1997 — 2001 |
Teplow, David B. |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Core--Protein Chemistry @ Brigham and Women's Hospital
(Adapted from the application) The core laboratory provides chemical synthesis, purification, and analysis services to each of the projects. By centralizing these activities, the project will make more efficient use of its resources and insure high quality and consistency in the starting materials for each section. Specifically, the core will synthesize wild type amyloid beta-proteins, truncated and elongated forms, as well as derivatives containing amino acid substitutions, D-isomers, unusual amino acids, and radioactive and stable atomic isomers. Peptide and non-peptide inhibitors of fibrillogenesis will be prepared. Cleaved and deprotected peptides will be purified (greater than 95%+) by HPLC to ensure homogeneous starting material. Chemical and isotopic purity will be confirmed by analytical HPLC, amino acid analysis, protein sequencing, and mass spectrometry. Proteins produced in in vitro expression systems will be purified and characterized by similar means.
|
0.907 |
1997 — 2001 |
Teplow, David B. |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Molecular Factors Controlling Ab Fibrillogenesis @ Brigham and Women's Hospital
(Adapted from the application) One approach to Alzheimer's disease therapy is inhibiting fiber formation. If efficacious therapeutic agents are to be developed, the molecular factors controlling fibril nucleation, fibril elongation, and fiber-fiber association must be identified. The applicant's have developed a powerful new paradigm for achieving this goal. Key elements of this paradigm include: 1) a model system for the reproducible growth of A beta fibers; 2) the use of quasielastic light scattering spectroscopy (QLS) to continuously, quantitatively monitor all phases of fibril growth, including prenucleation and nucleation, fiber elongation, and fiber-fiber association; and 3) a mathematical framework which operates upon the QLS data and provides numerical values for the parameters describing each phase of the fibrillogenesis process, including rates of nucleation and elongation. They propose to use this paradigm: 1) To determine the elements in the primary structure of A beta that control the kinetics of fibrillogenesis. They will study the kinetics displayed by wild type A beta peptides, by elongated and truncated A beta peptides, and by A beta molecules containing Glu22>Gln (Dutch), AIa21>GIy (Flemish), and Phe19>Pro substitutions. 2) To determine the effects of the biochemical milieu on the kinetics of A beta fibrillogenesis. The role of pH and ionic strength in controlling fibrillogenesis will be examined. To examine the energetics of fibril nucleation and elongation, they will study the temperature dependance of the rate for each process. 3) To determine the effects of AI3 variants and ApoE on the kinetics of wild type A beta fibrillogenesis. Three mixing strategies will be employed to assess the effects of structural variants of A beta, and of ApoE2, ApoEC, and ApoE4, on the nucleation and elongation stages of fibrillogenesis. 4) To determine the mechanisms of action of fibrillogenesis inhibitors. They will establish quantitatively how fibrillogenesis inhibitors affect A beta micellization, nucleation, and elongation. Four classes of inhibitors will be studied: amyloid dyes, peptides, sulfates, and surfactants. Based on initial results from these studies, as well as from the prior three aims and Project 1, systematic changes in the structures of the inhibitors will be made in order to identify the critical molecular elements mediating each type of fibrillogenesis inhibition.
|
0.907 |
1999 — 2001 |
Teplow, David B. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Structural/Kinetic Analysis of Abeta Fibrillogenesis @ Brigham and Women's Hospital
An increasingly diverse and compelling set of experimental observations supports the hypothesis that Abeta fibrillization starts a cascade of events which eventually results in AD. Although much is known about the primary structure of Abeta in plaques and about the core secondary structure of Abeta in fibrils, little is understand about the mechanisms by which nascent Abeta folds and assembles into fibrils. Identification and characterization of fibrillogenesis intermediates and determination of the kinetics of Abeta fibrillogenesis is crucial if thoughtful, focused efforts are to be made to develop therapeutic agents affecting the fibrillogenesis process. Informative studies in this area will not only significantly accelerate the pace of drug discovery for AD, but will also advance efforts to understand and treat other amyloidoses. Our long term goal is to test the above hypothesis in a series of three steps: 1) to elucidate in molecular detail the mechanisms in Abeta fibrillogenesis; 2) to develop chemical agents capable of blocking or reversing key steps in the fibrillogenesis pathway in vitro; and 3) to formulate and clinically test the efficacy of the agents developed in vitro. This proposal focuses on long term goal #1. The three specific aims proposed build on a number of exciting recent experimental findings. These include the discovery of a potential new therapeutic target, the amyloid protofibril: development of a powerful paradigm, using quasielastic light scattering spectroscopy, for the quantitative analysis of Abeta fibrillogenesis kinetics; and identification of a novel Abeta folding intermediate, the study of which could provide important information about the earliest stages of Abeta fibrillogenesis.
|
0.907 |
2002 — 2005 |
Teplow, David B. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Structural and Kinetic Analysis of Abeta Fibrillogenesis @ University of California Los Angeles
DESCRIPTION (Adapted from applicant's abstract): We hypothesize that amyloid B-protein (ABeta) assembly into oligomers and polymers is a seminal neuropathogenetic process in Alzheimer's disease (AD) and in cerebral amyloid angiopathy (CAA). Inhibiting formation of, or disrupting, ABeta assemblies thus could be of benefit in the treatment of these disorders. To test this hypothesis, detailed knowledge of the folding and assembly of ABeta is necessary. In this proposal, we seek to understand the structural and kinetic features of ABeta fibrillogenesis in order to facilitate later rational design and testing of therapeutic agents. Our prior work in this area has contributed to the discovery and characterization of heretofore-unrecognized conformational and structural intermediates in the assembly process, e.g., protofibrils. In fact, protofibrils have been found to be toxic to cultured neurons and new studies suggest that protofibrils may be key pathogenetic effectors of the Arctic form of familial Alzheimer's disease. Here, we have proposed an ambitious plan that seeks to eventually provide a rigorous structural and thermodynamic elucidation of the entire pathway through which nascent ABeta folds and assembles. Key areas of focus are the early oligomerization of ABeta monomer, the formation of an a-helix-containing conformational intermediate, protofibril assembly, assembly-dependent changes in the topology of amino acids in ABeta, and how the biophysical effects of AD- and CAA-linked amino acid substitutions correlate with the disease phenotype. These studies will contribute to our understanding of AD and CAA. In addition, because amyloid assemblies from most, if not all, of the -20 different kinds of amyloidoses share certain common structural features, the data generated here also should be relevant to these other disorders. Three Specific Aims are proposed: Aim 1. To characterize the conformational, morphologic, and assembly dynamics associated with key intermediates in fibril formation. Aim 2. To determine the topological organization of amino acids during ABeta folding and assembly. Aim 3. To determine the effects of novel AD- and CAA-associated amino acid substitutions on ABeta folding and assembly.
|
1 |
2002 — 2006 |
Teplow, David B. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
3d Structure of Amyloid Beta-Protein Assemblies @ Brigham and Women's Hospital
DESCRIPTION (provided by applicant): We hypothesize that amyloid B-protein (AB) assembly into oligomers and polymers is a seminal neuropathogenetic process in Alzheimer's disease (AD). A direct prediction of this hypothesis is that inhibiting formation of, or disrupting, AB assemblies would be of benefit in the treatment of AD. Detailed knowledge of the tertiary and quaternary structure of AB at each stage of fibril formation would facilitate the design of fibrillogenesis inhibitors. Unfortunately, the three-dimensional structure of AB, both in its monomeric and assembled states, has not been fully elucidated at the molecular level. This proposal seeks to determine the spatial interactions occurring among amino acid side-chains within prefibrillar and fibrillar AB assemblies and then to use these data as distance constraints to construct a dynamic, three-dimensional model of AB fibril assembly. The experimental approach proposed takes advantage of a novel method for "Photo-Induced Cross-linking of Unmodified Proteins" (PICUP). This method provides the means to construct a topological map of the interfaces among AB molecules, without pre factostructural modification of AB and under physiological conditions. In addition to providing valuable information relevant to AD, the results of the work could have broad import because fibrils from a variety of evolutionarily unrelated proteins and peptides appear to share a common core amyloid structure. Two primary aims and four subaims constitute this proposal: Aim 1. To identify interacting amino acids within monomeric AB, and within and between AB, molecules composing higher order AB assemblies. Aim 1A. To identify interacting amino acids within monomeric AB. Aim lB. To identify interacting amino acids within low order AB oligomers. Aim 1C. To identify interacting amino acids in AB protofibrils and AB-derived diffusible ligands (ADDLs). Aim ID. To identify interacting amino acids in AB fibrils. Aim 2. To construct an experimentally-based, dynamic, three-dimensional model of AB fibril assembly.
|
1 |
2003 — 2006 |
Teplow, David B. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Formation and Function of Prefibrillar Abeta Assemblies @ University of California Los Angeles
DESCRIPTION (provided by applicant): We hypothesize that amyloid Beta-protein (Abeta) assembly is a seminal neuropathogenetic process in Alzheimer's disease (AD) and in cerebral amyloid angiopathy (CAA). If so, controlling Abeta assembly could be of therapeutic value. Over the last decade, we have worked to elucidate the pathways of Abeta self-association in vitro, to identify assembly intermediates, and to evaluate the neurotoxic activities of these structures. This work led to the discovery of the amyloid protofibril, later found to be neurotoxic in vitro and in vivo, and to be linked to an "Arctic" form of AD. Recently, novel chemical cross-linking studies have revealed the existence of smaller oligomeric structures (paranuclei), which are formed rapidly by Abeta (1-42) but not by Abeta (1-40). The strong association of Abeta (1-42) with AD thus may result from Abeta (1-42)-specific assembly events occurring at the earliest stage of self-association, oligomerization. We also have described a novel helix-rich oligomeric assembly intermediate. We predicted that an Asp23->Asn amino acid replacement would affect the rate of formation of this intermediate and of fibrils. Interestingly, the importance of this site has been proven in humans through the discovery of an Iowa kindred suffering from an early onset form of CAA caused by this exact substitution. In this proposal, we will examine the thermodynamics and structural biology of early Abeta assembly reactions in order to understand the fundamental factors controlling these reactions and to characterize the structures formed. In concurrent experiments, we will examine the neurotoxic activities of the Abeta assemblies to establish which may be of most relevance pathobiologically. Our studies will provide a better understanding of the mechanisms of formation and the biological activities of Abeta assemblies and of general principles of amyloid formation and protein misfolding. Our experimental plan comprises four specific aims. Aim 1. To elucidate the thermodynamics of early Abeta assemblies and Abeta fibril formation. Aim 2. To determine the structural features of early Abeta assemblies. Aim 3. To determine the structure and mechanism of action of oligomeric Abeta fibrillogenesis inhibitors. Aim 4. To determine the biological activity of early intermediates.
|
1 |
2004 |
Teplow, David B. |
S10Activity Code Description: To make available to institutions with a high concentration of NIH extramural research awards, research instruments which will be used on a shared basis. |
Procise Clc Sequencing Syst: Alzheimer's Disease @ Brigham and Women's Hospital |
0.907 |
2004 |
Teplow, David B. |
S10Activity Code Description: To make available to institutions with a high concentration of NIH extramural research awards, research instruments which will be used on a shared basis. |
Procise Clc Sequencing System 2 Cart W/Pc @ Brigham and Women's Hospital
DESCRIPTION (provided by applicant): Support is requested for the purchase of an Applied Biosystems PROCISE (r) cLC Sequencing System. Applied Biosystems is the only manufacturer of protein sequenators. The PROCISE is a state-of-the-art instrument for automated Edman protein sequencing. The instrument will replace an obsolete Model 477A sequenator that originally was purchased 15 years ago. The PROCISE is approximately 100-fold more sensitive than the 477A, which is currently the sole protein sequenator in a core protein chemistry facility, the Biopolymer Laboratory (BPL), at Brigham and Women's Hospital (BWH). Since 1984, the BPL has provided standard sequencing services to users at BWH, Harvard Medical School and its affiliated teaching hospitals, and other institutions in the US and in foreign countries. In addition, the BPL is a world center for radiosequence analysis, which is done by relatively few laboratories. This technique uses pre.facto radiolabeled protein samples to identify positions of specific amino acids through the periodicity of radioactivity detected in each cycle of the Edman chemistry. This highly sensitive (attomolar) method allows sequence determination of samples not otherwise amenable to sequence analysis or to mass spectrometric characterization. Twelve NIH-funded projects comprise our "major user group." The number of projected sequencing samples, which will keep the new instrument running continuously, and the need for radiosequencing capabilities, make purchase of the PROCISE instrument necessary to achieve the aims of the NIH grants funding the user group. These aims address key questions related to important human diseases, including Alzheimer's disease, Parkinson's disease, multiple sclerosis, and osteoporosis. In addition to providing the means to study these diseases, the improved sensitivity of the PROCISE will open up new opportunities for researchers studying low abundance proteins and who were precluded from applying sequencing methods to their projects due to the relatively low sensitivity of the obsolete 477 instrument. Importantly, as has happened countless times following the introduction of new or improved instrumentation, we fully expect that unforeseen advances in the understanding and treatment of human disease will be enabled by the availability of the PROCISE instrument.
|
0.907 |
2004 |
Teplow, David B. |
S10Activity Code Description: To make available to institutions with a high concentration of NIH extramural research awards, research instruments which will be used on a shared basis. |
Procise Clc Sequencing Syst: Parkinson's Disease @ Brigham and Women's Hospital |
0.907 |
2004 |
Teplow, David B. |
S10Activity Code Description: To make available to institutions with a high concentration of NIH extramural research awards, research instruments which will be used on a shared basis. |
Procise Clc Sequencing Syst: Osteoporosis @ Brigham and Women's Hospital |
0.907 |
2004 |
Teplow, David B. |
S10Activity Code Description: To make available to institutions with a high concentration of NIH extramural research awards, research instruments which will be used on a shared basis. |
Procise Clc Sequencing Syst: Multiple Sclerosis, Experimental Autoimmune Encepha @ Brigham and Women's Hospital |
0.907 |
2006 — 2010 |
Teplow, David B. |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Protein Chemistry Core @ University of California Los Angeles
The "Pathologic Protein Folding and Human Disease" program project application (Program)[unreadable] proposes experimental and computational studies of protein folding and assembly. These studies[unreadable] have relevance to Alzheimer's disease (AD) and other diseases in which pathologic protein folding[unreadable] and aggregation are involved, including Parkinson's, Huntington's, and prion disease. The Peptide[unreadable] Chemistry Core (Core B) will provide chemical synthesis and analysis services to projects 1, 2, 3,[unreadable] and 5. The starting point for the majority of the experimental studies is the amyloid beta-protein (Abeta),[unreadable] which exists in vivo predominately as a 40-42 residue peptide. Core B will synthesize and[unreadable] characterize wild type Abeta peptides and a variety of structurally-related full-length analogues and[unreadable] peptide fragments. Syntheses will be done primarily using automated solid-phase peptide[unreadable] synthesis (SPPS) and FMOC chemistry. Peptide purification will be done using preparative reverse[unreadable] phase HPLC. To characterize the synthetic products, a combination of quantitative amino acid[unreadable] analysis, analytical HPLC, protein sequencing, and mass spectrometry will be used. In addition to[unreadable] direct provision of starting materials and expertise to projects 1, 2, 3, and 5, Core B will synthesize[unreadable] Abeta peptides in which structural modifications will be made to test hypotheses emerging from project[unreadable] 4. Project 4 will utilize in silico approaches to simulate Abeta folding and assembly. These peptides[unreadable] will be provided to the other projects, as appropriate, to perform actual, as opposed to "virtual,"[unreadable] experiments.[unreadable] Dr. Teplow (Director) has been involved in the development and application of methods for peptide[unreadable] and protein synthesis, analysis, and study for almost 25 years (1-55). Ms. Condron (Senior[unreadable] Research Associate) is equally experienced in peptide chemistry and instrumentation and has been[unreadable] working with Dr. Teplow for almost 15 years. In addition to the strictly technical contributions of[unreadable] Core B to the Program, Dr. Teplow, Ms. Condron, and other Core B personnel will collaborate with[unreadable] each of the five projects in the design of physical and virtual peptides for hypothesis testing and to[unreadable] address any technical problems that emerge during the execution of the aims of the projects.
|
1 |
2006 — 2010 |
Teplow, David B. |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Physical Biochemistry and Biology of Amyloid B-Protein @ University of California Los Angeles
In Alzheimer's disease (AD), progressive deposition of amyloid beta-protein (Abeta) fibrils, "amyloid plaques," occurs in the brain. However, neuronal dysfunction may occur prior to plaque formation. Structure-neurotoxicity studies have revealed progressively smaller toxic assemblies, including protofibrils, paranuclei, and ADDLs. In rodents, Abeta oligomers can inhibit long-term potentiation (LTP), a model for learning and memory. In AD patients, Abeta oligomers are present in levels significantly greater than in agematched normal individuals. Biophysical, cell culture, animal, and human studies thus all support the hypothesis that oligpmerization of Abeta is a key event in AD pathogenesis. If so, then the development of therapeutic strategies depends on elucidation of the mechanism(s) of pathologic protein folding, oligomerization, and higher-order assembly. Continuing efforts in our laboratory to understand the earliest steps in Abeta assembly, Abeta monomer folding and oligomerization, have revealed key structural features. These include turn formation in the Val24-Lys28 region of the unstructured Abeta monomer and interactions among the central hydrophobic cluster (Leu17-Ala21), N-terminus, and C-terminus. The four aims comprising this application seek to test mechanistic hypotheses emanating from these observations. Aim 1. To determine the mechanisms of turn formation in the Val24-Lys28 region of Abeta. a. To determine the role of hydrophobic interactions. b. To determine the role of electrostatic interactions. c. To determine the role of amino-acid turn propensity. Aim 2. To determine the mechanisms of intramolecular folding and early Abeta oligomerization. a. To determine the structural dynamics of central hydrophobic cluster (CHC)-C-terminus interactions. b. To determine the structural dynamics of CHC-N-terminus interactions. c. To determine the effects of alternative turn conformations on Abeta monomer structure and oligomerization. Aim 3. To use O?>N acyl migration chemistry to implement a new, quasisynchronous system for studies of Abeta42 folding and self-assembly. a. To synthesize 26-O-acyl-isoAbeta42 (26-AIAbeta42) and study the time-dependent changes in peptide secondary and quaternary structure following initiation of O?>N acyl migration. b. To use quasielastic light scattering spectroscopy to determine kinetic and thermodynamic parameters of Abeta42 self-assembly. c. To use ion mobility spectroscopy-mass spectrometry to monitor early oligomerization events in Abeta self-assembly. d. To synthesize and study the biophysical and biological behavior of Na-protected 26-AIAbeta42. Aim 4. To determine how structural features shown to be critical in controlling Abeta folding and oligomerization affect peptide neurotoxicity.
|
1 |
2006 — 2010 |
Teplow, David B. |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Pathologic Protein Folding and Human Disease @ University of California Los Angeles
DESCRIPTION (provided by applicant): Diseases caused by pathologic protein folding are among the most devastating suffered by the aged. These diseases include Alzheimer's, Huntington's, Parkinson's, familial amyloid polyneuropathy, and prion. Each is a fatal disorder causing progressive cognitive and physical decline. Each is linked to aberrant folding of proteins that results in protein dysfunction. Dysfunction comes both from intrinsic changes in protein monomer structure and protein self-association (aggregation). The latter phenomenon occurs frequently in the dementing illnesses, of which Alzheimer's disease (AD) is the most common. In AD, the amyloid beta-protein (Abeta) self-associates to form oligomeric structures that circulate in plasma and cerebrospinal fluid. As the disease progresses, deposits of Aa are found in increasing number and size in the brain. These deposits, termed "amyloid plaques," contain fibrillar polymers of Abeta. Most cases of AD are not linked to mutations in the cognate structural gene for Abeta. This raises the questions of why pathologic folding and assembly of Abeta occur and why disease incidence increases so sharply after the age of 65. AD is not unique in these respects, as other dementing illnesses also are "diseases of aging." The general problem area addressed by this Program Project (Program) is pathologic protein folding and assembly, as exemplified by Abeta. We hypothesize that conformational changes in Abeta lead to oligomerization and that the resulting oligomeric assemblies are the proximate neurotoxins causing AD. To test this hypothesis, we propose two long-term specific aims: 1. To understand, at the most fundamental biophysical and cellular levels, how aberrant folding of proteins produces human disease. 2. To translate this knowledge into the design and implementation of new therapeutic agents. To accomplish these aims, five principal investigators at four different American universities have joined together to create a tightly-integrated program comprising two administrative cores and five projects. Our long-term strategy seeks first to establish a cohesive and productive research enterprise focused on AD and Abeta. We make this choice because AD is the most common cause of late-life dementia, its incidence is predicted to increase significantly, and Abeta assembly has proven to be archetypal for amyloid proteins. This last point is important, because the Program is designed to advance our understanding of AD and provide new technologies applicable in studies of folding and assembly of other amyloid and non-amyloid proteins. We thus envision the significance of the Program extending beyond solely AD.
|
1 |
2007 — 2008 |
Teplow, David B. |
P41Activity Code Description: Undocumented code - click on the grant title for more information. |
Simulation of Amyloid Beta-Protein Folding and Assembly @ University of California Los Angeles |
1 |
2007 |
Teplow, David B. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Formation and Function of Prefibrillar Amyloid Beta Assemblies @ University of California Los Angeles
DESCRIPTION (provided by applicant): We hypothesize that amyloid Beta-protein (Abeta) assembly is a seminal neuropathogenetic process in Alzheimer's disease (AD) and in cerebral amyloid angiopathy (CAA). If so, controlling Abeta assembly could be of therapeutic value. Over the last decade, we have worked to elucidate the pathways of Abeta self-association in vitro, to identify assembly intermediates, and to evaluate the neurotoxic activities of these structures. This work led to the discovery of the amyloid protofibril, later found to be neurotoxic in vitro and in vivo, and to be linked to an "Arctic" form of AD. Recently, novel chemical cross-linking studies have revealed the existence of smaller oligomeric structures (paranuclei), which are formed rapidly by Abeta (1-42) but not by Abeta (1-40). The strong association of Abeta (1-42) with AD thus may result from Abeta (1-42)-specific assembly events occurring at the earliest stage of self-association, oligomerization. We also have described a novel helix-rich oligomeric assembly intermediate. We predicted that an Asp23->Asn amino acid replacement would affect the rate of formation of this intermediate and of fibrils. Interestingly, the importance of this site has been proven in humans through the discovery of an Iowa kindred suffering from an early onset form of CAA caused by this exact substitution. In this proposal, we will examine the thermodynamics and structural biology of early Abeta assembly reactions in order to understand the fundamental factors controlling these reactions and to characterize the structures formed. In concurrent experiments, we will examine the neurotoxic activities of the Abeta assemblies to establish which may be of most relevance pathobiologically. Our studies will provide a better understanding of the mechanisms of formation and the biological activities of Abeta assemblies and of general principles of amyloid formation and protein misfolding. Our experimental plan comprises four specific aims. Aim 1. To elucidate the thermodynamics of early Abeta assemblies and Abeta fibril formation. Aim 2. To determine the structural features of early Abeta assemblies. Aim 3. To determine the structure and mechanism of action of oligomeric Abeta fibrillogenesis inhibitors. Aim 4. To determine the biological activity of early intermediates.
|
1 |
2010 — 2014 |
Teplow, David B. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Structural Biology of Amyloid Beta-Protein @ University of California Los Angeles
DESCRIPTION (provided by applicant): We hypothesize that amyloid [unreadable]-protein (A[unreadable]) assembly into neurotoxic oligomers and polymers is a seminal neuropathogenetic process in Alzheimer's disease (AD). If so, assembly inhibition or dissociation of existing assemblies could be effective therapeutic approaches. To test our hypothesis, the structural biology of A[unreadable] must be elucidated in detail. What conformers form oligomers? By what mechanism? What are the structures of the oligomers thus formed? What is the relative toxicity of each oligomer species? Many, including ourselves, have striven to correlate structure with measures of biological activity. Recent work has suggested that dimeric or trimeric assemblies are important neurotoxins, but hexameric, nonameric, dodecameric, and larger oligomers also have been shown to be potent neurotoxins. The long-term goal of this proposal is to move past simple quaternary structure determination to elucidation of A[unreadable] monomer secondary and tertiary structure dynamics and the determination of mechanisms of monomer oligomerization. This means eventually understanding the interatomic interactions that control the dynamics, and in doing so, identifying therapeutic targets at atomic resolution. This "knowledge-based" approach is distinct from, but complementary to, high-throughput screening strategies. Both approaches should be executed to maximize the chances for identifying efficacious, disease-modifying therapeutic agents. We propose here to: (1) elucidate the physical biochemistry of A[unreadable] monomer folding and self-assembly;and (2) establish structure-neurotoxicity relationships of the A[unreadable] assemblies thus formed. To do so, we will chemically synthesize A[unreadable] peptides in which specific amino acids and chemical bonds are altered and then study the conformational dynamics and assembly of these peptides. The positions of these alterations, and the alterations themselves, have been chosen carefully so as to reveal the key structural features of the A[unreadable] molecule that control its assembly into structures that damage or kill neurons. We will identify, isolate, and structurally characterize specific types of assemblies and then determine quantitatively the toxic activity of each assembly by treating primary neurons in culture. The depth of understanding of the structures of the assemblies obtained in the first aim will be unprecedented. Thus the knowledge gained through this "structure-activity correlation" process is expected to provide the most accurate assessment of which assemblies, and which structures (at atomic resolution) on these assemblies, should be targeted therapeutically. In addition to its contributions to an improved understanding of AD and its treatment, results of the proposed project should have relevance for studies of other neurodegenerative diseases linked to aberrant protein assembly. These include Parkinson's, Huntington's, amyotrophic lateral sclerosis, familial amyloid polyneuropathy, and the prionoses. PUBLIC HEALTH RELEVANCE: This project will advance our understanding of how a protein, the amyloid-protein, causes Alzheimers disease. This understanding can be translated directly into the development of a new class of drugs that have the potential to modify or cure the disease. The project also will be of relevance to Parkinsons, Huntingtons, amyotrophic lateral sclerosis, familial amyloid polyneuropathy, the prionoses, and other neurodegenerative diseases of aging.
|
1 |
2011 — 2015 |
Teplow, David B. |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Physical Biochemistry and Biology of Amyloid Beta-Protein @ University of California Los Angeles
DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is a devastating disease of the aged. Two amyloid-forming proteins are associated with AD, the amyloid ?-protein (A?) and tau. Recent evidence supports an hypothesis, the amyloid cascade hypothesis, that posits that A? oligomers are the seminal neuropathogenetic agents in AD. The overall goal of this proposal is to understand the structural biology of A?, and fragments thereof, and to establish formal structure-activity relationships. In the long run, we seek to obtain an atomic-resolution determination of the structure of the proximate neurotoxins formed by A?, and in doing so, enable the development, for the first time, of disease-modifying AD treatments. A multidisciplinary strategy, employing complementary experimental and computational approaches, will be employed. This strategy has been used very successfully in the past, providing novel insights into the A? system. Three specific aims are proposed that systematically and logically progress from in vitro biophysical studies of A? and its oligomeric assemblies (Aim 1), to in vitro and in vivo studies of the biological activity of selected such assemblies (Aim 2), to determination of the effects of selected assemblies on differential gene expression in neurons (Aim 3). Taken together, these studies will provide the theoretical and experimental foundation for subsequent therapeutic compound development and clinical testing in humans. Aim 1. To determine the structural dynamics of A? and tau assembly. a. To use scanning Tyr mutagenesis to elucidate mechanisms of A? oligomerization. b. To determine the dynamics of intramolecular turn formation and its effects on A? assembly. c. To determine the effects of primary structure changes on the conformations and assembly dynamics of biologically relevant and theoretically important A? peptides. Aim 2. To determine the biological effects of A? assemblies. a. To determine the cytotoxic effects of A? assemblies on cultured neuronal cell lines and primary neurons. b. To determine the effects of A? assemblies on Drosophila eye development, locomotion, and longevity. Aim 3. To identify and validate AD-relevant genes using a bioinformatics approach that considers A? assembly structure, neuron type, and neuron senescence.
|
1 |
2011 — 2014 |
Teplow, David B |
P50Activity Code Description: To support any part of the full range of research and development from very basic to clinical; may involve ancillary supportive activities such as protracted patient care necessary to the primary research or R&D effort. The spectrum of activities comprises a multidisciplinary attack on a specific disease entity or biomedical problem area. These grants differ from program project grants in that they are usually developed in response to an announcement of the programmatic needs of an Institute or Division and subsequently receive continuous attention from its staff. Centers may also serve as regional or national resources for special research purposes. |
Design and Structure of Drugs Against Alzheimer's Disease @ University of California Los Angeles
Amyloid protein fibrils are associated with a group of devastating human diseases. The precise etiologic agents for these medical conditions remain undefined, but in several cases appear to be protein fibrils or pre-fibrillar oligomers. Currently there is no approved therapeutic agent that regulates the formation of amyloid fibrils and reverses the symptoms. Our working hypothesis is that interfering with amyloid fibrillation and oligomerization is of clinical benefit to patients suffering from Alzheimer's and other amyloid diseases. Amyloid proteins lack common sequence motifs; nevertheless, they display similar biophysical characteristics and a common 'cross-B spine' structure. The first fully objective atomic model of the common B-spine structure of a fibril-forming peptide was determined in our lab, and additional structures are already available. Based on these atomic structures, we are able to design inhibitors. Our recently designed peptide inhibitors of tau fibrils, based on the structure of the amyloid spines of the tau protein determined in our lab, interfere with fibrillation of tau. We plan to improve the bioavailability and potency of these inhibitors and to design similar peptide inhibitors against Amyloid-beta (AB) fibrils. In recent years several compounds were shown by others to inhibit fibrillation, although the molecular mechanism of this interference is not yet clear. We will determine crystal structures of the fibrils bound to various inhibitors that will advance our understanding of the mechanism of inhibition of fibrils and small oligomers by small molecule inhibitors. The structure determination will be coupled to a computational approach to detect non-toxic, specific and potent inhibitors that will cross the blood-brain-barrier and will bind strongly to fibrils and oligomers. Another important application of this study is to find compounds that could be useful as markers for fibrils in biochemical assays as well as in the diagnosis of fibrils in-vivo. Our project is consistent with the aims of The Therapeutic Imperative, and our proposal involves dose collaboration with members of the ADRC community.
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2011 |
Teplow, David B |
P50Activity Code Description: To support any part of the full range of research and development from very basic to clinical; may involve ancillary supportive activities such as protracted patient care necessary to the primary research or R&D effort. The spectrum of activities comprises a multidisciplinary attack on a specific disease entity or biomedical problem area. These grants differ from program project grants in that they are usually developed in response to an announcement of the programmatic needs of an Institute or Division and subsequently receive continuous attention from its staff. Centers may also serve as regional or national resources for special research purposes. |
Ucla Alzeheimer's Disease Research Center @ University of California Los Angeles
DESCRIPTION (provided by applicant): The UCLA Alzheimer's Disease Research Center (ADRC) supports cutting edge research in an environment that mentors junior investigators;attracts numerous researchers;hosts a pilot project program that allows investigators to gamer preliminary data for more advanced studies;collaborates with the National Alzheimer's Coordination Center (NACC) as well as other Alzheimer disease (AD) related investigators locally, nationally and internationally;extends AD research to women and minorities;and works closely with community and advocacy groups including the Alzheimer's Association. The UCLA ADRC is comprised of 6 cores and proposes 3 projects in this renewal application. Cores include: Administrative, Clinical, Data Management and Statistics, Neuropathology, Recruitment and Education, and Neuroimaging and Biomarkers. The three projects include an investigation of MRI techniques applicable to clinical trials and population studies (Project 1;Liana Apostolova;Junior Investigator), a study of the comparative information to be gained from FDDNP and Pittsburg Compound B molecular imaging and their relationship to CSF and plasma biomarkers (Project 2;Gary Small);and a study of antibodies that inhibit amyloid 6 and tau aggregation and may represent novel interventions in AD (Project 3;David Eisenberg). The Theme of the UCLA ADRC is The Therapeutic Imperative, emphasizing the urgency of developing new treatments for AD. Resource use, core organization, project selection, collaboration, and educational activities are prioritized according to their integration with the Center theme. Clinical Core is following 120 patients;Recruitment and Education Core sponsored lectures reaching 20,000 participants;Neuropathology Core performed autopsies on Center patients who died, and non-Center patients to augment tissue distribution. This application has a major emphasis on biomarkers as a key aspect of advancing new therapies for AD. A familial AD cohort is included in this renewal application and a new minority site (Harbor View Medical Center) has been added. Innovations in advancing research are proposed in each Core of this proposal. Each core has responded to criticisms and recommendations from the 2008 review in this renewal application.
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2012 — 2013 |
Teplow, David B |
P50Activity Code Description: To support any part of the full range of research and development from very basic to clinical; may involve ancillary supportive activities such as protracted patient care necessary to the primary research or R&D effort. The spectrum of activities comprises a multidisciplinary attack on a specific disease entity or biomedical problem area. These grants differ from program project grants in that they are usually developed in response to an announcement of the programmatic needs of an Institute or Division and subsequently receive continuous attention from its staff. Centers may also serve as regional or national resources for special research purposes. |
Ucla Alzheimer's Disease Research Center @ University of California Los Angeles
DESCRIPTION (provided by applicant): The UCLA Alzheimer's Disease Research Center (ADRC) supports cutting edge research in an environment that mentors junior investigators; attracts numerous researchers; hosts a pilot project program that allows investigators to gamer preliminary data for more advanced studies; collaborates with the National Alzheimer's Coordination Center (NACC) as well as other Alzheimer disease (AD) related investigators locally, nationally and internationally; extends AD research to women and minorities; and works closely with community and advocacy groups including the Alzheimer's Association. The UCLA ADRC is comprised of 6 cores and proposes 3 projects in this renewal application. Cores include: Administrative, Clinical, Data Management and Statistics, Neuropathology, Recruitment and Education, and Neuroimaging and Biomarkers. The three projects include an investigation of MRI techniques applicable to clinical trials and population studies (Project 1; Liana Apostolova; Junior Investigator), a study of the comparative information to be gained from FDDNP and Pittsburg Compound B molecular imaging and their relationship to CSF and plasma biomarkers (Project 2; Gary Small); and a study of antibodies that inhibit amyloid 6 and tau aggregation and may represent novel interventions in AD (Project 3; David Eisenberg). The Theme of the UCLA ADRC is The Therapeutic Imperative, emphasizing the urgency of developing new treatments for AD. Resource use, core organization, project selection, collaboration, and educational activities are prioritized according to their integration with the Center theme. Clinical Core is following 120 patients; Recruitment and Education Core sponsored lectures reaching 20,000 participants; Neuropathology Core performed autopsies on Center patients who died, and non-Center patients to augment tissue distribution. This application has a major emphasis on biomarkers as a key aspect of advancing new therapies for AD. A familial AD cohort is included in this renewal application and a new minority site (Harbor View Medical Center) has been added. Innovations in advancing research are proposed in each Core of this proposal. Each core has responded to criticisms and recommendations from the 2008 review in this renewal application.
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