Prabhas V. Moghe - US grants

9733007 Moghe The research program of this career award is to develop a technology to quantitatively evaluate matrix microstructure and the corresponding cellular activities within biodegradable tissue analog matrices. Specific aims are to: (1) develop a novel imaging methodology to quantitatively investigate topological and bulk microstructure of 3-D biodegradable porous tissue analogs under physiological conditions; (2) design novel microstructural configurations of polymeric matrices that promote the phenotype and functions of cultured liver cells; and (3) examine the effect of polymer degradation on the topological and bulk matrix microstructure and the local physico-chemical conditions to the physiology of liver cells. For the teaching component of this award, the specific aims are to: (1) develop an innovative university-wide curriculum in Tissue Engineering which reflects a focus on biomaterials engineering; (2) establish a new course for students from regional high schools with the goal of offering to selected students a broad study of bioengineering as a field combining life sciences with the physical sciences and mathematics; and (3) extend outreach to other universities and high schools by distance instruction through an internet- based network. ***

0201788
Moghe
Cardiovascular disease takes a staggering toll of casualties among adult Americans each year. Two of the significant vascular pathologies related to the abnormal accumulation of lipids are atherosclerosis (the hardening of arteries due to build-up of low density lipoproteins (LDL)), and macrovascular disease, typically correlated with insulin resistant diabetes, which claims a million lives each year globally. Much research has been directed at the molecular design of drugs to alleviate the disorders of lipid metabolism. However, such drugs can be toxic to the liver and kidneys, and fail to comprehensively treat lipoprotein transport and retention dynamics, particularly at peripheral vascular sites. Thus, a comprehensive approach to treating lipid-related vascular disease could involve use of molecules regulating lipid metabolism as well as molecules that are suitably lipoprotein-philic and serve as multifunctional carriers for processing lipoproteins in transit. Ultimately, such carriers could be engineered to (a) sequester lipoproteins from macromolecular depots such as proteoglycans that heighten atherogenic tendencies; (b) reduce lipoprotein oxidation (which leads to unregulated uptake of LDL by macrophages, transforming them into foam cells, the precursors to atherosclerosis); and (c) enhance lipoprotein transport and clearance of mildly oxidized lipoproteins (via macrophages, and the liver). However, to engineer such carriers, an understanding of the chemical and geometric determinants of lipoprotein-retentive carrier substrates is necessary. This proposal describes a major research initiative toward this goal.

The proteoglycans of the vascular intima are bulky, negatively charged molecules that present multimeric glycosaminoglycan (GAG) chains, which can co-operatively recruit low density lipoproteins, and encourage LDL hyperoxidation, which leads to foam cell formation during atherosclerosis. As a competitive strategy for LDL retention, the investigators propose to design novel diffusible, nanoscale carriers that can present GAG-mimetic chemistry and retain LDL with high affinity. To this end, two significant questions will be addressed: (a) Can the GAG-mimetic chemistry and nanoscale topography of model substrates be designed to synergistically recruit oxidized low density lipoproteins? (b) How can the insights derived in (a) be applied toward the use of mobile nanocarriers for LDL retention?

To address (a), the investigators will theoretically simulate and experimentally explore the ability of immobilized gold nanoparticles (model substrates to test the LDL-reactivity of various chemistries) and substrate arravs of gold/ZnO nanopillars. functionalized with alkanethiols terminating in negatively charged groups (-COOH, -OSO3H), to sequester LDL. The hypothesis is that at adequately high densities, and in topographic substrate configurations affording inter-pillar cooperativity, such chemistries can electrostatically sequester LDL through the positively charged aminoacid residues from the apolipoprotein B-100 of the LDL. To address (b), the investigators will explore the use of polymeric dendrimer-Iike hyperbranched nanocarriers to present the most LDL-retentive chemistry observed in (a), in various nanoarchitectural configurations, that is, by systematically manipulating the valency, branching, and tethering of the molecular bait for the lipoprotein.

This IGERT program at Rutgers University, focused on integratively engineered biointerfaces, will be an intimately collaborative effort of 32 selected faculty from graduate programs in Molecular Biosciences, Physical Sciences (Physics, Chemistry & Chemical Biology), and Engineering (Biomedical Engineering, Ceramics and Materials Engineering, Chemical and Biochemical Engineering, Mechanical and Aerospace Engineering).

Intellectual Merit: The program derives strength from the highly cross-disciplinary nature of over fifteen research project areas identified at the cutting edge of the field of biointerfaces, and programmatic partnerships with five strategic centers of excellence to promote cohesive access for the IGERT community to state-of-the-art research infrastructure. A wide range of thesis project themes is planned for the IGERT trainees, developed around three research and educational thrusts, (1) living cell-based interfaces, (2) microengineered and nanoengineered biointerfaces, (3) biosensing and bioresponsive interfaces. The five major partnering Centers for the IGERT program are: Keck Center for Collaborative Neuroscience, Center for Nanomaterials Research, New Jersey Center for Biomaterials, the Laboratory for Surface Modification, and the Rutgers Center for Computational Design. The educational core of the proposed IGERT program will intimately support the research program, and includes graduate courses in the integrative areas of biointerfacial engineering, as well as course modules on responsible conduct of research, technical communications, entrepreneurship and effective teaching/learning methods.

Broader Impact: The IGERT curriculum is designed to foster a community featuring the next generation of biointerfacial and biomaterials engineers by offering IGERT graduate fellows a range of interactive experiences at multiple levels: multi-disciplinary coursework, lab rotations in two cross-cutting research groups, biannual participation in symposia, and participation in a national/international conference resulting in a white paper. To maximize its impact, the IGERT program will offer varied programmatic pathways to promote diverse modes of professional development of IGERT graduate fellows: (1) Summer research internships at selected international sites for academically inclined students; and (2) Translational research and industrial summer internships for students interested in industrial and entrepreneurial careers. Through a partnership with the Robert Davis Learning Institute of the Rutgers Graduate School of Education Institute, the IGERT program will establish a COLTS (Community of Learners and Thought Shapers) program, inspired by communication-driven cognition models, to encourage IGERT fellows to develop as learners by dynamically communicating their research on integratively engineered biointerfaces.

IGERT is an NSF-wide program intended to meet the challenges of educating U.S. Ph.D. scientists and engineers with the interdisciplinary background, deep knowledge in a chosen discipline, and the technical, professional, and personal skills needed for the career demands of the future. The program is intended to catalyze a cultural change in graduate education by establishing innovative new models for graduate education and training in a fertile environment for collaborative research that transcends traditional disciplinary boundaries. In this sixth year of the program, awards are being made to institutions for programs that collectively span the areas of science and engineering supported by NSF.

0609000
Moghe

This proposal was received in response to Nanoscale Science and Engineering initiative, NSF 05-610, category NIRT. The objectives of this research are (1a) to design, fabricate, and characterize nanoparticles functionalized with matrix protein fragments, and (1b) to elucidate the role of nanoparticle size, ligand sequence and ligand loading on cell motility and matrix assembly; (2) to identify the key molecular signaling pathways that mediate nanoparticle internalization and increased cell activation; and (3) to develop imaging modalities to quantify nanoparticle trafficking and internalization dynamics. The approach involves the functionalization of albumin-derived nanoparticles with various fragments of fibronectin (ligand). The first phase of the study will examine how optimal configurations of substrates based on the ligand-nanoparticles can significantly alter the morphology and dynamics (motility, matrix assembly) of skin derived cells (keratinocytes, fibroblasts). The second phase will focus on identifying the underlying biologic mechanisms using protein and gene level signaling assays. The third phase will probe the nature and kinetics of nanoparticle-cell interactions using high resolution, two-photon microscopy and a magnetically responsive biosensor platform with nanoscale fidelity.

This research can help design improved nanomaterials for potential cell targeting applications in wound healing, tissue engineering, drug delivery, and cancer therapy.
The nanoparticles designed here are biodegradable, can be targeted to cells, and can be customized to cell functions by altering their size. Smaller nanoparticles can stimulate increased motility in epidermal cells (relevant for skin healing in burns, ulcers) while larger nanoparticles promote skin contractility and matrix assembly (relevant to wound repair). This project extends its outreach through the wide diversity network (graduate, postdoc, undergraduate) at Rutgers and beyond, and resonates with three new integrative graduate courses at Rutgers on engineering of cellular biointerfaces with biomaterials, a graduate training program on biointerfaces, and an international research collaboration.

This Integrative Graduate Education and Research Traineeship program (IGERT) renewal award establishes a training program focused on the science and engineering of stem cells. New cross-cutting thrusts for Ph.D. research, together with a new multidisciplinary graduate curriculum, will integrate stem cell biology with research in biomaterials, process engineering, and computational modeling. Trainees will participate in an IGERT Research Interchange Forum to develop their abilities to communicate across disparate disciplines. Professional development activities encompassing teaching, mentoring, and outreach will enable IGERT trainees to better realize the impact of their technological know-how. Each IGERT trainee will be guided by an advisory constellation of scholars drawn from over 30 faculty members from Engineering, Molecular Biosciences, Physical Sciences, Business, Public Policy, and Management. The IGERT program will leverage Rutgers' active "diversity infrastructure" to help broaden the participation of underrepresented minority students. In addition to providing research opportunities for visiting underrepresented undergraduates, the IGERT will offer two new initiatives: a teacher-student summer institute at Rutgers, and, a bridge-to-IGERT program. New outreach programs at the intersection of stem cell science and engineering with public policy and business include: (1) an initiative with the School of Management and Labor Relations to "bundle" IGERT research and curriculum into portable modules for scientific workforce training; (2) Rutgers Business School-mediated interactions with pharmaceutical management students and industry; (3) public policy workshops with policy makers, facilitated by the Eagleton Institute of Politics. IGERT trainees will acquire global perspectives through internships and workshops with leading stem cell researchers at over 15 sites in Europe and Asia. IGERT is an NSF-wide program intended to meet the challenges of educating U.S. Ph.D. scientists and engineers with the interdisciplinary background, deep knowledge in a chosen discipline, and the technical, professional, and personal skills needed for the career demands of the future. The program is intended to catalyze a cultural change in graduate education by establishing innovative new models for graduate education and training in a fertile environment for collaborative research that transcends traditional disciplinary boundaries.

This project is combining the theoretical perspectives of "communities of practice" and "situated learning" with the innovative research, curriculum, and community best practices developed in four Integrative Graduate Research and Education Traineeship (IGERT) projects. This is done horizontally across the institution's STEM graduate programs and centers, and vertically within the undergraduate research experience programs. This extensive program of activities is improving the quality of undergraduate and graduate students completing STEM degrees. Additionally, the project is using proven methods to increase the participation of groups underrepresented in STEM fields, including first generation, economically disadvantaged, and disabled students. With attention to addressing critical educational junctures, the senior leadership team is creating the Graduate Innovation and Integration Center (GIIC) to enhance the integration of the institution's and research and educational efforts. GIIC center is working with five existing interdisciplinary campus centers to build collaborations and synergistic outcomes. A comprehensive plan is evaluating each aspect of the project including critical junctures as well as the impact of the project on promoting institutional collaboration and synergy. Longitudinal studies are being planned that follow students for three years post graduation. The broader impact of this project is the synergy between a wide array of undergraduate and graduate programs; the broadened participation of undergraduate students in research experiences, particularly students from populations underrepresented in STEM disciplines; the improved retention of a diverse body of graduate students; and a model for student success that can be used at other institutions.

0923246
Bartynski
Rutgers U. New Brunswick

Technical Summary: X-ray photoelectron spectroscopy (XPS) is widely used as an analytical technique to determine the nature of the near-surface region of a material. Shifts in the core level binding energies of atoms at or near the surface of a material can reveal changes in oxidation state, surface potential or band bending, chemical or physical inhomogeneity, or dynamic response (i.e., screening) that are distinct from those of the bulk of the material. However, a growing number of modern applications employ materials in complicated structures that are laterally inhomogeneous and thus it is critical to perform XPS in a spatially resolved manner, along with high photon flux, and high energy resolution. Examples, that are currently active research areas at Rutgers include the study of: (i) transition metal ions and their diffusion in ZnO for room temperature spintronics, (ii) surface modification of organic single crystal surfaces, (iii) surface functionalization and characterization of novel nanocrystals used to enhance biomolecule imaging, (iv) surface characterization of plasma-treated and chemically-modified polymer films for cellular and related bioactivity studies, and (v) interface properties of nanoscale self-assembled solid state systems. The Rutgers Laboratory for Surface Modification (LSM) is a multidisciplinary research center that hosts a comprehensive set of facilities used to examine surfaces, interfaces, thin films, and nanoscale materials, and has strong collaborations with state-of-the-art research and development laboratories around the world. A gap in our suite of tools is lack of a modern high resolution XPS system that can adequately address the key issues in the study of modern materials systems. The state-of-the-art instrumentation requested in this proposal would replace a 20-year-old machine and will significantly strengthen our capabilities by enabling high energy resolution studies, in parallel with high resolution lateral imaging and depth profiling. These features are central to the diverse research and education activities both within Rutgers as well as the regional community.

Non-Technical Summary: Materials interact with their surroundings through their surfaces. Very often, the chemical or physical environment of surface atoms is significantly different from those of the bulk A powerful way to probe surface properties is to expose a material to X-rays of a specific wavelength and study the electrons that are emitted from surface. As only electrons that originate from the first one or two nanometers of the surface are able to escape the material, this technique is very surface sensitive. Moreover, these electrons escape with well-defined energies that not only depend upon the atomic species, but also exhibit small variations depending on the environment of the atom. The study of these electrons, known as X-ray photoelectron spectroscopy (XPS), enables one to determine the chemical and physical state of these near-surface atoms. In modern materials systems, such as nanoscale crystals used to enhance imaging of biological systems, or potentially revolutionary semiconductors made entirely of organic molecules, the atomic environment in one region of the surface can differ from that of another region. Therefore, it is critical to perform XPS studies in a spatially resolved manner. The Rutgers Laboratory for Surface Modification (LSM) is a multidisciplinary research center that hosts a comprehensive set of facilities used to examine surfaces, interfaces, thin films, and nanoscale materials, and has strong collaborations with state-of-the-art research and development laboratories around the world. A gap in our suite of tools is lack of a modern high resolution XPS system that can adequately address the key issues in the study of modern materials systems. The state-of-the-art instrumentation requested in this proposal would replace a 20-year-old machine and will significantly strengthen our capabilities by enabling high energy resolution studies, in parallel with high resolution lateral imaging and depth profiling. These features are central to the diverse research and education activities both within Rutgers as well as the regional community.

DESCRIPTION (provided by applicant): Pathologies in blood vessels arising from the uncontrolled build-up of oxidized lipids contribute to atherosclerosis, a severe cardiovascular disease, which underlies the most common cause of adult death in the U.S. (exceeding one million patients yearly). Few existing therapeutic strategies address the local management of atherogenesis (build-up of oxidized lipids in the blood vessel walls) and related inflammation. The overall goals of this study are to rationally design and characterize nanoscale biomaterials as a novel cell-targeted materials platform for investigating strategies to de-escalate the onset of atherogenesis and reduce accompanying inflammation. The proposed NIH R21 study involves three specific objectives to investigate nanoassembled amphiphilic polymers (NAPs) to maximally inhibit oxidized LDL uptake in human macrophages under physiologic conditions and exhibit potential for specific targeting to inflamed endothelia. Efforts in Aim 1 will involve investigation of innovative designs of nano-assembled amphiphilic polymers (NAP) composition and architecture to promote NAP binding to both SRA-1 and CD36 scavenger receptors on human THP-1 macrophages and thus inhibit uptake of oxidized low-density lipoproteins (oxLDL). New configurations of NAP will tested for improved lipid uptake inhibition in the presence of serum. Studies in Aim 2 will investigate the effect of NAP-scavenger receptor (SR) interactions on the downstream intracellular and cell-secreted intermediates regulating atherogenesis in macrophages, including cytokine secretion;cholesterol ester accumulation;matrix metalloproteinase secretion;and expression analysis of genes involved in pro-atherogenic signaling pathways. Aim 3 is concerned with design and evaluation of the potential of biofunctionalized NAPs to bind to and transport across activated endothelial cell cultures in vitro, thereby creating a simplified in vitro model of the rescue of macrophage cells involved in atherogenesis within the vascular intima. PUBLIC HEALTH RELEVANCE: The excessive uptake of modified forms of LDL in immune blood cells macrophages is one of the hallmarks of fat build-up and vascular disease within blood vessel walls, which can lead to blockage of blood flow, and cause heart disease or stroke. This study will investigate the design of nanoscale assembled polymers with specific architectures, charge displays, and chemistry so as to reduce the uptake of the most damaging forms of lipoproteins within macrophages. The goals of the study are to identify the most effective "nanolipoblocker" configurations that may prevent atherogenesis by targeting activated blood vessel cells and blocking foam cell formation.

Proposal: 0851831
PI Name: Charles M. Roth

"This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)."

This 3-year REU site program at Rutgers University will engage 10 undergraduate students each year in cutting-edge, cross-disciplinary research in the field of Cellular Bioengineering. Cellular Bioengineering encompasses a number of cutting edge research fields and articulates with nanotechnology, advanced materials, systems biology and stem cells. The research focus on Cellular Bioengineering From Biomaterials to Stem Cells will integrate these areas towards the establishment of a strong engineering base for the development of cell-based devices and therapeutics. An emphasis on stem cells is a relatively unique opportunity for an REU site.

The REU program will continue and expand upon an existing seed program called ISURF (Integrated Summer Undergraduate Research Frontiers). Students will receive training in fundamental laboratory techniques in a Cellular Bioengineering Boot Camp, will meet weekly for community building and exercises designed to develop their research communication skills, and will present their research results to their peers, faculty and visitors at a Summer Symposium. Another unique feature of this program is the Buddy System which will be employed to provide REU participants with near-peer mentors outside of their own laboratories, and participants across various years of the program will be linked via an E-Pals system and via an online blog and community called The Human Cell.

Recruitment efforts will be targeted to students from underrepresented groups and those from colleges without substantial graduate research and training. This REU program will train undergraduate students from diverse backgrounds in Cellular Bioengineering and motivate them to pursue graduate studies which will lead to a better trained workforce in this rapidly expanding field.

DESCRIPTION (provided by applicant): This study targets atherosclerosis, a chronic inflammatory disorder of the blood vessel wall, which underlies nearly 50% of all deaths in westernized countries and is the primary cause of mortality in patients with diabetes. This proposal utilizes rational molecular design approaches to a novel class of therapeutics - amphiphilic polymers that serve as athero-protective and anti-inflammatory therapeutics. The most innovative component is that molecularly designed polymers may have the potential to inhibit atherosclerosis by multiple scavenger receptor targeting and blockage during the early stages of atherogenesis. This binding behavior could be critical to blocking oxidized LDL uptake and more effectively abrogate the athero-inflammatory cascade, and retard the progression of atherosclerosis. The central hypothesis regarding the enhanced polymer structures is that combinations of strengthened hydrophobic features in conjunction with anionic charge and hydrophilic tails will yield polymers with optimal targeting to multiple scavenger receptors on both macrophages and endothelial cells, specifically SR-A, CD36 and LOX-1. To test this hypothesis, three specific aims are proposed. Aim 1 is focused on the molecular modeling (docking and scoring) and design of novel polymer classes for enhanced binding to multiple scavenger receptors. This effort will yield new polymer structures with a more rigid and space-filling backbone that, in conjunction with charge and hydrophilicity, enhance binding affinities to scavenger receptors - particularly under physiologic conditions. Aim 2 is focused on investigating the molecular mechanisms and polymer interactions with cultured macrophages and endothelial cells for inhibition of athero-inflammation in vitro. Aim 3 is focused on the evaluation of in vivo polymer efficacy in terms of binding to atherosclerotic lesions and degree of regression of athero-inflammatory markers using an animal model of accelerated atherosclerosis. At minimum, new insights will be obtained regarding multiple scavenger receptor blocking as a strategy to counteract the progression of atherosclerosis. The potential impact of this research proposal is high; the overall outcome may be a new approach to treating coronary artery disease - using enhanced polymers as multifunctional inhibitors.

DESCRIPTION (provided by applicant): Complex disease states such as metastatic cancers and acute atherosclerosis take a staggering toll on society in terms of mortality and health care costs. Current approaches to discriminate these pathologies are limited by their invasive nature, costs, and inability to target molecular features of the pathologies using real-time tracking methods. This exploratory R21 study is based on an innovative nanoscale concept to resolve and monitor tissue pathology using molecularly targeted optical imaging nanoprobes that emit infrared light. Infrared light is attractive because can be easily transmitted through thick biological tissues. Our nanoprobes consist of rare earth doped ceramic nanoparticles, which brightly emit infrared light in a novel window of emission. These particles are encapsulated with albumin nanoshells to impart cytocompatibility and aqueous dispersion and the albumin is conjugated with markers to target the disease of interest. The project proposes to develop a repertoire of rare earth-doped nanoparticles encapsulated in functionalized albumin nanoshells to establish nanoprobes with high biological availability in vivo, improved biocompatibility, and functional targeting to disease targets. A particularly innovative endpoint proposed is that of tracking disease phenotype progression through the in vivo imaging of intravenously injected cocktail of nanoprobes emitting across different infrared wavelengths and functionalized to four different markers of disease phenotypes. Two specific aims are proposed. In Aim 1, the project will investigate the role of nanoscale size and biofunctionalization of the rare earth nanoprobes on the biodistribution and accumulation at disease sites using a murine metastatic melanoma model. The in vivo distribution will be examined using infrared imaging of living animals and compared against conventional tissue profiles using high resolution inductively coupled mass spectrometry. Thus, optimal formulations of nanoprobes for in vivo imaging will be established. In Aim 2, the nanoprobe rare earth doped nanoparticles will be tailored to emit at different wavelengths and thus create a family of multi-chromatic nanoparticles. These will be functionalized to report simultaneously on four key markers for growth, invasiveness, and metastatic potential of tumors . The relative accumulation of the multiplexed nanoprobes will be used to track the progression/stabilization of tumors following established drug treatment. This will serve as a proof of concept for the foundations of this R21 project, and be the basis for developing this nanotechnology for a broader range of disease states of varying molecular phenotypes. PUBLIC HEALTH RELEVANCE: This project is concerned with the design of novel nanoscale probes based on biologically compatible rare earth-phosphors for optical imaging of biological tissues at near infrared wavelengths, which allows deeper penetration and real-time detection of pathologies. Outcomes will be insights into the role of size and biodistribution of the nanoprobes in vivo; design of multicolor emitting phosphors for identification of different molecular features of pathologies, and feasibility of molecular targeting to rapidly evolving disease states such as metastatic tumors. The overall health care applications are in diagnostic and multifunctional imaging of cardiovascular lesions/plaques, cancers, neurodegeneration, and infectious diseases.

This study proposes a novel nanomanufacturing paradigm for anisotropic micro- and nanoparticles, termed Janus particles. While this class of particles has received much attention recently, their formulations have not been compatible with biotechnology, where bio-benign polymers are called for, and staged delivery of multiple biotherapeutic factors is frequently desired but difficult to achieve. The key goal of this work is to create novel biodegradable and biocompatible anisotropic biphasic nano- particles (ABNPs) for large scale production, capable of dual compartmentalization and incorporation of two or more therapeutic drugs with staggered release profiles. Three specific aims will be pursued. Aim 1 is focused on the design and synthesis of a platform of ABNP/ABMPs with different combinations of polymeric and lipid materials that self-assemble into anisotropic particles. Aim 2 is focused on the modeling of polymer/polymer and polymer/lipid for the rational design of engineered ABNP/ABMP; Aim 3 proposes to demonstrate the broader functional impact of the nanomanufacturing paradigm of ABNP/ABMP on phased delivery of dual-combination drugs by applying these particles for difficult-to-treat cancers.

If successful this project will lead to significant advances in pharmaceutics, biomedicine, materials engineering and dispersion stabilization. Furthermore, multicompartmental, biocompatible particles will constitute a new avenue in the design of active and passive drug delivery vehicles, tissue engineering scaffolds, and biomimetic particles. The broader impacts are in the mechanisms for broadening participation of URM scholars; and developing curriculum and professional training mechanisms in the tools for nanoparticle manufacturing and drug delivery science and engineering. Four specific mechanisms for broadening participation are proposed: 1) Study modules will be created by the PI for all participating students to complete and discuss research findings in joint group meetings; 2) Results will be incorporated into teaching courses and laboratory modules; 3) Active participation in already established outreach programs at Rutgers; 4) Outreach to URM students and dissemination in workshops and conferences.

DESCRIPTION (provided by applicant): Rare Earth Nanoprobes for Optical Imaging and Disease Tracking PIs: Prabhas V. Moghe, PhD,; Richard E. Riman,PhD; Charles M. Roth, PhD. Collaborators: Mark Pierce, PhD., Shridar Ganesan, MD, PhD. This project aims to develop a library of new nanoscale imaging probes to identify and dynamically track microlesions that trigger the rapid spread of difficult to treat diseases like metastatic cancers and atherothrombosis. By causing mortality and morbidity as well as increasing health care costs, these diseases currently take a staggering toll on society. Technologies to treat these diseases are showing improvements, but clearly would have better outcomes when integrated with early diagnoses and rapid feedback on the effectiveness of pharmacotherapy. In this R01 proposal, we seek to harness the transformative potential of a hitherto undeveloped region of the optical spectrum, short wave infrared (SWIR)-based imaging, by advancing tunable SWIR emitting rare earth nanoprobes. Our team has demonstrated the potential of a new family of rare earth nanoprobes to image deeper within tissues than other modalities of optical fluorescence imaging, and with little autofluorescence and reduced scattering. The specific goals of this project are to develop new SWIR imaging probes and technology to detect and track microlesions in vivo and to identify their molecular phenotype. A novel panel of multispectral rare earth doped phosphor nanoprobes with high luminescence intensity will be synthesized and tested for their photonic properties. The ability of these probes to effectively label and resolve the SWIR-emission from engineered, sub-surface diseased cell clusters will be benchmarked in vitro (Aim 1). A suite of three convergent in vivo imaging tools for the SWIR probes will be optimized in Aim 2, including the design of biofunctionalized probes for targeting to metastatic lesions of breast cancer cells, SWIR macroscopic imaging of the lesions in the lymph nodes, and SWIR in situ confocal microscopy of lesions. In Aim 3, we will investigate the ability of the lesion-targeted probes to detect and track the development and regression of metastatic lesions in vivo following pharmacotherapy. Outcomes from this study will include new tools for more sensitively identifying and elucidating the molecular determinants of disease lesions in vivo. Potential applications include tissue- sparing imaging of lymph node tracking for early detection of cancer metastasis, as well as imaging of vascular lesions such as those in cardiovascular disease and atherothrombosis. The envisioned impacts of the R01 would be a first-in-class imaging probe/hardware framework, equivalent to an optical biopsy in vivo, to visualize the incidence, growth, and treatment of disease microlesions, stretching beyond the currently possible limits of spatial resolution (molecular phenotyping) and detection of sub-surface diseased tissues (depth penetration).

? DESCRIPTION (provided by applicant): This project aims to design innovative scaffolds that will integratively address two critical barriers for treating neurodegenerative diseases: (a) Cell Sourcing: support the maturation, specification, and function of reprogrammed human stem cell-derived neurons in vitro and (b) Subtype-specific Neuronal Transplantation: enable efficacious transplantation to treat neurodegenerative diseases in vivo. The central hypothesis is that 3D engineered microscale niches (EMNs) based on nanofibrous hydrogel scaffolds can support the induction and maturation of subtype specific neurons in vitro prior to transplantation and promote the survival and enhanced functional interaction with host tissue following transplantation. A specific application of interest to this project is the treatment of neurodegenerative diseases like Parkinson's disease (PD). To achieve our goal, two specific aims are proposed. The first aim is concerned with designing maturation-guiding EMNs of induced pluripotent stem cell (iPSC)-derived reprogrammed dopaminergic (DA) neurons. The 3-D EMNs will be based on transcription factor-transduced iPSCs cultured within nanofibrous hydrogels fabricated from self-assembling minimally immunogenic peptides. To guide the maturation and specification of the DA neurons, the EMNs will be functionalized with subtype specific cues. We will determine the emergent subpopulations of transplanted cells and examine changes in innervated host tissue through whole genome sequencing. The second aim will be focused on transplanting self-actuating EMNs of DA and excitatory neurons into the striatum of a mouse PD model. We hypothesize that a self-functioning E-DA mini neural circuitry within a microscaffold environment will provide sufficient excitatory drive to promote enhanced functional interaction of DA neurons with host tissue in vivo. We will transplant the self-actuating EMNs of E and DA neurons into the striatum of mice lacking DA innervations. We will evaluate the ability of self-actuating EMNs of E and DA neurons to improve functional deficits in the Parkinson's disease symptoms, and again examine maturation outcomes of transplanted cells and innervated host tissues. The overall outcomes from this study will help to address the critical barriers such as the functioning and survival of transplanted tissue in the field of cell-replacement therapies for neurodegenerative diseases.

PROJECT SUMMARY This project aims to design an innovative nanotherapeutic for treatment of synucleinopathies such as Parkinson's disease (PD), Parkinson's disease dementia (PDD), or dementia with Lewy bodies (DLB). These age-related neurodegenerative diseases are characterized by the deposition and aggregation of the protein alpha-synuclein (ASYN). Chronic and excessive ASYN accumulation can cause the recruitment of microglia cells that play the important role of clearance of ASYN. Sustained activation of microglia compromises the normal degradation pathways of ASYN, leading to a cycle of internal ASYN aggregation and neuroinflammation. Thus targeting microglial cells is a potentially viable approach for treating ASYN related neurodegeneration. The central hypothesis is that that by regulating ASYN interactions with microglial receptors, intracellular aggregation of ASYN can be inhibited, and that the progression of synucleinopathy can be halted. We aim to develop nanotherapeutics by designing synthetic nanoparticles (NPs) composed of amphiphilic molecules that can molecularly target specific microglial scavenger receptors (SRs). SRs mediate the uptake of ASYN and also catalyze ASYN oligomerization. In Aim 1, we propose a novel design for the NPs such that the NPs can serve as binding partners to SRs along with ASYN, but once internalized within microglia, the active core region of these nanoparticles will be exposed, disrupting intracellular aggregation of ASYN. In Aim 2, we will also investigate the efficacy of delivering an antioxidant payload to counteract microglial activation and thus ameliorate the activation of the pro-inflammatory M1 microglial phenotype. The effects of the NPs on microglial ASYN uptake and intracellular aggregation will be evaluated in vitro and in vivo. In addition to decreasing ASYN aggregation, targeting microglial activation states by suppressing the harmful effects of M1 microglia or switching activation states to the neuroprotective M2 phenotype can provide therapeutic benefits in neurodegenerative diseases. The overall outcomes from this study will help to address the critical barriers of maintaining microglial clearance of ASYN and decreasing the inflammatory effects of classically activated microglia within synucleinopathies and other neurodegenerative diseases.

Project Summary/Abstract The current NIBIB T32 postdoctoral training program, Training Without Borders: Translational Research in Regenerative Medicine (2012-2017), has focused on two innovative features: (1) the concept of a geographically dispersed training faculty to harness a unique team of mentors; and (2) a combination of conventional academic training (state-of-the-art science, proposal writing, responsible conduct of research) together with training in translation and commercialization. This has resulted in a geographically dispersed community of learning. The core strength of this program has been its interdisciplinary breadth, providing trainees with opportunities to conduct research on rationally designed biomaterials, bioactive microenvironments, cell profiling technologies, and regenerative biology. Based on this success the proposed training program (2018-2023) will retain the current structure, revise the mentor-institution network, and use a new paradigm - Bench-to-Business-to-Bedside (B3). While the term Bench-to-Bedside is often used to describe translation, we believe that the B3 model offers more relevant training in translation by providing trainees with mentoring constellations consisting of academic, industrial, and clinical subject matter experts. In addition to focusing on state-of-the-art science, postdoctoral training programs must improve the chances of their trainees finding employment across all sectors: academia, industry, and alternative career paths, in order to ultimately achieve clinical therapies. We have coined the term innovation-primed science to represent a range of breakthrough, innovation- generating scientific ideas that will offer our trainees a range of scientific projects to work on, from pioneering basic science to commercialization-ready projects, for which proof-of-concept has already been established. We will create a unique training infrastructure that is made up not only of the traditional academic mentors (scientific or clinical faculty who lead top laboratories), but also of a Translation Advisory Board (TAB) comprising experts in regenerative medicine and clinical practice, entrepreneurs, industrial scientists, investors, and leading subject-matter experts in translation and commercialization. The TAB will effectively fill the expertise gap among our academic faculty mentors, by completing a mentoring constellation of academic, industrial, and clinical experts for each trainee. A rich didactic program will combine both established approaches and new concepts to deliver a balance of experience to trainees while optimizing the expertise of 14 core mentors and 16 clinical, industrial and entrepreneurial advisors. Specific program elements are Individualized B3 Mentoring Constellations, Face-to- Face Interactions and Workshops, Online Interactions and Seminar Series, Defined and Monitored Integration of Didactic Elements during the Fellowship Term, Mentoring of Trainees and Professional Development, and Supporting Junior Mentors.