COMPARISON OF NEUROPROTECTIVE EFFECTS OF LEVETIRACETAM AND - - PowerPoint PPT Presentation

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COMPARISON OF NEUROPROTECTIVE EFFECTS OF LEVETIRACETAM AND PHENYTOIN IN TRAUMATIC BRAIN INJURY A NDREW R. P ETERSON D EPARTMENT OF P SYCHOLOGY , D AVIDSON C OLLEGE , NC 28035 SIGNIFICANCE AND BACKGROUND P OST - TRAUMATIC SEIZURES (PTS) ARE

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COMPARISON OF NEUROPROTECTIVE EFFECTS OF LEVETIRACETAM AND PHENYTOIN IN TRAUMATIC BRAIN INJURY

ANDREW R. PETERSON DEPARTMENT OF PSYCHOLOGY, DAVIDSON COLLEGE, NC 28035

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SIGNIFICANCE AND BACKGROUND

  • POST-TRAUMATIC SEIZURES (PTS) ARE SEIZURES THAT

RESULT FROM TRAUMATIC BRAIN INJURY (TBI).

  • RISK FACTOR FOR POST-TRAUMATIC EPILEPSY (PTE), A

RECURRENT SEIZURE DISORDER SECONDARY TO BRAIN INJURY FOLLOWING HEAD TRAUMA

  • SEIZURES EARLY AFTER TRAUMA INCREASE BRAIN DAMAGE

THROUGH:

  • HYPOXIA
  • EXCESSIVE RELEASE OF EXCITATORY NEUROTRANSMITTERS
  • INCREASED METABOLIC DEMANDS
  • INCREASED INTRACRANIAL PRESSURE
  • ANTICONVULSANT MEDICATIONS ARE ADMINISTERED TO

PREVENT FURTHER CELL DAMAGE FROM SEIZURES.

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SIGNIFICANCE AND BACKGROUND

  • COMMON ANTICONVULSANT MEDICATIONS ARE:
  • VALPROATE
  • PHENYTOIN (PHE)
  • PHENOBARBITAL
  • ALL OF THESE MEDICATIONS ARE USED BECAUSE OF THEIR

NEUROPROTECTIVE ABILITIES.

  • NO TREATMENT IS WIDELY ACCEPTED TO PREVENT THE

DEVELOPMENT OF SEIZURES AND EPILEPSY.

  • RECENT RESEARCH SHOWS THAT LEVETIRACETAM (LEV) MAY

BE A MORE EFFECTIVE TREATMENT THAN TRADITIONAL TREATMENTS, BUT WHY IS NOT CLEAR.

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SIGNIFICANCE AND BACKGROUND

ANTIEPILEPTIC LEV AS A NEUROPROTECTIVE AGENT

  • LEV REDUCED LESION VOLUME AND THE

OCCURRENCE OF NONCONVULSIVE SEIZURES IN EPILEPTIC RATS (CUOMO ET AL., 2013)

  • LEV PROTECTS CELLULAR STRUCTURES FROM

DAMAGE IN PTS (GIBBS, WALKER, AND COCK,

2006)

  • LEV ACTS AS A NEUROPROTECTIVE AGENT BY

ENHANCING HISTOLOGICAL, MOLECULAR, AND BEHAVIORAL ELEMENTS OF RECOVERY AFTER TBI

(ZOU ET AL., 2013)

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PURPOSE

COMPARE THE NEUROPROTECTIVE

EFFECTS OF THE ANTI-EPILEPTIC DRUGS LEV AND PHE IN ENHANCING AND FACILITATING NEURON RECOVERY AFTER TBI. THIS COMPARISON WILL BE DONE THROUGH BEHAVIORAL, MORPHOLOGICAL, AND IMMUNOHISTOCHEMICAL ASSESSMENT.

Frontal motor area damage in squirrel monkey

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DRUGS

Levetiracetam (LEV) Phenytoin (PHE) Brand Name Keppra Dilantin Uses Treat epilepsy:

  • Partial onset seizures
  • Tonic-clonic seizures
  • Myoclonic seizures

Treat epilepsy:

  • Partial onset seizures
  • Tonic-clonic seizures
  • Status epilepticus

Mechanism

  • f Action

(Anti- epileptic)

  • Exact mechanism not

known.

  • Binds to synaptic vesical

glycoprotein, SV2A.

  • Inhibits presynaptic

calcium channels

  • Voltage-dependent block of

voltage gated sodium channels--prevents sustained repetitive firing of action potentials.

  • Primary site of action is the

motor cortex. Treatment in T.B.I Primarily an adjunct therapy A standard treatment for post- traumatic seizures.

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SUBJECTS

  • 63 ADULT MALE SPRAGUE-DAWLEY RATS (HILLTOP, SCOTTSDALE, PA).
  • HOUSED IN THE DAVIDSON COLLEGE ANIMAL FACILITY ON A 12 HR LIGHT : 12HR DARK

CYCLE WITH ENRICHMENT AND AD LIBITUM ACCESS TO FOOD AND WATER.

  • SUBJECTS WILL BE HOUSED IN THE COLONY FOR A MAXIMUM OF 50 DAYS.
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DEPENDENT MEASURES- BEHAVIORAL

MOTOR FUNCTION

  • ROTOROD
  • TIME TAKEN TO FALL FROM

APPARATUS

SPATIAL LEARNING AND MEMORY

  • Y-MAZE
  • TIME TAKEN TO EXPLORE NOVEL

AREAS

  • MORRIS WATER MAZE
  • TIME TAKEN TO FIND HIDDEN

PLATFORM

Mouse on platform in Morris Water Maze

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DEPENDENT MEASURES- NEUROLOGICAL

  • HISTOLOGICAL ASSESSMENT
  • RATS PERFUSED AND STAINED WITH CRESYL

VIOLET.

  • AREAS OF NEURON DAMAGE WILL BE NOTED

AND THE AMOUNT OF NEURON DAMAGE WILL BE ANALYZED.

  • IMMUNOHISTOCHEMISTRY
  • INTERLUKIN1-B (IL-1B) AND GLT-1/EAAT2
  • CHARACTERIZE CHANGES IN INFLAMMATION

AND THE PRIMARY GLUTAMATE TRANSPORTER

GLT-1 IN REGIONS OF INTEREST.

Coronal slices stained with Luxol fast blue/Cresyl violet after intracerebral hemorrhage (Wang & Tsirka, 2005).

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TIMELINE

DAY 1: RATS ARRIVE DAY 7: BEGIN BEHAVIORAL PRE-TESTING DAY 14: SURGERY DAY AND BEGIN TREATMENT DAYS 15-20: ROTOROD BEHAVIORAL TEST

  • RATS WILL COMPLETE THREE TRIALS/DAY.

DAYS 21-26: Y-MAZE BEHAVIORAL TEST

  • RATS WILL COMPLETE THREE TRIALS/DAY.

DAYS 27-32: MORRIS WATER MAZE BEHAVIORAL TEST

  • DAYS 27-31: TIME TO LOCATE HIDDEN PLATFORM.
  • DAY 32: TIME TO LOCATE VISIBLE PLATFORM.
  • RATS WILL COMPLETE THREE TRIALS/DAY.

DAY 33: PERFUSE FOR CRESYL VIOLET DAY 34-35: WAIT PERIOD DAY 36-38: HISTOLOGICAL ASSESSMENT DAY 39-42: IMMUNOHISTOCHEMISTRY

High definition fiber-tracking map depicts neural connections broken in TBI and other disorders.

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CONTROLLED CORTICAL IMPACT (CCI) MODEL

A PNEUMATIC OR ELECTROMAGNETIC IMPACT

DEVICE DRIVES A RIGID IMPACTOR ONTO THE EXPOSED, INTACT DURA MIMICKING:

  • Cortical tissue loss
  • Acute subdural

hematoma

  • Axonal injury
  • Concussion blood
  • Brain barrier

dysfunction

  • Coma

CCI Surgical Apparatus A) TBI from CCI Model B) Coronal slice showing TBI

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BEHAVIORAL PROCEDURES

MOTOR FUNCTION

  • ROTOROD
  • MEASURES FORE- AND HIND LIMB

COORDINATION AND BALANCE.

  • ANIMALS ARE TRAINED TO WALK ON

THE ROD BEFORE BEING EXPOSED TO EXPERIMENTAL CONDITIONS. Rats in Rotorod testing apparatus

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BEHAVIORAL PROCEDURES

SPATIAL LEARNING AND MEMORY

  • MORRIS WATER MAZE TEST
  • ANIMALS ARE PLACED IN A POOL OF

WATER AND HAVE TO SWIM TO A HIDDEN ESCAPE PLATFORM.

  • Y-MAZE TEST
  • TO TEST IF RODENTS PREFER TO SPEND TIME

IN NEW OR FAMILIAR AREAS, ONE ARM OF THE Y-MAZE IS BLOCKED FOR A PERIOD OF TIME AND THEN REOPENED. THE TIME SPENT IN THE NEW AREA IS RECORDED.

Mouse in Morris Water Maze Test

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BEHAVIORAL ANALYSIS

10 20 30 40 50 60 Saline LEV PHE

Time (sec) Treatment Group Rotorod Latency

80 85 90 95 100 105 Saline LEV PHE

Time (sec) Treatment Group

Morris Water Test

20 40 60 80 100 120 Saline LEV PHE

Time (sec) Treatment Group

Y-Maze

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HISTOLOGICAL ANALYSIS

0.00% 10.00% 20.00% 30.00% 40.00% 50.00% 60.00% 70.00% Saline LEV PHE

% Intact Neurons (% contralateral) Treatment Group

Intact CA1 Neurons

5 10 15 20 25 30 Saline LEV PHE

Lesion Volume (mm3) Treatment Group

Cortical Contusion Volume

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IMMUNOCHEMICAL ANALYSIS

EFFECTS OF LEV AND PHE ON THE EAATS,

NEUROPLASTIC MARKERS, AND IL-IB IN THE IPSILATERAL FRONTAL CORTEX AND IPSILATERAL HIPPOCAMPUS

  • GLAST
  • GLT-1
  • EAAC1
  • GAP-43
  • SYNAPTOPHYSIN
  • IL-IB

0% 20% 40% 60% 80% 100% 120% 140% Saline LEV PHE

% Density Treatment Group

GLAST

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PREDICTIONS AND RESULTS

LEV WILL BE A MORE EFFECTIVE

NEUROPROTECTIVE AGENT THAN PHE.

THE MOST EFFECTIVE NEUROPROTECTIVE AGENT

WILL HAVE THE:

  • LEAST IMPAIRED BEHAVIORAL PERFORMANCE
  • HIGHEST PERCENTAGE OF NEURONS

PRESERVED

  • SMALLEST LESION VOLUME
  • LEAST CHANGE IN INFLAMMATION AND

REACTIVITY

MRI of patient after sever T.B.I.

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PROJECT BUDGET

Personnel Principal Investigator…………………………………………..….$ 60,000.00 (+ benefits) Research Technician…………………………………………….. $ 30,000.00 (+ benefits) Statistician (part-time; $55.00 x 200 hrs.)………………..….… $ 11,000.00 Section Total………………………………………………………. $ 101,000.00 (+ benefits) Equipment Computer w/software ($3,000 x 2)……………………..……...$ 6,000.00 Laptop w/software ($3,000 x3)………………………………….$ 9,000.00 Morris Water Maze…………………………………………………$ 2,000.00 Y-Maze…………………………………………………………...….$ 1,200.00 Rotorod………………………………………………………………$ 5,000.00 Section Total………………………………………………………… $ 23,200.00 Supplies Office Supplies…………………………………………………….. $ 2,000.00 Laboratory / Clinical Supplies…………………………………... $ 7,500.00 Drugs and Chemicals: Levetiracetam (LEV) (58.8 mg needed)……$ 218.00 Phenytoin (PHE) (58.8 mg)…………………….$ 57.00 Saline (PBS) (58.8 mg)……..…………………...$ 33.70 Drugs and Chemicals Total……………………………………… $ 308.70 Section Total………………………………………………………… $ 9,808.70 Other Travel to scientific meeting……………………………………….$ 2,000.00 Animal Purchase; Rats ($25.00 x 63)…………………………….$ 1,575.00 Animal Care; Rats ($0.50 x 63 x 50 days)………………………$ 1,575.00 Section Total………………………………………………………… $ 5,150.00 Total $ 139,158.70

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REFERENCES

CUOMO, O., RISPOLI, V., LEO, A., POLITI, G. B., VINCIGUERRA, A., RENZO, G. D., & CATALDI, M. (2013). THE ANTIEPILEPTIC DRUG LEVETIRACETAM SUPPRESSES NON-CONVULSIVE SEIZURE ACTIVITY AND REDUCES ISCHEMIC BRAIN DAMAGE IN RATS SUBJECTED TO PERMANENT MIDDLE CEREBRAL ARTERY OCCLUSION. PLOS ONE, 8(11). DOI:10.1371/JOURNAL.PONE. 0080852 GIBBS, J. E., WALKER, M. C., & COCK, H. R. (2006). LEVETIRACETAM: ANTIEPILEPTIC PROPERTIES AND PROTECTIVE EFFECTS ON MITOCHONDRIAL DYSFUNCTION IN EXPERIMENTAL STATUS EPILEPTICUS. EPILEPSIA, 47(3), 469-478. DOI:10.1111/J.1528-1167.2006.00454.X ZOU, H., BRAYER, S. W., HURWITZ, M., NIYONKURU, C., FOWLER, L. E., & WAGNER, A. K. (2013). NEUROPROTECTIVE, NEUROPLASTIC, AND NEUROBEHAVIORAL EFFECTS OF DAILY TREATMENT WITH LEVETIRACETAM IN EXPERIMENTAL TRAUMATIC BRAIN INJURY. NEUROREHABILITATION AND NEURAL REPAIR, 27(9), 878-888. DOI: 10.1177/1545968313491007