Sonin D.L.,Medical College of Wisconsin |
Sonin D.L.,Blood and Endocrinology Center |
Wakatsuki T.,Invivosciences, Inc. |
Routhu K.V.,Medical College of Wisconsin |
And 4 more authors.
Journal of Cardiovascular Pharmacology and Therapeutics | Year: 2013
Purpose: Fibroblast activity promotes adverse left ventricular (LV) remodeling that underlies the development of ischemic cardiomyopathy. Transforming growth factor-b (TGF-b) is a potent stimulus for fibrosis, and the extracellular signal-regulated kinases(ERK) 1/2 pathway also contributes to the fibrotic response. The thrombin receptor, protease-activated receptor 1 (PAR1), has been shown to play an important role in the excessive fibrosis in different tissues. The aim of this study was to investigate the influence of a PAR1 inhibitor, SCH79797, on cardiac fibrosis, tissue stiffness and postinfarction remodeling, and effects of PAR1 inhibition on thrombin-induced TGF-b and (ERK) 1/2 activities in cardiac fibroblasts. Methods: We used a rat model of myocardial ischemia-reperfusion injury, isolated cardiac fibroblasts, and 3-dimensional (3D) cardiac tissue models fabricated to ascertain the contribution of PAR1 activation on cardiac fibrosis and LV remodeling. Results: The PAR1 inhibitor attenuated LV dilation and improved LV systolic function of the reperfused myocardium at 28 days. This improvement was associated with a nonsignificant decrease in scar size (%LV) from 23+% in the control group (n = 10) to 16%+5.5% in the treated group (n = 9; P = .052). In the short term, the PAR1 inhibitor did not rescue infarct size or LV systolic function after 3 days. The PAR1 inhibition abolished thrombin-mediated ERK1/2 phosphorylation, TGF-b and type I procollagen production, matrix metalloproteinase-2/9 activation, myofibroblasts transformation in vitro, and abrogated the remodeling of 3D tissues induced by chronic thrombin treatment. Conclusion: These studies suggest PAR1 inhibition initiated after ischemic injury attenuates adverse LV remodeling through late-stage antifibrotic events. © The Author(s) 2012.
Invivosciences, Inc. | Date: 2014-08-12
An automated method for culturing stem cells using a robotic liquid handling system including a translatable bed and a movable multichannel pipette. The method includes the steps of: locating a first multi-well cell culture plate and a multi-trough plate on the bed; placing a suspension of stem cells in at least one trough of the multi-trough plate; using the multi-channel pipette, transferring a portion of the suspension of stem cells to each well of the first multi-well cell culture plate such that at least two of the wells of the first multi-well cell culture plate have different densities of stem cells; selecting a well of the first multi-well cell culture plate having a desired stem cell density; locating a second multi-well cell culture plate; and using the multi-channel pipette, transfer ring the cells of the selected well to a plurality of wells of the second multi-well cell culture plate.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 177.72K | Year: 2016
Project Summary Abstract Cardiovascular drugs are primarily prescribed to older people andgt years of age since the prevalence of chronic heart failure and fibrillation increases with age The incidence of other diseases including cancer and neurological disorders increases with age too Any medications hold potential risks to cause adverse events Therefore the risks of medications in older people also increase with age While cardiac safety of drugs is one of the primary concerns among drug developers and regulatory agencies potential new drugs are tested using animal models of normal age Limited progress has been made in developing a tool to predict potential cardiac safety issues in aging heart representing a significant unmet need in drug discovery industry The United States will have million aging population andgt years of age in The development of a computational model that can predict potential cardiac safety reactions is a plausible step towards preparing the drug discovery industry for a rapidly approaching era of longevity This Phase I proposal will validate and improve a computational model that incorporate potential age related changes in regulators of excitation contraction coupling ECC of cardiomyocytes CMs and contribution of fibroblasts FBs that are known to increase their population in aging hearts The specific Aims are to develop and refine a computational model representing coupled FBs and CMs to simulate electrophysiology and calcium handling of aging human heart tissue to establish human engineered heart tissues by incorporating aging cardiac FBs and applying environmental stress that known to induce cardiac aging These Aims are proposed to optimize a computational model Wisdom Heart WH that mimics age associated changes in cardiac ECC of myocardium with increasing number of FBs The quality of WH model will be evaluated and improved against experimental data obtained from myocardium models The established model will be shared with the consortium organized by HESI hesiglobal org with FDA industry and academia that is trying to improve cardiac safety issues The model will be incorporated to our services for conducting contract research projects of cardiac safety assessments and drug discovery for heart failure We have active contracts with FDA research scientists at the National Center for Toxicological Research One of the largest market of pharmaceutical industry is aging Americans that are expected to reach million in In the drug discovery research and development model systems are often used but are not taking aging into account Our computational aging heart model that will be validated through this proposal is designed to predict potential pharmaceuticalsandapos safety and efficacy concerns which should contribute to develop safer and more effective treatments for aging Americans
Invivosciences, Inc. | Date: 2012-06-06
Engineered cardiac tissues are provided herein. The tissues include cardiomyocyte cells derived from a pluripotent cell, fibroblast cells and extracellular matrix components. Methods of using the tissues described herein are also provided.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 758.66K | Year: 2015
DESCRIPTION provided by applicant We propose to demonstrate the strength of combining our engineered tissue based drug screening approach and a systematically organized mixture based chemical library This approach could become an ultimate tool to develop treatments for complex diseases such as cardiac fibrosis without known drug targets initially Heart failure is the most common cause of hospitalization among Americans or older The heart failure with preserved ejection fraction HFPEF is becoming an epidemic among increasing population of aging Americans Existence of activated fibroblasts i e myofibroblasts differentiated from fibroblasts endothelial cells and non muscle cell is a hallmark of myocardium with HFPEF and they stiffen myocardium to progress HFPEF We developed a disease model for the human cardiac fibrosis using myofibroblast containing engineered tissue constructs that recapitulate connective tissues of myocardium with fibrosis Using our phenotypic screening system we identified TPI compound mixture after analyzing a scaffold scaffold ranking library each scaffold sample contains mixtures of compounds which is equivalent to screening andgt million compounds While our final goal is to identify a novel drug for treating cardiac fibrosis the proposed Phase I project will focus on performing a secondary sample screening of a compound mixture in the chemical scaffolds and identifying individual compounds to pick hit compounds to be tested and optimized in Phase II study In a parallel study we will analyze mechanisms of action of TPI in fibrosis relieving phenotypes and identify potential drug targets by performing affinity free target identification method stabilizing target molecules with individual hit compounds Successful demonstration of combing engineered tissue based drug discovery system and scaffold ranking library with positional scanning technology will be extended to other drug discovery projects for age related dieses such as osteoarthritis PUBLIC HEALTH RELEVANCE Heart injuries such as heart attaches leave non healing wounds and develop stiff scars that hamper healthy pump function While almost all heart failure patients aged years develop this condition cardiac fibrosis no effective treatment exists Our novel drug screening system using human grown wound tissues screens a chemical library that is systematically designed to identify drug candidates running fewer tests
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 2.09M | Year: 2012
DESCRIPTION provided by applicant Current NIGMS SBIR funding supported InvivoSciences LLCandapos s IVS launch of several product lines in IVS generated revenues from the sales of three dimensional D cell culture tools MC TM and IVS InsertsTM that can grow various hydrogel tissues without any support layers The culture tools enable Palpator TM and Tissue StretcherTM to stretch the hydrogel tissues for biomechanical measurements and mechanical conditioning e g cyclic stress applications respectively IVS also performed contract research services using our tools and devices for industry and academic laboratories for profiling compound induced effects on cell and tissue physiology To further demonstrate our ability to screen drug candidates especially for drug developers the market demands benchmark studies against compounds and drugs whose pharmacological functions including toxicity information have been well characterized To fully commercialize our current start up activities IVS will improve its rapid drug screening system that uses engineered heart tissues EHTs to monitor the effects of test compounds on cardiac contractility and associated regulatory molecules In Aim EHTs will be developed using cardiomyocytes derived from human induced pluripotent stem iPS cells to commercialize a drug screening system using human samples Using this system we will determine the beneficial and toxic effects of a panel of drugs based on the drug induced changes in the cardiac functions of EHTs as well as the signal transduction pathways that underlie their activities In Aim we will establish ISO specified requirements for a quality management system so that we may more confidently provide contract research services for drug developers In addition using a list of well known cardio effective and toxic drugs compounds we will measure drug induced cardiac function changes using EHTs to establish the benchmark In Aim we will identify mechanisms of cardiotoxicity and will demonstrate the ability of the EHTs to predict cardiotoxicity in vitro without the need for establishing animal studies Our approach will advance drug target identification and optimization as well as biomarker discovery critical for diagnosing cardiotoxicity As a demonstration of the ability of our approach to elucidate a mechanism of cardiotoxicity we will use as an example genetic knockdown with shRNA and drugs to inhibit mTOR mammalian target of rapamycin Successful completion of our aims will prove the ability of our in vitro system to predict drug induced cardiotoxicity in humans clearly benefiting early stage drug discovery Existing cardiotoxicity testing in vitro is not sufficient to accurately predict drug induced cardiotoxicity The proposed project will establish a comprehensive cardiotoxicity assessment system using engineered heart tissues fabricated with cardiomyocytes derived from human induced pluripotent stem cells With the new technology drug developers can predict potential drug induced cardiotoxicity at the early stages of drug discovery so will reduce late stage attrition and protect patients from developing cardiac failure
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 225.00K | Year: 2015
Invivosciences, Inc. | Date: 2012-08-21
Clinical and laboratory analyzers for measuring, testing and analyzing biological tissues; cell culture apparatus for clinical and laboratory use, namely, cell culture chambers and cell culture supports. Clinical and laboratory services related to preparation of biological tissues for others, namely, three-dimensional tissues from cells; Clinical and laboratory services for others, namely, using three-dimensional biological tissues for use in developing screening assays, and diagnostic assays, and in product development.
Invivosciences, Inc. | Date: 2013-10-23
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 2.01M | Year: 2010
DESCRIPTION (provided by applicant): Heart disease frequently leads to cardiac fibrosis. In almost all cases of chronic heart disease, the myocardium exhibits fibrosis developed by activated cardiac fibroblasts. These fibroblasts are quiescent in the healthy heart. Interstitial fibrosis due to extracellular matrix deposition by fibroblasts increases the stiffness of the tissue and impairs cardiac relaxation. Discovering pharmaceutical treatments that can reverse fibrosis is a critical unmet need; no such drugs currently exist. This project will develop a novel high-throughput screening platform for drug discovery that measures the physiological properties of live, engineered tissue samples, and their controls, cultured in 96-well plates(the PalpatorTM system). This platform will measure drug-induced changes in the physiological properties of engineered tissues. The Phase I project focuses on completing the development of the PalpatorTM screening system and obtaining feedback from academic and industrial collaborators. In addition, the algorithm used for the data analysis software will be modified to reduce failure rates. The modified software will be beta tested for its ability to obtain meaningful values for the physiological parameters used to indicate that treatment with the panel of chemical compounds has altered the properties of the engineered tissues. The final packaging of the project-related software will be outsourced for its launch. The Phase II project focuses on scaling up the engineered tissue-based screening system to make it amenable to high-throughput applications in industry. The tissue culture consumables for growing engineered tissues in 96-well plates will be produced in collaboration with Engineering Industries, Inc. (Verona, WI). The scaled-up engineered tissue production will significantly improve screening efficiency. This highly efficient Palpator screening system will be used to profile the effects of 50 commonly prescribed cardiovascular drugs on engineered heart tissues. Although the engineered heart tissues are constructed to mimic the physiological properties of native heart muscles, profiling known cardiovascular drugs will validate the utility of employing engineered heart tissues in drug discovery and toxic compound testing. To further validate the engineered tissue model, a library of compounds with known cardiovascular effects will be screened using the Palpator system. A novel phenotypic screening protocol that employs both engineered tissues cultured with highly contractile fibroblasts to mimic the fibrotic heart and 'normal' engineered heart tissues will be used to identify chemical compounds that reduce the contractility of the fibrotic engineered tissues but yet maintain the healthy contractile activities of normal engineered heart tissues. The combination of the engineered tissue models and the Palpator screening device will accelerate drug discovery and reduce the need (and associated costs) of extensive animal studies. PUBLIC HEALTH RELEVANCE: In the United States today, about 5 million people suffer from heart disease, one of the most prevalent chronic conditions and the number one complication of heart attacks. This proposal describes an entirely new method for testing potential drugs to learn if they can help repair the heart or if they are toxic to the body, by measuring whether a drug changes how a heart tissue model behaves. This research may provide a breakthrough in accelerating drug discovery and reducing costs, because it is rapid, high-throughput, and may reduce the need for animal testing.