Slide 1: Dual SGLT1 and 2 inhibition with sotagliflozin ameliorates adverse cardiac remodeling in HCM mice
Dual SGLT1 and 2 inhibition with sotagliflozin ameliorates adverse cardiac remodeling and diastolic dysfunction in mice with HCM due to tropomyosin E1806 mutation Fuzhong Qin, MD, PhD; David R. Pimentel, MD; Tomas Baka, MD, PhD; Aifeng Zhang, Ivan Luptak, MD, PhD; Wilson S. Colucci, MD. Cardiovascul
Slide 2: Disclosure
Disclosure * Supported in part by an Investigator Initiated Grant from Lexicon Pharmaceuticals.
Slide 3: Background
Background Hypertrophic cardiomyopathy (HCM) is characterized by adverse cardiac remodeling associated with myocardial hypertrophy, diastolic dysfunction, dysregulated calcium handling and fibrosis. The dual SGLT1 and 2 inhibitor sotagliflozin (Sota) is being tested in patients with obstructive and
Slide 4: Methods
Methods 1 Male and female E1806 and non-carrier (NC) mice 7-8 weeks old Randomized to Sota formulated in the chow (30 mg/kg chow) or control diet (CD) for 8 weeks Cardiac structure and function were assessed by echocardiography at baseline, 4 and 8 weeks of treatment Myocardial fibrosis was assessed
Slide 5: Sota prevents myocardial hypertrophy and atrial enlargement in E180G HCM mice
Sota prevents myocardial hypertrophy and atrial enlargement in E180G HCM mice = o r HW/TL (mg/mm) LAW and RAW /TL HWI/TL 4+ a) * E * v y v # = 34 vY «BE = - Pru FE 4 site vv ‘a £2) a aad e 7 a“~a > E e — J y S$ SR me FF th Q > S cs & > oe 3? e & se oF sal ae s° e ) \ o Y Px ¢ gs *P<0.05 vs NC CD #P<
Slide 6: Total wall thickness
Total wall thickness Sota prevents myocardial hypertrophy in E180G HCM mice Total wall thickness -o- NC CD (n=6) 2.04 —* NC sota (n=3) 22- -- E180G CD (n=10) in . —® £180G sota (n=11) 7) # 18-4 2 2.04 % aS & — * OS o- 1.8+ * * E 1.64 se 0 < | Ea El PER ES 1.44 Bou 3 : |? r=) e FE 1.2 T T T 1.2-— T T
Slide 7: E/A ratio
E/A ratio Sota prevents diastolic dysfunction in E180G HCM mice —o- NC CD (n=6) —* NC sota (n=3) ++ E180G CD (n=10) —® £180G sota (n=11) Baseline 4 8 Weeks Values are means + SEM; n=3-11 *P<0.05 vs E180G CD group 30-5 s— = - 205 * * 49-4-2- NC CD (n=6) —* NC sota (n=3) ++ E180G CD (n=10) —® £180G so
Slide 8: Myocardial fibrosis is increased in E180 HCM mice and markedly decreased by Sota
Myocardial fibrosis is increased in E180 HCM mice and markedly decreased by Sota #5) . a] 3 & ey. & nol G2 g 7 2 \. | Wh i Bee NC CD NC Sota E180GCD €E180G Sota
Slide 9: Summary
Summary 1 Sota treatment for 8 weeks prevented the development of: LV hypertrophy LV diastolic dysfunction LV fibrosis Atrial enlargement
Slide 10: Background 2: Mitochondrial dysfunction and calcium dysregulation in HCM
Background 2: Mitochondrial dysfunction and calcium dysregulation in HCM HCM is associated with Van ysteine 674 m mitochondrial dysfunction which Low RNOS High RNOS may decrease ATP production Reversible Irreversible and increase the generation of Stimulation ater ary reactive oxygen species (ROS) a
Slide 11: Methods
Methods 2 * Myocardial oxidative stress was assessed by immunohistochemical staining for 4-OH-nonenal (HNE) * Programs encoding mitochondrial genes were assessed by RNA sequencing * SERCA oxidative sulfonylation at cysteine 674 (SERCA C674 SO3H) was assessed by immunohistochemical staining with a si
Slide 12: Oxidative stress, as reflected by 4-HNE, is increased in E180G mice and decreased by Sota
Oxidative stress, as reflected by 4-HNE, is increased in E180G mice and decreased by Sota Bar =25um 35 ime * n 2 8 2 B F = 1 vt 0 NC CD NC Sota E180G CD E180G Sota *P<0.05 vs NC CD #P<0.05 vs E180G cD
Slide 13: SERCA oxidation at C674 is increased in E180G mice and prevented by Sota
SERCA oxidation at C674 is increased in E1806 mice and prevented by Sota B. x . rs) oo mM £8 or oo oe a Ww oo 0 Bar = 25 um *P<0.05 vs NC CD or NC sota ##P<0.05 vs E1806 CD
Slide 14: Ten most decreased gene programs in E180G myocardium relate to mitochondrial structure and function
Ten most decreased gene programs in E1806 myocardium relate to mitochondrial structure and function HCM, sota vs CD CD, HCM vs WT Gene Set Name v Size v |NES v |p y FOR y|NES~i p(y FDRG ¥ GOCC MITOCHONDRIAL PROTEIN CONTAINING COMPLEX 260 -3.11 0.0000 GOCC_ MITOCHONDRIAL MATRIX 290 -2.97 0.0000 GOBP
Slide 15: Ten most decreased gene programs in E180G myocardium relate to mitochondrial structure and function
Ten most decreased gene programs in E1806 myocardium relate to mitochondrial structure and function HCM, sota vs CD CD, HCM vs WT Gene Set Name v Size v |NES v |p y FDRG v|NES-'p ¥ FDRG ¥ GOCC MITOCHONDRIAL PROTEIN CONTAINING COMPLEX 260 3.21 0.0000 -3.11 0.0000 GOCC_ MITOCHONDRIAL MATRIX 290| 2.37
Slide 16: Ten most increased gene programs are related to fibrosis
Ten most increased gene programs are related to fibrosis HCM, sota vs CD CD, HCM vs WT Gene Set Name |v |Sizev|NESv|p | FDRq y|NES-!\p |v FDRG y|I ¥GOCC COLLAGEN CONTAINING EXTRACELLULAR MATRIX 367 2.92 0.0000 1GOMF EXTRACELLULAR MATRIX STRUCTURAL CONSTITUENT 136) 2.89 0.0000 ¥GOCC EXTERNAL ENCAPSUL
Slide 17: Ten most increased gene programs are related to fibrosis
Ten most increased gene programs are related to fibrosis HCM, sota vs CD CD, HCM vs WT Gene Set Name |v |Sizev|NES |p |» FDRq y|NES-!\p |v FDRG y|I ¥GOCC COLLAGEN CONTAINING EXTRACELLULAR MATRIX 367 1.44 0.0021 0.2518} 2.92 0.0000: 1GOMF EXTRACELLULAR MATRIX STRUCTURAL CONSTITUENT 136, 1.39 0.0164 0
Slide 18: Summary
Summary 2 * HCM was associated with increased oxidative stress (4HNE) and oxidative modification of SERCA at C674, both of which were prevented by Sota. * HCM was associated with decreased expression of genes for oxidative metabolism, and increased expression of genes for myocardial fibrosis. * Sota
Slide 19: Conclusions
Conclusions In E1806 HCM mice, dual SGLT1 and 2 inhibition with Sota attenuated myocardial hypertrophy, diastolic dysfunction, oxidative stress, SERCA oxidation and fibrosis. Mitigation of SERCA oxidation and fibrosis by Sota may contribute to improved diastolic function. Sota-induced up-regulation