Translational Neuroscience Social Hour · Clinical results, without neuroimaging BD participants received Uridine 500mg by mouth twice daily for 6 findings, were previously reported

Translational Neuroscience Social Hour

May 23, 2013

Mike Economo Matt Wachowiak

Imaging sensory-evoked activity in the

olfactory bulb in vivo

Fluorescent reporters

of neural activity…

Widefield imaging

using

Synapto-pHluorin

Two-photon

imaging with

GCamp3.3

…targeted to genetically-

defined populations

Periglomerular

interneurons

Mitral/tufted

neurons

…monitored with

single-cell resolution.

Spontaneous activity in a

single mitral cell over

several minutes

Markus Marta Daniela Tristan Ryan

Toward a naturalistic model of migraine aura. Volodymyr Bogdanov, Brennan Lab.

Aura is a neurological condition of temporary sensory deficit preceding migraine attack. Migraine aura ~ cortical spreading depression (CSD). Migraine aura can be induced by sensory stimulation. Can CSD be facilitated by peripheral stimulation of sensory cortex?

bregma lambda

10 sec 20 sec

30 sec 40 sec

0 sec

Future directions:

Measure intracortical [K+]

with ion sensitive electrodes during sensory induction of CSD.

Optogenetic vascular control (Myh-Chr2 mice) to evaluate effects on CSD induction.

Record CSD induction at cellular resolution with two photon microscopy.

Under superfusion of cranial window with 20 mM K+ CSD can be induced by sensory stimulation in some experimental animals.

Baseline evoked potentials DC potential decline during CSD

Intrinsic optical imaging of CSD

Right click on MPRAGE:

Export via DICOM to INC_Research3

Wait 10 minutes…

Images show up on PACS

Go ahead and try this at home…

Ziyad Khaleel, Anderson Lab, Neuroradiology

We enrolled 22 depressed adolescents with BD and 24 HC. There were no medication-related Serious Adverse Events, psychiatric hospitalizations, suicide attempts or suicides in the BD group. As shown in FIGURE 3, the mean CDRS-R raw score at study entry for was 63.6 (SD=6.18). The mean CDRS-R score for BD participants completing 6 weeks of treatment with uridine was 32.3 (SD=6.30), a decrease of 49%.

Neuroimaging data was acquired on 14 BD and 24 HC. No intracranial abnormalities were detected on anatomical MRIs. There were no significant between-group differences between BD and HC in age, educational level, or handedness.

The BD participants included both unmedicated (n=8) & medicated (n=6) adolescents. Four unmedicated BD participants were medication-naïve; the remaining 4 unmedicated BD participants had a mean medication-free period of 70 + 47 weeks (range, 28-116 weeks) at study entry.

31P-MRSI: To assess for differences in mean cerebral metabolite concentrations between the 3 participant groups, ANOVA was performed on the baseline 31P-MRSI metabolite data (TABLE 1). Post-hoc Tukey-Kramer HSD pairwise comparisons were then performed on the frontal lobe phosphorus-31 metabolites that were significant using ANOVA (PCr and Pi).

In unnmedicated BD participants, we found decreased baseline Pi compared to both HC (p=0.035), and medicated BD (p=0.021).

1H-MRSI: Consistent with MRSI studies of adult BD (Yuksel and Ongur,

2010), our adolescent participants with bipolar depression demonstrated elevated Glx at baseline, compared with HC (Effect Size = 0.86) (data not shown).

TABLE 2 displays the 31P-MRSI data from the baseline (Week 0) and follow-up (Week 6) scans of HC and BD. Between-group differences in high-energy phosphate levels were noted in BD vs. HC: uridine-treated BD participants demonstrated changes in phosphocreatine (PCr) (p=0.01) and β-NTP (~ATP) (p=0.007) compared with untreated HC. (See FIGURE 4)

1.Based on these open-label results, further study of Uridine as a treatment for depressed adolescents with BD is warranted.

2.Our results support the view that frontal lobe mitochondrial function is altered in adolescent BD, and may have implications for the use of Pi and Glx as biomarkers.

3.Phosphorus-31 magnetic resonance spectroscopic imaging (31P-MRSI) has the potential to become a valuable translational research tool in child & adolescent psychiatry. This work was supported by a NARSAD Young Investigator Award to Dr. Kondo from the BRAIN & BEHAVIOR RESEARCH FOUNDATION’S Research Partners Program; by NIMH grant MH058681 to Dr. Renshaw; and by the Utah Science Technology and Research initiative (USTAR).

Gore et al. (2011) Global burden of disease in young people aged 10-24 years: a systematic analysis. Lancet 377(9783): 2093-2102. Kondo et al. (2011) Open-label uridine for treatment of depressed adolescents with bipolar disorder. J Child Adolesc Psychopharmacol 21(2):171-5. Patel et al. (2008) Neurochemical alterations in adolescent bipolar depression: a proton magnetic resonance spectroscopy pilot study of the prefrontal cortex. J Child Adolesc Psychopharmacol 18:623-627. Quiroz et al. (2008) Mitochondrially mediated plasticity in the pathophysiology and treatment of bipolar disorder. Neuropsychopharmacology 33:2551-2565. Sanacora et al. (2008) Targeting the glutamatergic system to develop novel, improved therapeutics for mood disorders. Nat Rev Drug Discov 7:426-437. Vitiello, Correll et al. (2009) Antipsychotics in children and adolescents: increasing use, evidence for efficacy and safety concerns. Eur Neuropsychopharmacol 19:629-635. Yuksel & Ongur (2010) Magnetic resonance spectroscopy studies of glutamate-related abnormalities in mood disorders. Biol Psychiatry 68:785-794. .

This study had three objectives: (1) to initiate testing of the nutritional supplement uridine as a treatment for depressed adolescents with bipolar disorder with an open-label study; (2) to measure the changes in participants’ neurochemistry associated with uridine treatment with proton (1H-MRSI) and phosphorus (31P-MRSI) magnetic resonance spectroscopic imaging; and (3) to compare the concentrations of specific brain chemicals measured with 1H-MRSI and 31P-MRSI between adolescents with bipolar depression and healthy controls.

The World Health Organization ranks bipolar disorder (BD) as the 4th most disabling condition among persons 10-24 years of age (Gore et al. 2011). Adults with BD commonly report their symptoms and impairment began in adolescence. Experts including Husseini Manji have implicated both mitochondrial dysfunction (Quiroz et al., 2008), and the neurotransmitter glutamate (Sanacora et al., 2008), in the pathophysiology of BD.

Current pediatric BD drugs include mood stabilizers and 2nd generation antipsychotics (SGAs). Adherence is poor, and Vitiello and Correll have articulated the contrast between the rising number of SGA prescriptions to youth, and concerns regarding both efficacy and safety (2009).

Despite the fact that more than 50% of pediatric BD patients experience a major depressive episode, there are no FDA-approved treatments for bipolar depression in youth.

Uridine is an endogenous pyrimidine required for normal brain function. It is found in human mother’s milk, and is an ingredient in commercial infant formulas. Some patients with inborn errors have been treated with oral uridine for > 20 years. Triacetyluridine and cytidine are pyrimidines with positive studies in adult bipolar depression. Pyrimidines’ mechanism-of-action includes increased synthesis of monoamines, improved mitochondrial function, increased pH, decreased levels of Glutamate/Glutamine/γ-Aminobutyric acid (Glx) and improved phospholipid metabolism.

Magnetic Resonance Spectroscopic Imaging (MRSI) is a safe, non-invasive method for in vivo measurement of the brain chemicals that reflect both mitochondrial and glutamatergic function. MRSI has been used to discover differences in brain chemistry between pediatric bipolar depression and healthy controls, including N-acetyl aspartate (NAA), choline, and the creatine/phosphocreatine ratio (Patel et al., 2008).

We administered uridine, a supplement that may alter both the mitochondrial and glutamatergic systems, as an open-label treatment for adolescent bipolar depression. As shown in Figure 2, in addition to standardized clinical instruments, we used 1H-MRSI and 31P-MRSI to assess treatment response.

Clinical results, without neuroimaging findings, were previously reported for the first 7 participants (Kondo et al, 2011). We now report results for 23 depressed adolescents with BD and 25 healthy controls.

The University of Utah IRB approved the study. Written consent & assent were both obtained prior to study procedures. A Data Safety and Monitoring Board (DSMB) was established.

Inclusion criteria: females and males ages 13-18 with a primary diagnosis of BD I, II or NOS; and Children’s Depression Rating Scale-Revised raw score > 40. Medication-free and medicated adolescents were enrolled.

Exclusion criteria: primary Axis I diagnosis other than BD; Young Mania Rating Scale (YMRS) score > 10; psychotic symptoms; high risk for suicidal behavior; positive urine pregnancy test or drug screen; developmental disability; or unstable medical condition.

Lab studies including complete blood count, metabolic panel, lipid profile, TSH, and urinalysis were obtained at baseline, then repeated after 6 weeks, to prospectively identify abnormalities associated with uridine administration.

Healthy control (HC) adolescents were recruited for brain scans, to enable comparison with BD participants.

BD participants received Uridine 500mg by mouth twice daily for 6 weeks. Study visits included: CDRS-R; Clinical Global Impressions (S/I); Columbia-Suicide Severity Rating Scale (C-SSRS); and adverse events. Change in CDRS-R score was the primary outcome measure; magnetic resonance (MR) brain chemistry measurements were secondary outcomes.

A two-dimensional chemical shift imaging free induction decay (2D CSI FID) pulse sequence with an Fourier voxel resolution of 25×25×25mm3; Field of View=200x200x25mm3; TR/TE=3000/2.3ms; vector size=1024; bandwidth =2500Hz; data collection time=11.2 minutes; and number of averages=24 was implemented to collect 2D CSI FID data. The high-resolution localization images of CSI data were acquired with an inversion recovery magnetization prepared rapid gradient echo (MP-RAGE) pulse sequence with isotropic 1mm3 resolution. The imaging parameters were: TR/TE=2000/3.37ms, FOV = 256x192x144mm3; matrix size=256x192x144; total acquisition time=4.8 minutes. These data were analyzed using the jMRUI software package (JMRUI VERSION 4.0, EUROPEAN COMMUNITY). Nine voxels from a 25mm slice located at the corpus callosum, anterior commissure and posterior commissure were summed following 2D FFT. Each voxel FID was apodized with a 10 Hz exponential line broadening before zero filling and FFT. Zero-order and first-order phase correction was performed in all spectrums.

Signal amplitudes for individual proton-1 and phosphorus-31 metabolites were calculated with the Advanced Method for Accurate, Robust and Efficient Spectral fitting of MRS data (AMARES) algorithm in JMRUI.

Depressed Adolescents with Bipolar Disorder Treated with Open-Label Uridine: a Proton

(1H-MRSI)

and Phosphorus (31P-MRSI) Magnetic Resonance Spectroscopic Imaging Study Douglas Kondo MD,1,2,3 Kristen K. Fiedler BS,1 Tracy L. Hellem RN,1 Xianfeng Shi PhD,1 Young-Hoon Sung MD,1,2 Rebekah S. Huber MS1 and Perry F. Renshaw MD, PhD,

MBA1,2,3

The Brain Institute,1 Department of Psychiatry,2 VISN 19 Mental Illness Research, Education and Clinical Center (MIRECC),3 University of Utah School of Medicine, Salt

Lake City, Utah

BACKGROUND

METHODS

CONCLUSIONS

OBJECTIVES

UtahBrain.org

FIGURE 1 Frontal Lobe Region of Interest

RESULTS: Clinical Measures

BASELINE

NEUROIMAGING: 31P-MRSI & 1H-

MRSI

FIGURE 2 Study Design Incorporating

Neuroimaging Into

Treatment Studies of Pediatric Mood

Disorders

ACKNOWLEDGEMENTS

REFERENCES

Dr. Kondo, Ms. Fiedler, Ms. Hellem, Dr. Shi, Dr. Sung and Ms. Huber report no real or potential conflict of interest, and did not receive travel support or honoraria from a commercial business to attend this meeting. Dr. Renshaw is an inventor on a patent application that describes the use of uridine as a treatment for bipolar disorder. This patent application is assigned to McLean Hospital, and is licensed to the Repligen Corporation. The University of Utah manages conflicts of interest related to intellectual property. As part of this management, Dr. Renshaw did not evaluate any study participant at any point in time.

FIGURE 3 CDRS-R Scores During 6 Weeks of Open-Label

URIDINE

in Depressed Adolescents with Bipolar Disorder

*Start of treatment **End of

treatment

REPEATED

MEASURES

NEUROIMAGING: 31P-MRSI

FIGURE 4

ΔPCr (LEFT PANEL) and

Δβ-NTP (RIGHT PANEL)

from repeated

measures 31P-MRSI

brain scans of BD and

HC, performed 6 weeks

apart. BD received

Uridine 500mg twice

daily; HC were

untreated.

- PCr Effect Size = 1.2

- β-NTP Effect Size =

1.0

Microglia and Astrocyte Reactivity in Repeated

Methamphetamine Exposure

SS MS

MM SM

SS MS

MM SM

Microglia Reactivity

Astrocyte Reactivity

Future Questions:

•Glutamate transporter

•Extracellular levels of glutamate

• Blood brain barrier

•Gap junctions

(McCann et al., J. Neurosci., 1998)

(Hanson et al., Eur J Pharmacol. 2009)

Danielle Friend, Keefe Lab

Biomarker-controlled deep brain stimulation for Parkinson’s disease

Mark Lehmkuhle

Major Translational Questions We Are Working on at the

Center for Alzheimer’s Care, Imaging and Research

● Why does Alzheimer’s disease (AD) and FTD sometimes affect the brain asymmetrically? How can this be defined?

● What are the initiates clinical expression of AD?

● What are the predictors of cognitive decline in AD?

● What accounts for the delay between amyloid deposition and disease expression?

● Are there true escapees – those with AD pathology who will never get symptoms? If so how does this occur?

● What accounts for the selective vulnerability of neurons to amyloid pathology? FTD pathology?

● What is the relationship between amyloid and tangle pathology?

● What is the role of inflammation, glia, and cytokines in AD?

● What happens when amyloid is removed from diseased brain?

Norman Foster, CACIR

In vivo cerebrovascular characterization of CCM in a mouse model Khaled M. Krisht, M.D.

• Collaborating labs: Dr. Kevin Whitehead and Dr. KC Brennan

• Objective: To elucidate the vascular morphology and function of cavernous malformations and nearby cortical vessels in CCM mice in an in vivo setting.

• CCM : Cerebrovascular malformation secondary to endothelial cell dysfunction. • Spontaneous • Familial (AD) • 3 main genes responsible for CCM: Krit1, CCM2, Pdcd10

• What we know? • Krit1 and CCM2 are believed to act through similar molecular mechanisms: Prevent the activation of the GTPase Rho-A which is

believed to be responsible for endothelial cell dysfunction and breakdown of cell-cell interaction. • Inhibition of RhoA and down stream Rho-Kinase with simvastatin and Fausadil, respectively, reversed many of the cell permeability and

cytoskeletal aberrations observed with CCM KO mice: Improved cell resistance, improved endothelial cell alignment to laminar flow, and permeability.

• Dr. Whitehead’s lab has early results showing that the structural and functional deficits observed with their CCM2 KO mice can be rescued with Tempol, a superoxide scavenger.

• What we are trying to do? • Expand our work to include in vivo characterization of the vascular defects of CCM mice using 2-photon microscopy and IOS/IOI.

• SSER in arteries/arterioles

• CSD induction threshold

• Vasodilatory effects of Ach

• Permeability studies

• Neurovascular mapping

• Attempt to rescue with .tempol and simvastatin.

ATXN2-Luc SCA2

qHTS w/ NCGC ATXN2 ASOs w/ Isis

Braak et al. Staging of brain pathology related to sporadic Parkinson’s

disease. Neurobiology of Aging 24 (2003) 197–211.

MOVEMENT DISORDERS RESEARCH COLLABORATION

David Shprecher, DO MS Director, Sleep and Movement Disorders Division, Dept of Neurology

Neurodegenerative Disease

Biorepository

• DNA samples from over 300

subjects so far

• ALS, PSP, PD, HD, ET

Huntington Disease Research

• ENROLL-HD

• HD Registry study enrolling 40 to 60 families per year

in Utah. 10,000 worldwide at 200 centers

• CREST- HD (Phase III neuroprotection trial of

creatine monohydrate in early stage HD)

Progressive Supranuclear Palsy

Research • Genetic and Environmental Risk Factors

• 18 patient-caregiver-control trios in Utah, 300

nationally

• Phase II/III RCT neuroprotection study of

davunetide

• 10 participants in Utah, 300 internationally

Parkinson Disease Research

• Interventional trials for neuroprotection, psychosis,

mild cognitive impairment, DBS

• Observational Research

– Parkinson’s Genetic Risk Study

• At least 12 families in Utah, 300 nationally

– Biomarker Study of de novo PD and RBD

• Transcriptomics, metabolomics, proteomics

– Enteric pathology as a predictor of neurodegenerative

disease risk

Mitochondrial Membrane Potential Dynamics in a Whole-Heart Model of Sudden Cardiac Arrest

Paul Venable, Zaitsev Lab, CVRTI Department of Bioengineering

Background •Mitochondrial membrane potential (ΔΨm) loss during SCA leads to defibrillation failure and arrhythmias upon reperfusion • ΔΨm loss -> [ATP] -> KATP channel opens -> action potential shortens, asystole occurs

Goal •Investigate ΔΨm depolarization and its relation to electrical activity •New analysis method based on Spatial Fourier Transform of confocal fluorescent images of cationic dye TMRM

Imaging Mitochondrial Potential in Whole-hearts

Initial Results: ΔΨm depolarization late and after asystole

Ctrl

ΔΨm

Uncoupler

FCCP

Single Cell TMRM Fluorescence

30 min

Ischemia

70 min

Ischemia

Cadherins In Circuit Formation Raunak Basu, Megan Williams Lab

Learning

Memory

Navigation

Adult stem cells

General Synapse

Axon Spine

Future Directions

• Spine analysis in KO

• Signaling- prospect for drug development

• Behavior analysis

Mechanical Ventilation:

It Doesn’t Do a Body Good!

Preterm infants

Preterm lambs

Fetal Reference PT 3d MV

Preterm lambs

Focus on Cognitive Outcome after

Pediatric TBI – Michelle Schober ◦ 7 day old and 35 day old rat pups

Controlled cortical impact (mod-severe)

Gene expression; MRI, DLM/NOR/MWM; Histology; effect

EPO/DHA on outcome

◦ PICU patients (mod-severe)* CSF/serum bank creation and prospective study of

demographics/clinical, biomarkers, EEG & 6 month outcome

◦ Community: mild TBI in high school athletes*

◦ Funding: K12+two intramural grants* & third* pending

◦ GOALS: Multi-PI RO1 for bench to bedside

study.

Epileptogenesis

Novel Therapies

Dudek Lab – Jay Spampanato Epilepsy

Excitation Inhibition

CA1

Diphtheria Toxin Receptor - GFP

Gad67

CA1

Mechanisms of Seizure Generation

?

a1 Ivermectin Receptor - GFP

CA1

1mM ivermectin

Intrinsic structural covariance map S1 S2 S3 S4 S5 S6 S7 Seeley et al., 2009

• Functional connectivity mapping (fcMRI, MEG, EEG) • Structural connectivity analysis (scMRI, DTI, DSI)

Zielinski, et al. PNAS, 2010

DMN

(radar detector)

SN

(socio-emotional)

red = autism, blue = controls. p = 0.001 (FWE)

Zielinski, et al. PLoS One, 2012

5-8 yrs 9-11 yrs 12-14 yrs 16-18 yrs

Brandon A. Zielinski, MD, PhD

Division of Child Neurology

University of Utah

Describing Structure-Function Relationships

In Developing Brain Networks

Alcohol blocks Kir2.1 to cause FAS

Katz, 1999.

Wattendorf, 2005

Fetal Alcohol

Andersen-Tawil

P. Aryal, 2009

Dahal; 2012

Bates; 2013

alcohol

Basal

Ba++ Wild type Kir2.1 KO

Emily Bates, BYU

The Cre-dependent GCaMP5 Reporter Mouse

Petr Tvrdik Human Genetics

Main Features: Targeted to Polr2a (not ROSA26); FLP-removable Tomato reporter (Mike Gee)

Spontaneous activity waves in P2 cortex (Markus Rothermel, Mike Economo)

Calcium transients in PFC (Mike Economo) [adult Emx1-Cre x Pol2-GCaMP5 mouse]

Vascular loading in TBI Blast Injury

Stewart Yeoh (Ken Monson Lab)

Regulated Arc expression modulates trafficking

of AMPA receptors and stabilization of LTD/LTP

Jason Shepherd

Bonomi et al. (2001) Ophthalmologica 215:34-38.

bausch.com/en/Eye-concerns/eye-diseases-and-disorders/Glaucoma

Intraocular pressure (mm Hg)

Vision loss in glaucoma

Relationship between glaucoma and intraocular pressure (IOP)

Brn3a (RGCs) TRPV4

vision.arc.nasa.gov

Daniel Ryskamp Neuroscience Program PhD Candidate HHMI Med into Grad Scholar

Mouse retina section

Human retina section

Christensen and Corey (2007) Nature Reviews: Neurosci

TRPV4 is a mechanosensitive ion channel

Protecting the retina from pressure

GSK = TRPV4 agonist HC = TRPV4 antagonist

([C

a2+] R

GC) RGCs respond to < 1% elongation

Hypo

Seeing is believing: Neural representations of illusory

speech perception

“VA”

/BA/

E. Smith, S. Duede, S. Hanrahan,

T. Davis, P. House, B. Greger

“BA” - video

/BA/ - audio

“VA” - video

/BA/ - audio

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