Research

All projects and Cores integrate with each other through the sharing of research results and nanotechnology platforms. This integration allows the TCCN to achieve clinical translation of its research breakthroughs, and aggressively manage the risks that are naturally associated with any highly innovative program at a rapid pace. To fuel translation to the clinic, several TCCN investigators have successfully developed spin-off companies based upon their research. Collectively, with a combination of synergistic projects supported by cores that provide sen/ices to each project and a track record of successful bench-to-bedside translation, the TCCN is uniquely positioned to bring forth highly effective nanotechnology platforms for prevention, therapy and monitoring of ovarian and pancreatic cancers.

The TCCN aims to utilize innovative nanotechnologies for new therapeutic strategies, methodologies for reliable monitoring of therapeutic efficacy, early detection approaches from biological fluids and advances in imaging, and cancer-prevention protocols for ovarian and pancreatic cancers. The TCCN will apply a diverse array of nano-platforms to achieve these aims. While the primary emphasis In the TCCN is on ovarian and pancreatic cancers, it is likely that the new approaches will have applications for many other malignancies.

Project 1: Multistage Vectors for Ovarian Cancer Therapeutics

(Lopez, Ferrari, Decuzzi, Ozpolat,Tanaka, West)

Ovarian cancer is the leading cause of death from gynecologic cancers, and the fifth leading cause of cancer deaths in women in the United States. As seen from these statistics there is an urgent need for the development of novel therapeutic agents and strategies beyond conventional chemotherapy for the Program Director/Principal Investigator (Last, FirsL Middle); Ferrari, Mauro; Sood, Anil; Lopez-Boresteln, Gabriel treatment of advanced ovarian cancer, Nanotechnology holds tremendous promise in the development of novel therapeutics to address these long standing clinical unmet needs. The fundamental goal of Project 1 is to demonstrate the feasibility for translation of a novel biocompatible nano-delivery system for efficient in vivo siRNA and drug delivery to achieve control of tumor growth and angiogenesis for a prolonged period. The Innovative "multi-stage" approach, using biodegradable porous silicon particles as a carrier of therapeutic nanoparticles, will offer a paradigm shift from conventional nanotechnology based drug delivery by improving unfavorable pharmacokinetics of nanoparticles. Beyond the therapeutic needs, a persistent void in the emergence of efficient ways to monitor therapeutic responses is evident Here the investigators present how novel nanotechnologies offer unequaled solutions to such undisputable necessity. The investigators will develop nanochip technologies to establish proteomlc profiles to effectively monitor therapeutic responses. We will complement these with the development of sensitive gold-gold nanoshells to image tumor microvasculature with superb resolution to monitor tumor vasculature response to therapy. Moreover, Project 4 will utilize corecross linked polymeric micelles to detect and image active apoptosis in vivo, indicative of response to most anti-cancer therapies. The proposed work will greatly contribute to fill the existing gap between discovery of basic cancer biology and nanotechnology.

The overall goal of Project 1 is to demonstrate the feasibility for clinical translation of biocompatible nanoparticles with favorable pharmacokinetics and tissue distribution for the highly efficient delivery of novel anti cancer therapeutic agents. This project is supported by a substantial body of preliminary data and our track record in solving the "delivery problems" of existing drugs. For example, selective delivery of sIRNA or small molecule inhibitors into tumor tissues is highly desirable, but currently represents an unmet need. Nonetheless, we have developed neutral nanoliposomes that can effectively transport and deliver siRNA and small molecule inhibitors to orthotopic tumor sites in experimental animals.

In this project, the investigators will develop additional delivery tools that achieve great efficiency in delivery through different mechanisms, to address different delivery challenges. For efficient and sustained delivery, the investigators have developed a multistage approach: a biodegradable silicon-based nanoporous particle that transports sIRNA or small molecule inhibitors incorporated in nanoparticles. While the multistage vehicle degrades in the body to innocuous silicic acid, the therapeutic nanoparticles are steadily released. By achieving sustained release rates of the nanoparticles, the therapeutic levels of siRNA and small molecule Inhibitors will be maintained, thereby enhancing their efficacy against ovarian tumors. Beyond presenting novel therapeutic strategies, we are also interested in offering solutions to the patent need for monitoring therapeutic response. To address this need, we present innovative Imaging nanotechnologies focused on apoptosis as a marker of response to therapy. Furthermore, we develop nanochips to generate proteome profiles that can determine whether tumors are responding to therapy. Project 1 's central hypothesis is that multistage vectors (MSVs) will result in prolonged and sustained release of anticancer agents for efficient treatment of ovarian cancer, and the therapeutic response can be effectively monitored by nanochip profiling and nanoimaging.

Project 2: Nanotechnology Platforms for Targeting Ovarian Cancer Vasculature 

(Sood, Decuzzi, Gorenstein, Li, Tanaka, West)

The progressive growth of ovarian cancer and associated metastases is dependent on an adequate blood supply (angiogenesis). Despite advances in surgery and chemotherapy, ovarian cancer remains the most deadly gynecologic malignancy. Therefore, new treatments are urgently needed. Targeting angiogenesis is a particularly attractive strategy because of the presumed genetic stability of endothelial cells.*^ This is best illustrated by recent successes of anti-angiogenic therapy (e.g., bevacizumab) in patients with solid tumors. However, despite initial responses, most patients eventually develop tumor progression resulting in their demise. Therefore, new anti-angiogenesis therapeutic strategies are needed. The overall goal of Project 2 is to develop novel nanoparticle-based strategies to target the tumor vasculature specifically. We propose to utilize two types of biocompatible therapeutic nanoparticles (chitosan and gold nanoshell nanoparticles) forthe delivery of therapeutic payloads (e.g., siRNA) or near-infrared (NIR) laser mediated thermal ablation. These platforms are supported by integrated approaches for selective delivery into the tumor vasculature using either rationally designed multi-stage carriers or surface ligands (thioaptamers) selected from screening libraries based on selective binding. Using genomic approaches, the investigators have identified novel candidate target genes in ovarian cancer vasculature that will be targeted using RNAi approaches (Aim 1) because many are difficult to inhibit with small molecules or monoclonal antibodies. In our preliminary findings, the investigators have identified thiophosphate oligonucleotide aptamers (thioaptamers) that selectively bind to tumor, but not to normal endothelial cells based on counter selection strategies using freshly Isolated endothelial cells from human ovarian cancer or normal ovaries. In Aim 1, we will develop thioaptamer-targeted nanoparticles for selective delivery of therapeutic sIRNA. On the basis of preliminary findings regarding the critical role of size and shape in vascular localization of nanoparticles, we will pursue rational design of nanoparticles for targeting the tumor vasculature in Aim 2. Gold-based nanoshells offer unique opportunities for thermal ablation using NIR light In Aim 3, the investigators will develop and characterize novel approaches for thermal ablation of ovarian cancer vasculature using targeted gold nanoshells. All three aims are complementary to each other and findings of this study should allow the design and translation of new therapeutic approaches for women with ovarian cancer.

Project 3: Nanotechnology Platforms for the Prevention and Personalized Therapy of Pancreatic Adenocarcinoma

(Logsdon, Decuzzi, Menter, Hwang, West)

Project 3 will focus on the application of nanotechnology to the unique problems of pancreatic adenocarcinoma (PAC). Unlike what is observed in other solid tumors, pancreatic cancer cells exist as foci embedded in an abundant dense hypovascular fibrotic stroma. Development of this dense stroma begins with early premalignant disease (pancreatic Intraepithelial neoplasia (PanIN)) and continues through tumor progression. A primary goal of this project will be to develop nanotechnologies that can either penetrate or accumulate in the abundant stroma that characterizes pancreatic cancer. The ability to specifically deliver therapies to the pancreatic tumors will also allow the use of therapies directed at appropriate targets in individual patients, opening the door to individualized therapies.

Unfortunately, the current preclinical tumor models for PAC do not develop stroma and thus do not accurately represent the microenvironment found in the human disease. Recently, the investigators have developed novel preclinical mouse models, both xenografts and genetic, which accurately mimic this fibrotic microenvironment. Project 3 will use these novel and unique model systems to develop optimized nanotechnologies that target and accumulate within pancreatic stromal networks for therapy, imaging,   and prevention of PAC. Thus, the investigators will effectively turn what was a barrier to therapeutic delivery into a reservoir for preventive and therapeutic agents. Alternatively, we will suppress the biological activity of the stromal cells to prevent cancer progression or to render the cancer more amenable to cancer cell directed therapies. Together these studies will, for the first time, utilize nanocarriers to penetrate the barrier surrounding pancreatic tumors to achieve clinically significant goals.

Project 4: Multifunctional Nanoassemblies for Ligand-Directed Imaging and Therapy of Endocrine Pancreatic Tumors 

(Arap, Pasqualini, Decuzzi, Libutti)

The development of tools for the targeted delivery of imaging-probes and therapeutic agents has become the focus of intense efforts in the context of many human diseases. Over the past years, the investigators have developed an in vivo screening method in which phage can be selected from engineered combinatorial peptide libraries for their ability to target specific vascular beds. This work has uncovered a vascular address system that allows specific angiogenesis-related targeting to blood vessels in cancer. The investigators have expanded on this phage display-based targeting by the direct assembly of gold (AuNP) and multistage nanoporous silicon (SINP) nanoparticles onto phage for nanomedical applications. Through exploiting the nanodimensions of the phage particle as a molecular network Project 4 investigators generated biologically active nanoassemblies (NAs) with concomitant unique and tunable chemical and physical properties. These properties include near-infrared (NIR) radiation conversion to heat, enhancement of fluorescent signals, NIR surface enhanced Raman scattering (SERS) and the ability to conjugate and incorporate therapies or Imaging-tracers. Furthermore, "multi-stage" approach biodegradable porous silicon carrier for tumor and targeted nanoparticles will the innovative by using particles as a tumor-associated offer a paradigm shift from conventional nanotechnology based drug delivery by improving unfavorable pharmacokinetics of nanoparticles. These nanoparticles targeted through phages network to tumor vasculature are fashioned considering the distinct biology of the tumors to be treated to adequately target malignant tissue while sparing considerably normal tissue to reduce non-specific toxicity. Specifically, SiNP are rationally designed to better marginate in the bloodstream, are decorated with Au-phage networks with specific recognition of tumor vascular endothelium and loaded with smaller nanoparticles, e.g. nanoliposomes bearing cytotoxic agent or contrast agents which can be released from the SiNP in a pre-programmed fashion. This tuning capability combined with the programmable tissue targeting affords the integration of multiple functionalities into a single NA and serves as a complementary and non-mutually exclusive tool among different applications, including chemotherapy targeting and molecular imaging. In this proposal the aim is to develop the ligand-directed Si particles- phage- Au particles NAs as novel systems for targeted imaging and therapy in endocrine pancreatic tumors. These efforts will be translated into wide-ranging clinical applications.

Core 1: Biomathematics Core 

(Decuzzi, Ferrari, Mueller, Almeida, Cristini, Macklin)

The Biomathematics Core has three main objectives: (1) To employ the mathematical tools core investigators developed, to rationally design therapeutic vectors and predict the impact of nanoparticle-based therapy on tumor growth; (ii) To provide statistical tools for the design of experiments and post-processing of data; (iii) To provide web-based infrastructures and informatic tools for managing and sharing within the TCCN and with the wider scientific community the data and Information generated. These objectives will be pursued through three components of the Core: the BioSimulation component (Decuzzi, Cristini and a post-doctoral fellow); the Biostatistics component (Mueller and Basset) and the Data Management component (Silva Almeida and a post-doctoral fellow). All four projects of the proposed TCCN involve the use of sut)-mlcrometer and nanometer particle systems (nPSs) for the delivery of therapeutic and imaging agents, and for performing physical therapy. The Core 1 will provide the mathematical tools to predict (i) the accumulation of intravascularly injected nPSs within the tumor mass (Project 2 to 4); (ii) the spatiotemporal dynamics of tumor growth upon controlled release of therapeutic agents (Project 1, 3 and 4) and photothermal ablation (Project 2 and 4) from the nPSs. The core will also provide service for the biostatistical analysis and design of experiments (all Projects), as well as the management and dissemination of the data and analytical procedures within the Center and to the wider scientific community in a controlled fashion.

Core 2: Targeting Core

(Gorenstein, Pasqualini, Arap)

The Targeting Core focuses on the rapid identification of robust protein-specific targeting ligands for the TCCN nanoparticles that show high affinities, specificities and biological stability. Traditionally, these ligand requirements have been addressed with the use of antibodies (Abs). Antibodies however often have significant problems. Including high cost, selection difficulties, selectivity problems, preparation difficulties, stability and immunogenicity. This Core's objective is to provide the TCCN projects with sets of targeting reagents for the nanoparticles that not only include "gold standard" antibodies but novel aptamers and peptides to both endothelial, cancer and stem cells. All projects will use the resources from the Targeting Core. In particular, the Targeting Core aims to (i) Develop X-aptamers for targeting nanoparticles to CD44, E-Selectin, VGFR, EGFR, and others (Dr. Gorenstein, co-Director); (ii) Develop peptides and antibodies via phage display for targeting (Dr. Pasqualini, co-Director); (iii) Provide conjugation of targeting ligands and all micro/nanoparticles.

Core 3: Nanoengineering Core 

(Ferrari, Liu, West)

The TCCN comprises investigators with a long and successful track record of development, production/synthesis, characterization and validation of nanoparticles.