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3. Safe therapeutic effects on post-infarct cardiac dysfunction and remodeling following low-dose oral triiodo-L-thyronine in rats Viswanathan Rajagopalan, Ph.D. Dept. of Biomedical Sciences NYIT College of Osteopathic Medicine Old Westbury, NY

4. Background A large body of evidence suggests that the alarmingly high incidence of overt hypothyroidism, subclinical hypothyroidism and low Triiodo-L-thyronine (T3) syndrome in cardiac patients has a negative impact on outcomes. Recent rodent studies show that cardiovascular diseases, including Myocardial Infarction (MI), activate cardiac D3 deiodinase contributing to low tissue T3 and Heart Failure (HF) (reviewed in Gerdes AM. AJP Heart Circ 2015). We previously showed that 3 days of T3 treatment in rats following MI resulted in increased left ventricular (LV) contractility and decreased myocyte apoptosis (Chen YF et al. J Mol Cell Cardiol 2008). In addition, a supraphysiologic dose of Thyroxine (T4) improved long-term LV function and myocyte survival (Chen YF et al. J Transl Med 2013). However, a clinically translatable TH treatment/monitoring protocol for MI demonstrating safe therapeutic benefits has not been clearly established.

5. A therapeutic T3 treatment/monitoring protocol (e.g. translatable to humans) will achieve safe cardioprotective effects in rats following MI Hypothesis

6. Adult female Sprague–Dawley rats underwent MI by ligation of the left anterior descending coronary artery (n=16-20/group). Treatment of T3 with a dose of ~6 μg/kg/day in drinking water was begun after MI and continued for 2 months (Mo). The other group of MI rats and sham controls received vehicle treatment – MI-Veh and Sham-Veh respectively. Appropriate one-way ANOVA or statistical t-tests were performed, data presented as mean±SE and p<0.05 considered statistically significant. Methodology

7. Morphometrics * p<0.05 vs. Sham ** p<0.01 vs. Sham §§ p<0.01 vs. MI-Veh N= 6-11 Sham+VehMI+VehMI+T3 Body weight Pre-op (g)211±11224±12234±24* Body weight Post-op (g)253±16275±15*262±21 HW (mg) 1±0.091.4±0.3**1.4±0.22** LVW+SepW (mg) 0.66±0.060.77±0.05**0.68±0.07 §§ RVW (mg) 0.18±0.010.30±0.13*0.32±0.11* HW/BW (mg/g) 4.0±0.35.0±0.9*5.3±0.8** (LVW+SepW)/BW (mg/g) 2.7±0.22.8±0.22.6±0.2 RVW/BW (mg/g) 0.7±0.11.1±0.41.2±0.4*

8. Serum TH levels were not significantly altered with T3 treatment

9. ** p<0.01; *** p<0.001 vs. Sham at respective time-point ## p<0.01 vs. MI-Veh at corresponding time-point N = 6-10 per group Cardiac MRI – Improved EF following T3

10. * p<0.05 ** p<0.01; *** p<0.001 vs. Sham at respective time-point # p<0.05 vs. MI-Veh at corresponding time-point N = 6-10 per group Cardiac MRI – Improved LV Volumes following T3

11. Representative EKGs with burst/tachy atrial pacing

12. # p<0.05 vs. MI-Veh; N = 9-13 per group Oral T3 reduced atrial arrhythmias

13. * # Oral T3 improved contractility without change in Heart Rate

14. MI-Veh MI-T3 Representative Histology sections

15. p=0.068 Improvement in infarct area with oral T3

16. * * * -N=8 per group -*p<0.05 vs. Sham Restoration of thyroid gene expression following oral T3

17. ** # * * p=0.077 p=0.076 * # ** # -N=8 per group; *p<0.05, **p<0.01 vs. Sham; #p<0.05 vs MI+V Restoration of cardiac gene expression following oral T3

18. # ** * ## -N=8 per group -*p<0.05, **p<0.01 vs. Sham -#p<0.05, ##p<0.01 vs. MI+V Restoration of ion channel gene expression following oral T3

19. ** # * p=0.05 * * * * * -N=8 per group -*p<0.05, **p<0.01 vs. Sham -#p<0.05 vs. MI+V Restoration of ECM gene expression following oral T3

20. p=0.078 # * # p=0.06 p=0.08 ** * * # * * * * N=8 per group; *p<0.05, **p<0.01 vs. Sham; #p<0.05 vs. MI+V Restoration of mitochondrial gene expression following oral T3

21. Summary 1.For the first time, a safe and effective post-MI/HF T3 treatment strategy that dramatically improves LV function, atrial arrhythmogenesis, and tissue remodeling, with restoration of gene expression and no adverse effects has been demonstrated. 2.This study provides an effective treatment/monitoring protocol for T3 treatment of MI that should be readily translatable to humans.

22. Acknowledgments Dr. Martin Gerdes and laboratory, Old Westbury, NY Dr. Youhua Zhang, NYITCOM, Old Westbury, NY Dr. Kaie Ojamaa, The Feinstein Institute, Manhasset, NY Dr. Yuefeng Chen, NYITCOM, Old Westbury, NY Dr. Alessandro Pingitore, CNR Institute, Italy Dr. Rheal Towner, OMRF, Oklahoma City, OK Dr. Eduardo Dedkov, NYITCOM, Old Westbury, NY

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