Document text
Principal Investigator: Kasey Lynn Brida
Organization: UNIVERSITY OF ALABAMA AT BIRMINGHAM
Fiscal Year: 2024
Award: $42,629
Funding agency: National Institute on Drug Abuse
PROJECT SUMMARY
Substance use disorder (SUD) is a national health crisis, with over 93,000 deaths due to overdose in 2020.
Currently, no FDA-approved therapeutics exist for SUD, with targeted therapeutics out of reach due to
incomplete understanding of the neurobiology of addiction. Our lab has identified expression of Reln mRNA as
necessary for cocaine-responsiveness in NAc Drd1+ medium spiny neurons (MSNs). Reelin is a secreted
glycoprotein that plays a critical role in cortical cell migration during brain development, with mutations leading
to lissencephaly, defined by a smooth-brain phenotype. In the adult brain, Reln expression remains high in the
cerebellum, hippocampus and striatum. Brain-wide Reelin signaling blockade attenuates response to
methamphetamines, indicating a critical role for Reelin in the striatum. Despite high striatal Reln expression
and evidence suggesting Reelin modulates behavioral output of the striatum, Reelin’s role in the striatum
remains under-studied. Using RNA single-molecule fluorescent in-situ hybridization (smFISH), I have validated
our single-cell data showing Reln marks cocaine-sensitive Drd1+ cells. I have also found Drd1+ cells are
enriched in Reln compared to Drd2+ cells in both rat and human brain tissue. My smFISH data show Reln
expression is restricted to the dorsal striatum and nucleus accumbens (NAc) core and absent in the NAc shell.
As these regions have distinct functions, this pattern may reflect the functional relevance of Reelin signaling. I
have assessed the role of Reelin signaling using a CRISPR-interference (CRISPRi) strategy to knockdown
Reln. NAc whole-cell patch clamp demonstrates that CRISPRi at Reln significantly impairs MSN excitability.
Critically, in vivo CRISPRi-knockdown of striatal Reln attenuates cocaine preference in a rat model.
Given these findings, I hypothesize that Reelin facilitates the dopamine-induced neuronal adaptations that
follow drug exposure and promote drug-seeking behavior. The proposed study will take a multidisciplinary
approach to rigorously investigate the following aims: (1) Reelin is required for cocaine-related behavioral
adaptations and (2) Reelin regulates membrane excitability to facilitate dopamine signaling. The proposed
studies will deepen our understanding of the molecular underpinnings of SUD, providing avenues for
therapeutic exploration for a disease completely lacking targeted treatments. Under this award, I will master
intravenous self-administration and locomotor sensitization assays to assess the relevant consequences of
genetic manipulations. I will hone patch clamp electrophysiology skills to assess mechanisms behind
behavioral adaptations. These techniques will aid my success as a physician-scientist studying mechanisms of
and engineering therapeutics for brain disorders.
Terms: <21+ years old><AD dementia><Action Potentials><Adult><Adult Human><Alzheimer Type Dementia><Alzheimer disease dementia><Alzheimer sclerosis><Alzheimer syndrome><Alzheimer's><Alzheimer's Disease><Alzheimers Dementia><Ammon Horn><Architecture><Assay><Attenuated><Award><Behavior><Behavioral><Bioassay><Biological Assay><Brain><Brain Diseases><Brain Disorders><Brain Nervous System><COVID crisis><COVID epidemic><COVID pandemic><COVID-19 crisis><COVID-19 epidemic><COVID-19 era><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 pandemic><COVID-19 period><COVID-19 public health crisis><COVID-19 years><CRISPR><CRISPR activation><CRISPR activator><CRISPR approach><CRISPR based activation><CRISPR based approach><CRISPR gene activation><CRISPR interference><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR transcription activation><CRISPR transcriptional activation><CRISPR-CAS-9><CRISPR-Cas-9-mediated gene activation><CRISPR-based gene activation><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR-dCAS9 Activator><CRISPR-dCas9-mediated repression><CRISPR-mediated transcriptional activation><CRISPR/CAS approach><CRISPR/CAS9 activation><CRISPR/CAS9 gene activation><CRISPR/Cas method><CRISPR/Cas system><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><CRISPR/dCas9 activation><CRISPR/dCas9 interference><CRISPR/dCas9-based transcriptional activation><CRISPR/dCas9-mediated transcriptional inhibition><CRISPRa><CRISPRi><Cas nuclease technology><Cell Body><Cell Communication and Signaling><Cell Locomotion><Cell Migration><Cell Movement><Cell Signaling><Cells><Cellular Matrix><Cellular Migration><Cellular Motility><Cerebellum><Cessation of life><Clustered Regularly Interspaced Short Palindromic Repeats><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats interference><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Cocaine><Cocaine use disorder><Common Rat Strains><Complex thinking><Cornu Ammonis><Corpus Striatum><Corpus striatum structure><Critical Thinking><Crystal Meth><Crystal methamphetamine><Cytoskeletal System><Cytoskeleton><Data><Data Set><Death><Death Rate><Deoxyephedrine><Desoxyephedrine><Development><Dimensions><Disease><Disorder><Dissociation><Dopamine><Dopamine Receptor><Dorsal><Drug Exposure><Drugs><Economic Burden><Electrophysiology><Electrophysiology (science)><Encephalon><Encephalon Diseases><Engineering><Engineering / Architecture><Evaluative Thinking><FDA approved><FISH Technic><FISH Technique><FISH analysis><FISH assay><Fluorescence In Situ Hybridization><Fluorescent in Situ Hybridization><Genetic Alteration><Genetic Change><Genetic defect><Glycoproteins><Health><Health Care Costs><Health Costs><Healthcare Costs><Hippocampus><Human><Hydroxytyramine><Impairment><Induced Neurons><Injections><Intracellular Communication and Signaling><Intracranial CNS Disorders><Intracranial Central Nervous System Disorders><Intravenous><Investigation><Learning><Link><Mediating><Medication><Membrane><Messenger RNA><Methamphetamine><Methylamphetamine><Modeling><Modern Man><Molecular><Motivation><Mutation><N Methyl D aspartic Acid><N methyl D aspartate><N-Methyl-D-aspartate><N-Methylamphetamine><N-Methylaspartate><NMDA><Nerve Cells><Nerve Unit><Nervous System Diseases><Nervous System Disorder><Neural Cell><Neurobiology><Neurocyte><Neurologic Disorders><Neurological Disorders><Neurons><Neurophysiology / Electrophysiology><Non-Polyadenylated RNA><Nucleus Accumbens><Output><Overdose><Pathogenesis><Pathway interactions><Pattern><Pharmaceutical Preparations><Phenotype><Physical Dependence><Physicians><Physiologic><Physiological><Physiology><Play><Primary Senile Degenerative Dementia><Proteins><Publishing><RNA><RNA Gene Products><Rat><Rats Mammals><Rattus><Regulation><Rewards><Ribonucleic Acid><Role><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 pandemic><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><Saccharose><Schizophrenia><Schizophrenic Disorders><Scientist><Self Administered><Self Administration><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 pandemic><Signal Transduction><Signal Transduction Systems><Signaling><Single-Nucleus Sequencing><Slice><Stimulus><Striate Body><Striatum><Substance Use Disorder><Substance abuse problem><Sucrose><Synapses><Synaptic><Techniques><Testing><Therapeutic><Time><Ventral Tegmental Area><Work><abuse of substances><abused drug><abused drugs><activating CRISPR technology><addiction><addictive disorder><adulthood><adverse consequence><adverse outcome><agyria><attenuate><attenuates><behavior influence><behavior response><behavioral influence><behavioral response><biological signal transduction><brain tissue><cell cortex><cell motility><cocaine behavior><cocaine motivated behaviors><cocaine related behaviors><cocaine reward><cocaine self-administration><cocaine sensitization><cocaine-associated behaviors><conditioned place preference><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global pandemic><coronavirus disease 2019 health crisis><coronavirus disease 2019 pandemic><coronavirus disease 2019 public health crisis><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease pandemic><coronavirus disease-19 global pandemic><coronavirus disease-19 pandemic><cost><dementia praecox><developmental><drug abused><drug induced behavior><drug of abuse><drug reward><drug seeking behavior><drug/agent><drugs abused><drugs of abuse><electrophysiological><experience><experiment><experimental research><experimental study><experiments><gene induction><gene manipulation><genetic manipulation><genetically manipulate><genetically perturb><genome mutation><hippocampal><iNeuron><in vivo><induction of genes><innovate><innovation><innovative><insight><interdisciplinary approach><intracellular skeleton><knock-down><knockdown><lissencephaly><lissencephaly pachygyria><mRNA><membrane structure><meth><mortality><mortality rate><mortality ratio><multidisciplinary approach><neurobiological><neurological disease><neuronal><neuronal excitability><neuropsychiatric disease><neuropsychiatric disorder><novel><overdose death><overdose fatalities><overexpress><overexpression><pandemic><pandemic disease><patch clamp><pathway><place conditioning><preference><prevent><preventing><primary degenerative dementia><programs><reeler gene product><reelin gene product><reelin protein><reln gene product><repressing CRISPR-dCas9 system><response><sNuc-Seq><schizophrenic><self-administer cocaine><senile dementia of the Alzheimer type><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><sex><single molecule><single nucleus RNA-sequencing><single nucleus seq><single-nucleus RNA-seq><skills><snRNA sequencing><snRNA-seq><social role><striatal><substance abuse><substance use and disorder><success><synapse><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><treatment strategy><trend><ventral tegmentum>