Identifies Potential Therapeutic Compounds to Treat BDNF- Related Neurological Diseases
This phenotypic high-throughput screening (HTS) assay identifies potential therapeutic compounds for treating brain-derived neurotrophic factor (BDNF)-related neurological disorders. Brain-derived neurotrophic factor (BDNF)is a secreted growth factor that binds to TrkB receptor tyrosine Kinases, inducing its dimerization and activation. It activates many signaling cascades, cooperatively promoting neuronal survival, regulating the development of neural circuits, stimulating synapse formation, enhancing synaptic transmission, and facilitating synaptic plasticity in many brain regions. These physiological functions implicate BDNF deficiencies, linked to several neurological and psychiatric disorders, including Alzheimer's and Huntington's disease, major depressive disorder, anxiety, eating disorders, and Rett Syndrome. Identifying compounds able to boost neuronal BDNF synthesis is attractive for serving as potential therapeutics.
Current approaches to increase BDNF-TrkB signaling include recombinant BDNF, small molecule or antibody TrkB agonists, and viral expression of BDNF. However, these approaches have limitations. Short half-life, poor blood-brain barrier penetration, and brain parenchyma penetration hinder the recombinant BDNF. The small molecule TrkB agonists face challenges in inducing TrkB dimerization. While monoclonal antibodies may successfully bind or activate TrkB, few can cross the blood-brain barrier. Also, both small molecule and antibody TrkB agonists struggle to replicate the synapse-specific and time-sensitive effects of endogenous BDNF. Lastly, using viral vectors to express BDNF in the brain is highly invasive. As a progressing disease spreads to multiple brain regions, multiple virus injections are necessary to produce adequate BDNF. The safety of the protocols is dubious, with no good way to turn off viral BDNF expression in case of side effects. Western Blot or ELISA measures BDNF levels in tissues or cultured neurons, but the detection methods are too slow and expensive for high-throughput screening (HTS). A sensitive and quantitative BDNF assay for high-throughput screening is necessary.
Researchers at the University of Florida have developed a sensitive phenotypic, nano luciferase-linked brain-derived neurotrophic factor (BDNF) high-throughput screening assay for identifying BDNF-enhancing compounds. These compounds can be used as potential therapeutic compounds to combat BDNF-linked neurological disorders. The phenotypic assay has the advantages of speed, cost, and throughput over measuring BDNF levels traditionally.
Application
Nano-luciferase (NLuc)-based high-throughput screening (HTS) assay for identifying BDNF-boosting compounds for developing potential therapeutics for neurological disorders
Advantages
- Provides a mechanism for identifying BDNF-boosting compounds, enabling the development of therapeutics to combat neurological disorders
- The BDNF-NLuc assay achieves a reproducible Z score greater than 0.5, indicating its suitability for high-throughput screening
- Addresses the complexity of poorly understood diseases, favoring speed, cost, and throughput over measuring BDNF levels in a traditional manner
Technology
This nano-luciferase (NLuc)- based high-throughput screening (HTS) assay enables the detection of novel brain-derived neurotrophic factor (BDNF)-boosting compounds. The new mouse BDNF allele, BDNF-NLuc, is generated by inserting the sequence encoding of nano-luciferase into the BDNF locus. This BDNF-NLuc protein functions like BDNF, achieving a reproducible Z score greater than 0.5. The assay successfully scales and screens against the 1280-compound Library of Pharmacologically Active Compounds (LOPAC). The screening identifies several previously known BDNF-boosting compounds, including Bay K8644, an L-type voltage-gated calcium channel (L-VGCC) agonist. This indicates this phenotypic BDNF-NLuc neuronal assay approach is ready for high-throughput screening to identify BDNF-boosting compounds.
