Harry V. Vinters, MD - US grants

The long-term objectives of the project are to examine the biochemical nature, immunocytochemistry, and pathogenesis of cerebral amyloid angiopathy (CAA), a microvascular change in human brain that correlates with non-traumatic intracerebral hemorrhage, and is found with high frequency in patients with Alzheimer's disease (AD). CAA is likely to increase in importance as a public health problem because at least one common form of CAA increases in the brain with aging. The study will utilize brain tissue derived from patients with CAA-related syndromes to study the following parameters: (1) Biochemical nature of amyloid peptides in affected cerebral vessel walls, to be examined using isolation of microvessels involved by CAA, SDS polyacrylamide gel electrophoresis of CAA proteins, and their characterization using high performance liquid chromatography, determination of amino acid composition and sequence. (2) Immunocytochemical characterization of CAA, CAA-related vasculopathies and consequences of these for the brain parenchyma, by use of antibodies against peptides with the sequence(s) of the isolated proteins described above, done at light microscopic and ultrastructural levels using (respectively) an avidin-biotin peroxidase technique and protein A-gold methods. Immunohistochemistry of other AD hallmarks (e.g. senile plaques, neurofibrillary tangles) will be simultaneously examined. (3) Presence of CAA peptides or precursors in blood or cerebrospinal fluid (CSF) from patients, by use of the above derived antibodies and the Western blotting procedure--to be carried out on human serum, CSF, extracts of brain arterioles and capillaries, and microvessel-free brain tissue of patients at risk for CAA and appropriate controls. (4) Detailed immunocytochemical study of AD-related CAA using synthetic peptides representing portions of the AD amyloid precursor. (5) Studies of interaction of the AD amyloid precursor with isolated microvessels (capillaries and arterioles from AD or control brain), and cultured cells derived from human brain microvessels. All phase of the application address the issues of the pathogenesis of subtypes of CAA, the relationship of CAA to blood-brain barrier breakdown and AD.

The Neuropathology (NP) Core of the UCLA-Alzheimer's disease Center (ADC) has accomplished its original goals and formulated new and continuing specific aims for this proposal. Over 100 brains have been accessioned to the UCLA ADC bank. In the renewal phase, the NP will perform complete or 'brain only' autopsies on patients enlisted in the Autopsy Program, and will serve as a data and tissue/fluid resource for individuals performing innovative research on AD/SDAT, non-AD, dementias, and a broader spectrum of scientists interested in neurodegenerative conditions. Brain tissues from appropriate autopsies will be triaged at the time of necropsy in order to optimize their subsequent use for structural and molecular/pharmacologic research. The tissue harvesting method will be one developed over the initial grant funding period, i.e. materials will be stored frozen, paraformaldehyde fixed, or in paraffin blocks. Tissue studies in the NP Core will include systematic blocking of fixed brain tissue, routine and specialized silver stains that demonstrate microscopic AD-related lesions (seline plaques and neurofibrillary tangles), and immunohistochemical staining using both commercially available antibodies and reagents developed in the NP Core or with collaborating investigators. Thus tissues/fluids provided to investigators will have been carefully assessed for AD and related disease processes. Other functions of the NP Core will include (a) interactions with the Clinical and Imaging Cores to examine correlative data, (b) morphologic/immunohistochemical support of ongoing AD-related studies by direct collaboration or provision of key reagents, ~ liaison with community pathologists who are called upon to assess brain tissue from dementia patients, (d) discussion with lay groups of the importance of autopsies on demented patients, and (e) assessing ApoE genotypes in ADC patients.

DESCRIPTION (provided by the applicant): This Core will serve as a vital resource center for blood and tissue-based studies integral to the completion of investigational projects, related to aggressive, interventional treatment of cerebral hemorrhage and ischemic infarcts. It will bring to analyses of blood and tissues derived from study patients a wealth of expertise in morphoanatomical techniques pertinent to stroke pathogenesis, and provide access to key investigators of samples of brain tissue derived from biopsy and autopsy specimens of study patients. The core will support Project 1, the MR RESCUE trial, in which intravascular thrombus material will be removed in the course of therapeutic thrombectomy. The core will assist investigators studying the biochemical pathogenesis of thrombus formation, and its possible importance in evolution of cerebral infarcts, and will assist investigators in correlating their findings with light microscopic/ultrastructural features of the thrombi. The core will also provide support to Project 2, the HEME Surgery trial, in which brain tissue originating from 'clot' evacuations carried out in patients with spontaneous intracerebral (parenchymal) hemorrhage will be carefully evaluated for both the likely etiology of hemorrhage and the tissue response to intracerebral blood, and readied for studies of differential gene expression employing qualitative, real-time RT-PCR and cDNA arrays. The Core will store blood samples from appropriate patients for DNA analysis, and provide these to researchers following appropriate review of their investigational goals and projects. The Core will serve as an important 'blood and brain tissue' resource to SPOTRIAS investigators at UCLA, and a larger network of investigators with interests in the treatment of acute ischemic and hemorrhagic stroke.

The newly reconfigured Neuropathology and Molecular Genetics (NPMG) Core of the UCLA AIzheimer's Disease Research Center (ADRC) will perform autopsies and genetic studies on patients with neurodegenerative diseases (and appropriate controls) and will serve as a tissue/fluid and DNA resource for individuals performing innovative research on Alzheimer's disease (AD) and non-AD dementias. The specific aims of the Core are to carry out comprehensive postmortem examinations; harvest and store relevant tissues and fluids; assess lesions semi-quantitatively using special stains and immunohistochemistry; provide definitive diagnoses; prepare tissue microarrays for protein expression studies; expand the genetic resource to include apolipoprotein E (apoE) determinations, DNA storage, and preparation of cDNA on patients with known or suspected mutations; and support other Cores and Projects. Project 2 will use brain tissue from the NPMG Core. The autopsy and tissue harvesting/sampling method has evolved and been optimized in conjunction with interactions with the National Alzheimer's Coordinating Center (NACC). Genetic and autopsy information will be maintained in the ADRC database and transmitted to the ARCC database. The Core has performed comprehensive autopsies on 98 patients characterized in Clinical Core; and supplies tissue to investigators locally and nationally. NPMG Core interacts with Clinical Core to provide autopsy confirmation of clinical diagnoses; with Imaging Core to supply autopsy confirmed diagnoses with patients studied in neuroimaging; with the Data Management and Statistics Core through the ADRC database; with the Education/Information Transfer Core through monthly clinicopathological conferences and other educational activities. NPMG Core accomplished its specific aims in the past funding period and has added innovative new technologies such as the "molecular autopsy" approach involving routine use of microarray techniques to study gene and protein expression in appropriate autopsy cases.

Acute stroke is a common and devastating disorder, the 3rd leading cause of death and leading cause of adult disability in the US. This application for renewal of the UCLA Specialized Program of Translational Research in Acute Stroke Center proposes an integrated research/training program to develop innovative therapies for acute ischemic and hemorrhagic stroke. Five research projects are proposed. Project 1, the Therapeutic Aqueous Oxygen (TAO) Trial is a 50 patient trial to evaluate the feasibility and safety of endovascular delivery of supersaturated, aqueous oxygen to ischemic human cerebral tissues. This trial will be the first formal study of endovascular infusion of neuroprotective agents, a highly promising complementary strategy to endovascular recanalization. Project 2, the Intraoperative CT and Endovascular-Guided Surgery for Intracerebral Hemorrhage (ICES) Trial is a critical phase 2 trial of a new, minimally invasive surgical technique for the evacuation of primary intracerebral hemorrhage, using mobile intra-operative CT and neuroendoscopy. In addition, studies interrogating the fundamental pathophysiology of brain injury in human ICH are proposed delineating differential gene expression, tissue morphometrics, and multimodal MR signatures in the perihematomal region. In Project 3, Telestroke in the Field: Improving Prehospital Stroke Care with Mobile Broadband Telemedicine Technologies, will evaluate the benefit upon prehospital diagnostic accuracy and informed consent of physician-patient interaction in the field employing emerging video handset technologies. Project 4, The MR and Recanalization of Stroke Clots Using Embolectomy (MR RESCUE) Trial, is an international, multicenter pilot trial to determine whether diffusion-perfusion MRI can identify acute cerebral ischemia patients who will benefit substantially from mechanical embolectomy with the Merci Retriever up to 8 hrs from symptom onset. Project 5, the Counterpulsation to Upgrade Forward Flow in Stroke (CUFFS) Trial will assess the tolerability, feasibility, and safety of increasing blood flow to ischemic cerebral fields through external counterpulsation in 30 patients with persisting arterial occlusion. Five Cores support the Projects: Administrative, Patient Access, Blood/Tissue Specimen, Biostatistics/Data Management, and Neuroimaging. A Career Development Program will train the next generation of translational scientists in neurovascular disease at the premedical, medical student, and fellowship level.

The Neuropathology Core (NPC) of the ADRC will perform autopsies on individuals evaluated in the Center, and other consented patients with neurodegenerative diseases who expire at UCLA Medical Center, or whose bodies are transferred here for that purpose. Patients examined will include 'controls'followed in the ADRC and control subjects that come through the UCLA-CHS autopsy service;in the latter group, a brief telephonic questionnaire will be administered to family members/next of kin to assess whether an autopsied individual had been cognitively normal during life (this is a new initiative of the core). In addition to providing appropriate diagnoses on these patients (based upon state-of-the-art immunohistochemical studies), tissues and CSF from all possible autopsies will be rapidly frozen, stored, and made available to investigators doing research in Alzheimer disease and related disorders. The NPC is committed to providing research materials to Centers doing excellent research, but who lack availability of well-characterized autopsy tissues/fluids. These resources also will be distributed to local investigators requiring access to tissues. Autopsy brain specimens from patients with AD, non-AD dementias and controls are an important resource to provide a 'gold standard'diagnosis in a given patient, and can assist in the evaluation of treatment efficacy or unwanted complications. As biomarkers of disease progression are increasingly used to monitor disease progression during life, as well as responses to therapy, understanding the morphoanatomic correlates of these biomarkers (as measures of disease severity) becomes crucial. One group of biomarkers, viz. new neuroimaging techniques that label brain amyloids, needs to be validated in autopsy brain specimens, and NPC is providing valuable material for such a study. The NPC has a wide tissue distribution network and disseminates tissue to many investigators. The NPC supports projects at UCLA, collaborates on national studies (e.g., GWAS), and encourages international collaborative studies. The NPC is committed to developing new methods (e.g. tissue micro-array) to facilitate high throughput analysis and expression profiles of new molecules relevant to neurodegeneration, which are discovered in experimental systems, e.g. those using Drosophila and rodents. Finally, the NPC is committed to supporting other Cores and Projects within the ADRC through assistance with developing protocols, teaching, conferences, and providing tissue and neuropathologic data.

DESCRIPTION (provided by applicant): This application seeks support for a conference entitled Cerebral Amyloid Angiopathy and Related Cerebral Microvasculopathies to be held November 12-14, 2009, at the Ronald Reagan UCLA Medical Center and David Geffen School of Medicine in Los Angeles, CA. The conference will seek to recruit clinical and basic scientists currently active in studying cerebral amyloid angiopathy as well as leading investigators from related fields such as the non-amyloid cerebral microangiopathies (including cerebroretinal vasculopathies, CADASIL) and amyloid immunotherapy, including individuals with expertise in biomarkers. A special effort will be made to include promising clinical and basic junior scientists. The Cerebral Amyloid Angiopathy and Related Cerebral Microvasculopathies conference will focus on recent developments in areas such as amyloid angiopathyrelated microbleeds and other biomarkers, animal models, the non-amyloid angiopathies, and the relation between amyloid angiopathy and anti-amyloid immunotherapeutic approaches to Alzheimer disease.

Core 1: Biospecimen and Pathology Core (BiPC) SUMMARY/ABSTRACT The Biospecimen and Pathology Core (BiPC, Core 1), a biorepository and multi-faceted research resource led by experienced neuropathologists and cell biologists with ongoing active research expertise, will provide critical support for the translational prognostic, diagnostic, and therapeutic studies proposed in the UCLA SPORE in Brain Cancer. Working closely with the Biostatistics, Bioinformatics, and Data Management Core (BBD, Core 3), BiPC will maximize optimal biospecimen collection/storage and efficient disbursement of consented, clinically annotated, and molecularly characterized biospecimens. This Core will support all four SPORE projects with 24/7 tissue procurement, clinical and family history annotation, and molecular characterization of tissues, plus research support in gliomasphere/neurosphere cultures, tissue microarray, and patient-derived orthotopic xenografts (PDOX). The Biospecimen and Pathology Core Co-Directors and Investigators have a long collaboration history with each other and with all the SPORE Project Leaders, as shown in joint publications, grants, and consortia. In addition to an excellent program for the collection, processing, storage, and dissemination of important biospecimens in support of the proposed SPORE research projects, the BiPC agenda goes well beyond these core functions to the innovative development of new scientific approaches for the maximization of rare/sparse specimen collection and the development of new animal models for therapeutic drug discovery. As part of this core, brain cancer cells grown in animals will be specially adapted to have a fluorescent label so that growth or shrinkage of the tumor can be serially followed in vivo when experimental treatments are given to the animals. The BiPC will develop and provide these materials, resources, and information to SPORE investigators and others, both inside and outside of UCLA. An outstanding team of experienced, accomplished, determined, and productive investigators has been assembled for this Biospecimen and Pathology Core. It also has strong institutional commitment in an excellent collaborative environment, which will help to ensure the success of the overall goals of this SPORE proposal.