Gillian Murphy, Ph.D.
Professor, Department of Oncology, University of Cambridge, UK
A novel strategy for targeting metalloproteinases in cancer
Epithelial tumours evolve in a multi-step manner, involving both inflammatory and mesenchymal cells. Although intrinsic factors drive malignant progression, the influence of the micro-environment of neoplastic cells is a major feature of tumorigenesis. Extracellular proteinases, notably the metalloproteinases, are key players in the regulation of this cellular environment, acting as major effectors of both cell-cell and cell-extracellular matrix (ECM) interactions. They are involved in modifying ECM integrity, growth factor availability and the function of cell surface signalling systems, with consequent effects on cellular differentiation, proliferation and apoptosis.This has made metalloproteinases important targets for therapeutic interventions in cancer and small molecule inhibitors focussed on chelation of the active site zinc and binding within the immediate active site pocket were developed. These were not successful in early clinical trials due to the relative lack of specificity and precise knowledge of the target proteinase(s) in specific cancers. We can now appreciate that it is essential that we understand the relative roles of the different enzymes (of which there are over 60) in terms of their pro and anti tumour activity and their precise sites of expression The next generations of metalloproteinase inhibitors need the added specificity that might be gained from an understanding of the structure of individual active sites and the role of extra catalytic domains in substrate binding and other aspects of their biology. We have prepared scFv antibodies to the extra catalytic domains of two membrane metalloproteinases, MMP-14 and ADAM17, that play key roles in the tumour microenvironment. Our rationale and experiences with these agents will be presented in more detail.
Nader Pourmand, Ph.D.
Director, UCSC Genome Technology Center,University of California, Santa Cruz
Biosensor and Single Cell Manipulation using Nanopipettes
Approaching sub-cellular biological problems from an engineering perspective begs for the incorporation of electronic readouts. With their high sensitivity and low invasiveness, nanotechnology-based tools hold great promise for biochemical sensing and single-cell manipulation. During my talk I will discuss the incorporation of electrical measurements into nanopipette technology and present results showing the rapid and reversible response of these subcellular sensors to different analytes such as antigens, ions and carbohydrates. In addition, I will present the development of a single-cell manipulation platform that uses a nanopipette in a scanning ion-conductive microscopy technique. We use this newly developed technology to position the nanopipette with nanoscale precision, and to inject and/or aspirate a minute amount of material to and from individual cells or organelle without comprising cell viability. Furthermore, if time permits, I will show our strategy for a new, single-cell DNA/ RNA sequencing technology that will potentially use nanopipette technology to analyze the minute amount of aspirated cellular material.
Lalitha Subramanian, Ph.D.
Chief Scientific Officer & VP, Services at Scienomics, USA
Nanoscale Simulations – Tackling Form and Formulation Challenges in Drug Development and Drug Delivery
Lalitha Subramanian, Dora Spyriouni, Andreas Bick, Sabine Schweizer, and Xenophon Krokidis Scienomics
The discovery of a compound which is potent in activity against a target is a major milestone in Pharmaceutical and Biotech industry. However, a potent compound is only effective as a therapeutic agent when it can be administered such that the optimal quantity is transported to the site of action at an optimal rate. The active pharmaceutical ingredient (API) has to be tested for its physicochemical properties before the appropriate dosage form and formulation can be designed. Some of the commonly evaluated parameters are crystal forms and polymorphs, solubility, dissolution behavior, stability, partition coefficient, water sorption behavior, surface properties, particle size and shape, etc. Pharmaceutical development teams face the challenge of quickly and efficiently determining a number of properties with small quantities of the expensive candidate compounds. Recently the trend has been to screen these properties as early as possible and often the candidate compounds are not available in sufficient quantities. Increasingly, these teams are leveraging nanoscale simulations similar to those employed by drug discovery teams for several decades. Nanoscale simulations are used to predict the behavior using very little experimental data and only if this is promising further experiments are done. Another aspect where nanoscale simulations are being used in drug development and drug delivery is to get insights into the behavior of the system so that process failures can be remediated and formulation performance can be improved. Thus, the predictive screening and the in-depth understanding leads to experimental efficiency resulting in far-reaching business impacts.
With specific examples, this talk will focus on the different types of nanoscale simulations used to predict properties of the API in excipients and also provide insight into system behavior as a function of shelf life, temperature, mechanical stress, etc.
John Stanley, Satheesh Babu, Ramacahandran T and Bipin Nair
Pt-Pd decorated TiO2 nanotube array for the non-enzymatic determination of glucose in neutral medium
Rapidly expanding diabetic population and the complications associated with elevated glycemic levels necessitates the need for a highly sensitive, selective and stable blood glucose measurement strategy. The high sensitivity and selectivity of enzymatic sensors together with viable manufacturing technologies such as screen-printing have made a great social and economic impact. However, the intrinsic nature of the enzymes leads to lack of stability and consequently reduces shelf life and imposes the need for stringent storage conditions. As a result much effort has been directed towards the development of ‘enzyme-free’ glucose sensors (Park et al. 2006). In this paper, a non-enzymatic amperometric sensor for selective and sensitive direct electrooxidation of glucose in neutral medium was fabricated based on Platinum-Palladium (Pt–Pd) nanoparticle decorated titanium dioxide (TiO2) nanotube arrays. Highly ordered TiO2 nanotube arrays were obtained using a single step anodization process (Grimes C A and Mor G K 2009) over which Pt–Pd nanoparticles where electrochemically deposited. Scanning Electron Microscopy (SEM) analysis revealed the diameter of the TiO2 nanotubes to be approximately 40 nm. Elemental analysis after electrochemical deposition confirms the presence of Pt–Pd. Electrochemical characterization of the sensor was carried out using cyclic voltammetry technique (−1.0 to +1.0V) in phosphate buffer saline (PBS) pH 7.4. All further glucose oxidation studies were performed in PBS (pH 7.4). The sensor exhibited good linear response towards glucose for a concentration range of 1 μM to 20mM with a linear regression coefficient of R = 0.998. The electrodes are found to be selective in the presence of other commonly interfering molecules such as ascorbic acid, uric acid, dopamine and acetamidophenol. Thus a nonenzymatic sensor with good selectivity and sensitivity towards glucose in neutral medium has been developed.