Retrograde Tracers with Suicide Enzyme?Prodrug to Study Specific Neural Circuits

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$595,044.00
Award Year:
2009
Program:
SBIR
Phase:
Phase I
Contract:
1R43NS065555-01
Award Id:
94067
Agency Tracking Number:
NS065555
Solicitation Year:
n/a
Solicitation Topic Code:
n/a
Solicitation Number:
n/a
Small Business Information
2633 MARTINEZ DR, BURLINGAME, CA, 94010
Hubzone Owned:
N
Minority Owned:
N
Woman Owned:
N
Duns:
160127655
Principal Investigator:
XINMIN XIE
(650) 784-5658
SIMONXIE@AFASCI.COM
Business Contact:
XINMIN XIE
() -
simonxie@afasci.com
Research Institution:
n/a
Abstract
DESCRIPTION (provided by applicant): Tracing techniques have long been used to analyze specific neural pathways but do not reveal their function. Lesion techniques have also long been employed to analyze the function of neural pathways but are not very sel ective. Current lesion methods use excitotoxic, electrolytic or mechanical lesions, and transgenic expression of a toxic gene product. Here we propose to develop a system utilizing retrogradely transported suicide enzyme or recombinant adenovirus and local ly injected prodrug that will enable the specific ablation of neurons based upon the location of their cell bodies and the projection of their axons. If successful, this innovative technique would significantly advance our ability to lesion particular path ways in the nervous system and characterize their function. The first approach employs a conjugate of a retrograde tracer cholera toxin B subunit (CTB) linked to a suicide enzyme such as nitroreductase (NTR), purine nucleoside phosphorylase (PNP), or horse radish peroxidase (HRP). The second approach uses a replication-defective adenovirus containing genes encoding the suicide enzyme as an efficient tool to deliver this enzyme into neurons via retrograde transport. The conjugate, or the viral vector stereota xically injected into a specific region of interest, is taken up by local axon terminals, and retrogradely transported through the axons to the distant cell bodies of neurons that innervate this brain area. Following 2-5 days after injection, the nontoxic substrate of the enzyme, so called 'prodrug' is injected into one or more selected areas in the brain. This substrate is then converted into a highly toxic product by the suicide enzyme within the retrogradely labeled cell body. Consequently, the toxin ind uces degeneration of only those neurons that contain the suicide enzymes. The Specific Aims of the proposed project are to: 1. Make conjugates of CTB-HRP, CTB-NTR or CTB-PNP that can effectively transport the suicide enzymes from axonal terminals into thei r cell bodies and construct recombinant adenoviral vectors containing the suicide enzyme cDNAs which can move efficiently by retrograde transport into neurons. 2. Determine the conjugated suicide enzyme activity in vitro and investigate the selectivity and neurotoxicity produced by the 'tracer-suicide enzyme-prodrug' systems in rats in vivo Our innovative 'tracer-suicide enzyme-prodrug' approach should produce selective lesions, destroying only those neurons that contain retrogradely transported conjugates or adenoviral construct, while sparing other neurons in the brain. We believe that our user-friendly complete toolkits will be of high importance to the neuroscience community. Our ultimate goal is to develop novel research toolkits to facilitate studies o f neural circuits and discovery of novel drug targets in the neurological and metal health areas. PUBLIC HEALTH RELEVANCE: Lesion techniques have been used to study structure and function of neural circuits in the brain. But the current methods are less se lective and time-consuming. We propose to develop a novel toolkit that allows investigators to rapidly analyze the structure and function of a particular neuroanatomical pathway or a specific neural circuit. This innovative research toolkit will facilitate research and drug discovery in the neurological and metal health areas.

* information listed above is at the time of submission.

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