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Principal Investigator: Stephanie Kristin Seidlits
Organization: UNIVERSITY OF TEXAS AT AUSTIN
Fiscal Year: 2024
Award: $221,843
Funding agency: National Institute on Drug Abuse
Project Summary
There is an urgent need to address the current public health crisis of opioid abuse and overdose deaths through
development of effective treatments for opioid use disorder (OUD) and non-addictive therapeutics to manage
pain. Ultimately, achievement of these goals would be greatly facilitated by the existence of novel ex-vivo models
that recapitulate key features of neurobiology underlying the addictive process using human cells and advanced
imaging systems that can monitor the interactions between multiple neurotransmitter actions driving opioid
responses and reward pathways. This proposal aims to develop new technologies to create such ex-vivo models
and imaging systems, aiming at probing dynamic behaviors within the models at high spatial and temporal
resolution. Aim 1 will focus on recapitulating neuronal pathways in the ventral tegmental area (VTA) and nucleus
acccumbens core (NAc), which have long been recognized to form the basis of substance abuse disorders.
Specifically, we propose an additive manufacturing approach based on 3D bioprinting of human induced
pluripotent stem cells (hiPSCs) to establish 3D cultures within brain-mimetic scaffolds. Moreover, use of an
innovative, self-healing biomaterial as a printing medium and 3D culture scaffold enables “stitching” of unique
neuronal tissues constructs into integrated, yet regionally defined, complex networks. For example, we propose
to integrate tissue modules representing the GABAergic/dopaminergic circuitry of the VTA with tissue modules
representing the GABAergic medium spiny neurons on the NAc. In addition, these tissue-engineered networks
are optically transparent and easy to image, making them great models to study neural networks driving opioid
responses and reward pathways. In Aim 2, we will separately develop a TIRF (Total Internal Reflection
Fluorescence) microscopy technique to probe agonist-dependent dimerization of MOR (human µ-opioid
receptor) at the single-molecule level and a hyperspectral and lifetime imaging system to monitor the dynamics
of dopamine, GABA, glutamate and Ca2+ simultaneously. Together, these new imaging methods will enable
dynamic monitoring of the effects of opioids, and other small-molecule therapeutics, on the neuronal circuitry
underlying addictive processes. Overall, we expect these technological innovations to provide crucial tools for
development of new therapeutics which can effectively combat the opioid crisis.
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Matrix><Cells><Complex><Consumption><Dangerousness><Detection><Development><Dimerization><Disease><Disorder><Dopamine><Duragesic><ECM><Encephalon><Engineering><Extracellular Matrix><FLIM imaging><Fentanest><Fentanyl><Fentyl><Fluorescence><GABA><Glutamates><Goals><HPLC><High Performance Liquid Chromatography><High Pressure Liquid Chromatography><High Speed Liquid Chromatography><Human><Hydroxytyramine><Image><Investigators><L-Glutamate><Laser Electromagnetic><Laser Radiation><Lasers><Maps><Measurement><Mesencephalon><Methods><Microscopy><Mid-brain><Midbrain><Midbrain structure><Model System><Modeling><Modern Man><Monitor><Nerve Cells><Nerve Transmitter Substances><Nerve Unit><Neural Cell><Neural Transmission><Neurobiology><Neurocyte><Neurons><Neurotransmitters><Nucleus><Nucleus Accumbens><Opiate Receptors><Opiates><Opioid><Opioid Receptor><Optics><Pain Control><Pain Therapy><Pain management><Pathway interactions><Patients><Phentanyl><Prevalence><Printing><Process><Production><Protein 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pain><response><reward circuitry><sample collection><scaffold><scaffolding><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><single molecule><small molecule therapeutics><specimen collection><substance abuse><synapse><tailored medical treatment><tailored therapy><tailored treatment><technological innovation><temporal measurement><temporal resolution><therapy development><three dimensional><three dimensional cell culture><three dimensional printing><time measurement><tool development><transmission process><treat chronic pain><treatment development><two-dimensional><unique treatment><ventral tegmentum><γ-Aminobutyric Acid>