translatable results to progress drug candidates to the clinic.

GW29-e0752 Study on the Early Evaluation and Screening Method of Heart Failure Model after Myocardial Infarction from the Correlation Analysis of Electrocardiogram and Ultrasonic Cardiogram

To establish a new method for early evaluation and screening of heart failure model after myocardial infarction and provide basis for early random grouping on drug intervention study. SD Rats were randomly divided into model and sham group. The MI models were established by ligating left anterior

Improving the quality of preclinical research echocardiography: observations, training, and guidelines for measurement.

Informal training in preclinical research may be a contributor to the poor reproducibility of preclinical cardiology research and low rates of translation into clinical research and practice. Mouse echocardiography is a widely used technique to assess cardiac structure and function in drug intervention studies using disease models. The interobserver variability of clinical echocardiographic measurements has been shown to improve with formalized training, but preclinical echocardiography lacks similarly critical standardization of training. The aims of this investigation were to assess the interobserver variability of echocardiographic measurements from studies in mice and address any technical impediments to reproducibility by implementing standardized guidelines through formalized training. In this prospective, single-site, observational cohort study, 13 scientists performing preclinical echocardiographic image analysis were assessed for measurement of short-axis M-mode-derived dimensions and calculated left ventricular (LV) mass. Ten M-mode images of mouse hearts acquired and analyzed by an expert researcher with a spectrum of LV mass were selected for assessment and validated by autopsy weight. After the initial observation, a structured formal training program was introduced, and accuracy and reproducibility were reevaluated. Mean absolute percentage error for expert-calculated LV mass was 6 ± 4% compared with autopsy LV mass and 25 ± 21% for participants before training. Standardized formal training improved participant mean absolute percentage error by ~30% relative to expert-calculated LV mass ( P < 0.001). Participants initially categorized with high-range error (25-45%) improved to low-moderate error ranges (<15-25%). This report reveals an example of technical skill training insufficiency likely endemic to preclinical research and provides validated guidelines for echocardiographic measurement for adaptation to formalized in-training programs. NEW & NOTEWORTHY The informal training common to academic/research institutions may be a contributor to the relatively poor reproducibility observed for preclinical cardiac research. In our observation of echocardiography analysis in murine models, we present evidence of moderate interobserver variability in standard preclinical research practice at an Australian heart research institute. These observations give rise to our recommendations for practical guidelines for echocardiography analysis in an adaptable approach to general preclinical research skill training. Listen to this article's corresponding podcast at https://ajpheart.podbean.com/e/preclinical-echocardiography-training-and-guidelines/ .

‘Ice Rushes’, Data Shadows and Methylamphetamine Use in Rural Towns: Wastewater Analysis

Australia's world-class drug monitoring systems have difficulty gathering metrics in rural communities for reasons due to, among other things, the size of the country and problems with recruiting sufficient sample sizes. Some rural communities in data shadows (where few metrics on substance use exist) may benefit from wastewater analysis (‘WWA’) as a means of estimating per capita drug consumption. Wastewater analysis could be employed when debates rise about the consumption of particular drugs in certain communities. Other ways to use WWA are examined, including long-term monitoring of community drug consumption and intervention studies to test the effectiveness of health or law enforcement drug strategies. To explore the utility of WWA, this article references media coverage of methylamphetamine consumption in a small Tasmanian town, Smithton, and presents the results of the first Tasmanian WWA pilot study of methylamphetamine consumption, conducted in 2014–15.

Applying metabolomics to cardiometabolic intervention studies and trials: past experiences and a roadmap for the future.

Metabolomics and lipidomics are emerging methods for detailed phenotyping of small molecules in samples. It is hoped that such data will: (i) enhance baseline prediction of patient response to pharmacotherapies (beneficial or adverse); (ii) reveal changes in metabolites shortly after initiation of therapy that may predict patient response, including adverse effects, before routine biomarkers are altered; and( iii) give new insights into mechanisms of drug action, particularly where the results of a trial of a new agent were unexpected, and thus help future drug development. In these ways, metabolomics could enhance research findings from intervention studies. This narrative review provides an overview of metabolomics and lipidomics in early clinical intervention studies for investigation of mechanisms of drug action and prediction of drug response (both desired and undesired). We highlight early examples from drug intervention studies associated with cardiometabolic disease. Despite the strengths of such studies, particularly the use of state-of-the-art technologies and advanced statistical methods, currently published studies in the metabolomics arena are largely underpowered and should be considered as hypothesis-generating. In order for metabolomics to meaningfully improve stratified medicine approaches to patient treatment, there is a need for higher quality studies, with better exploitation of biobanks from randomized clinical trials i.e. with large sample size, adjudicated outcomes, standardized procedures, validation cohorts, comparison witth routine biochemistry and both active and control/placebo arms. On the basis of this review, and based on our research experience using clinically established biomarkers, we propose steps to more speedily advance this area of research towards potential clinical impact.

GW26-e4459 Cellular Electrophysiological Mechanisms and Drug Intervention Study of the CACNA1C Gene Mutation Related to Early Repolarization

GW26-e4459 Cellular Electrophysiological Mechanisms and Drug Intervention Study of the CACNA1C Gene Mutation Related to Early Repolarization

Abstract 1850: Evaluation of Pim-1 kinase inhibition in a preclinical model of mouse prostate cancer

Abstract Pim-1 is a serine/threonine kinase that is in involved in the transcription of genes that regulate apoptosis and cell cycle progression. Pim-1 has been shown to be overexpressed in a variety of cancers and has been implicated in human prostate tumorigenesis. The aim of our study was to clarify the role of Pim-1 during the prostate tumorigenesis and determine its potential as a molecular target for therapy. To accomplish this goal, we carried out a series of experiments using a genetically engineered mouse model of prostate cancer driven by the conditional inactivation of PTEN. High levels of Pim-1 were observed during the early stages of the tumorigenesis process which decreased as tumors progressed. Expression of Pim-1 was localized in the nucleus and cytoplasm of prostatic intraepithelial neoplasia (PIN), well-differentiated, and moderately-differentiated tumors. In poorly-differentiated tumors, Pim-1expression was heterogeneously expressed at lower levels and was predominantly localized to the cytoplasm of cancer cells. Levels of Pim-1 expression were increased in tumors from mice after surgical castration. To evaluate the therapeutic potential of Pim-1as a target for prostate cancer, we tested the antitumor effects of AZD1208, a highly potent Pim kinase inhibitor, on a panel of cell lines derived from castration-naïve and castration-resistant mouse prostate cancer. AZD1208 inhibited the tumor cell growth of several cell lines and demonstrated higher sensitivity to those derived from castration resistant tumors. In vivo, drug intervention studies show that treatment with AZD1208 reduced tumor growth of castration-naïve and castration-resistant prostate tumors by 12.1 (P = 0.162) and 24.3% (P&amp;lt;0.001), respectively. These findings provide further support for a role of Pim-1 during prostate tumorigenesis and could thus serve as a rational target for the development of novel combination strategies. Citation Format: Marco A. De Velasco, Takashi Oki, Yurie Kura, Naomi Ando, Emiko Fukushima, Barry R. Davies, Dennis Huszar, Yutaka Yamamoto, Yuji Hatanaka, Nobutaka Shimizu, Kazuhiro Yoshimura, Masahiro Nozawa, Kazuhiro Yoshikawa, Kazuto Nishio, Hirotsugu Uemura. Evaluation of Pim-1 kinase inhibition in a preclinical model of mouse prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1850. doi:10.1158/1538-7445.AM2015-1850

Commentary: dissecting the PNPLA3 association with liver fat and stiffness, and interaction with diet

We read the article by Shen et al. with great interest, and highlighted some of the key features of this study.1 The authors conducted a prospective cohort study and reported that the rs738409 GG variant in patatin-like phospholipase domain-containing protein 3 (PNPLA3) genotype was associated with an increased risk of non-alcoholic fatty liver disease (NAFLD) by magnetic resonance spectroscopy (MRS) of the liver, and the presence of gene effect on trait was not explained by the diet history recorded by the food frequency questionnaire. In multivariable-adjusted analyses, metabolic syndrome was a much stronger correlate of presence of hepatic steatosis than PNPLA3 genotype. Incremental net reclassification rate (although not shown) by including PNPLA3 genotype in addition to clinical risk factors of NAFLD appeared modest, and not clinically significant, when these results are seen in context of the broader themes in the field at this time. This study provides validation that PNPLA3 genotype is associated with increased risk of NAFLD in an urban Asian population, predominantly of Chinese origin, residing in Hong Kong. The investigators also found that liver stiffness measured by transient elastography did not reveal any association with PNPLA3 genotype. These data suggest that liver stiffness in NAFLD is not affected by PNPLA3 genotype, and possibly other genetic variants may be associated with hepatic fibrosis and/or liver stiffness. Previous studies have shown that the association of the GG variant and hepatic lipid content is associated with the presence of other risk factors, such as severe obesity,2 visceral adiposity,3 increased intake of sugars4 and omega-6 poly-unsaturated fatty acids.5 However, the results presented by Shen and colleagues do not find an association. Most of the prior association studies were conducted in Caucasian populations in contrast to the study by Shen et al., which is derived from a biogeo-graphically distinct population compared with previous studies. However, the association between NAFLD and various metabolic traits was consistent across all previous studies, as well as this study, suggesting some generalis-ability between cohorts.6–10 Overall, the study provides four key messages. First, it confirms the association between PNPLA3 genotype and liver fat content by MRS in a Chinese population residing in Hong Kong. Second, PNPLA3 genotype effect on trait was not found to be influenced by diet in this study. Third, presence of metabolic syndrome trumps PNPLA3 genotype in it's association with the trait (i.e., NAFLD). Fourth, PNPLA3 genotype did not influence liver stiffness. These findings underscore the need to better understand the molecular mechanisms by which the GG variant is linked to fat accumulation in the liver, and further research is warranted to unravel the physiological role of the PNPLA3 in times of starvation and caloric excess. Improved understanding of the role of PNPLA3 could open new areas of investigations and may provide novel avenues for drug development and intervention studies to reverse NAFLD.