Tau protein-binding DNA aptamers with capture/detection and therapeutic utilities in tauopathy-related neurodegenerative disorders
Inventors
WANG, Ka W. • Tan, Weihong • Li, Xiaowei • YADIKAR, Hamad • TENG, I-Ting
Assignees
US Department of Veterans Affairs • University of Florida Research Foundation Inc
Publication Number
US-10584340-B2
Publication Date
2020-03-10
Expiration Date
2037-09-29
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Abstract
DNA aptamers that bind to tau protein at its phosphorylatable sites are identified. These disclosed DNA aptamers not only recognize tau at specific phosphorylatable sites, but also demonstrate inhibitory effects on phosphorylation and oligomer formation of tau protein. Molecular probes based on these disclosed DNA aptamers can be used as capture or detection agents to detect the levels of tau and phosphor-tau in cerebrospinal fluid as well as for cell-based or in vivo brain imaging in live animals or human. Compositions comprising these disclosed DNA aptamers can be used to arrest or treat the progression of tauopathy associated neurodegenerative disorders.
Core Innovation
The invention identifies DNA aptamers that specifically bind to tau protein at its phosphorylatable sites. These DNA aptamers recognize tau at designated phosphorylatable sites and demonstrate inhibitory effects on phosphorylation and oligomer formation of tau protein. Molecular probes utilizing these aptamers can be used as capture or detection agents to measure tau and phosphor-tau levels in cerebrospinal fluid and for cell-based or in vivo brain imaging in living animals or humans. Compositions comprising these aptamers can be used to arrest or treat progression of tauopathy-associated neurodegenerative disorders.
The problem addressed arises from the pathological role of tau protein hyperphosphorylation, which leads to tau dissociating from microtubules, forming aggregates and filaments that cause neuron death, a phenomenon termed tauopathy. Tauopathy is implicated in neurodegenerative diseases including Alzheimer's disease and chronic traumatic encephalopathy. Current detection methods for total tau and phosphorylated tau levels, such as mass spectrometry, PET imaging, and western blotting combined with immunoprecipitation or HPLC, require large sample amounts, are costly, or tedious, and no tau-binding DNA aptamers are commercially available. Thus, there remains a need for specific binding agents to recognize and inhibit tau phosphorylation and aggregation.
Claims Coverage
The patent claims multiple independent inventive features related to DNA aptamers capable of specifically binding tau protein at phosphorylatable sites, their truncated forms, compositions including these aptamers, and methods for detection, imaging, and treatment of tauopathy-related diseases.
DNA aptamers comprising 66 nucleotide sequences binding tau at phosphorylatable sites
A DNA aptamer comprising a nucleic acid sequence of 66 nucleotides selected from SEQ ID NO: 1 through SEQ ID NO: 7, capable of specifically binding tau protein at phosphorylatable sites.
Truncated DNA aptamers from SEQ ID NO: 1
A DNA aptamer comprising nucleic acid sequences selected from SEQ ID NO: 8, 9, 10, and 11, which are truncated fragments of SEQ ID NO: 1, comprising 100 nucleotides or less, binding tau protein at phosphorylatable sites.
Truncated DNA aptamers from SEQ ID NO: 3
A DNA aptamer comprising nucleic acid sequences selected from SEQ ID NO: 17, 18, and 19, which are truncated fragments of SEQ ID NO: 3, comprising 100 nucleotides or less, binding tau protein at phosphorylatable sites.
Truncated DNA aptamers from SEQ ID NO: 4
A DNA aptamer comprising nucleic acid sequences selected from SEQ ID NO: 20, 21, and 22, which are truncated fragments of SEQ ID NO: 4, comprising 100 nucleotides or less, binding tau protein at phosphorylatable sites.
Truncated DNA aptamers from SEQ ID NO: 5
A DNA aptamer comprising nucleic acid sequences selected from SEQ ID NO: 23, 24, and 25, which are truncated fragments of SEQ ID NO: 5, comprising 100 nucleotides or less, binding tau protein at phosphorylatable sites.
Truncated DNA aptamers from SEQ ID NO: 6
A DNA aptamer comprising nucleic acid sequences selected from SEQ ID NO: 26 and 27, which are truncated fragments of SEQ ID NO: 6, comprising 100 nucleotides or less, binding tau protein at phosphorylatable sites.
Compositions comprising DNA aptamers with specified sequences
Pharmaceutical or other compositions comprising DNA aptamers capable of specifically binding tau protein at phosphorylatable sites, wherein the aptamers include sequences of 66 nucleotides selected from SEQ ID NO: 1 through SEQ ID NO: 7 or sequences selected from the truncated forms SEQ ID NO: 8 through SEQ ID NO: 27, comprising 100 nucleotides or less, optionally including pharmaceutically acceptable carriers or excipients.
The claims cover DNA aptamers binding tau protein at phosphorylatable sites in full-length and truncated forms, compositions containing these aptamers, and their use in detection, imaging, and treatment related to tauopathy neurodegenerative diseases.
Stated Advantages
DNA aptamers provide high specificity and affinity for tau protein at phosphorylatable sites.
Aptamers inhibit tau phosphorylation and oligomer formation, offering potential therapeutic effects to arrest tauopathy progression.
Aptamers can be produced with high quantity and fidelity, are smaller than antibodies, and can be chemically modified for improved properties, making them superior detection and therapeutic agents compared to antibodies.
Aptamer-based molecular beacons allow for direct, sensitive, and potentially rapid detection of tau protein levels in biological fluids and imaging in cells or in vivo.
Documented Applications
Detecting levels of tau and phosphorylated tau in cerebrospinal fluid, blood, and other biofluids using DNA aptamers as capture or detection agents in molecular beacon or sandwich assay formats.
Cell-based imaging of phosphorylated tau, non-phosphorylated tau, and total tau by introducing DNA aptamer conjugates linked with cell-penetrating peptides and detectable moieties such as fluorophores into cultured cells.
In vivo imaging of tau protein in subjects by administering DNA aptamer conjugates linked with signaling moieties (fluorophores, radioisotopes for PET, SPECT, or MRI contrast agents) for brain imaging in live animals or humans.
Therapeutic treatment of tauopathy-associated neurodegenerative disorders, including Alzheimer's disease and chronic traumatic encephalopathy, by administering compositions comprising DNA aptamers that inhibit tau phosphorylation and oligomerization in the brain.
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