SHAPE Society

1. Heart dialysis systemHeart dialysis system
 Studying the effect of systemic apoA-1 treatmentStudying the effect of systemic apoA-1 treatment
on carotid atherosclerotic plaqueon carotid atherosclerotic plaque
 Studying the effect of LDL aphaeresis on LDL,Studying the effect of LDL aphaeresis on LDL,
plasma inflammatory markers, and cytokinesplasma inflammatory markers, and cytokines
 comparing the effect of local high dose of apoA1comparing the effect of local high dose of apoA1
on atherosclerotic plaque versus systemic therapyon atherosclerotic plaque versus systemic therapy
 Comparing the effect of single therapy versusComparing the effect of single therapy versus
multiple therapymultiple therapy

2. Animal studyAnimal study
 25 Rabbits25 Rabbits
 Cholesterol fed diet (1%)Cholesterol fed diet (1%)
 Inducing vascular injury in carotid arteriesInducing vascular injury in carotid arteries
 Systemic LDL aphaeresisSystemic LDL aphaeresis
 Systemic apoA-1 administrationSystemic apoA-1 administration
 Local apoA-1 administrationLocal apoA-1 administration
 Studying the effect of apoA-1 on plaque asStudying the effect of apoA-1 on plaque as
well as lipid profilewell as lipid profile
 Studying the effect of LDL aphaeresis onStudying the effect of LDL aphaeresis on
lipid profilelipid profile

3. Heart Dialysis System In RabbitsHeart Dialysis System In Rabbits
25 (wattanabe) rabbits
1%Cholesterol fed diet
Control group:
No treatment
N#10
Systemic treatment:
LDL aphaeresis
Systemic apoA1
Local treatment:
of apoA1
Systemic LDL aphaeresis
Single session therapy
N#5
Sacrified after 48 h
N#5
Sacrified after 6th
w
Weekly for 5 w
Sacrified at 6th
w
Sacrified at 6th
w
N#10N#5

4. Treatment modalitiesTreatment modalities
 Apo A-1m 40 mg/doseApo A-1m 40 mg/dose
 Phosphatidylcholine 120 mg/dosePhosphatidylcholine 120 mg/dose
 LDL aphaeresisLDL aphaeresis

5. Measured indicesMeasured indices
Biochemical measurementsBiochemical measurements
Lipid profileLipid profile
LDL, HDL, and TGLDL, HDL, and TG
CRPCRP
Lp(a)Lp(a)
FibrinogenFibrinogen
PlasminogenPlasminogen
Anti-thrombinAnti-thrombin
C3, C4, C1 inhibitorC3, C4, C1 inhibitor
Interlukin-1Interlukin-1
TNF alphaTNF alpha
EdotoxinsEdotoxins
FerritinFerritin
CD 14CD 14
Tissue factorTissue factor
Before, 1 h, 6h, 24h after treatmentBefore, 1 h, 6h, 24h after treatment
Histological studyHistological study
Intimal thichnessIntimal thichness
Intima/medail ratioIntima/medail ratio
Lipid content of the plaqueLipid content of the plaque
Macrophage content of theMacrophage content of the
plaqueplaque
Smooth muscle cell of polaqueSmooth muscle cell of polaque

6. Local infusion of ApoA-1m in toLocal infusion of ApoA-1m in to
carotid artery of rabbitcarotid artery of rabbit
 25 rabbits25 rabbits
 1.5% cholesterol diet for 90 days1.5% cholesterol diet for 90 days
 Perivascular electrical injuryPerivascular electrical injury
 G1:salineG1:saline
 G2:phospholipid vesicleG2:phospholipid vesicle
 G3,4,5:Apo A-1m :250-500-1000 mgG3,4,5:Apo A-1m :250-500-1000 mg
 Local delivery through IVUS catheterLocal delivery through IVUS catheter
 Ninety minutes infusionNinety minutes infusion
Giulia ChiesaGiulia Chiesa

7. plaque lipid contentplaque lipid content
Right carotid Left carotidRight carotid Left carotid
Saline 85.30±8.06 80.70±7.14Saline 85.30±8.06 80.70±7.14
DPPC 83.74±6.19 71.34±15.16DPPC 83.74±6.19 71.34±15.16
AIM 250 75.45±7.83 70.62±10.87AIM 250 75.45±7.83 70.62±10.87
AIM 500 64.21±2.60 61.56±4.01AIM 500 64.21±2.60 61.56±4.01
AIM 1000 53.30±10.18 51.82±6.87AIM 1000 53.30±10.18 51.82±6.87
Giulia ChiesaGiulia Chiesa

8. Study protocolStudy protocol
IVUS study 0, 30m, 60m, 90mIVUS study 0, 30m, 60m, 90m
Blood sample 0, 45,90m , 6,24,48,72hBlood sample 0, 45,90m , 6,24,48,72h
Histologic examination on both carotidsHistologic examination on both carotids
Giulia ChiesaGiulia Chiesa

9. Plasma HDL 72 h after infusionPlasma HDL 72 h after infusion
Giulia ChiesaGiulia Chiesa

10. Plaque areaPlaque area
Giulia ChiesaGiulia Chiesa

11. Inhibition of carotid intimal thickening by apoA-1mInhibition of carotid intimal thickening by apoA-1m
in rabbitsin rabbits
 New Zealand white rabbitsNew Zealand white rabbits
 1% cholesterol-fed1% cholesterol-fed
 Flexible collar application at 21Flexible collar application at 21stst
dayday
of dietof diet
 EPC+A-1m(119+40mg)EPC+A-1m(119+40mg)
EPC(119mg)EPC(119mg)
Maurizio and SomaMaurizio and Soma

12. Study protocolStudy protocol
Maurizio andMaurizio and
SomaSoma
NZWR
1% cholesterol diet
EPC+A-!m EPC control
Significant
intimal
thickening
Drug before injury
Sig decrease in intima
Drug the day of injury
No sig change
No decrease in intima

13. Effect of various regimensEffect of various regimens
Maurizio and SomaMaurizio and Soma
I/M ratio
Control
0.63+/-o.11
EPC treated
0.50+/-0.14
A-1m treated
0.26+/-0.19

14. Reduction of intima thickening inReduction of intima thickening in
hypercholesterolemic rabbithypercholesterolemic rabbit
 1% Cholesterol fed rabbits1% Cholesterol fed rabbits
 Balloon injured ileofemoral arteriesBalloon injured ileofemoral arteries
 G1: apoA-1m+phospholipidG1: apoA-1m+phospholipid
 G2: phospholipidG2: phospholipid
 G3: controlG3: control
PK Shah et allPK Shah et all

15. Intimal thicknessIntimal thickness
PK Shah et allPK Shah et all
Control
1.69+/-0.43
mm2
Carrier treated
1.14+/-0.38
mm2
A-1m treated
o.49+/-0.29
mm2

16. Intima-Media RatioIntima-Media Ratio
 Control:2.1+/-0.1Control:2.1+/-0.1
 Carrier :1.5+/-0.5Carrier :1.5+/-0.5
 Apo1-A:0.7+/-0.2Apo1-A:0.7+/-0.2
PK Shah et allPK Shah et all

17. Intimal lesion covered by macrophagesIntimal lesion covered by macrophages
 Carrier treated:59.3+/-12.3%Carrier treated:59.3+/-12.3%
 ApoA-1 treated:25.3+/-17%ApoA-1 treated:25.3+/-17%
PK Shah et allPK Shah et all

18. conclusionconclusion
Apo A-1m significantlyApo A-1m significantly
Reduces intimal thickeningReduces intimal thickening
Reduces macrophage contentReduces macrophage content
PK Shah et allPK Shah et all

19. ApoA-1m in EKO miceApoA-1m in EKO mice
 EKO mice fed with high cholesterolEKO mice fed with high cholesterol
 G1:Control(saline treated)G1:Control(saline treated)
 G2:Carrier (Phosphatidylcholine)G2:Carrier (Phosphatidylcholine)
 G3:Apoa-1 treated+phospholipidG3:Apoa-1 treated+phospholipid
PK ShahPK Shah

20. ResultsResults
 1.6-fold increase in tissue cholesterol1.6-fold increase in tissue cholesterol
mobilization in apoA-1 groupmobilization in apoA-1 group
 40%-50% decrease in plaque lipid40%-50% decrease in plaque lipid
content in apoA-1 groupcontent in apoA-1 group
 29%-36% decease in macrophage29%-36% decease in macrophage
content in apoA-1 groupcontent in apoA-1 group
PK Shah et allPK Shah et all