COMPARISON OF NEUROPROTECTIVE EFFECTS OF LEVETIRACETAM AND - PowerPoint PPT Presentation

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 SEIZURES THAT RESULT FROM TRAUMATIC BRAIN INJURY (TBI). • R ISK FACTOR FOR POST - TRAUMATIC EPILEPSY (PTE), A RECURRENT SEIZURE DISORDER SECONDARY TO BRAIN INJURY FOLLOWING HEAD TRAUMA • S EIZURES EARLY AFTER TRAUMA INCREASE BRAIN DAMAGE THROUGH : • H YPOXIA EXCESSIVE RELEASE OF EXCITATORY NEUROTRANSMITTERS • INCREASED METABOLIC DEMANDS • INCREASED INTRACRANIAL PRESSURE • • A NTICONVULSANT MEDICATIONS ARE ADMINISTERED TO PREVENT FURTHER CELL DAMAGE FROM SEIZURES .

SIGNIFICANCE AND BACKGROUND • C OMMON ANTICONVULSANT MEDICATIONS ARE : • V ALPROATE • P HENYTOIN (PHE) • P HENOBARBITAL • A LL OF THESE MEDICATIONS ARE USED BECAUSE OF THEIR NEUROPROTECTIVE ABILITIES . • N O TREATMENT IS WIDELY ACCEPTED TO PREVENT THE DEVELOPMENT OF SEIZURES AND EPILEPSY . • R ECENT RESEARCH SHOWS THAT L EVETIRACETAM (LEV) MAY BE A MORE EFFECTIVE TREATMENT THAN TRADITIONAL TREATMENTS , BUT WHY IS NOT CLEAR .

SIGNIFICANCE AND BACKGROUND A NTIEPILEPTIC LEV AS A N EUROPROTECTIVE A GENT • LEV REDUCED LESION VOLUME AND THE OCCURRENCE OF NONCONVULSIVE SEIZURES IN EPILEPTIC RATS (C UOMO ET AL ., 2013) • LEV PROTECTS CELLULAR STRUCTURES FROM DAMAGE IN PTS (G IBBS , W ALKER , AND C OCK , 2006) • LEV ACTS AS A NEUROPROTECTIVE AGENT BY ENHANCING HISTOLOGICAL , MOLECULAR , AND BEHAVIORAL ELEMENTS OF RECOVERY AFTER TBI (Z OU ET AL ., 2013)

PURPOSE C OMPARE THE NEUROPROTECTIVE EFFECTS OF THE ANTI - EPILEPTIC DRUGS LEV AND PHE IN ENHANCING AND FACILITATING NEURON RECOVERY AFTER TBI. T HIS COMPARISON WILL BE DONE THROUGH BEHAVIORAL , MORPHOLOGICAL , AND IMMUNOHISTOCHEMICAL ASSESSMENT . Frontal motor area damage in squirrel monkey

DRUGS Levetiracetam (LEV) Phenytoin (PHE) Brand Name Keppra Dilantin Uses Treat epilepsy: Treat epilepsy: • Partial onset seizures • Partial onset seizures • Tonic-clonic seizures • Tonic-clonic seizures • Myoclonic seizures • Status epilepticus Mechanism • Exact mechanism not • Voltage-dependent block of of Action known. voltage gated sodium (Anti- • Binds to synaptic vesical channels--prevents sustained epileptic) glycoprotein, SV2A. repetitive firing of action • Inhibits presynaptic potentials. calcium channels • Primary site of action is the motor cortex. Treatment in Primarily an adjunct A standard treatment for post- T.B.I therapy traumatic seizures.

SUBJECTS • 63 ADULT MALE S PRAGUE -D AWLEY RATS (H ILLTOP , S COTTSDALE , PA). • H OUSED IN THE D AVIDSON C OLLEGE A NIMAL F ACILITY ON A 12 HR LIGHT : 12 HR DARK CYCLE WITH ENRICHMENT AND AD LIBITUM ACCESS TO FOOD AND WATER . • S UBJECTS WILL BE HOUSED IN THE COLONY FOR A MAXIMUM OF 50 DAYS .

DEPENDENT MEASURES- BEHAVIORAL M OTOR F UNCTION • R OTOROD • T IME TAKEN TO F ALL FROM A PPARATUS S PATIAL L EARNING AND M EMORY • Y-M AZE • T IME TAKEN TO EXPLORE NOVEL AREAS • M ORRIS W ATER M AZE • T IME TAKEN TO FIND HIDDEN Mouse on platform in Morris Water Maze PLATFORM

DEPENDENT MEASURES- NEUROLOGICAL • H ISTOLOGICAL A SSESSMENT • R ATS PERFUSED AND STAINED WITH C RESYL VIOLET . • A REAS OF N EURON DAMAGE WILL BE NOTED AND THE AMOUNT OF NEURON DAMAGE WILL BE ANALYZED . • I MMUNOHISTOCHEMISTRY • I NTERLUKIN 1-B (IL-1 B ) AND GLT-1/EAAT2 • C HARACTERIZE CHANGES IN INFLAMMATION AND THE PRIMARY GLUTAMATE TRANSPORTER Coronal slices stained with Luxol fast blue/Cresyl violet after intracerebral hemorrhage (Wang & GLT-1 IN REGIONS OF INTEREST . Tsirka, 2005).

TIMELINE D AY 1: R ATS A RRIVE D AY 7: B EGIN BEHAVIORAL PRE - TESTING D AY 14: S URGERY D AY AND BEGIN TREATMENT D AYS 15-20: R OTOROD BEHAVIORAL TEST • R ATS WILL COMPLETE THREE TRIALS / DAY . D AYS 21-26: Y-M AZE BEHAVIORAL TEST • R ATS WILL COMPLETE THREE TRIALS / DAY . D AYS 27-32: M ORRIS W ATER M AZE BEHAVIORAL TEST • D AYS 27-31: TIME TO LOCATE H IDDEN PLATFORM . • D AY 32: TIME TO LOCATE VISIBLE PLATFORM . • R ATS WILL COMPLETE THREE TRIALS / DAY . D AY 33: PERFUSE FOR CRESYL VIOLET High definition fiber-tracking map depicts D AY 34-35: W AIT P ERIOD neural connections broken in TBI and other disorders. D AY 36-38: H ISTOLOGICAL A SSESSMENT D AY 39-42: I MMUNOHISTOCHEMISTRY

CONTROLLED CORTICAL IMPACT (CCI) MODEL A PNEUMATIC OR ELECTROMAGNETIC IMPACT DEVICE DRIVES A RIGID IMPACTOR ONTO THE EXPOSED , INTACT DURA MIMICKING : • Cortical tissue loss • Concussion blood • Acute subdural • Brain barrier hematoma dysfunction • Axonal injury • Coma CCI Surgical Apparatus A) TBI from CCI Model B) Coronal slice showing TBI

BEHAVIORAL PROCEDURES M OTOR F UNCTION • ROTOROD • M EASURES FORE - AND HIND LIMB COORDINATION AND BALANCE . • A NIMALS ARE TRAINED TO WALK ON THE ROD BEFORE BEING EXPOSED TO EXPERIMENTAL CONDITIONS . Rats in Rotorod testing apparatus

BEHAVIORAL PROCEDURES S PATIAL L EARNING AND M EMORY • M ORRIS W ATER M AZE T EST • A NIMALS ARE PLACED IN A POOL OF WATER AND HAVE TO SWIM TO A HIDDEN ESCAPE PLATFORM . • Y-M AZE T EST • T O TEST IF RODENTS PREFER TO SPEND TIME IN NEW OR FAMILIAR AREAS , ONE ARM OF Mouse in Morris Water Maze Test THE Y- MAZE IS BLOCKED FOR A PERIOD OF TIME AND THEN REOPENED . T HE TIME SPENT IN THE NEW AREA IS RECORDED .

Morris Water Test BEHAVIORAL ANALYSIS 105 100 Time (sec) Rotorod Latency 95 60 90 85 50 80 Saline LEV PHE Treatment Group 40 Time (sec) Y-Maze 120 30 100 80 Time (sec) 20 60 40 10 20 0 0 Saline LEV PHE Saline LEV PHE Treatment Group Treatment Group

HISTOLOGICAL ANALYSIS Cortical Contusion Volume Intact CA1 Neurons 70.00% 30 % Intact Neurons (% contralateral) 60.00% 25 Lesion Volume (mm3) 50.00% 20 40.00% 15 30.00% 10 20.00% 5 10.00% 0 0.00% Saline LEV PHE Saline LEV PHE Treatment Group Treatment Group

IMMUNOCHEMICAL ANALYSIS GLAST E FFECTS OF LEV AND PHE ON THE EAAT S , 140% NEUROPLASTIC MARKERS , AND IL-IB IN THE 120% IPSILATERAL FRONTAL CORTEX AND I PSILATERAL HIPPOCAMPUS 100% • GLAST % Density 80% • GLT-1 • EAAC1 60% • GAP-43 40% • S YNAPTOPHYSIN • IL-IB 20% 0% Saline LEV PHE Treatment Group

PREDICTIONS AND RESULTS LEV WILL BE A MORE EFFECTIVE NEUROPROTECTIVE AGENT THAN PHE. T HE MOST EFFECTIVE NEUROPROTECTIVE AGENT WILL HAVE THE : • L EAST IMPAIRED BEHAVIORAL PERFORMANCE • H IGHEST PERCENTAGE OF NEURONS PRESERVED • S MALLEST L ESION VOLUME • L EAST CHANGE IN INFLAMMATION AND REACTIVITY MRI of patient after sever T.B.I.

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

REFERENCES C UOMO , O., R ISPOLI , V., L EO , A., P OLITI , G. B., V INCIGUERRA , A., R ENZO , G. D., & C ATALDI , M. (2013). T HE A NTIEPILEPTIC D RUG L EVETIRACETAM S UPPRESSES N ON -C ONVULSIVE S EIZURE A CTIVITY AND R EDUCES I SCHEMIC B RAIN D AMAGE IN R ATS S UBJECTED TO P ERMANENT M IDDLE C EREBRAL A RTERY O CCLUSION . PL O S ONE, 8 (11). DOI :10.1371/ JOURNAL . PONE . 0080852 G IBBS , J. E., W ALKER , M. C., & C OCK , H. R. (2006). L EVETIRACETAM : A NTIEPILEPTIC P ROPERTIES AND P ROTECTIVE E FFECTS ON M ITOCHONDRIAL D YSFUNCTION IN E XPERIMENTAL S TATUS E PILEPTICUS . E PILEPSIA , 47 (3), 469-478. DOI :10.1111/ J .1528-1167.2006.00454. X Z OU , H., B RAYER , S. W., H URWITZ , M., N IYONKURU , C., F OWLER , L. E., & W AGNER , A. K. (2013). N EUROPROTECTIVE , N EUROPLASTIC , AND N EUROBEHAVIORAL E FFECTS OF D AILY T REATMENT W ITH L EVETIRACETAM IN E XPERIMENTAL T RAUMATIC B RAIN I NJURY . N EUROREHABILITATION AND N EURAL R EPAIR , 27 (9), 878-888. DOI : 10.1177/1545968313491007