Andrew Baird - US grants

This proposal is a continuation of a program project that, over the course of the last sixteen years, has dealt with problems in fundamental and applied neuroendocrinology as they apply to the fields of diabetes, pituitary and brain function and the biology of cell proliferation. The long-term objectives are to understand the role played by key proteins in the onset and progression of diabetes and its complications. It is proposed (1) to establish how angiogenic factors are regulated and identify the mechanisms which promote and/or limit their bioavailability; (2) to determine how dysfunction of these regulatory elements in disorders such as diabetes might mediate microvascular complications; (3) to characterize novel growth inhibitors and use these molecules to study diseases of cell proliferation; (4) to study the molecular biology of fibroblast growth factor; (5) its receptor; and (6) the regulation of their expression. These specific aims are to be supported by the presence of state-of-the-art cores of immunotechniques, nucleotide synthesis, peptide synthesis and sequencing, which serve on one hand as the backbone uniting a comprehensive research program in diabetes, pituitary, and growth factor biology and on the other as the necessary resource to develop and import novel and/or improved techniques and technology to the research projects to ensure that the program remains at the forefront of science.

biomedical equipment purchase;

growth factor receptors; neurotrophic factors; fibroblast growth factor; brain mapping; receptor expression; gene expression; brain injury; disease /disorder model; in situ hybridization; laboratory rat; molecular cloning; polymerase chain reaction;

DESCRIPTION (provided by applicant): Certain proteins, like growth factors have significant potential to treat neurodegenerative disease. Accordingly, it should be possible to stimulate the growth of endogenous progenitor stem cells so that they could enhance the natural course of injury repair. Unfortunately, the blood brain barrier forms a significant hurdle that limits the clinical utility of most peptides. We propose to apply principles of medicinal biology and combinatorial pharmacology to improve the delivery of growth factors to the brain and describe a comprehensive strategy to address a major problem in the development of CNS biotherapeutics: drug delivery to the brain.

DESCRIPTION (provided by applicant): Technologies originally developed in one field of research (e.g. cancer), is sometimes slow to be introduced in other fields (e.g. wound healing). The overall objective of this application is to apply one such advance, in vivo combinatorial biology, to wound healing (WH) so as to create and modify potential WH biotherapeutics. We will screen for new therapeutics with 4 technologies: (1) Biopanning to identify targeting peptides for the ischemic wound for better drug delivery;(2) SNAAP screening to create pharmacologically optimized growth factor chimera with improved pharmacokinetics for wound repair;(3) LIVE recoveries to evolve a gene delivery vector, genetically optimized for the WH milieu and (4) RBT to identify cell surface biomarking signatures on bone marrow-derived precursor cells that can migrate to and engraft the wound. Innovation: Combinatorial technologies have never been applied to a wound healing paradigm. Because phage display can answer research questions like no other technique;it will open new venues of WH research. It will increase basic understanding of WH mechanisms, identify intrinsically useful biomarking signatures and create new biotherapeutic agents. Need for a multidisciplinary team: Phage display fails as a "molecular biology kit" and multidisciplinary skills in cores and projects provide (1) specific experience to create, optimize and analyze libraries (QA/QC, inventory), (2) access to prototypic wound healing models, (3) the ability to design, develop and test ideal prototypic screens and (4) the hit to lead transformation for the wound healing community. Impact on wound healing: Combinatorial techniques could be applied to virtually any WH paradigm (e.g. trauma burns). It will generate unique research reagents, preclinical candidates, pharmacologically improved gene vectors and a better understanding of progenitor, stem and stromal cell targeting to WH. It can have ancillary applications in all kinds of different WH paradigms. Relevance to public health: Progress in ambulatory care increases pressures to accelerate normal WH, minimize reconstructive surgery, modify scar formation and return the patient to the workplace. Technologies originally developed for other disciplines, generates completely new ways to enhance, modify and understand the WH response in a fashion that can impact all kinds of different kinds of wounds.

[unreadable] DESCRIPTION (provided by applicant): In order to fully exploit knowledge garnered from sequencing the human genome, there is a dire need to deliver biologically active drugs and biopharmaceuticals into the CNS for stroke therapies and the treatment of neurodegenerative disease. The goal of this exploratory R21 feasibility research proposal is to identify peptides capable of targeting the choroid plexus (CP) so that they can be evaluated for their capacity to deliver drugs to the CP or across the CP into cerebrospinal fluid (CSF). The anatomical and therapeutic rationale for the proposed research originates with the observation that the CP is a structurally unique interface between blood and CSF that acts as a heretofore unrecognized focal gateway that can deliver naturally occurring agents like ascorbic acid into the brain, and specifically into CSF. Accordingly, in addition to drug targeting the CNS, CP-targeting would also open the possibility of creating new CP-based neurotherapeutics that either (1) modify CP drug transport functions directly, (2) alter its capacity to secrete CSF or (3) modulate the composition and concentration of factors (e.g. growth factors) that it places into CSF. Rather then apply rational design to create drug delivery agents for CP and CSF therapeutics however, we propose to exploit the power of combinatorial biology and genetic selection to mine libraries of peptides for those capable of targeting CP. Specifically, we propose to identify (1) peptides that target CP endothelial and stromal cells (2) peptides that target CP epithelial cells and (3) peptides that can translocate across the CP and into CSF. At the conclusion of this project, we will have identified 3 classes of peptides capable of specifically targeting the blood (basolateral) and CSF (apical) interfaces of the CP for CP and CSF-targeted for drug delivery to the brain during neurodegeneratrive disease. [unreadable] [unreadable] [unreadable]