tbuckley/cabot-search huggingface dataset:
Abstract Mo138: Renal NPFFR2-mediated increase in blood pressure is associated with downregulation of cAMP signaling and upregulation of Na <sup>+</sup> /K <sup>+</sup> -ATPase protein expression in the renal proximal tubule
Authors: Bibhas Amatya, Jacob Polzin, Jun Feranil, Laureano Asico, Ines Armando, Robin Felder, Pedro Jose, Hewang Lee
Journal: Circulation Research (2024)

Neuropeptide FF (NPFF), an octapeptide originally found in the brain, participates in the regulation of blood pressure because it is present with its receptors, NPFFR1 and NPFFR2, in the cardiovascular regulatory center in the hypothalamus. However, the role of NPFF and its receptors in the kidney on blood pressure regulation is not known. In both the human and mouse renal proximal tubule, NPFFR2, rather than NPFFR1, is the predominant NPFF receptor determined by RNA in situ hybridization and immunofluorescence microscopy. In mouse renal proximal tubule cells, AC263093 (10 -6 M, 15 min), a specific NPFFR2 agonist and NPFFR1 antagonist, inhibited the forskolin-stimulated cAMP production (Vehicle: 100±6.14%, n=6; AC263093: 72.4±6.53%, n=6, p&lt;0.05), which was reversed by pretreatment with RF-9, an antagonist of both NPFFR1 and NPFFR2 (98.2±9.10%, n=6); RF-9, by itself, had no significant effect (100.8±11.44%, n=6), indicating linkage of NPFFR2 to the inhibitory G protein Gai in renal proximal tubule cells. In human renal proximal tubule cells, NPFF also increased Na + /K + -ATPase protein expression, determined by immunoblotting, in a time- and concentration-dependent manner. Protein expression of Na + /K + -ATPase was decreased in NPFFR2 -deficient human renal proximal tubule cells caused by specific NPFFR2 siRNA (Mock: 100±1.71%, NPFFR2 siRNA: 58.4±4.93%, n=4; P &lt; 0.05). Furthermore, blood pressure, measured by telemetry, was increased and sodium excretion was decreased in conscious C57Bl/6 mice infused with NPFF (9.25 mmol, 0.5 mL/hr/7 days) underneath the renal capsule. The effect was probably via NPFFR2 because the blood pressure of mice was increased by the intraperitoneal injection of AC263093 (20 mg/kg/day, 7 days) and fed a high salt diet (4% NaCl), which is consistent with the decrease in blood pressure (98±4 vs 113±3 mmHg; P &lt; 0.05; n=3-5/group) by the renal subcapsular infusion of Npffr2 siRNA in C57Bl/6 mice also fed the high salt diet. Taken together, NPFFR2 activation increases blood pressure and decreases sodium excretion that is associated with downregulation of cAMP signaling and upregulation of Na + /K + -ATPase protein expression in the renal proximal tubule.
Abstract Mo025: Noninvasive cardiac phenotyping in patients treated with chemo- and immunotherapy for breast cancer
Authors: Fatemeh Khashami, Ivan E Dimitrov, Maximilian Fuetterer, Sebastian Kozerke, Crystal Harrison, Mai Huynh, Jae Mo Park, Zoltan Kovacs, Craig Malloy, Anke Henning, Nisha Unni, Vlad Zaha
Journal: Circulation Research (2024)

Current breast cancer diagnosis and treatment protocols achieve survival rates of&gt;80% at 10 years.However, long-term cancer survivors have&gt;4-fold higher incidence of cardiovascular complications, including heart failure.Early potentially reversible changes that may underlie downstream irreversible loss of mechanical function are unknown.Therefore, we aimed to test the feasibility of a noninvasive cardiac phenotyping protocol in patients undergoing treatment for breast cancer with doxorubicin, without or with immune checkpoint inhibitors, including multiparametric proton cardiac magnetic resonance(CMR) imaging and novel metabolic carbon13MR. A comprehensive set of studies included conventional 1HCMR,as well as metabolic 13CCMR using hyperpolarized(HP)[1-13C] pyruvate, following an oral glucose load. 13CCMR with HP[1-13C] pyruvate allows the detection of downstream metabolites of injected pyruvate within the myocardium: bicarbonate from mitochondrial oxidation and lactate from anaerobic cytosolic metabolism.HP[1-13C]pyruvate was prepared using dynamic nuclear polarization in a commercial polarizer.Imaging and spectroscopy(MRS) of hyperpolarized pyruvate was conducted using a 13C transmit/receive Helmholtz loop-pair coil on a 3Tscanner.13C data were acquired from a short-axis slice, ECG-triggered in end-systole. ProtonCMR images were analyzed using Circle, and 13CCMR images were analyzed using MATLAB. The left ventricular mid-myocardium and blood pool were segmented for metabolite quantitation. Baseline cardiac studies were performed in a consecutive series of 25 women treated for breast cancer. Of these 14(56%) were Caucasian, 3(12%) were Hispanic, 5(20%) Black, and 3(12%) Asian, with ages ranging 46±11 years.Patients received standard-of-care treatment plans based on the tumor type: 11 patients were treated with dose-dense anthracycline and cyclophosphamide, and 14 also received pembrolizumab. The comprehensive baseline phenotyping protocol is planned to be repeated after standard of care therapy in all patients. Baseline data are presented in Table1, with values reported as mean±standard deviation. Noninvasive cardiac phenotyping, including dynamic detection of metabolic substrate utilization is feasible,without showing significant adverse effects,and is well tolerated in a diverse group of patients.Completion of this protocol post-oncological therapy may allow to detect early changes in cardiac function due to chemo-and immunotherapy in susceptible patients.
Abstract Mo121: Sarcomere activation biosensor reveals key functional differences in live cell active states between cardiac and skeletal muscle
Authors: Ashley Martin, Joseph Metzger
Journal: Circulation Research (2024)

The sarcomere is the functional unit of striated muscle contraction. Previous studies have provided valuable information about the sarcomeric proteins and modulators that are necessary for activation of contraction through inter-myofilament signaling. This work has revealed key differences in contraction kinetics and paradigms between cardiac, slow skeletal, and fast skeletal muscle. We seek here to expand these studies through the use of myofilament incorporated Förster Resonance Energy Transfer (FRET) fluorescent biosensor in live, intact muscle. This biosensor fuses fluorescent donor and acceptor proteins to the N- and C-terminals of fast skeletal and cardiac (slow) troponin C, respectively, and allows for the monitoring in real-time of the stepwise physiological mechanism of muscle sarcomere activation in an intact system which preserves significant features of the muscle, including excitation-contraction coupling and load. This biosensor has been designed and validated in both cardiac and skeletal muscle and using intact muscles from transgenic animals we have demonstrated FRET fluorescence in cardiac papillaries, extensor digitorum longus, and soleus muscles. This biosensor has shown unique sarcomere activation properties between cardiac and skeletal muscle, notably the emergence of a prolonged return to baseline of the FRET signal in loaded fast and slow skeletal muscle, which is evidence of a “primed state” of myofilament activation unique to skeletal muscles under load. Elucidation of the “primed state” of sarcomere activation is evidence of a transient memory of recent activity enabling the enhanced contraction properties of skeletal muscle. This finding advances previous understanding of skeletal muscle activation, including summation and tetanic contraction, which is not possible in cardiac muscle. The implementation of this biosensor provides a new approach to interrogate the essential physiological features of muscle sarcomere activation and contraction and adds a valuable new tool for small molecule and gene-based discoveries for muscle disease remediation.
Abstract Or125: Numb Family Proteins regulate left ventricular compaction by modulating autophagic process
Authors: Luqi Zhao, jun li, Lianjie Miao, Yangyang Lu, Anika Nusrat, Sung Yun Jung, Sylvia Evans, Mingfu Wu
Journal: Circulation Research (2024)

Background: Left ventricular noncompaction (LVNC) is a type of cardiomyopathy, which patients manifest clinical features of heart failure, arrhythmias, thrombotic events, and sudden death. We previously found that the ablation of Numb(Nb) , and its homologue Numbl(Nl) at early stage, causes LVNC defects resulting in embryonic lethality. In this study, we found that mice with Nb and Nl deletion at later stage survive to adulthood and develop toLVNC cardiomyopathy. Method: We applied aMHC-Cre to delete and generate Nb and Nl double knockout (ADKO); examine the structure and functions of the ADKO hearts via echocardiography (ECHO); determine the heart arrhythmia via telemetry; determine the Nb subcellular localization via a Nb overexpression line (R26-Flag:mCherry:Numb); and determine the mechanistic function of NFPs in autophagy via autophagic reporter, electron microscopy, and molecular and biochemical assays. Results: The cTnT-Cre mediated Nb and Nl deletion causes LVNC and embryonic lethality, while the ADKO can survive to adulthood and died between eight- and twelve-month-old. The ADKO hearts display LVNC by two months of age, a reduced systolic function by four months old, and left ventricular remodeling and dilatation starting at six months old. ADKO hearts manifest major clinical features of LVNC, including arrhythmias and thrombosis. ADKO hearts exhibit significant accumulation of damaged mitochondria, autophagosomes and autophagolysosomes compared to controls. Biochemical analyses reveal elevated levels of p62 and a higher ratio of LC3II/LC3I in ADKO hearts. NFP null cells and ADKO hearts exhibit abnormal autophagic flux, characterized by arrested at autophagolysosome, increased expression of Lamp1 and Lamp2, but reduced levels of cathepsin D. Mass spectrometry analyses reveal Numb's interaction with proteins involved in endocytosis and autophagy. Consistently, Numb localizes to autophagosomes and autophagolysosomes. Conclusion: NFPs regulate left ventricular compaction at both embryonic and postnatal stages, suggesting that LVNC can be both congenital and acquired. As LVNC mice progress, they develop dilated cardiomyopathy (DCM) at later stage, suggesting that LVNC can occur independently at early stage, and is associated with DCM at later stage. Deletion of NFPs in cardiomyocytes disrupts endocytosis and autophagic flux. Our studies suggest that NFPs mediated endocytosis and autophagy are essential for left ventricular compaction.
Abstract Or123: Increasing ketone supply to the heart has beneficial effects on the source of cardiac ATP production in heart failure with preserved ejection fraction (HFpEF)
Authors: Qiuyu Sun, Cory Wagg, Nathan Wang, Ezra Ketema, Liyan Zhang, Gary Lopaschuk
Journal: Circulation Research (2024)

Introduction: There is considerable confusion as to what changes in cardiac energy metabolism occur in heart failure with preserved ejection fraction (HFpEF). In HFpEF mice hearts, we found a marked decrease in cardiac glucose oxidation, which is accompanied by an increase in fatty acid oxidation. Ketones have been proposed as an extra source of fuel for the failing, heart, although it remains unclear if ketones may adversely affect glucose and fatty acid oxidation. Methods: 13-month-old C57BL/6N female mice were administered a 60% high-fat diet and L-NAME (0.5g/L/day) for 6 weeks to induce HFpEF. Control mice were fed with regular low-fat diet and regular drinking water. Hearts of the mice were excised and perfused in the isolated working mode aerobically with 5 mM glucose, 0.8 mM palmitate, 100 µU/ml insulin, with either low (0.6 mM) or high (1 mM) levels of β-hydroxybutyrate. Metabolic rates of the hearts were measured with radiolabelled [U- 14 C] glucose, [9,10- 3 H] palmitate, and [3- 14 C] β-hydroxybutyrate. Results: In HFpEF mouse hearts, glucose oxidation was significantly decreased with a parallel increase in fatty acid oxidation. As a result, the HFpEF hearts showed overall preserved ATP production rates (79 ± 12 vs. to 106 ± 13 μmol . g dry wt -1. min -1 control vs. HFpEF). Increasing β-hydroxybutyrate levels from 0.6 mM to 1 mM resulted in a rise in ketone oxidation rates in HFpEF hearts (from 692 ± 47 to 952 ± 90 nmol . g dry wt -1. min -1 ). Interestingly, this increase in ketone oxidation did not further compromise the rates of glucose oxidation in HFpEF hearts, but actually increased glucose oxidation rates (from 95 ± 13 to 190 ± 17 nmol . g dry wt -1. min -1 ), and had minor effects on fatty acid oxidation rates (from 796 ± 123 to 678 ± 92 nmol . g dry wt -1. min -1 ). This resulted in an increase in the contribution of ketone and glucose oxidation to ATP production from 15 % to 28 % in HFpEF hearts. Conclusion: In HFpEF mice hearts overall ATP production is not impaired, but rather the heart switches from glucose oxidation to fatty acid oxidation as the main source of ATP production. Increasing ketone supply to the heart increases ketone oxidation and results in a favourable shift in ATP production in HFpEF.
Abstract Mo029: Angiotensin Receptor Neprilysin Inhibitor improves cardiac hemodynamics and arrhythmogenesis in mitral regurgitation-induced heart failure—from bench to bedside approach
Authors: Wei-Ting Chang, Sudeshna Fisch, Ping-yen Liu
Journal: Circulation Research (2024)

Background: An effective pharmacological therapy yet exists to mitigate mitral regurgitation (MR) progression before surgical interventions. Angiotensin receptor-neprilysin inhibitor (ARNI) has shown benefits in patients with heart failure (HF) with reduced ejection fraction but whether it can reduce degenerative MR induced HF remains unknown. Aim: To test the efficacy of ARNI in improving cardiac function in degenerative MR -induced HF. Methods and Results: Using a novel rat model of degenerative MR, we found that in degenerative MR rats treated with ARNI, left ventricular function and atrial fibrillation (AF) inducibility significantly improved compared with those treated with valsartan only (fractional shortening 61.78±7.26% v.s. 55.07±6.63%). Also, the AF inducibility dropped from 83% to 25% in MR rats treated with ARNI than to 58.3% in those treated with valsartan. ARNI treatment demonstrated significant attenuation of fibrosis, apoptosis and expressions of endoplasmic reticulum (ER) stress and inflammatory-associated proteins in left ventricular tissues compared to valsartan. In a shear stress model, the treatment of ARNI on cardiomyocytes mitigated the flow triggered activities of both ER released calcium signaling and store-operated calcium entry. Parallelly, we prospectively included 65 patients with degenerative MR receiving valsartan or ARNI and followed Holter electrocardiography and echocardiography. Notably, compared with degenerative MR patients receiving valsartan, those prescribed with ARNI demonstrated significant improvement of AF burden and left ventricular function. Conclusions: Degenerative MR-induced cardiovascular dysfunction including fibrosis and ER stress, were more effectively attenuated by ARNI treatment than by valsartan alone.
Abstract Mo132: Pouring Salt in the Wound: Sex-specific Cardiometabolic Responses to Increased Dietary Fat, Sugar, and Salt.
Authors: Emily Schulman, Avery Kendall, Helen Collins
Journal: Circulation Research (2024)

Introduction: The general population is becoming more obese, and diabetes rates continue to rise, with overnutrition and consumption of processed foods likely contributing factors. Many people consume diets high in fat, sugar, and salt; however, this triad of factors is rarely examined in unison in studies. In addition, studies that examined diets high in fat and sugar in the context of heart failure with preserved ejection fraction and cardiometabolic outcomes have often performed short dietary interventions, with many studies performed in only male mice, and dietary salt content is often not considered or manipulated. Study Goal: This study aimed to assess the long-term impact of high-fat, high-sucrose diets with differing levels of salt (NaCl) on systemic and cardiovascular physiology in male and female mice. Methods and Results: 12-week-old C57BL/6J male and female mice ( n = 4/group) were fed one of the following diets for 25 weeks: normal chow (NC), low-fat diet (LFD; 10 kcal%), high-fat high sucrose (HFHS; 60 kcal%) with 0.33% NaCl, HFHS with 1% NaCl (HFHS1%), and HFHS with 8% NaCl (HFHS8%). Mice were subjected to basal tail blood metabolite assessment and weighed weekly. In week 24, mice were subjected to glucose tolerance testing, dual-energy X-ray absorptiometry followed by postmortem gravimetric, blood metabolite, histological, and biochemical analyses at 25 weeks. We found that irrespective of sex, body weight was significantly greater in mice receiving HFHS ( p &lt;0.001) and HFHS1% ( p &lt;0.001) versus the other diets. By 25 weeks, male mice were significantly larger than female mice with all diets except HFHS1% ( p &lt;0.05), which correlated with increased percent body fat ( p &lt;0.01), impaired glucose tolerance ( p &lt;0.05) and increased ketone levels ( p &lt;0.05) in female mice subjected to HFHS1% versus males on the same diet. There was also increased heart weight and atrial weights in female mice subjected to HFHS and HFHS1% versus the other females, which was not observed in male mice. Conclusions: Although previous short-term dietary studies have shown that males have altered cardiometabolic responses more than females, our findings together show that long-term dietary interventions with HFHS diets with salt may adversely impact female systemic and cardiovascular physiology.
Abstract Mo015: Myeloid Specific PD-L1 Deletion Promotes Cardiac Dysfunction And Myocardial Inflammation
Authors: Angelica Toro Cora, Rohan Sethi, qinkun zhang, Hind Lal, Tousif Sultan
Journal: Circulation Research (2024)

Background: Immunotherapy, particularly PD-1/PD-L1 blockers, is effective in cancer treatment but can cause rare, but potentially fatal heart issues. Despite the significant impact of these adverse events, there is limited research into the underlying mechanisms of immune checkpoint inhibitor (ICI)-mediated cardiotoxicity. PD-L1, which is abundantly expressed in cardiac tissue, particularly exhibits high expression levels over myeloid cells. However, the precise role of myeloid-specific PD-L1 signaling in mediating autoimmune cardiac disease pathology remains unclear. Methods and Results: To study the role of myeloid cell-specific PD-L1 (myeloid-PD-L1) in cardiac pathophysiology, we generated myeloid-specific PD-L1 KO mice, using Lyz2tm1(Cre) Ifo systems. As expected, PD-L1 expression was significantly down-regulated in Mφs from myeloid-PD-L1 KOs. The cardiac function was followed by echocardiography. Controls and myeloid-PD-L1-KO hearts had comparable ventricular function at 1 month of age. Interestingly, as early as 2 months of age, myeloid-PD-L1-KOs displayed marked cardiac dysfunction as reflected by reduced LVEF, LVFS, and HF markers. Immune profiling was conducted whether chronic inflammation contributes to cardiac dysfunction and remodeling at a later age as observed at 2 and 3 months of age. Interestingly, excessive systemic inflammation and substantial immune infiltration (innate and adoptive), activation, and a pro-inflammatory cytokine storm were detected in the KO hearts. Furthermore, myeloid-PD-L1 KO hearts showed increased recruitment of pro-inflammatory CCR2+ monocytes and macrophages. Conclusion: Our data with myeloid-PD-L1 KOs suggest that deleting PD-L1 in myeloid lineage promotes myocardial and systemic inflammation through innate and adaptive immune cell activation and recruitment of CCR2+ MΦs. Subsequently, this leads to impaired cardiac pathophysiology. Thus, these translational studies will directly evaluate the potential of targeting myeloid-PD-L1 signaling to alleviate cardiac damage.
Abstract Mo129: Elevation in male sex hormone metabolites with increased risk factors for inflammation and clotting in female kidneys following chronic binge drinking
Authors: Prasanth Puthanveetil
Journal: Circulation Research (2024)

Background: Binge drinking is a major systemic health concern. Binge drinking could alter renal metabolome and induce markers of injury as per our previously published work. Further investigation into the top regulated pathways revealed alterations sex hormone biosynthesis pathway was altered in a sex specific manner following 6 weeks of binge drinking. Our goal is to unveil individual metabolite mediators belonging to sex hormone biosynthesis pathway and to investigate the connecting link to cardiorenal health in both sexes following chronic binge drinking. Materials and Methods: For this work, n=3 male and female mice belonging to either control (water fed) or ethanol (20% fed in water) treatment groups were included. At the age of 8- 10 weeks, mice underwent water or binge (20% ethanol) drinking from Monday to Thursday for 4 hours, in dark cycle. This cycle was repeated for 6 weeks. Renal tissues from our experimental groups were collected at the end of 6 weeks and subjected to untargeted metabolomics. In brief, Ultra Performance Liquid Chromatography (UPLC) with Time-of-flight Mass Spectrometry (ESI-TOF-MS) followed by Mass spectrometry based separation was used to identify significantly altered metabolites. Results: Steroid hormone biosynthesis was the one among the top regulated pathways. Metabolites belonging to male hormone biosynthesis were the significantly altered one but specifically in females. Dehydroepiandrosterone, and it’s downstream 11-hydroxytestosterone were upregulated only in females and not in males. Female binge drinking group demonstrated an increase of 4.24 (fold) and 2.21 (fold) for dehydroepiandrosterone, and 11-hydroxytestosterone respectively as per the Log 2 Fold change values with a statistical significance of P&lt;0.05 . Interestingly, this increase in male hormone metabolites we observed in females were also accompanied by an increase in metabolites of proinflammatory and prothrombotic mediators, thromboxane B3 and 5- Oxo- 12 HETE, of 2.3 (fold) and over 1.6 (fold) respectively as per Log 2 Fold change values and with a statistical difference of P&lt;0.05 . Conclusion: Our data clearly demonstrate a sex specific effect following chronic binge drinking, where females have an increase in male hormones accompanied by an increase in prothrombotic and proinflammatory factors.