Methods for the diagnosis and treatment of neurological and neurodegenerative diseases, disorders and associated processes
Inventors
West, Tim • Paoletti, Andrew C.
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12253529-B2
Publication Date
2025-03-18
Expiration Date
Abstract
The invention generally relates to methods for the diagnosis and treatment of neurological and neurodegenerative diseases, disorders, and associated processes. Specifically, the present invention is directed toward methods to determine prognosis, diagnosis or efficacy of a therapeutic regimen by using a detectable label to measure levels of alpha-synuclein.
Core Innovation
The invention provides an in vitro method of identifying an agent that increases alpha-synuclein clearance or decreases alpha-synuclein synthesis in a cell. The method contacts a first neuronal cell with a labeled amino acid and an agent, produces labeled and detectable alpha-synuclein in the first cell culture media, and determines metabolic incorporation of the labeled amino acid into alpha-synuclein using a stable isotope labeling kinetics (SILK) assay.
The SILK assay determines the amount of labeled alpha-synuclein and unlabeled alpha-synuclein in an alpha-synuclein polypeptide selected from SEQ ID NOs: 4 to SEQ ID NO: 13 in the first cell culture media, and provides a first ratio of labeled alpha-synuclein to unlabeled alpha-synuclein polypeptide. A second metabolic incorporation is determined in a second cell culture media collected from a second cell not contacted with the agent, providing a second ratio of labeled alpha-synuclein to unlabeled alpha-synuclein polypeptide.
The first and second cells are obtained from a subject and are neuronal cells from the same subject. An agent is identified as increasing alpha-synuclein clearance or decreasing alpha-synuclein synthesis when the first ratio is less than the second ratio.
Claims Coverage
The partial content identifies one independent claim. It has four main inventive features centered on SILK-based labeled-to-unlabeled alpha-synuclein ratios, treated and matched control neuronal cells from the same subject, and an identification criterion where the treated ratio is lower than the control ratio.
SILK-based agent identification by labeled-to-unlabeled alpha-synuclein ratio comparison
Contacting a first neuronal cell with a labeled amino acid and an agent, collecting alpha-synuclein from the first cell culture media, and determining a first ratio of labeled alpha-synuclein to unlabeled alpha-synuclein polypeptide using a stable isotope labeling kinetics (SILK) assay measuring alpha-synuclein polypeptides selected from SEQ ID NOs: 4 to SEQ ID NO: 13.
Matched control metabolic incorporation measurement in a second cell culture media
Determining a second ratio of labeled alpha-synuclein to unlabeled alpha-synuclein polypeptide in a second cell culture media collected from a second neuronal cell contacted with the labeled amino acid but not contacted with the agent, wherein the first and second cells are obtained from a subject.
Agent identification when treated incorporation ratio is lower than control
Identifying the agent as an agent that increases alpha-synuclein clearance or decreases alpha-synuclein synthesis when the first ratio is less than the second ratio.
Agent selection and neuronal-cell context from the same subject
Requiring the agent to be selected from a group including small molecule inhibitor, antibody, sirtuin 2 inhibitor, proteosome inhibitor, L-DOPA, CEPT inhibitor, metalloprotease inhibitor, cholinesterase inhibitor, NMDA receptor antagonist, hormone, neuroprotective agent, or cell death inhibitor, and requiring the first cell and the second cells to be neuronal cells from the same subject.
Overall, the independent claim coverage centers on SILK assay measurement of labeled-versus-unlabeled alpha-synuclein polypeptides in treated versus matched control neuronal cells from the same subject, and identifying the agent when the treated labeled-to-unlabeled ratio is lower than the control ratio.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
Interested in licensing this patent?