Lee E. Goldstein - US grants

CBET-0821304
Goldstein

This NSF Major Research Instrumentation project requests funding for a femtosecond laser ablation (LA) solid sampling system for analytical coupling ("hyphenation") to a newly-acquired magnetic sector field inductively-coupled plasma mass spectrometer (SF-ICP-MS). The grant hyphenated LA-SF-ICP-MS system is intended for high-resolution two-dimensional elemental and isotopic profiling in a diverse range of materials and research applications. The system will greatly expand the analytical capabilities of the recently-established Center for Biometals and Metallomics (CBM), a unique multidisciplinary analytical facility at the Boston University School of Medicine and College of Engineering. The CBM is the only dedicated facility of its kind in the nation. The centerpiece of the facility is a stateof- the-art SF-ICP-MS instrument acquired through a Shared Instrument Grant funded by the National Center for Research Resources, NIH. This powerful instrument is the gold standard for definitive ultra-trace elemental and isotopic analyses in a wide variety of matrices. Deployment of the proposed hyphenated analytical instrumentation will provide researchers locally and throughout the region with an unparalleled analytical resource that will accelerate lead-edge discovery in a wide range of fields.

Funds are requested for instrument acquisition to hyphenate the SF-ICP-MS instrument with a powerful and complementary technology, high-resolution laser ablation solid sampling. The proposed system couples definitive elemental and isotopic analysis with high-resolution femtosecond laser-assisted spatial sampling for wide-ranging application involving a variety of material substrates. The application proposes to address new questions with new technologies coupled in new ways and capitalizes on presently available analytical capabilities and interdisciplinary expertise. Collaborations that will benefit through use of the hyphenated analytical instrumentation include the Museum of Fine Arts, Boston, Lawrence Berkeley National Laboratory, Museum of Ireland, Harvard and Boston University Schools of Public Health, and the US Geological Service, amongst others. Access will be open to qualified students, staff, and researchers throughout the region who require this advanced analytical instrumentation for scientific inquiry. Senior scientists, postgraduate researchers, graduate students, undergraduates, and facility staff will be directly involved in utilizing the requested instrumentation.

The hyphenated system will be managed as a shared-collaborative user resource allied to other significant institutional analytical facilities, including the Boston University Mass Spectrometry Resource. A strong emphasis will be placed on interdisciplinary interaction as well as education and training. Facility staff will develop training courses, facilitate analytical protocol development, assist collaborating scientists with experimental design and execution, and organize internal and external conferences to promote the use of advanced mass spectrometry instrumentation and allied hyphenated technologies for application in a wide array of research programs and applications. Educational activities at the high school, undergraduate, graduate, and post-graduate levels will be integrated with ongoing analytical operations to facilitate both formal and informal interaction amongst students, trainees, senior investigators, and resource staff. Regional, national, and international scientific exchange is already operative and is expected to expand with project funding. Novel utilization, pilot experimentation and technology development will be encouraged to open new and expanded research opportunities using LA-ICP-MS instrumentation.

Intellectual Merit
Reactive oxygen species (ROS) and oxidative stress are major contributors to the pathogenesis of important neurodegenerative diseases including Alzheimer's disease, Parkinson's disease, and cerebrovascular disease. The central nervous system comprises the most oxidatively active organ system in the body. Under normal physiological conditions, brain activity- and the neuronal and synaptic processes underpinning this activity- generates free radical species that progressively damage essential biomolecules (nucleic acids, lipids, proteins). Under a variety of pathological states, ROS-mediated oxidative damage is dramatically accelerated and leads to irreversible brain damage, cerebral dysfunction, cognitive decline, and death. An overwhelming body of scientific evidence now points to ROS-mediated oxidative damage as a key pathogenic pathway involved in the earliest stages of many neurodegenerative diseases. Technology to quantitatively detect and monitor ROS is critical for understanding and treating these disorders, Currently available ROS assay systems (1) detect only a single (or at most a limited number) of biological relevant species, (2) chemically interact with the species under analysis, (3) require complex, time-consuming, labor-intensive analytical processing, and (4) are temporally disconjugate with respect to the short half-lives of most biologically relevant ROS species. This last point is especially important and frequently overlooked. By the time analytical measurements are initiated using conventional methods, significant loss of signal has accrued due to decomposition. For all of these reasons, available ROS detection technology does not meet the analytical standards required for modern biomedical research. A transformative research program to develop an innovative nanotechnology-based toolkit for measuring ROS in biological systems is proposed. A non-enzymatic probe (nanoceria), integrated sensor components, and simplified detection procedure will enable sequential analytical operation on a small, inexpensive chip. An outstanding merit of the proposed approach is the use of a versatile nanoparticle detector array that generates a detectable amperometric signal following oxidation state alterations induced by interaction with ROS. The proposed technology development program will enable fundamental studies of neurodegenerative disease pathogenesis that have not been previously possible.

Broader Impact
The outcome of this research is linked to high-impact national healthcare priorities. The program will establish an interdisciplinary collaboration between the University of Central Florida; Boston University School of Medicine, College of Engineering, and Photonics Center; and the National Institutes of Health (NIH)-funded Alzheimer's Disease Center at Boston University. Education and training are essential components and leverages graduate and undergraduate teaching opportunities, coursework (including a highly successful internet-based off-site access program), and summer research programming. Additional emphasis will focus on minority and K-12 students who participate in on-campus interdisciplinary educational programs at both institutions. The proposed research and educational activities will provide a unique cross-dimensional (nano-to-meso scale) and cross-disciplinary (materials, electrochemistry, fluid mechanics, electrical engineering, neurobiology) approach for development of innovative non-enzyme biosensors with far-ranging biomedical impact. This research will deepen understanding of electrochemical and biochemical reactions at the nanoscale and affords significant potential to provide new insights into the pathogenic role of ROS in human neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, and stroke.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project is investigating the protein-protein interactions between the Alzheimer's peptide A-beta and alpha-B crystallin, which may mediate neurotoxicity in Alzheimer's disease. Western blots have provided some evidence for colocalization of A-beta and alpha-B-crystallin in AD deposits. Preliminary evidence indicates that alpha-B crystallin blocks A-beta fibril formation and potentiates the neurotoxicity of recombinant human A-beta 1-42. Experiments are now underway to identify Ab-aB crystallin crosslink signature(s) in vitro using purified proteins and mass spectrometry. If this proves successful, later experiments will explore the occurrence of such crosslinks in human tissue samples.

DESCRIPTION (provided by applicant): Funds are requested for a femtosecond infrared laser ablation (FIRLA) platform for coupling (hyphenation) to the NCRR-funded high-resolution sector field ICP mass spectrometer (SF-ICP-MS) located at the Center for Biometals and Metallomics at Boston University. The hyphenated FIRLA-SF-ICP-MS system will enable unprecedented ultra-trace elemental and isotopic mapping studies conducted by over forty innovative research teams representing leading institutions throughout the region and across the globe. The hyphenated system will be the first such dedicated biometallomic resource of its kind in the nation. Femtosecond laser ablation technology provides unprecedented spatial resolution with greatly improved analytical accuracy and precision with minimal collateral thermal damage, thus affording for the first time ultra-trace elemental and isotopic mapping studies in biological specimens at micron resolution. The requested shared resource will facilitate a wide range of innovative research conducted by leading interdisciplinary teams at Boston University School of Medicine, Boston University School of Public Health, Harvard Medical School, Harvard School of Public Health, Massachusetts General Hospital, Brigham &Women's Hospital, United States Geological Service, Armed Forces Radiobiology Research Institute, Lawrence Berkeley National Laboratory, Museum of Ireland, Queen's University, and the University of Melbourne. Deployment of the proposed hyphenated instrument at the NCRR-supported Center for Biometals and Metallomics will accelerate lead-edge discovery in Alzheimer's disease, Parkinson's disease, traumatic brain injury, ocular disorders, environmental toxicology, cancer biology, micronutrient physiology, sepsis and inflammation, biometallomic osteoarcheology, forensic pathology, military medicine, and nanotherapeutics. The requested instrument will be managed as a shared resource allied to other regional resources at Boston University, including the NCRR Mass Spectrometry Resource, NIA Alzheimer's Disease Center, NCRR Clinical and Translational Science Institute, the Framingham Study, and the Boston University Photonics Center. The CBM is staffed by an expert analytical team with over two decades of experience in biomedical metallomic analyses, laser ablation, laser-induced breakdown spectroscopy, elemental and isotopic ICP-MS, and optical emission spectroscopy. The Principal Investigator and Advisory Committee will promote equitable access and cost-effective utilization of this unique analytical resource. This shared instrument and allied core resource will facilitate establishment of a National Center of Excellence in metallomics and catalyze scientific discovery at the forefront of this important new field. PUBLIC HEALTH RELEVANCE: Funds are requested for an ultra-fast laser for metallomic tissue mapping at the NCRR-funded Center for Biometals &Metallomics, Boston University. This shared resource will support pioneering research in Alzheimer's disease, Parkinson's disease, traumatic brain injury, lung cancer, environmental toxicology, and nanomedicine.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is investigating the protein-protein interactions between the Alzheimer's peptide A-beta and other prtoeins, such as alpha-B crystallin, which may mediate neurotoxicity in Alzheimer's disease, and with metals. Western blots have provided some evidence for colocalization of A-beta and alpha-B-crystallin in AD deposits. Preliminary evidence indicates that alpha-B crystallin blocks A-beta fibril formation and potentiates the neurotoxicity of recombinant human A-beta 1-42. Experiments are now underway to identify Ab-aB crystallin crosslink signature(s) in vitro using purified proteins and mass spectrometry. If this proves successful, later experiments will explore the occurrence of such crosslinks in human tissue samples. Recent experiments have begun to use 2D-MALDI-TOF MS/MS to explore the spatial distribution of proteins in mouse brain which has already been analyzed by ICP-MS to locate metal distributions.

ABSTRACT More than half of residents in nursing home communities suffer from cognitive impairment with Alzheimer?s disease (AD) or AD related dementia (ADRD), and one-third of all US COVID-19 deaths are long- term care facility residents. The COVID-19 pandemic places AD patients at a greater risk of respiratory failure and mortality. Hypertension, which is prevalent among elderly populations, results in more severe COVID-19 symptoms. The goal of this supplement NIH application is to examine whether AD patients vulnerable to infection by severe acute respiratory syndrome coronavirus (SARS-CoV-2) can improve clinical outcomes of COVID-19 with angiotensin converting enzyme inhibitors (ACEI). This supplement application fits within the scope of our NIA-funded parent project, ?Big data and small molecules for Alzheimer?s disease (RF1- AG063913).? The hypothesis from the parent project was that tauopathy and related neurodegenerative disease pathologies can be suppressed in mice treated with ACEI and statins. The hypothesis for this supplement project is that ACEI reduces the susceptibility, severity, and improves outcomes of SARS- CoV-2 infection in AD patients. This supplement research shares the same goal of the parent project, which aims to meet an urgent need to identify and fast-track new AD therapies (ACEI) with a clear efficacy readout. The scientific premise for our approach is strong. First, angiotensin II is elevated in both COVID-19 and AD patients, making ACE (converting angiotensin I to II) the prime target for inhibition. Second, SARS-CoV-2 binds to its target cells through ACE homolog ACE2. Third, treating human cell organoids with recombinant ACE2 reduces the viral load of SARS-CoV-2, and treating patients with ACEI up-regulates ACE2 in those with hypertension. Using a national database, we have reported significantly longer preclinical (asymptomatic) intervals before AD onset in subjects treated with ACEI and statins compared to those taking neither drug. We have identified ~350,000 subjects on an ACEI, with sufficient power to determine whether there is an association between ACEI and COVID-19 among AD patients. We propose to achieve three Specific Aims. Aim 1. To determine the susceptibility of AD to SARS-CoV-2 infection. This is a unique Aim that supplements the parent grant by using the original data set extended with data on COVID-19 and other variables including geographic regions. Aim 2. To determine the association of individual ACEI with the reduced occurrence of COVID-19 in medicated AD patients. We will divide all ten prescribed ACEIs into blood-brain barrier (BBB) crossing and non-crossing ACEIs and determine which group of/individual ACEIs reduce the occurrence of COVID-19 in AD patients. Aim 3. To determine the association of ACEI therapies with the severity of COVID- 19 symptoms in AD patients. The severity of COVID-19 will be defined by self-quarantine, hospitalization, intensive care unit admission, use of mechanical ventilators, as well as mortality.

Biomarkers provide important information for early detection, differential diagnosis, and disease monitoring of Alzheimer?s disease (AD) and AD-related dementias (ADRDs), including chronic traumatic encephalopathy (CTE). The newly established Biomarker Core of the Boston University Alzheimer?s Disease Research Center (BU ADRC) will be led by three clinician-scientists with experience in biomarker development and validation: Lee Goldstein, M.D., Ph.D. and Wendy Qiu, M.D., Ph.D., who will serve as co-leaders, and Ronald Killiany, Ph.D., who will lead the neuroimaging component. The Biomarker Core will leverage their expertise and existing institutional resources to focus on fluid biospecimen and neuroimaging biomarkers relevant to Alzheimer?s disease (AD) and related dementias (ADRD), including chronic traumatic encephalopathy (CTE). The goal of the Biomarker Core is to collaborate with other cores and centralize biomarker initiatives to support the BU ADRC mission to improve early and accurate diagnosis, differentiation, and monitoring of AD and ADRDs, including CTE. The new Biomarker Core will bank, distribute, and analyze fluid biospecimens and neuroimaging data for shared use by investigators within and outside the BU ADRC and allied national consortia. We will focus on established biofluid and neuroimaging biomarkers (Amyloid-Tau-Neurodegeneration A/T/N NIA-AA Research Framework) with the goal of identifying differences between AD and ADRDs. The Core will also conduct discovery-based development of novel emerging biofluid and neuroimaging biomarkers to enable earlier and more accurate detection, differential diagnosis, staging, and tracking of AD and ADRDs across the disease spectrum. Four Specific Aims are proposed. Aim 1: Process, bank, and distribute fluid biospecimens and neuroimaging data. Aim 2: Measure and analyze established biomarkers (A/T/N NIA-AA Research Framework). Aim 3: Conduct discovery and development of emerging biomarkers, including analysis of blood-derived exosomes and multimodal computational strategies for neuroimaging data. Aim 4: Train next-generation AD biomarker and neuroimaging research leaders. We anticipate that the new Biomarker Core will strengthen BU ADRC research, promote sharing of fluid and neuroimaging biomarker resources, harmonize with efforts to advance national AD/ADRD initiatives, and provide new new insights and biometrics for personalized medicine approaches to diagnose, treat, and prevent AD and ADRD.

Project Summary The path to effective treatment and prevention of Alzheimer's disease (AD) depends on disease detection that occurs before it is too late to reverse progression. Amyloid beta (Aß) is the widely accepted gold standard biomarker of AD and current methods for measuring this biomarker rely on positron emission tomography (PET) scans and/or analysis of cerebrospinal fluid (CSF). These AD biomarker acquisition methods, however, are expensive, invasive, and difficult to scale and reliance on these approaches have exacerbated racial and ethnic disparities in AD research. Digital technologies offer an alternative method for clinical phenotyping that can detect AD-related changes well before the threshold of clinical symptom severity meets diagnostic criteria. Further, digital phenotyping makes possible the identification and validation of digital biomarkers by determining digital indices that correlate highly with more widely-accepted biological biomarkers. Within this context, this application seeks to capitalize on the opportunistic timing of the Framingham Heart Study (FHS) middle-aged Generation 3 and Omni Generations 2 cohorts as participants return for their NHLBI-funded 4th health examination. The NHLBI funding, however, only covers costs associated with about 20% of the health exam components. The remaining 80% of the health exam will be determined by ancillary studies such as the project proposed here. This project aims to add two new components to the Gen 3/OmniGen 2 health exam. Aim 1 proposes conducting a novel lens A? eye scan that pairs a topically-applied fluorescent A?-binding ligand with a specialized spectroscopic eye scanner that can detect Aß deposition in the lens of the eye and has demonstrated higher sensitivity and specificity to detect early AD-related A? pathology compared to amyloid-PET brain scans. Aim 2 seeks to use a smartphone application to collect 3 years of longitudinal cognitive metrics from which to characterize those with stable cognition versus declining cognition. Proposed analyses across these two aims will test the overall hypothesis that novel digital cognitive profiles that are unique combinations of digital features (e.g., item-specific responses, latencies, error rates, acoustic and linguistic measures) can detect those who are lens A? positive and/or at high AD risk (e.g., high cardiovascular risk, ApoE4+, family history of dementia, women, age >60+). Aim 3 will further apply traditional a priori and novel data-driven machine learning computational tools to construct multi-marker profiles that are highly predictive (AUC > .85) of stable cognition and cognitive decline. We posit that machine learning methods will generate more highly predictive models specific to digital cognitive profiles as compared to a priori methods.