Dr. Ganesh Halade’s investigation in how “good’’ fats repair the heart could enhance treatment of cardiovascular disease.

TAMPA, FL (April 11, 2022) – New breakthrough research by a University of South Florida lab team describing how certain fats can harm or repair the heart after injury has been accepted by a journal of the American Physiological Society.

A manuscript by Ganesh Halade, PhD, an associate professor of cardiovascular sciences at the USF Health Morsani College of Medicine and a researcher in the USF Health Heart Institute, appears  in the American Journal of Physiology-Heart and Circulatory Physiology, published March 25.

Dr. Ganesh Halade.

Dr. Halade’s research article is titled “Metabolic Transformation of Fat in Obesity Determines the Inflammation Resolving Capacity of Splenocardiac and Cardiorenal Networks in Heart Failure.’’

A key message of the manuscript is how a certain type of healthy fat known as docosahexaenoic acid (DHA) – which is present in Omega-3 fish oil, as found in salmon and tuna – works in tandem with enzymes from the spleen to clear the inflammation in a damaged heart. The spleen plays an important role because it sends immune cells with bags of healthy fat that operates cardiac repair after major injury such as a heart attack.

“So the fat intake needs to be of optimal quality and used by the right enzyme of immune cells,’’ Dr. Halade said. “This is all about cardiac repair and the inflammation clearing molecules (resolution mediators) involved in that repair. It’s essential to the resolution process.’’

Another key message is more about prevention and the genesis of cardiovascular disease: How a chronic and surplus dietary intake of safflower oil (SO, omega-6) can lead to residual inflammation of spleen, kidney, heart, and biosynthesis of pro-inflammatory mediators after an ischemic event. SO is a type of fat commonly used in processed and fast foods that drives chronic inflammation.

“The big question for most people is whether a fat is good or bad, or is omega-3 helpful for heart health?’’ Dr. Halade said. “Everyone is dealing with this question. We’re thinking beyond that by looking at how fat is used in the body after a heart attack and in what forms.’’

“All fats are not created equal,’’ he added, “and despite the extensive literature, the effect of fat intake is the most debated question in obesity, cardiovascular, and cardiorenal research.’’

In his research, Dr. Halade and his team put 100 mice on a 12-week diet of processed (SO) foods to develop residual inflammation and then 50 mice randomized on a primarily DHA-enriched diet for next eight weeks before subjecting to ischemic surgery in mice.

The team made sure both diets had same quantity of calorie per gram of diet. The surplus and chronic intake of SO increased inflammation along with a dysfunctional cardiorenal network. In contrast, DHA increased survival following such heart damage (heart attack).

A result of the study was that the alignment of immune cell enzymes from the spleen and DHA fats are essential to cardiac repair. These so-called “resolution mediators (a family of specialized pro-resolving mediators) is the body’s natural defense process without a negative impact on the body’s physiological response,’’ Dr. Halade said.

Among the key findings in the study:

• DHA supplement improved survival after experimental heart attack to mice
• DHA boost safe clearance of inflammation (resolution) from an injured heart without change in the acute phase of the inflammatory response (day 1), with increased expression of Arg-1, MRC-1, and YM-1 in spleen and infarcted area. These agents are resolution and reparative markers of immune response.
• DHA, along with the body’s natural enzymes, enhanced the ability for the spleen and heart to work together in repairing damage.
• SO primed the spleen and kidney to induce pro-inflammatory pathways and renal inflammation.

“Our next step is to determine the enzymatic machinery or immune responsive enzymes that biosynthesize resolution mediators after ischemic (decreased blood flow commonly called a heart attack) event,’’ Dr. Halade said.

Part of Dr. Halade’s research focuses on how unresolved chronic inflammation and immune responsive metabolic dysregulation contributes to ischemic and non-ischemic heart failure. He is involved in studies of heart failure etiology with an integrative approach focusing on splenic leukocytes and heart, as well as the measurement of inflammatory mediators that impair cardiac repair and resolving lipid mediators that facilitate cardiac repair after a heart attack.

Related story on Dr. Halade’s heart research at USF: https://hscweb3.hsc.usf.edu/blog/2021/05/10/blocking-lipoxygenase-leads-to-impaired-cardiac-repair-in-acute-heart-failure/

Dr. Halade hopes his latest work can shed new light on controlling chronic inflammation and treating heart failure — a progressively debilitating condition in which weakened or stiff heart muscle cannot pump enough blood to meet the body’s demand for nutrients and oxygen.

It has become a growing public health problem, fueled in part by an aging population, poor diet and obesity epidemic. About 6.2 million adults in the U.S. suffer heart failure, and nearly have died within five years of diagnosis, according to the Federal Centers for Disease Control and Prevention.

The American Physiological Society (APS), which publishes the journal, is a nonprofit devoted to fostering education, scientific research, and dissemination of information in the physiological sciences.

“The editors commend you on your outstanding contribution to the journal,’’ the accepting team wrote to Dr. Halade. “We would like to thank you for contributing this novel and important article.’’

The USF Health study was supported by grants from the National Center for Complementary and Integrative Health (NCCIH, formerly known as National Center for Complementary and Alternative Medicine; (NCCAM), and the National Heart, Lung and Blood Institute.

Written by Kurt Loft

USF Health-UT Southwestern Medical Center preclinical study suggests inhibiting PPP1R3G/PP1γ may protect against tissue damage from heart attacks, other diseases linked to inflammation

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TAMPA, Fla. (Feb. 16, 2022) – Cell death plays an important role in normal human development and health but requires tightly orchestrated balance to avert disease. Too much can trigger a massive inflammatory immune response that damages tissues and organs. Not enough can interfere with the body’s ability to fight infection or lead to cancer.

Zhigao Wang, PhD, associate professor of cardiovascular sciences at the University of South Florida Health (USF Health) Morsani College of Medicine, studies the complex molecular processes underlying necroptosis, which combines characteristics of apoptosis (regulated or programmed cell death) and necrosis (unregulated cell death).

During necroptosis dying cells rupture and release their contents. This sends out alarm signals to the immune system, triggering immune cells to fight infection or limit injury. Excessive necroptosis can be a problem in some diseases like stroke or heart attack, when cells die from inadequate blood supply, or in severe COVID-19, when an extreme response to infection causes organ damage or even death.

A new preclinical study by Dr. Wang and colleagues at the University of Texas Southwestern Medical Center identifies a protein complex critical for regulating apoptosis and necroptosis — known as protein phosphatase 1 regulatory subunit 3G/protein phosphatase 1 gamma (PPP1R3G/PP1γ, or PPP1R3G complex). The researchers’ findings suggest that an inhibitor targeting this protein complex may help reduce or prevent excessive necroptosis.

The study was reported Dec. 3, 2021, in Nature Communications.

Zhigao Wang, PhD, associate professor of cardiovascular sciences, in his laboratory at the USF Health Heart Institute. Images on the monitor depict two types of cell death: apoptosis (left) and necroptosis. — Photo by Allison Long, USF Health Communications

“Cell death is very complicated process, which requires layers upon layers of brakes to prevent too many cells from dying,” said study principal investigator Dr. Wang, a member of the USF Health Heart Institute. “If you want to protect cells from excessive death, then the protein complex we identified in this study is one of many steps you must control.”

Dr. Wang and colleagues conducted experiments using human cells and a mouse model mimicking the cytokine storm seen in some patients with severe COVID-19 infection. They applied CRISPR genome-wide screening to analyze how cell function, in particular cell death, changes when one gene is knocked out (inactivated).

Receptor-interacting protein kinase (RIPK1) plays a critical role in regulating inflammation and cell death. Many sites on this protein are modified when a phosphate is added (a process known as phosphorylation) to suppress RIPK1’s cell death-promoting enzyme activity. How the phosphate is removed from RIPK1 sites (dephosphorylation) to restore cell death is poorly understood. Dr. Wang and colleagues discovered that PPP1R3G recruits phosphatase 1 gamma (PP1γ) to directly remove the inhibitory RIPK1 phosphorylations blocking RIPK1’s enzyme activity and cell death, thereby promoting apoptosis and necroptosis.

Dr. Wang (back) and laboratory associate Ken Chen. — Photo by Allison Long, USF Health Communications

Dr. Wang uses the analogy of a car brake help explain what’s happening with the balance of cell survival and death in this study:  RIPK1 is the engine that drives the cell death machine (the car). Phosphorylation applies the brake (stops the car) to prevent cells from dying. The car (cell death machinery) can only move forward if RIPK1 dephosphorylation is turned on by the PPP1R3G protein complex, which releases the brake.

“In this case, phosphorylation inhibits the cell death function of protein RIPK1, so more cells survive,” he said. “Dephosphorylation takes away the inhibition, allowing RIPK1 to activate its cell death function.”

The researchers showed that a specific protein-protein interaction – that is, PPP1R3G binding to PP1γ — activates RIPK1 and cell death. Furthermore, using a mouse model for “cytokine storm” in humans, they discovered knockout mice deficient in Ppp1r3g were protected against tumor necrosis factor-induced systemic inflammatory response syndrome. These knockout mice had significantly less tissue damage and a much better survival rate than wildtype mice with the same TNF-induced inflammatory syndrome and all their genes intact.

Overall, the study suggests that inhibitors blocking the PPP1R3G/PP1γ pathway can help prevent or reduce deaths and severe damage from inflammation-associated diseases, including heart disease, autoimmune disorders and COVID-19, Dr. Wang said. His laboratory is working with Jianfeng Cai, PhD, a professor in the USF Department of Chemistry, to screen and identify peptide compounds that most efficiently inhibit the PPP1R3G protein complex. They hope to pinpoint promising drug candidates that may stop the massive destruction of cardiac muscle cells caused by heart attacks.

The research was supported by grants from the Welch Foundation and the National Institute of General Medical Sciences, a part of the National Institutes of Health.

Graphic created with Biorender app by Zhigao Wang, USF Health Heart Institute.

USF Health research discovers APC limits heart damage by preventing excessive loss of endothelial protein C receptors on the cardiac muscle cell membrane

TAMPA, Fla. (Jan. 31, 2022) — A University of South Florida Health (USF Health) preclinical study offers molecular insight into how activated protein C (APC) may improve aging patients’ tolerance to reperfusion injury – a potentially adverse effect of treatment for ischemic heart disease.

The research, published online Dec. 21 in Circulation Research, suggests that drugs derived from APC may limit ischemia and reperfusion-induced heart damage (reperfusion injury for short) and thereby help preserve cardiac function in older hearts.

Advanced age is a major risk factor for ischemic heart disease, often caused by a buildup of plaques in coronary arteries that narrows the vessels and restricts the supply of oxygenated blood to the heart. This “hardening of the arteries” can eventually trigger a heart attack.

Blood thinners, clot-buster medications, and other drugs, as well as procedures such as coronary artery bypass surgery and balloon angioplasty, are commonly used to restore blood flow to oxygen-starved (ischemic) heart muscle tissue. Paradoxically, especially in older patients, these necessary revascularization treatments can worsen cellular dysfunction and death around the site already damaged by a heart attack, or coronary artery disease. No effective treatments currently exist to prevent age-related reperfusion injury.

“Our research focuses on trying to determine why older hearts are at greater risk for reperfusion injury than younger hearts,” said lead author Di Ren, PhD, a research associate in the Department of Surgery, USF Health Morsani College of Medicine. “Our goal is to find targeted therapeutic strategies to help older people improve their resistance to the pathological condition of ischemia and reperfusion stress.”

“The preliminary evidence in this paper suggests that treatment with activated protein C has the potential to strengthen the cardiac tolerance of aging patients to reperfusion injury from surgery, minimally invasive procedures, or drugs, and (thereby) increase heart attack prevention or survival,” said the study’s principal investigator Ji Li, PhD, a professor of surgery at the USF Health Heart Institute.

Di Ren, PhD, a research associate in the USF Health Department of Sugery, was the Circulation Research paper’s lead author.

APC, a protein circulating in blood, has both anticoagulant (blood clot prevention) and anti-inflammatory functions that can help protect cells from disease and injury. Endothelial protein C receptor (EPCR) – located both on cells lining blood vessels and on the surface of cell membranes, including heart muscle cells – is associated with increased APC production and regulates APC’s subsequent cell signaling (or cell communication).

In this mouse model study, the researchers analyzed how APC exerts cardiac protection during ischemia and reperfusion. The groups of mice observed included young and old “wild-type” mice with all their genes intact, and young “knock-in” EPCR R84A/R84A mice genetically modified to make their EPCR receptors incapable of interacting with the APC protein as well as their wild-type littermates without the EPCR R84A/R84A mutation.

Naturally occurring APC or one of two laboratory-engineered APC derivatives were administered to the mice with heart attack-induced ischemia before reperfusion. One derivative (compound APC-2Cys) selectively activated a signaling pathway to promote cell protection without inhibiting blood clotting (coagulation). The other derivative (compound APC-E170A) selectively triggered a signaling pathway promoting only anticoagulation.

Ji Li, PhD, a professor of surgery at the USF Health Heart Institute, was the study’s principal investigator. — Photo by Allison Long, USF Health Communications

Among the team’s key preclinical findings:

— The stress of Ischemia and reperfusion injury induced “shedding” of EPCRs in young and old wild-type mice – that is, a greater number of these receptors were cut from the heart muscle cell membrane and then moved into the bloodstream. This EPCR shortage (deficiency) in the heart can impair activated protein C signaling critical for favorably regulating energy metabolism and anti-inflammatory responses, preventing cell death, and stimulating other activities needed to protect cardiac muscle cells.

— While the hearts of the old and young wild-type mice both showed EPCR shedding, older hearts experienced a more severe EPCR deficiency and decline in APC signaling activity in response to reperfusion injury. No APC signaling was detected in the EPCRR84A/R84A mice, because APC was blocked from binding to the cell membrane receptor.

— Administering APC or its derivatives helped reduce heart damage inflicted by ischemia and reperfusion, particularly in the old mice. Digging deeper, the researchers discovered that by stabilizing (maintaining) EPCR on the cardiac cell membrane, APC strengthens the aging heart’s resistance both to heart attack-related ischemia and to injury associated with restoring coronary artery blood flow.

— Furthermore, APC and the APC-2Cys signaling derivative, but not the APC-E170A anticoagulant-selective signaling (a potential bleeding risk), helped preserve cardiac function. All cardioprotective effects of APC were weaker in young mice in which EPCR was eliminated; their hearts looked and performed like that of older mice.

— The researchers detailed how APC treatments improve cardiac function by regulating both acute (short-term) and chronic (longer-term) metabolic pathways. They demonstrated that enzyme AMPK (AMP-activated protein kinase) mediates an acute adaptive response to cardiac stress immediately following heart attack, while enzyme AKT (protein kinase B) regulates chronic metabolic adjustments to reperfusion stress over time. APC treatment led to better enzyme activity and more efficient energy balance needed to contract cardiac muscle cells and pump blood from the heart to the rest of the body.

“APC is beneficial for ischemia-reperfusion injury both in the acute and chronic stages, so appropriate APC derivatives might be used both as preventive and therapeutic drugs,” Dr. Li said.

Activated protein C (green) interacts with endothelial protein C receptors (red) to form APC/EPCR binding complex (yellow) and stabilize subsequent EPCR-regulated signaling in heart muscle cells under hypoxia-reperfusion stress.  Image courtesy of Ji Li Laboratory, USF Health; first appeared in Circulation Research; DOI: 10.1161/CIRCRESAHA.121.319044

The USF Health Heart Institute researchers collaborated with scientists from Scripps Research Institute, McMaster University (Canada), and the Oklahoma Medical Research Foundation.

Their work was funded by grants from the National Institutes of Health, both the NIGMS and NHLBI.

USF Health-led research validates crucial role of the spleen in cardiac healing, suggests targeting lipid mediator S1P may offer a promising heart failure treatment

Tampa, FL (Aug. 23, 2021) — Although we can survive without a spleen, evidence continues to mount that this abdominal organ plays a more valuable role in our physiological defenses than previously suspected.

“The spleen holds a whole army of immune cells and signaling molecules that can be rapidly mobilized to respond whenever a major injury like a heart attack or viral invasion occurs,” said Ganesh Halade, PhD, an associate professor of cardiovascular sciences at the University of South Florida Health (USF Health) Morsani College of Medicine.

Principal investigator Ganesh Halade, PhD, is an associate professor of cardiovascular sciences at the USF Health Morsani College of Medicine and a member of the college’s Heart Institute. | Photo by Allison Long, USF Health Communications

Dr. Halade led a new preclinical study that analyzed the interactions of the lipid mediator sphingosine-1-phosphate (S1P) in the spleen and heart during the transition from acute to chronic heart failure. The researchers discovered new cardiac repair mechanisms to help shed light on spleen-heart coordination of physiological inflammation in a mouse model of heart failure.

The study appeared online August 20 in the American Journal of Physiology- Heart and Circulation.

“Simply put, we showed that the spleen and the heart work together through S1P for cardiac repair,” said principal investigator Dr. Halade, a member of the USF Health Heart Institute. “Our study also suggests that early detection of little or no S1P levels after a heart attack and targeted activation of this bioactive lipid mediator may provide a cardioprotective treatment for patients at high risk of heart failure.”

Dr. Halade and colleagues have defined connections between fatty acids, dysfunctional inflammation control, and heart failure. His laboratory focuses on discovering ways to prevent, delay or treat unresolved inflammation after a heart attack. In this latest study, the researchers turned their attention to where S1P is produced and its role in cardiac repair.

S1P is a lipid mediator dysregulated during inflammatory responses, including heart failure. Moreover, several groups have demonstrated the potential significance of this signaling molecule as a treatment target for heart failure triggered by heart attack and ischemia-reperfusion injury.

People can survive without a spleen, a fist-sized abdominal organ that helps fight infection. But its removal (due to abdominal trauma, or certain medical conditions) has been linked to an increased risk of death from ischemic heart disease.

The USF Health study captured time-dependent movement of S1P from the spleen through circulating blood plasma to the heart. The work was the first to quantify interactions between S1P and S1P receptor 1 (S1PR1) during the progression from acute to chronic heart failure, Dr. Halade said.

The researchers defined S1P/S1PR1 signaling in both mice and humans with heart failure after a heart attack. The otherwise young, healthy “risk-free” mice had no variable cardiovascular risk factors such as obesity, diabetes, hypertension, and aging commonly seen in a clinical setting. The researchers correlated the physiological data from the cardiac-repair mouse model experiments with what they observed in pathologically failing human hearts.

Among their key findings:

• Cardiac-specific S1P and S1PR1 levels were reduced in patients with ischemic heart failure.
• In the risk-free mice, physiological cardiac repair was facilitated by activation of S1P in the heart and the spleen. S1P/S1PR1 signaling increased in both organs from acute through chronic heart failure, helping to promote cardiac repair after heart attack.
• Increased plasma S1P indicates cardiac repair in the acute phase of heart failure.
• Selective activation of the S1P receptor in macrophages (immune cells that that help clear inflammation and guide tissue repair) suppressed biomarkers of inflammation and accelerated biomarkers of cardiac healing in mouse cells.

“This study provides another example that the spleen should not be underestimated, because it contributes to the foundation of our immune health as well as the root cause of inflammatory diseases, including cardiovascular disease,” Dr. Halade said.

The research was supported by grants from the National Institutes of Health and the U.S. Department of Veterans Affairs.  The University of South Florida team worked with collaborators at the University of Alabama at Birmingham and Hokkaido University, Japan.

USF Health preclinical study suggests boosting cardiac SIRT1/SIRT3 levels in older heart attack patients may help protect against ischemia-reperfusion injury

Tampa, FL (Aug. 20, 2021) — Sirtuins are a family of anti-aging proteins that help regulate cellular lifespan, metabolism, and resistance to stress. The potential protective effect of these sirtuin enzymes in age-related diseases, including cardiovascular diseases, remains an area of intense investigation.

Principal investigator Ji Li, PhD, is a professor of surgery and member of the USF Health Heart Institute at the USF Health Morsani College of Medicine. | Photo by Allison Long, USF Health Communications

Now, a new preclinical study led by University of South Florida Health (USF Health) researchers has determined that sirtuin 1 (SIRT1) and sirtuin 3 (SIRT3) levels decline in aging hearts, disrupting the ability of cardiac muscle cells (cardiomyocytes) to contract in response to ischemia-reperfusion injury (also known as reperfusion injury). Furthermore, age-related SIRT1 and SIRT3 deficiency can impair cardiac function by altering mitochondrial dynamics, which play an important role in metabolic health and inflammatory response, the researchers report.

The findings were published online July 3 in Aging Cell.

“We discovered that age-related changes in mitochondrial dynamics are caused by SIRT1/SIRT3 deficiency, specifically in the cardiomyocytes,” said principal investigator Ji Li, PhD, professor of surgery in the USF Health Morsani College of Medicine. “You need a strong presence of SIRT1 and SIRT3 to keep mitochondrial dynamics healthy in the heart. Otherwise, the heart’s pumping function becomes weak.”

Diastolic functions assessment of a mouse heart imaged with ultrasound echocardiography | Photo by Allison Long, USF Health Communications

Mitochondria produce the energy needed to drive nearly all processes in living cells. Cardiac muscle cells contain more mitochondria than any other cells, because the heart needs large amounts of energy to constantly pump blood throughout the body. Stabile mitochondrial dynamics maintain a healthy balance between the constant division (fission) and merging (fusion) of mitochondria and help ensure the quality of these specialized structures known as the “powerhouse” of the cell.

Reperfusion, a common treatment following acute heart attack, restores blood flow (and thus oxygen) to a region of the heart damaged by a blood clot blocking the coronary artery. Paradoxically, in some patients this necessary revascularization procedure triggers further injury to heart muscle tissue surrounding the initial heart attack site. No effective therapies currently exist to prevent reperfusion injury.

Research associate Di Ren, PhD (left) works with the heart perfusing system in the Department of Surgery physiology laboratory as USF undergraduate student Julia Fedorova watches. | Photo by Allison Long, USF Health Communications

To help analyze the response of cardiac mitochondria to ischemia-reperfusion stress, the USF Health researchers deleted SIRT1 or SIRT3 in cardiac muscle cells of mouse hearts, and examined the mitochondrial response to ischemic stress by restricted blood flow. They found that the mitochondria in mouse hearts lacking cardiomyocyte SIRT3 were more vulnerable to reperfusion stress than the mouse hearts with SIRT3 intact. The cardiac mitochondrial dynamics (including shape, size, and structure of mitochondria) in these knockout mice physiologically resembled that of aged wildtype (normal) mice retaining cardiac SIRT3.

Furthermore, the young mice with SIRT1 or SIRT3 removed had measurably weaker cardiomyocyte contractions and exhibited aging-like heart dysfunction when ischemia-reperfusion stress was introduced. In essence, without SIRT1/SIRT3 the hearts of these otherwise healthy young mice looked and behaved like old hearts.

“We started this study trying to understand why older people have higher incidences of heart attacks than younger people, and why they die more often even if they receive maximum treatment. Younger people are much more likely to recover from heart attacks and less likely to suffer from ischemia-reperfusion injury,” said Dr. Li, a member of the USF Health Heart Institute. “Our research suggests that one reason could be that both SIRT1 and SIRT3 are downregulated with aging. Younger people have higher levels of these proteins needed to make mitochondrial dynamics healthier.”

Dr. Li’s research team (pictured here) focuses on understanding the molecular mechanisms of coronary artery disease, the most common cause of age-related heart disease. | Photo by Allison Long, USF Health Communications

The study also suggests that, before surgically opening blocked coronary arteries to restore blood flow in older patients, administering a treatment to “rescue” (improve) their diminished SIRT1/ SIRT3 levels may increase tolerance to cardiac muscle reperfusion stress, thereby reducing heart attack complications and deaths, Dr. Li said. Such a cardioprotective treatment might apply a genetic approach to increase SIRT1/SIRT3 production, or an agonist (drug) to activate SIRT1/ SIRT3, he added.

If their mouse model findings translate to human hearts, Dr. Li’s group wants to work with companies interested in developing and testing SIRT1/SIRT3 activators to mitigate heart attack-related reperfusion injury.

“Our ultimate goal is to identify ideal targets for the treatment of heart attack, especially in older patients,” said Dr. Li, whose research is supported by grants from the National Heart, Lung, and Blood Institute, the National Institute on Aging, and the National Institute of General Medical Sciences.

USF Health-UAB study indicates that lipid mediators may offer new targets for more personalized heart failure diagnosis and treatment

The new study profiled bioactive lipids in blood samples collected from hospitalized black and white patients soon after a severe heart attack.

TAMPA, Fla (May 4, 2020) — Black men and women have higher incidences than whites of developing advanced heart failure following a heart attack. Despite racial disparities in heart attacks (a leading contributor to heart failure), and rehospitalizations and deaths caused by heart disease, the underlying physiology accounting for worse cardiovascular outcomes among blacks is poorly understood.

A new study published May 4 in ESC Heart Failure profiles bioactive lipids in blood samples from hospitalized black and white patients soon after a severe heart attack. The preliminary research was conducted by a team at the University of South Florida Health (USF Health) Morsani College of Medicine and the University of Alabama at Birmingham. The researchers wanted to delineate potential differences in the immune-responsive processes needed to safely clear (resolve) acute inflammation after heart attack-induced tissue injury, with the aim of finding more individualized therapies for heart failure.

“Metabolic and leukocyte-responsive signaling control the acute inflammation needed for timely cardiac repair after a heart attack. But inflammation that is not cleared and remains long-term plays a key role in the pathology leading to heart failure,” said lead author Ganesh Halade, PhD, associate professor of cardiovascular sciences at the Morsani College of Medicine and a member of the USF Health Heart Institute.

“Understanding race and sex-based differences in inflammation and its resolution will help us develop more personalized diagnoses and treatments to delay or prevent heart failure.”

A mouse model study published by Dr. Halade last month discovered that heart repair occurs faster in female mice than males after a heart attack, which improves survival and delays cardiac failure.

In this human study, the researchers collected blood plasma from 53 patients, grouped by race and sex, within 24 to 48 hours after a heart attack. Baseline acute injury caused by the heart attack was similar in all the patients, and so were their ages and body mass indexes. No significant sex-or race-specific differences were detected in total cholesterol, HDL, LDL or triglyceride levels – all indicators (biomarkers) currently used by clinicians to help predict risk and manage cardiovascular disease. Measures of various subtypes of leukocytes (cells that regulate immune fitness) were similar across all patients.

Lead author Ganesh Halade, PhD, associate professor of cardiovascular sciences, USF Health Morsani College of Medicine

Looking for distinct bioactive lipid “signatures,” or inflammatory biomarkers, that might predict poorer cardiovascular outcomes after heart attack, the researchers measured three major polyunsaturated fatty acids: arachidonic acid (AA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA). These omega fatty acids circulate in blood and depend upon what people eat. Also analyzed were dozens of specific proresolving mediator (SPM) indicators and a few other signaling molecules that form when these fatty acids metabolize in response to immune activation.

Overall, black patients showed higher concentrations of the three activated fatty acids after a heart attack than white patients, the researchers found. The comparative analyses of SPMs showed that resolvin E1, a potent proresolving mediator of inflammation derived from the fatty acid EPA, was significantly lower in black men and women than in whites. An earlier major clinical trial linked EPA with reduced ischemic events such as heart attack and stroke in patients with high risk for, or existing, cardiac disease, and another showed that high levels of EPA significantly decreased the risk of heart failure.

The researchers conclude that bioactive lipids are key for diagnosis and treatment of cardiac repair after heart attack to delay heart failure.

Randomized controlled clinical trials will be needed to definitively determine whether distinct SPM signatures can be used to predict, diagnose, treat or prevent heart failure following a heart attack, Dr. Halade said. “If we can stratify risk among larger patient groups to determine who is deficient in SPMs critical for cardiac repair, we may be able to restore those targeted SPMs to improve outcomes.”

The study was supported by grants from the National Institutes of Health.

Heart failure affects about 6.5 million adults nationwide and leads to one in 8 deaths each year, according to the Centers for Disease Control and Prevention. The condition usually develops as the heart gradually loses its ability to pump enough blood through the body.

A University of South Florida-led study of mouse cardiac healing may lead to more precise, sex-specific therapies for heart failure, a leading cause of death

TAMPA, Fla. (April 17, 2020) –  The short-term acute inflammatory response triggered to mend injured cardiac tissue following a heart attack can lead to weakening of the heart’s pumping function if the inflammation remains active over the long-term. Heart failure associated with this unresolved chronic cardiac inflammation has become a leading cause of death in the U.S. and worldwide, yet little is known about the differences in cardiac repair and safe clearance of inflammation between men and women.

Ganesh Halade, PhD, an associate professor of cardiovascular sciences at the University of South Florida Health (USF Health) Morsani College of Medicine and Heart Institute, looks for ways to delay or prevent heart failure — including targeted therapies that may account for potential physiological sex differences.

Ganesh Halade, PhD

Dr. Halade’s team delves into the details of metabolic and leukocyte responsive signaling that facilitate cardiac repair during acute inflammation after injury (like a heart attack) and the resolution thereafter. In particular, he studies how unresolved inflammation driven by a deficiency in fatty acid-derived signaling molecules influences heart failure. Known as specialized proresolving mediators (SPMs), these molecules are naturally made by the body (endogenous).

Now, a new study led by Dr. Halade has investigated the molecular and cellular processes underlying cardiac repair in male and female mice after a severe heart attack. The USF Health study, conducted with collaborator Charles N Serhan, PhD, DSc, at Harvard Medical School, reports that females showed improved heart failure survival characterized by differences in cardiac functional recovery and structure, more reparative immune cells and higher levels of epoxyeicosatrienoic acids (EETs), signaling molecules with anti-inflammatory effects.

The findings were published April 16 in the Journal of the American Heart Association.

“We discovered heart repair happens faster in the female mice than the males after heart attack, that improves survival and delays cardiac failure,” said Dr. Halade, the paper’s senior author.

His ongoing translational work may have applications for the development of sex-specific and other more precise heart failure therapies with fewer side effects due to endogenous nature of bioactive signaling molecules. Currently, men and women receive the same standard first-line medications (angiotensin converting enzyme/receptor inhibitors, diuretics, and beta-blockers) to manage mild-to-severe forms of heart failure.

For this study appearing in JAHA, the researchers used “risk-free” young, healthy mice to control for variable cardiovascular risk factors — such as obesity, insulin resistance, diabetes, hypertension and aging, — common in a clinical setting. They compared the risk-free male and female mice who underwent a procedure to induce severe heart attack with those that did not.

To frame the study, it helps to know that physiological inflammation after tissue injury has two steps — an acute response, where white blood cells rush to the heart to remove dead cardiac tissue, and a resolving phase, where inflammation is cleared with the help of macrophages that arrive to repair the damage, and form stable scar. Both responses are governed by ‘get in’ and ‘get out’ signals to leukocytes (a type of immune cell) infiltrating at the site of the heart muscle injured by the heart attack.

Among the key USF Health research findings:

• Following a heart attack, leukocyte infiltration to clear diseased cardiac muscle cells is coordinated by the production of SPMs that resolve inflammation and promote timely cardiac repair.
• Female mice showed better recovery of the heart’s capacity to pump blood compared to males. “Improvement in heart functional recovery is believed to be enabling the female mice to ‘bounce back’ and survive at a significantly higher rate than male mice after myocardial infarction (heart attack),” the authors wrote.
• Less post-MI scarring and adverse structural remodeling of heart muscle in female mice helps explain their improved recovery of cardiac function and survival in acute and chronic heart failure.
• While both male and female mice equally produced SPMs in response to massive heart attacks, the females generated higher levels of a particular lipid signaling molecule known as epoxyeicosatrienoic acid (EET) to facilitate healing after a heart attack.

“The beauty of these SPM and EET molecules is that they are endogenously biosynthesized and can be useful for clearing harmful inflammation in asthma and other diseases, not just heart failure,” Dr. Halade said.

The USF Health researchers plan to study these bioactive lipid signaling molecules after heart attack in men and women, and consider human-related variable factors absent in mice, such as race.

The study was supported by grants from the NIH’s National Institute of Heart Lung and Blood Institute (Dr. Halade) and the National Institute of General Medical Sciences (Dr. Serhan).

USF Health’s Hua Pan, PhD, MBA, won first place for her moderated poster presentation titled “Anti-thrombin nanoparticles limit ischemia-reperfusion injury and no-reflow in myocardial infarction” on Sept. 2 at the annual European Society of Cardiology Congress in Paris. The Congress draws some 35,000 scientists from more than 100 countries over five days.

As part of the highly competitive presentation moderated by two chairpersons, Dr. Pan, a biomedical engineer and assistant professor of cardiovascular sciences at the USF Health Morsani College of Medicine, was required to present a 5-minute talk on the research poster in front of a group of audiences.  Her prize – free registration at next year’s Congress – was awarded for the poster session covering innovations in cardiac magnetic resonance Imaging.

USF Health Heart Institute’s Hua Pan, PhD, MBA

Dr. Pan and colleagues at the USF Health Heart Institute developed antithrombosis perfluorocarbon nanoparticles, which act as “smart Band-Aids” to find and stay only at the injured area in the heart to deliver treatment. These nanoparticles were evaluated in a rat model for heart attack and their treatment effect was visualized by MRI.  The study showed that the treatment limited further vascular damage from ischemia-reperfusion injury (IRI), a common complication following acute treatment of heart attacks.

Ironically, when blood flow is restored to the region of the heart injured by a blood clot that blocks the coronary artery, this blood reflow can expand injury to tissue surrounding the initial heart attack and lead to congestive heart failure. In explaining IRI, Dr. Pan compares an obstructed blood vessel to a clogged water pipe, already weakened by prior damage, which may leak once the pipe is unclogged and water (blood) flows freely again.

“The antithrombin nanoparticles we developed acted locally to preserve the blood vessels (pipes) in the heart, so that the restored blood reached areas where the treatment was needed, without leaking into areas where it could cause more harm,” Dr. Pan said.  The precision nanoparticle treatment calmed unnecessary inflammation as well as inhibiting the thrombin from forming more blood clots leading to blood vessel obstructions in the heart, she added. It did so without causing the bleeding risk associated with existing anticoagulant drugs.

Because the antithrombin nanoparticles “do not prolong bleeding times or coagulation parameters beyond approximately 30 to 60 minutes after injection, yet maintain prolonged surveillance against activated thrombin locally in the injured area, they represent a potentially useful therapy for cardiac IRI,” Dr. Pan and her poster co-authors concluded.

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Leaning forward, Julio Robaina intently watched as USF Health cardiologist Bibhu Mohanty, MD, sketched a series of shapes on a piece of notebook paper. As Dr. Mohanty pulled his hand away, the sketches were revealed. It was a heart, Robaina’s heart.

Minutes earlier, Dr. Mohanty smiled and asked what had prompted Robaina to come to see him for a second opinion. In tears and obviously frightened about his health, the 57-year-old explained that days earlier he had suffered his second heart attack in seven years. Again, a stent had been placed in one of his coronary arteries, further testing had been performed, and for reasons he did not understand, his cardiologist had recommended surgery.

Dr. Mohanty then pointed to his sketch and shook his head no.

“You could tell right away he knew what he was talking about,” Robaina said.

Julio Robaina, went to USF Health cardiologist Bibhu Mohanty, MD, for a second opinion after suffering his second heart attack in seven years.

Robaina’s eyes were laser focused as his doctor pointed to the heart on the piece of paper and explained what was going on inside his chest. “‘Dr. Mohanty told me, ‘This is your heart, these are your arteries and here is where the cardiologist placed the stents after your first heart attack’,” Robaina recalled.

Then came the great news: with some necessary dietary modifications, a guided exercise program, adherence to medication and regular follow up about any changes, Dr. Mohanty told Robaina he could beat this problem without additional procedures. Robaina was relieved — a huge load was lifted off his already taxed shoulders, and he wholeheartedly agreed to making the changes.

For Robaina, the prospect of another procedure added a lot more anxiety to an already stressful situation.  Robaina had lived in Florida for 14 years after residing most of his life with his extended family in northern New Jersey.

“His fear, anxiety, lack of understanding, lifestyle and issues with medications all would have rendered any additional procedure futile, no matter how technically correct,” said Dr. Mohanty, an assistant professor in the USF Health Morsani College of Medicine Department of Cardiovascular Sciences. “This is why we devised a plan to address him — the patient. First, maximize his health status, and see where that landed us. If he needed surgery in the future, he’d be a vastly different, and better-suited candidate.”

It wasn’t easy to change his lifestyle, but with the support of wife Magaly (pictured here) and Dr. Mohanty, Robaina continues working to optimize his health to reduce the risk of future heart attacks. Dr. Mohanty developed the patient-centered care plan in consultation with Robaina.

Dr. Mohanty detailed the nutritional elements that needed to be a part of the changes. Some choices were easy: cut out sodas. Some weren’t as easy: cut out the high-fat meals, like fast food, which he grabbed on the run while working his busy customer service job.

Changing his lifestyle wasn’t easy, but with the support of Dr. Mohanty and his wife, Magaly, Robaina stuck with it. “The hardest part was getting accustomed to the different foods and getting rid of the salt shaker.  Trying to get the stress out of my life is an ongoing battle,” Robaina said.

Along with eating better, Robaina took Dr. Mohanty’s advice and began to take more time for himself.  “I’m in the customer service business so it’s not always easy. These days people want everything yesterday. The hours I work are getting longer,” Robaina said. “But we like to garden. We are doing a lot more of that.  It’s the way I relieve stress on my time off work.”

Better nutrition and guided exercise, along with medication adjustments as needed, have helped Robaina improve his blood pressure numbers.

Dr. Mohanty also recommended Robaina begin doing cardiovascular rehabilitation. It is offered to any patient with prior heart attack, stenting or bypass surgery. In the past, it was very expensive, but 36 sessions are now covered by most insurance companies. It’s tailored to the patient’s metabolic needs and ability. Blood pressure and heart rate are monitored as patients are taken through a series of exercises that pushes them but does notovertax their abilities.

Because of past knee issues, Robaina’s twice-weekly cardio sessions included a lot of time on the recumbent bike. “Exercising has become very important for me and I enjoy it,” Robaina said. “It’s helped my blood pressure and I feel a lot better. I want to live to see my four grandkids get married and go to college — that’s the whole idea.”

“Many patients going through this, that I have met, even a young “fit” 45-year-old who had a heart attack out of the blue, love it because they find that while they are moved along slowly, they reach further in their fitness goals than they otherwise would have,” Dr. Mohanty added. “In clinical studies, few of the drugs we use have as much mortality and future heart attack risk reduction than cardiac rehab. And this brings us back to Mr. Robaina – and why lifestyle change was so important.”

Dr. Mohanty, an assistant professor in the USF Health Department of Cardiovascular Sciences, specializes in structural interventional cardiology.

Robaina had suffered with high blood pressure issues for years. Following Dr. Mohanty’s advice, Robaina has seen his blood pressure numbers improve. He also made sure to report changes in blood pressure or side effects quickly to Dr. Mohanty, and they made adjustments together that have continued to yield good results.

In addition to receiving excellent medical care, Robaina also enjoys Dr. Mohanty’s sense of humor. He recalled a recent incident at the hospital cafeteria. “I had a healthy plate of food and was drinking water.  My wife Magaly had a soda,” Robaina said. “Dr. Mohanty walked in and saw the orange drink, and said ‘What? There’s a soda in the room!’”

“‘That’s my wife’s!’ I told him,” Robaina said. “Dr. Mohanty laughed and started busting her chops instead.”

“That’s the kind of guy he is. Dr. Mohanty makes you feel real comfortable — like you are one of the family. I love him,” Robaina said. “He’s going to go far in life. I want him to be around for as long as I’m around. I definitely want to keep him as my physician.”

Robaina enjoys gardening at his home, and is doing more it to help relieve the stress that accompanies his high-intensity customer service job.

-Video and photos by Torie M. Doll, USF Health Communications and Marketing

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After suffering his first heart attack 22 years ago, David Gilberg has endured more heart procedures than anyone could count on both hands. Fourteen in fact.  But today, the 72-year-old Minnesota native has a new lease on life, thanks to a procedure performed by cardiologist Bibhu Mohanty, MD, of the USF Health Department of Cardiovascular Sciences.

While the procedure to open a narrowed coronary artery in Gilberg’s heart was fairly routine as far as percutaneous coronary intervention (PCI) goes, the real magic occurred when he felt someone listened to his concerns at long last.

Gilberg spent his entire life farming dairy cattle in northern Minnesota.  A humble and spiritual man, Gilberg worked an area of the country that he termed as having “real, real cold weather.” After visiting Florida for years to escape the frigidity, he moved permanently to Sun City in 2013.  Solidifying the lifestyle change, Gilberg sold his farm last year.  “I had a hard time letting it go. When you’ve been a farmer all of your life, that soil is in your hands. It’s just something that is a part of you.”

Patient David Gilberg of Sun City Center with USF Health cardiologist Bibhu Mohanty, MD.

For more than a year, Gilberg dealt with a severe dysfunction, including shortness of breath.  At times Gilberg could hardly get from bed to bathroom. A quick trip to the grocery store was dreaded.

“I had to know exactly what I wanted and where it was,” he said.  I hurried through the aisles, sometimes having to sit down. After just a few minutes, I returned to the car out of breath and exhausted.”

Gilberg’s ongoing heart issues stemmed from heart disease on both sides of his family. Before seeing Dr. Mohanty, Gilberg took as many as 15 nitroglycerine pills daily. The pain in his arms and chest grew stronger every day.  When he complained to his cardiologist of five years, the doctor said he was fine.

In December, he’d finally had enough.  On a recommendation, he made an appointment with Dr. Mohanty for a second opinion.

When Gilberg met Dr. Mohanty in Sun City, he was concerned that the doctor would diagnose his problem and it would be the last time he saw him.

At the initial visit, the USF cardiologist wondered if one of the stents was possibly clogging up.

“From the beginning, Dr. Mohanty assured me he would be with me throughout my hospital stay.  He said he’d be right there and he was,” Gilberg said with a big smile.  “He came to my hospital room, checked in several times, and always had a very positive attitude.  His bedside manner is second to none. He is the nicest, most concerning fella.”

Not only is Dr. Mohanty’s bedside manner on point, “He saved my life!” said Gilberg, after Dr. Mohanty placed a 10th stent in his heart in January.  Just one month later, Gilberg is walking up to a mile and a half each day.

Dr. Mohanty speaks with Gilberg during an office follow-up visit.

“Being strictly adherent to an optimal medication regimen for coronary artery disease is critical,” Dr. Mohanty said. “When we met, his regimen was not optimal for his history, and we made some significant adjustments.”

With a new vigor, Gilberg is now making plans to visit Minnesota, and might even travel to Sweden to visit relatives in his homeland.  With a daughter, Jenny, in Sarasota and another, Molly, still in Minnesota, he has a lot of love and support.  Molly’s family will welcome their eighth child in July.  Gilberg now plans to visit them twice a year to celebrate his grandchildren’s birthdays.

Gilberg smiles big when he talks about visiting family in Minnesota, an adventure he never dreamed possible a couple of months ago.  “No matter where you go in this world, there is a place that’s home.  ‘Home’ sometimes is the things you have – what you bring with you that kind of makes it feel like home.  But home is always where you were born and raised. And home for me is Minnesota.”

Gilberg’s grandkids are a bright spot as well.   Prior to Hurricane Irma, they wanted to Facetime every day, and nervously asked him about impending Hurricane Irma.   “You know Papa,” 10-year old grandson George said at the time, “I want to see you a few more times — not just in heaven, so you have to take care of yourself and go to a shelter if the storm comes, ok?”

Thanks to Dr. Mohanty, Gilberg will be much more than a just a face on a phone screen. His grandkids will now be able to play with “Papa” in person for a long time.

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Dr. Mohanty attended Duke University Medical School and also did his residency at Duke.  He completed three fellowships: Cardiovascular Disease at Mouth Sinai School of Medicine, Interventional Cardiology at Johns Hopkins Medical Center, and Structural Intervention at Dartmouth Medical School.

To learn more about life-changing cardiovascular research and patient care at USF, visit the USF Health Heart Institute website.

-Story and photos by Michelle Young, USF Health Communications and Marketing