Yong Huang, M.D., Ph.D. - US grants

DESCRIPTION (provided by applicant): Use of human primary cells (from health donors or patients) as physiology- or disease-relevant models provides significant advantages in basic research and therapeutic development for understanding and treating various human diseases, including cancer, cardiovascular and inflammatory diseases. Unfortunately, utility of precious primary cells is largely hindered by lack of effective means for transfection with minimal side effects, particularly in high-throughput mode. Our proposal is specifically designed to address this critical unmet need. Our Phase I studies have demonstrated that our technology is superior than existing methods in transfecting primary cells, especially fully differentiated primary cell monolayers. Phase II work is proposed to develop a 96-well high-throughput transfection system, and to further validate its applicability in extended primary cultures, including both adherent and suspension cells. Success in this project would have notable impact on basic research and therapeutic development by expanding the utility of primary cells, which are the best in vitro models to human. Success in this project will benefit public health by advancing basic research in decoding the functional roles of ~25,000 human genes, and by impacting therapeutic development for fighting diseases like cancer and inflammation.

The studies will characterize the absoprtion, distribution, metabolism, and elimination of potential medications for drug abuse treatment.

DESCRIPTION (provided by applicant): RNA interference (RNAi), a biological process for modulating gene expression using short interfering RNA (siRNA) molecules, has emerged as one of the most promising technologies for treating a wide range of difficult-to-cure diseases, including liver diseases, cancer, neurodegenerative diseases, etc. However, lack of effective means to deliver siRNA into targeted organs in the human body significantly hinders clinical applications of RNAi technology, particularly in liver therapy. Our proposal is specifically designed to address this critical challenge. Phase I work will focus on development of a patent-pending targeted siRNA delivery technology with real-time optimization capability. Applying the technology, we will optimize siRNA delivery to ex vivo liver tissue and demonstrate its clinical relevance by delivering specific siRNA to functionally knockdown certain therapeutic targets of liver diseases. Successful Phase I development will lead to extensive future animal study to demonstrate the utility of this novel siRNA delivery technology in controlling liver diseases, such as liver cirrhosis and alcoholic liver, which could potentially benefit millions of patients suffering from these formidable liver diseases. PUBLIC HEALTH RELEVANCE: Success in this project will accelerate the progress of transferring advanced siRNA based therapeutic technology into clinical application. It will benefit public health by revolutionizing clinical treatment of a broad range of difficult-to-cure diseases such as cancers, liver diseases, viral infection, and cardiovascular diseases, etc.

The Division of Pharmacotherapies and Medical Consequences of Drug Abuse (DPMCDA) of NIDA supports research and development of new medications for the treatment of drug addictions. A critical aspect of this development involves studies on the metabolism, pharmacokinetics and clinical pharmacology of new medications. Biological specimens from these studies require quantitative determination of the study medication and its metabolites and the abused substances by highly sensitive and specific methodology. The purpose of this contract is to support the Institute's efforts in these areas by providing a centralized analytical facility capable of carrying out analyses on a wide variety of compounds, e.g., illicit drugs, potential medications and their metabolites, as well as endogenous substances, in biological fluids in support of medications development.

This contract provides a resource for conducting preclinical pharmacology, pharmacokinetic, and metabolism studies with candidate therapeutic agents selected for development by the Division of Cancer Treatment, NCI. Most of the compounds selected are potential anticancer agents, but some are putative therapeutics for a wide range of prevalent and orphan human diseases. Compounds selected for study under this contract arise from peer-reviewed applications submitted to NCI by academic investigators, pharmaceutical companies, or others. Work carried out under the contract may include: (1) development of analytical methods to quantify compounds in plasma, urine, and other biological matrices; (2) plasma stability and protein binding studies; (3) pharmacokinetic characterization following administration to animals by various routes and schedules; (4) quantification and identification of drug metabolites generated in vivo and in various in vitro systems; and (5) assessment of pharmacodynamic effects in tumor or surrogate tissues and correlation of these effects with drug levels and/or total drug exposures. The characterizations obtained from these pharmacokinetic investigations will be used in the prioritization and selection of agents for further development; in the design of preclinical efficacy and toxicology studies; and in regulatory submissions in support of clinical trials of selected agents.

DESCRIPTION (provided by applicant): Transporters are a class of more than 1000 membrane proteins that regulate how nutrients and drugs enter and leave biological cells. Transporters have validated clinical significance and affect every aspect of drug ADME (Absorption, Distribution, Metabolism, Elimination), and they also mediate drug-drug interactions (DDIs) that can lead to serious Adverse Drug Reactions (ADRs). Utilizing the full potentials of transporters opens new possibilities of designing safer and more efficacious drugs, through improving drug bioavailability, achieving more targeted tissue distribution and reducing and/or managing ADRs due to DDI. Despite that many transporters are known to influence drug levels, suitable cell lines, specific transporter inhibitors, and predictive computational models are not generally available. In the proposed Phase II studies we will address these unmet needs by developing transporter assays for 27 important drug transporters in humans and rats, using the Opti-Expression technology developed in Phase I. In addition, through comprehensive screening of 1000 prescription drugs against 6 high priority drug uptake transporters, this research will provide a basis for identification of potential transporter-mediated DDIs in the liver and kidney. Importantly, these screens represent the largest and most complete screen of compounds for inhibition of transporters. Compounds identified as potential transporter inhibitors can be tested in follow-up clinical DDI studies. The research will lead to the discovery of specific and general inhibitors that can be used as tools in in vitro and pre-clinical (as well as clinical) in vivo studies to identify transporters involved in drug absorption and disposition, and to determine the mechanisms that underlie clinical DDIs. Currently, there is no database or knowledgebase that houses specific large-scale information on transporter mediated DDIs. This study will facilitate the establishment of a transporter inhibition database, which is based on the data obtained in the proposed screening studies. This database will be an important tool for scientists in academia, industry and at the FDA. PUBLIC HEALTH RELEVANCE: Success of this project will benefit public health by facilitating discovery and development of more efficacious drugs for treating various formidable diseases, particularly CNS diseases and cancer, and by improving drug safety through reducing adverse drug effects due to unwanted tissue distribution and drug-drug interactions.

This research and development contract will provide bioanalytical chemistry resources to support NIDA's medications development program. This contract will also provide as a centralized bioanalytical facility to carry out quantitative and/or qualitative assays for a variety of compounds, including but not limited to, new medications and/or their metabolites, abused drugs/metabolites, potential concomitant medications/compounds and molecular biomarkers (small molecules, peptides and proteins).

DESCRIPTION: CNS Transporters have crucial roles on neurophysiology and etiology. Inhibition of these critical transporters can have therapeutic benefits or cause serious side effects in the CNS. Development of an assay platform and a comprehensive profile of drugs' inhibitory effects on important CNS transporters can have enormous impact on drug discovery and development, such as elucidating drugs' potential side effects in the CNS, and advancing development of new drugs to treat various CNS diseases, such as Alzheimer's disease, Parkinson's diseases, epilepsy etc., The overall goal of the proposed studies is to further the knowledge and available tools to study drug interactions with membrane transporters localized through-out the CNS. Built upon successful Phase I studies, the Phase II studies will result in the most comprehensive collection of CNS transporter assays (25+) commercially available to academic and industrial researchers, removing a critical assay bottleneck in transporter research and CNS drug development. The screening studies (350 CNS and peripherally acting against 20+ key CNS transporters) represent the largest effect to date on in vitro inhibition screen, characterization and in vitro-in vivo extrapolation (IVIVE) of prescription drugs against a panel of key CNS transporters. This innovative approach will provide critical information fundamental to CNS transporter pharmacology, moreover, it could potential lead to discovery of new therapeutic indications for approved drugs.

This contract provides a resource for conducting nonclinical pharmacology, pharmacokinetic, and metabolism studies with candidate therapeutic agents selected for development by the Division of Cancer Treatment, NCI. Candidate therapies selected for study under this contract arise from peer-reviewed applications submitted to NCI by academic investigators, pharmaceutical companies, or others. Work carried out under the contract includes: (1) development of analytical methods to quantify the levels of test agents in biological matrices; (2) plasma stability and protein binding studies; (3) pharmacokinetic characterization following administration by various routes and schedules; (4) quantification and identification of metabolites generated; and (5) assessment of pharmacodynamic effects in tumor or surrogate tissues and correlation of these effects with levels of agent and/or total exposures. The characterizations obtained from these pharmacokinetic investigations are used in the prioritization and selection of agents for further development; in the design of preclinical efficacy and toxicology studies and in regulatory submissions in support of clinical trials of selected agents.

The Contractor shall conduct non-clinical Absorption, Metabolism, Distribution, and Elimination (ADME) studies, protein binding studies, and analytical services in support of NIDA medication development program.

This award supports a workshop to bring together additive manufacturing researchers in the broad field of health. Additive manufacturing describes a collection of manufacturing processes that produce parts layer-by-layer, controlled by a computer, and without the use of expensive tools or dies. This manufacturing strategy has been an enabling technology for a large number of existing and emerging biomedical research and medical industry applications, and the approach has become widespread in recent years. Additive manufacturing is expected to fundamentally change the nature of advanced manufacturing and revolutionize many areas of research in the 21st Century. The workshop will help realize the full potential of additive manufacturing technology for various health-related applications and products. First, the increased awareness of additive manufacturing for health will increase the level of interest in manufacturing research in general and additive manufacturing in particular. Any research thrusts initiated subsequently based on workshop recommendations and a better understanding of additive manufacturing for health are expected to increase the further adoption of additive manufacturing in various medical research and development efforts.

The workshop will last 1.5 days, and will include thought leaders from industry and academe to address the future of additive manufacturing in health as well as the research accomplishments to date and the basic research needs. Invited speakers will make presentations regarding the future applications of additive manufacturing in health. Further invited speakers will address funding perspectives, both in terms of needs and the prognosis for funding support. Speakers will be recruited from industry, academe, and government in order to obtain a broad perspective. The objectives of the workshop are to: (1) review the state-of-the-art in basic research in additive manufacturing for health; (2) examine future prospects of additive manufacturing for health; (3)share perspectives on additive manufacturing for health from funding agencies such as DoD, NIH, NIST, and NSF; (4) identify needs, gaps, and challenges facing additive manufacturing for health, for both existing products and new enabled technologies; and (5) formulate recommendations for basic research initiatives.

DESCRIPTION (provided by applicant): Discovery and development of a drug from early to late stages on average cost 2 billion dollars, and require many different types of simulation and modeling tools in order to help scientists to better understand the drug disposition mechanisms and predict possible outcomes within various experimental settings. Importantly, because modeling approach involves assumptions and reductions of certain components while new concepts and knowledge should validate or replace the old ones, the approach should be built on the fundamentally strong algorithms and modules that can be easily refined, rather than curve-fitting and using fudge factors. Here, we will develop a mechanistic modeling platform to make better predictions in pharmacokinetics and cellular drug disposition for drug-like molecules in human using primarily in vitro data. We use mathematical approach to simulate drug clearance in a well-defined in vitro system constructed with our patented technology that allows us to tailor the expression of drug transporters in MDCK cell monolayers, and measure intrinsic parameters of individual component of transcellular drug transport. Thus our novelty comes from measuring and incorporating the true, intrinsic kinetic parameters at the site of action int the model as opposed to apparent observations. The models for drug-drug interaction, as well as transporter-metabolizing enzyme interplay which affects the temporal dynamics of disposition will undergo a series of rigorous validation processes tailored to each specific example (rosuvastatin-rifampicin, metformin-cimetidine, and erythromycin-CYP3A4 metabolite). Each models will incorporate new concepts, such as, intracellular unbound concentration, intrinsic efflux kinetic parameters (Vmax,intrinsic and Km,intrinsic), and steady-state concentration-dependent SLC transporters generally discussed among International Transporter Consortium and regulatory agencies. Although selected test models represent the liver and renal transporter-mediated disposition, the intrinsic parameters and our in vitro expression systems together provide fundamental building blocks in the scope of systems pharmacology, allowing future modification to other scenarios and upgrades (e.g. brain or intestine, etc.), and promote application of modeling and simulation in the field of science and biotechnology business.

The National Institute on Drug Abuse (NIDA) supports research and development of new medications for the treatment of addiction. Medication development involves in vitro evaluations, non-clinical pharmacology and toxicology studies, pharmaceutical development, and clinical evaluations of potentiaal new medications for treating addiction. A critical aspect of the non-clinical evaluations involves characterizing the pharmacokinetics (PK) of the medication under development. Pharmacokinetics provides an assessment of the Absorption, Distribution, Metabolism, and Excretion (ADME) properties, which is critical in all phases of a fully integrated medication development program. The ADME studies provide supportive information to understand pharmacological effects and toxciological effects of a potential medication and its metabolites. Furthermore, the data generated from the ADME studies provide a basis for the selection of a proper animal model for pharmacological/toxicological evaluations, the selection of promising medications for clinical development, and rational design of dosage forms and dosage regimens. The non-clinical ADME studies for this contract typically include conducting the following: Bioavailability and pharmacokinetic studies, mass balance studies, tissue distribution studies, and toxicokinetic analysis of new potential medications. Additionally, this contract involves conducting analytical services which include structure identification of major metabolites by mass spectrometry or nuclear magnetic resonance(NMR) and the development of sensitive and specific assays for a potential medication and its metabolites in biological matrices using methods such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), liquid chromatography with tandem mass spectrometry (LC/MS/MS), and gas chromatograph mass spectrometry (GCMS).