1. Role of microRNAs in heart diseases
miRNA induced myocardial interstitial fibrosis
Baofeng Yang, Zhiguo Wang, Yanjie Lu, Zhenwei Pan
Department of Pharmacology, Harbin Medical University
State-Province Key Laboratories

2. Background
In China, about 3 million people died each year of cardiovascular disease, approximately 45% of the total mortality.
About 600,000 cardiogenic sudden death occurred each year, more than 90% induced by malignant arrhythmias.
Myocardial interstitial fibrosis can result in structural remodeling, and induce lethal arrhythmias.
Harbin Medical University

3. Fibrosis is a characteristic of all forms of cardiac diseases of different origins
Ventricular tachycardia
Myocardial fibrosis
Structural remodeling
Ischemia
Hypertrophy
Atrial fibrillation
Heart failure

4. Mechanism of myocardial interstitial fibrosis?
Electrical
remodeling ?
Heart diseases
Structural
remodeling
Myocardial fibrosis

5. Induction of atrial fibrosis by nicotine in dogs
Control
Nicotine (50)
Collagen content
Fibrosis

6. Induction of atrial fibrosis by nicotine in dogs
TGF-β1
TGF-βR2
CTGF
Nicotine (50)
Nicotine (5)
Nicotine (50)
Nicotine (5)
Nicotine (5)
Nicotine (50)
Control
Control
Control
25kDa
70kDa
38kDa
GAPDH
GAPDH
GAPDH

7. Intracardiac pacing induced fibrosis
Collagen content
Control
Pacing
Fibrosis

8. Intracardiac pacing induced fibrosis

9. Myocardial ischemia induced fibrosis
Infarction area
Collagen content
Sham (LV)
MI (Border zone)
Fibrosis

10. Myocardial ischemia induced fibrosis
NIZ IZ
NIZ IZ

11. How fibrosis related signaling pathway was regulated ?

12. Alteration of expression of miRNAs in different fibrosis models
Canine A-TP
Canine nicotine
Rat MI

13. Alteration of expression of miRNAs in different cardiac fibrosis patients
Atrial fibrillation patients
Smoking patients
CAD patients

14. miR-133 and miR-590 regulate TGF-β1 and TGF-βRII protein expression

15. miR-133 and miR-590 target TGF-β1 and TGF-βRII 3’UTR
miR-133 & TGF-β1 3’UTR
miR-590 & TGF-βRII 3’UTR

16. Cardiac-specific overexpression of miR-328 causes fibrosis and cardiac dysfunction
Collagen content WT TG
Fibrosis

17. Knockdown of miR-328 protective against fibrosis induced by MI
MI Border zone
Infarction area WT
miR-328 Knockdown WT
Fibrosis
miR-328 Knockdown

18. MiR-328 regulates collagen production

19. Elevated TGF-β signals in TG mice heart
miR-328
M-miR-328
+AMO-328 NC
TGF-β1
12KDa
GAPDH
36KDa

20. Elevated TGF-β signals in TG mice heart
P-smad2/3
miR-328
+AMO-328
M-miR-328 NC
p-smad2/3
60KDa
GAPDH
36KDa

21. MiR-328 targets directly to TGFBRIII
TGF-βRIII

22. MiR-328 targets directly to TGFBRIII
Luciferase: miR-328 targets TGFBRIII 3’UTR

23. miR-328 overexpression incuced fibrosis and triggered AF
Canine atrium
Mice ECG

24. miR-328 targets at CACNA1C and CACNB1

25. miR-328 overexpression caused cardiac fibrosis
Ischemia
Pacing
Fibrosis
miR-328
Collagen AF
TGF-β1
TGFBRIII
TGFBRI/II
Under certain pathological conditions, level of miR-328 is increased and triggers down-regulation of its target gene TGFBR3 in cardiac fibroblast, which is leading to production of collagens and formation of cardiac fibrosis.
Harbin Medical University

26. miRNAs and cardiac fibrosis
Hypertrophy Heart Failure
Myocardial Infarction
Diabetic
Cardiomyopathy
Atrial Fibrillation
miR-328
miR-133
miR-590
Calcineurin
Cav1.2/ICa
CACNAB1/Cavβ1
TGF-βRIII
TGF-βRII
TGFβ1
repolarization
structure
Ca2+ handling
dispersion
collagen
Fibrosis
Circulation, 2010; Cardiovasc Res, 2009a; Hypertension, 2009; BJP, 2009a, 2009b
Harbin Medical University

27. Problems in clinical application of miRNAs
Effective drug delivery way
Specific action on the target tissue
Selection of miRNAs carriers
纳米技术：最近美国科学家通过试验发现，可以用一种纳米微粒来包裹小分子RNA，再在其外包裹一层传递蛋白，这样就可以帮助的顺利的进入人体细胞。蛋白质外壳会在细胞内环境下自行溶解，释放出携带抗病RNA的纳米微粒。使用这种技术，能够有效的治疗白鼠感染的尤文氏肉瘤。
锁核苷酸技术：丹麦制药公司Santaris Pharma近日宣布SPC3649(LNA-antimiRTM-122)开始进入人体第一阶段的临床试验，这是世界上首个在人体中试验的microRNA药物。SPC3649是由Santaris Pharma开发的丙型肝炎的新型治疗手段，
胆固醇：针对癌基因性质的miRNA（如miR-155）合成互补的AMO，可能使致癌性miRNA失活，减缓肿瘤生长。AMO经2’-O-甲基修饰稳定性增加,毒性降低。目前正在进行小鼠体内与胆固醇结合的AMO的多器官注射实验，它成为新的miRNA抑制性治疗药物。

28. Advances in clinical application of miRNAs
Nanotechnology：American scientists used nano-particles to wrap miRNAs, and then passes on the outer package layer of protein. This technique can effectively treat mice with the Ewing's sarcoma.
Locked nucleic acids technique：Santaris Pharma, a Danish company has commenced a Phase I human volunteer trial of SPC3649 (LNA-antimiRTM-122, developed as a new approach to treat hepatitis C infection), the world's first miRNA medicine to be tested in man.
Cholesterol-conjugated AMO：Complementary AMO targeted oncogene miRNA (eg miR-155) may cause carcinogenic miRNA inactivation, slow tumor growth. AMO combined with cholesterol was taken as a new treatment way to inhibit oncogene miRNA in mice.
纳米技术：最近美国科学家通过试验发现，可以用一种纳米微粒来包裹小分子RNA，再在其外包裹一层传递蛋白，这样就可以帮助的顺利的进入人体细胞。蛋白质外壳会在细胞内环境下自行溶解，释放出携带抗病RNA的纳米微粒。使用这种技术，能够有效的治疗白鼠感染的尤文氏肉瘤。
锁核苷酸技术：丹麦制药公司Santaris Pharma近日宣布SPC3649(LNA-antimiRTM-122)开始进入人体第一阶段的临床试验，这是世界上首个在人体中试验的microRNA药物。SPC3649是由Santaris Pharma
胆固醇：针对癌基因性质的miRNA（如miR-155）合成互补的AMO，可能使致癌性miRNA失活，减缓肿瘤生长。AMO经2’-O-甲基修饰稳定性增加,毒性降低。目前正在进行小鼠体内与胆固醇结合的AMO的多器官注射实验，它成为新的miRNA抑制性治疗药物。

29. Lab members Yanjie Lu Zhiguo Wang Deli Dong Xiaobin Luo Xu Gao
Hongli Shan
Jing Ai
Baoxin Li
Guofen Qiao
Chaoqian Xu
Wenfeng Chu
Benzhi Cai
Yong Zhang
Zhiguo Wang
Xiaobin Luo
Zhenwei Pan
Jiening Xiao
Huixian Lin
Ning Wang
Lihua Sun
Yan Liu
Yunlong Bai
Dongfang Gu
Ying Zhang
Harbin Medical University