Multiple System Atrophy News - March 2002

[Guest guest]

Multiple System Atrophy News - March 2002

Table of Contents

1. MSA Annual Patient Conference Rescheduled in Boston May 3 - 5, 2002

2. MSA Donations

3. Announcing the European Multiple System Atrophy Study Group

4. Boston Globe article on MSA

5. New fact sheet explaining Olivopontocerebellar atrophy (OPCA)

6. News from the Center for Neurodegenerative Disease Research

7. NIH Funded Studies involving Alpha-Synuclein (2002)

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

1. MSA Annual Patient Conference Rescheduled in Boston May 3 - 5, 2002

(Contributed by Don Summers donsums@...)

We're back on track for Boston!!!!!

The Boston Meeting for MSA Patients, Families & Researchers has been

re-scheduled for

May 3rd thru 5th, 2002

As initially scheduled, the meeting will take place at the Holiday

Inn,

Logan Airport. Again, a block of rooms has been reserved in

the name of " THE SDS/MSA SUPPORT GROUP " .

Reservations may be made by calling either the hotel direct

at or the Holiday Inn toll free number, .

When requesting room reservations, please use the following

identifier code: 2SDS

The rooms are reserved at $129.00 per night. The facility is within

a mile

of the airport and offers free shuttle service to and from the

airport. Call

the Holiday Inn, Logan Airport from the free phone near the baggage

claim

area. Handicapped rooms are available. Please inform the staff of

your special needs when you register and when you arrive.

GENERAL MEETING SCHEDULE

Friday, May 3rd

7:00 PM -- Registration and Social Hour

Saturday, May 4th

8:00 AM -- Buffet Breakfast

9:00 AM -- Meeting begins

12:00 Noon to 1:15 -- Lunch

1:15 to 5:00 -- Meetings Continue

Sunday, May 5th

9:00 AM to 10:00 AM -- Continental Breakfast

10:00 AM to 12:00 Noon -- Wrap-up session and general business meeting

(A detailed schedule will be available at registration.)

Dr. Roy Freeman of Beth Israel-Deaconess Medical Center will be our

host

and has arranged for several guest speakers.

There is no cost for the meeting. Scheduled meals are provided by the

SDS/MSA Support Group. Your lodging, transportation and other meals

are at

your expense.

Please e-mail me don.summers@...

if you plan to attend. In your e-mail please include the following:

1. Your name

2. Your home city

3. The number of people attending

This information is urgently needed to finalize food services!

Don Summers

President

SDS/MSA Support Group

1-866-SDS-4999

email: don.summers@...

website: http://www.shy-drager.com

-----------------------------------------

2. MSA Donations

These are two of the most popular options for MSA donations in the

USA.

If you need suggestions for donations in other countries let me know.

a. The SDS/MSA Support Group

The SDS/MSA Support Group now has memorial envelopes

available. These envelopes are pre-printed with the

address of the Support Group and have a space inside

to enter the name of the person the gift is intended

to be allocated to. They are tastefully done and are

available in any number to survivors.

Please notify me if you wish to distribute these

envelopes at the memorial services for your loved

ones.

Don Summers

President, The SDS/MSA Support Group

email: Don.Summers@...

Toll free:

The Shy-Drager Syndrome/Multiple System Atrophy

Support Group is a full non-profit organization

approved by the Internal Revenue Service.

Donations fund the entire operation of the Group.

Services include:

- a toll-free number to call for information and

physician referral

- the maintenance of the group website

http://www.shy-drager.com

- access to the Shydrager mail list

- an annual patient/caregiver/family member meeting

with prominent physicians.

Please visit the website at www.shy-drager.com

Don Summers can be reached at this toll-free number,

Contributions of any amount may be mailed to:

The SDS/MSA Support Group

2004 Lane

Austin, Texas 78728

b. Vanderbilt Shy-Drager Research Fund

Family members and friends of patients with Multiple

System Atrophy or Shy-Drager syndrome have

occasionally expressed an interest in making a

financial contribution that can be used for research

on this condition. Accordingly, we have established a

fund for this purpose. All gifts are very much

appreciated.

Donations for Multiple System Atrophy or Shy-Drager

syndrome research can be sent to:

Vanderbilt Shy-Drager Research Fund

Vanderbilt University Medical Center

AA-3228 Medical Center North

Nashville, TN 37232-2195

-----------------------------------------

3. Announcing the European Multiple System Atrophy Study Group - EMSA-

SG

" Dear Pam,

With interest I read you mail about the MSA/SHY-Drager " online "

support

group. This is just to inform you that there exists a European MSA

Study

Group, initiated from the Department of Neurology, University of

Innsbruck (study coordinator: Prof. Werner Poewe, head of the

Department

of Neurology, assistant coordinator: Prof. Gregor Wenning).

It would be of great help for us if you could inform your members,

especially those from Europe, about this initiative. More details you

can find on our homepage under

http://www.emsa-sg.org

In case you need more information please let us know.

Kindest regards and many thanks for your help in advance. "

Ursula Knapp

EMSA-SG secretariat

Department of Neurology, University of Innsbruck

Head: Prof. Werner POEWE, MD

A-6020 Innsbruck, Anichstraße 35

Telefon: ++43/512/504-3850

Fax: ++43/512/504-3852

Email: werner.poewe@...

gregor.wenning@...

EMSA-SG Secretariat: ursula.knapp@...

-----------------------------------------

4. Boston Globe article on MSA

(Contributed by Carol Langer cblanger@...)

This appeared in the Boston Globe as a side bar in an article about

brain

pacemakers. Tony Swartz-Lloyd is a member of the Boston area MSA

support group.

---

The Boston Globe

January 1, 2002

" For MSA Patients, Remedies Are Elusive "

By: Judy Foreman

Despite the promise of deep brain stimulation for a number of

neurologic

problems, there are some conditions for which it doesn't seem to help,

including a baffling - and devastating - condition called multiple

system

atrophy, or MSA, one of several diseases loosely termed " Parkinson's

Plus. "

MSA, a neurodegenerative disease that is often misdiagnosed as

Parkinson's,

affects an estimated 25,000 to 100,000 people.

" It's like a thief in the night, " said Tony Swartz-Lloyd, 65, a

longtime

vice president at Beth Israel Deaconess Medical Center who is now

retired

and coping with MSA. " It's a weird and elusive disease. . . . It

takes a

little piece of you here, a little piece there. You don't realize

what's

missing at first. "

MSA, for which there is no long-term effective treatment, often

starts, like

Parkinson's, with a loss of dopamine-producing cells in the brain. But

unlike Parkinson's, dopamine-boosting drugs don't seem to help for

more than

a couple of years.

Early symptoms of MSA include loss of balance and coordination,

difficulty

speaking, a drop in blood pressure upon standing up but high blood

pressure

while lying down, stiffness and slowness of movement. Patients often

develop

other problems, such as impotence and difficulty urinating, that are

triggered by degeneration in the autonomic nervous system, which

controls

involuntary bodily functions.

Despite the gloomy prognosis that many MSA patients face - gradual

loss of

many bodily functions and death within six to eight years - there are

some

bright spots, notably research suggesting that an underlying problem

appears

to be abnormal deposits (on brain cells) of a protein called

alpha-synuclein.

Researchers are also studying neuroprotective drugs to keep brain

cells from

dying in diseases such as MSA and Parkinson's, and other agents to

help new

brain cells grow.

Copyright © 2002 Globe Newspaper Company

Record Number: 0201010293

-----------------------------------------

5. New fact sheet explaining Olivopontocerebellar atrophy (OPCA):

Finally they've rewritten this topic to include more up to date

information.

Note that OPCA can fall under the heading of:

1. MSA - Multiple System Atrophy (sporadic/non-hereditary)

or

2. SCA - Spinocerebellar Ataxia (hereditary)

See: http://www.emedicine.com/neuro/topic282.htm

-----------------------------------------

6. News from the Center for Neurodegenerative Disease Research.

(Contributed by Steve Crawford bigdeebe@...)

" Dear Mr. Crawford,

We have come a long way since the discovery of MSA and this has been

driven

by exciting work in the last 4 years on the protein (known as

alpha-synuclein) that forms the toxic aggregates in brain cells of MSA

patients. These new insights into MSA are prerequisites for

identifying

targets for novel drug discovery efforts for MSA patients, but this

still

requires substantial investments in time and resources. However, I am

pleased to say Penn and our Center for Neurodegenerative Disease

Research

(CNDR; with which Dr. Hurtig and his colleagues work very closely) is

now a

leader in research on this subject, so you should know that many of

us feel

we have turned an important corner on efforts to develop better

therapies

for MSA. Please visit our CNDR website at the address below for more

information.

http://www.uphs.upenn.edu/cndr/

Finally, as an indication of our optimism about moving forward in

understanding MSA, I can tell you that we have a grant pending now

that will

enable experts from across the USA to work together in

multidisciplinary

studies of MSA and we are hopeful it will be funded in the near future

thereby enabling us to ramp our activities further in this area of

research. "

Sincerely,

Q. Trojanowski, M.D., Ph.D.

Center for Neurodegenerative Disease Research

Division of Anatomic Pathology

Department of Pathology and Laboratory Medicine

HUP, Maloney 3rd Floor

36th and Spruce Streets

Philadelphia, PA 19104-4283 USA

Tel: ; Fax:

E-mail: trojanow@...

-----------------------------------------

7. National Institutes of Heath - Funded Studies involving Alpha-

Synuclein

(2002)

These studies are all investigating possible treatments which might

prevent

alpha-synuclein aggregation and thus have potential for Multiple

System

Atrophy as well. See: https://www-commons.cit.nih.gov/crisp/

----

Grant Number: 5P01AG009215-12

PI Name: TROJANOWSKI, JOHN Q.

PI Email: trojanow@...

PI Title: LABORATORY MEDICINE

Project Title: MOLECULAR SUBSTRATES OF AGING AND NEURON DEATH

Abstract: Mechanisms of brain dysfunction and death due to

neurodegenerative

diseases of the central nervous system (CNS) are poorly understood,

but the

emergence of profound cognitive and/or motor impairments in these

heterogeneous diseases is a manifestation of the progressive and

massive

degeneration of selectively vulnerable populations of neurons that

distinguishes neurodegenerative diseases from normal aging.

Additionally,

intracellular filamentous inclusions are neuropathological hallmarks

of many

neurodegenerative diseases despite their heterogeneity. For example,

abnormal alpha-synuclein filaments aggregate to form Lewy bodies

(Lbs) in

neurons and they are signature lesions of Parkinson's disease (PD),

dementia

with Lbs (DLB) and an Alzheimer's disease (AD) subtype known as the LB

variant of AD (LBVAD), while multiple system atrophy (MSA) is

characterized

by glial cytoplasmic inclusions (GCIs) composed of alpha-synuclein

filaments

aggregate to form Lewy bodies (Lbs) in neurons and they are signature

lesions of Parkinson's disease (PD), dementia with Lbs (DLB) and an

Alzheimer's disease (AD) subtype known as the LB variant of AD

(LBVAD),

while multiple system atrophy (MSA) is characterized by glial

cytoplasmic

inclusions (GCIs) composed of alpha-synuclein filaments. Although

these

inclusions often were regarded as epiphenomena unrelated to

mechanisms of

brain degeneration, this view has undergone dramatic revisions after

the

discoveries that: mutations in the alpha- synuclein cause familial PD

in

rare kindreds, alpha-synuclein is a major component of Lbs and GCIs,

and

wild type, as well as mutant alpha- synuclein form filaments in vitro

similar to those in Lbs and GCIs. Moreover, the pressure or abundant

alpha-synuclein from filaments in vitro similar to those in Lbs and

GCIs.

Moreover, the presence of abundant alpha-synuclein Lbs in the most

common

variant of sporadic AD (i.e., LBVAD), >60% of familial AD brains and

>50% of

Down's syndrome brains with AD provides an opportunity to elucidate

mechanisms of the enigmatic, but frequent overlap of AD and PD.

Finally, we

also have shown that beta- and gamma-synucleins accumulate in

dystrophic

overlap of AD and PD. Finally, we also have shown that beta- and

gamma-synucleins accumulate in dystrophic hippocampal processes in PD

and

DLB. Thus, we hypothesize that accumulations of synuclein filaments or

aggregates play a mechanistic role in neurodegenerative diseases

characterized by abundant synuclein pathology, and this Program

Project

grant describes four complementary Projects to test this hypothesis

with the

support of an Administrative,. Clinical, and Neuropathology Core. The

Projects are highly synergistic and pursue research conducted on

disease

brains with authentic human synuclein pathology (Project 1), as well

as on

mechanisms of synuclein pathologies using in vitro (Project 2),

transgenic

fly (Project 3) and transgenic mouse (Project 4) models. This Program

Project will provide new insights into mechanisms of synuclein

pathologies

and their role in brain degeneration, which are likely to accelerate

efforts

to improve the diagnosis and therapy of these and other

neurodegenerative

disorders characterized by filamentous brain lesions.

Thesaurus Terms:

Alzheimer's disease, Lewy body, Parkinson's disease, aging, alpha

synuclein,

histopathology, molecular pathology, neural degeneration

Institution: UNIVERSITY OF PENNSYLVANIA

1 COLLEGE HALL

PHILADELPHIA, PA 19104

Fiscal Year: 2002

Department: PATHOLOGY AND LAB MEDICINE

Project Start: 01-AUG-1990

Project End: 30-APR-2005

ICD: NATIONAL INSTITUTE ON AGING

IRG: ZAG1

----

Grant Number: 1R43AG020031-01

PI Name: LEBOWITZ, MICHAEL S.

PI Email: msl@...

PI Title:

Project Title: Alpha-Synuclein: A Drug Target for Parkinson's Disease

Abstract: DESCRIPTION (provided by applicant): Parkinson's disease

(PD)

affects 1 percent of the population over 60 years of age and is one

of the

most common motor disorders. A pathologic hallmark of PD is the

deposition

of intracellular protein inclusions known as Lewy bodies in the

neurons of

affected individuals. The protein alpha-synuclein is a primary

component of

Lewy bodies and recent evidence has implicated alpha-synuclein

oligomerization as a key step in Lewy body formation and in PD

neuropathology. The ultimate goal of this work will be the

development of

pharmaceuticals that can inhibit alpha-synuclein oligomerization and

thus

Lewy body formation and neurodegeneration. In this phase I grant we

will

identify and characterize peptide-based inhibitors of alpha-synuclein

oligomerization. We will study the alpha-synuclein and

alpha-synuclein-peptide interactions and identify the

structure/activity

relationships involved in the peptide inhibition of alpha-synuclein

oligomerization. Finally, we will verify the plausibility of using

a-synuclein oligomerization inhibitors in a cell culture model of

Lewy body

formation. This work will lay the foundation for a phase II grant in

which

we will identify small molecule drug candidates that act as a-

synuclein

oligomerization inhibitors, and begin pre-clinical testing of both

the small

molecule and peptide-based drug candidates in animal models of Lewy

body

disease. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

Thesaurus Terms:

Lewy body, Parkinson's disease, alpha synuclein, antiparkinson drug,

drug

design /synthesis /production cell aggregation, inhibitor /antagonist,

neural degeneration, protein protein interaction, protein structure

function

Institution: PANACEA PHARMACEUTICALS, INC.

9700 GREAT SENECA HWY

ROCKVILLE, MD 20850

Fiscal Year: 2002

Department:

Project Start: 01-AUG-2001

Project End: 30-APR-2002

ICD: NATIONAL INSTITUTE ON AGING

IRG: ZRG1

----

Grant Number: 1R01AG018440-01A1

PI Name: MASLIAH, ELIEZER

PI Email: emasliah@...

PI Title: PROFESSOR

Project Title: B-Synuclein as a Treatment for Lewy Body Disease

Abstract: The new title of this revised application: " beta- synuclein

as a

treatment for Lewy body disease " reflects the modified and sharper

focus on

our scientific approach in response to the reviewers' major concern

that

synuclein might be more relevant for understanding Lewy body disease

(LBD).

We now propose a new concept where beta-synuclein, a naturally

occurring

anti-aggregation molecule and non-amyloidogenic homologue of

alpha-synuclein, might prevent the neurotoxic effects of alpha-

synuclein

and could be a suitable target for the development of an alternative

treatment for LBD. In this context, we propose the following Specific

Aims:

1) To characterize the mechanisms by which beta-synuclein blocks

alpha-synuclein aggregation. We hypothesize that beta-synuclein may

interact

with alpha-synuclein through specific beta-synuclein domains, leading

to

inhibition of alpha-synuclein aggregation. To test this hypothesis,

we will

study the effects of mutant recombinant beta-synucleins and synthetic

beta-synuclein derived peptides on alpha-synuclein aggregation, using

immunoblotting analysis, Congo red/Thioflavine-S staining, and

electron

microscopy. Additional studies of synuclein binding will be performed

by

immunoblotting with His-tagged alpha- and beta-synuclein 2) To

determine if

the anti-aggregation effect of beta-synuclein is protective in

neuronal cell

lines expressing alpha-synuclein. We hypothesize that beta-synuclein

may be

neuroprotective by blocking alpha- synuclein aggregation. To test this

hypothesis, alpha-synuclein- overexpressing GT1-7 and B103 neuronal

cells

will be co- transfected with beta-synuclein GT1-7 cells will be

evaluated

for cell viability, mitochondrial function, oxidative stress

conditions,

GnRH secretion, altered mitochondria morphology and inclusion body

formation. B103 cells will be evaluated by analysis of neurite

formation and

cell adhesion. Immunoblotting experiments will be performed with cells

co-transfected with c- myc-tagged beta-synuclein to assess binding to

alpha-synuclein. To determine potential novel treatments, alpha-

synuclein-

overexpressing GT1-7 and B103 cells will be treated with beta-

synuclein-expressing recombinant adeno-associated viral vector

(rAAV). 3) To

determine if beta-synuclein blocks alpha-synuclein aggregation and

neurodegeneration in in vivo model systems of LBD. We hypothesize that

beta-synuclein may inhibit alpha- synuclein aggregation in vivo. To

test

this hypothesis, we will cross alpha-synuclein tg mice with either

beta-

synuclein tg or knockout mice where murine alpha- or beta-synuclein

gene is

deleted. Mice will undergo detailed behavioral, neurochemical and

neuropathological examination to determine if beta-synuclein

expression

affects the functional and structural alterations promoted by

alpha-synuclein and might act as a basis for treatment of LBD. To

assess the

potential for treatment development, alpha-synuclein tg mice will be

treated

with a beta- synuclein-expressing rAAV. In summary, we will utilize a

multi-system approach (a cell-free, cell culture and tg mouse

systems) to

ascertain the anti- aggregation potential of beta-synuclein as a

therapeutic

target for development of novel treatments for LBD.

Thesaurus Terms:

Lewy body, alpha synuclein, antitoxin, inhibitor /antagonist,

neuroprotectant, protein folding, protein structure function

axon, cell adhesion, cytotoxicity, dendrite, inclusion body, mutant,

nervous

system disorder therapy, protein binding, protein protein interaction

adeno

associated virus group, electron microscopy, gene targeting,

immunoprecipitation, laboratory mouse, staining, tissue /cell culture,

transfection /expression vector, transgenic animal, western blotting

Institution: UNIVERSITY OF CALIFORNIA SAN DIEGO

GILMAN & LA JOLLA VILLAGE DR

SAN DIEGO, CA 92093

Fiscal Year: 2002

Department: NEUROSCIENCES

Project Start: 01-SEP-2001

Project End: 31-AUG-2006

ICD: NATIONAL INSTITUTE ON AGING

IRG: ZRG1

----

Grant Number: 1R21NS043661-01

PI Name: MORGAN, DAVID G.

PI Email: dmorgan@...

PI Title:

Project Title: A vaccine approach to Parkinson's disease

Abstract: DESCRIPTION (provided by applicant) Recent work evaluating

gene

defects leading to Parkinson's Disease (PD) suggests that

accumulation of

alpha-synuclein may be a critical step in the pathogenic mechanisms

leading

to Lewy body Parkinsonism. One of us (RM) has developed a rat model of

alpha-synuclein over-expression which results in long term elevations

of

synuclein production and concomitant degeneration of tyrosine

hydroxylase

neurons in substantia nigra. Because the model uses intracranial

gene-transfer with viral vectors, it has considerable versatility

compared

to transgenic models, where over-expression is constant throughout the

lifespan. Vaccines, while traditionally viewed as prophylactic

approaches to

disease, are increasingly being viewed as therapeutic adjuncts in

cancer and

cardiovascular disease. Others of us (DM and KU) have found that

immunization of transgenic mouse models of amyloid over-expression

with the

All peptide, is surprisingly effective in reducing the Alzheimer

phenotype,

both pathologically and behaviorally, that develops in this model of

the

disease. This application will use the new rat model of alpha-

synuclein over

expression to test the hypothesis that vaccination against alpha-

synuclein

could diminish the toxicity of this agent towards dopamine producing

neurons. We will use immunization with whole recombinant protein and

synthetic peptides and immunization with DNA vaccines. Initial

studies will

verify that these vaccination regimens can indeed produce high

antibody

titers in rats, and determine the active immunization regimen(s) that

leads

to high stable titers. We will next test the hypothesis that

antibodies

against synuclein produced by these vaccines can arrest the

neurotoxicity of

dopaminergic neurons caused by alpha-synuclein over expression.

Additionally, we will examine direct injections of anti-synuclein

antibodies

into the brain, bypassing the blood-brain barrier, to evaluate the

effectiveness of this approach in rescuing dopaminergic neurons.

Success

will encourage further development of a vaccine as a therapeutic

agent in

PD.

Thesaurus Terms:

Parkinson's disease, alpha synuclein, nonhuman therapy evaluation,

synthetic

vaccine, vaccine development, vector vaccine

Lewy body, antiantibody, disease /disorder model, dopamine, neuron,

passive

immunization, recombinant protein, synthetic peptide biotechnology,

cell

sorting, computer program /software, enzyme linked immunosorbent

assay,

green fluorescent protein, immunocytochemistry, laboratory rat

Institution: UNIVERSITY OF SOUTH FLORIDA

4202 E FOWLER AVE

TAMPA, FL 33620

Fiscal Year: 2002

Department: PHARMACOLOGY AND THERAPEUTICS

Project Start: 01-FEB-2002

Project End: 31-JAN-2004

ICD: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE

IRG: ZNS1

----

Grant Number: 1R01NS041799-01

PI Name: MEREDITH, GLORIA E.

PI Email: meredithg@...

PI Title:

Project Title: Synaptic Proteins, Trophic Factors and

Neurodegeneration

Abstract: Description (Provided by applicant): One of the most

fundamental

questions related to the progressive nature of neurodegeneration in

human

disease is how neurons die. Protecting nerve cells against

morphological

decline and death requires blocking intrinsic factors that inhibit

neural

repair. In the present proposal, we offer an innovative approach to

study

those factors that are active in Parkinson's disease (PD) in a new

mouse

model that shows synaptic loss and irreversible nigrostriatal

degeneration.

We propose to track changes of a key synaptic protein, a-synuclein,

both in

its native environment at presynaptic terminals and under neurotoxic

conditions, when it becomes insoluble and accumulates. We will further

correlate those changes with altered neurotrophic support. We have

established an animal protocol by treating C57/bl mice with a combined

regimen of 10 doses of probenecid at 250mg/kg and MPTP at 25mg/kg for

5

weeks. These mice show a slow, progressive loss of nigrostriatal

dopaminergic function for at least 6 months, that mimics PD, with no

signs

of recovery. Three weeks after drug treatment, there is a significant

reduction in the number of substantia nigra (SN) cells and dramatic

changes

in the subsynaptic distribution and density of a-synuclein-

immunoreactive

terminals. These changes could signal the beginning of a chain of

events

that leads to cell death. In this proposal, we will focus on the

progressive

deterioration of dopaminergic neurons in the SN and their inputs, and

present three specific aims to be addressed through a series of

hypotheses.

Specifically, we plan to 1) ascertain the origin and neurochemical

phenotype

of synapses in the SN that contain a-synuclein and to establish

whether MPTP

+ probenecid treatment leads to their degeneration; 2) determine, in

the

MPTP+P model, the temporal relationships between cell death and

a-synuclein-positive synapses, decline in dopamine function and

behavior;

and 3) ascertain whether changes in a-synuclein expression and

production

are precipitated by altered neurotrophic support. The overall

objective of

our research is to understand the relationship between the synaptic

protein,

a-synuclein, neurotrophic support, especially brain-derived

neurotrophic

factor (BDNF) and their respective roles in the PD form of

neurodegeneration. The findings of this research should shed light on

target

areas where neuroprotection strategies can be implemented.

Thesaurus Terms:

Parkinson's disease, alpha synuclein, brain derived neurotrophic

factor,

neural degeneration, protein structure function, substantia nigra,

synapse

Lewy body, cell death, neurochemistry, neuron electron microscopy,

fluorescence microscopy, image processing, immunocytochemistry, in

situ

hybridization, laboratory mouse, methylphenyltetrahydropyridine,

polymerase

chain reaction, statistics /biometry

Institution: UNIVERSITY OF MISSOURI KANSAS CITY

5100 ROCKHILL RD

KANSAS CITY, MO 64110

Fiscal Year: 2002

Department: BASIC MEDICAL SCIENCES

Project Start: 01-MAY-2001

Project End: 30-APR-2005

ICD: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE

IRG: ZNS1

----

Grant Number: 7R01AG017984-02

PI Name: SIERKS, MICHAEL R.

PI Email: sierks@...

PI Title:

Project Title: INHIBITING AGGREGATION WITH PHAGE DISPLAY ANTIBODIES

Abstract: Altered protein processing including misfolding and

aggregation is

a frequent occurrence in aging neurons compared to younger cells. A

number

of neurological diseases such as Alzheimer's Disease (AD) and

Parkinson's

Disease (PD) have been connected with increased misfolding and

aggregation

of specific proteins or peptides. Beta-amyloid (Abeta) is involved in

the

progression of AD through formation of extracellular amorphous

plaques and

neurotoxic fibrils, while alpha-synuclein is involved in the

progression of

PD through formation of intracellular fibrillar aggregates. Different

variants and morphologies of Abeta and alpha-synuclein have been

correlated

with increased formation of the neurotoxic aggregates. Early

detection and

subsequent inhibition of neurotoxic aggregate formation can slow or

stop the

progression of such diseases. The long term goal of this project is to

develop antibody fragments which can be used to identify critical

protein

morphologies which promote neurotoxic aggregate formation; and to

engineer

these antibodies so they can inhibit formation of these aggregates in

vivo

as a potential treatment. Phage display antibody libraries will be

utilized

to isolate pools of single chain antibody fragments (scFvs) which

bind to

particular morphologies of Abeta and alpha synuclein. The specific

aims of

this proposal are to use phage display antibody libraries to; 1)

isolate

scFv antibody fragments specific to various lengths, conformations and

morphologies of Abeta, 2) isolate scFvs specific to various

morphologies of

wild-type and mutant alpha-synuclein proteins, 3) identify which of

these

scFv antibodies can inhibit either Abeta or alpha-synuclein

aggregation and

fibril formation in vitro, which of these scFv antibodies can inhibit

either

Abeta or alpha-synuclein aggregation and fibril formation in vitro,

and 4)

increase the specificity of these antibodies as needed for imaging and

inhibiting aggregation of Abeta or alpha- synuclein under in vivo

conditions. Antibody specificity will be increased by subjecting the

isolated parent antibody to random mutagenesis of targeted antibody

blinding

regions. From this pool of second generation antibodies, scFv

fragments with

increased specificity will be isolated. These high-specificity

antibodies

will be tested for their ability to inhibit formation of neurotoxic

aggregates under in vivo conditions. Protein aggregation will be

monitored

using different techniques including Thioflavin T staining, atomic

microscopy, and electron microscopy.

Thesaurus Terms:

alpha synuclein, amyloid protein, antibody, inhibitor /antagonist,

peptide

library, protein engineering, protein structure

Alzheimer's disease, Parkinson's disease, antibody specificity,

chemical

aggregate, gene mutation, molecular pathology, protein binding atomic

force

microscopy, electron microscopy

Institution: ARIZONA STATE UNIVERSITY

TEMPE, AZ 85287

Fiscal Year: 2002

Department: CHEMICAL & MATERIALS ENGINEERG

Project Start: 01-MAY-2000

Project End: 30-APR-2004

ICD: NATIONAL INSTITUTE ON AGING

IRG: ZAG1