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Article ; Online: Measuring the Contractile Kinetics of Isolated Myofibrils from Human-Induced Pluripotent Stem Cell Derived Cardiomyocyte (hiPSC-CM) Models of Cardiomyopathy.

Mohran, Saffie / Steczina, Sonette / Mandrycky, Christian / Kao, Kerry / Regnier, Michael

Methods in molecular biology (Clifton, N.J.)

2023 Volume 2735, Page(s) 213–233

Abstract: Isolated myofibrils provide biomechanical data at the contractile organelle level that are independent of cellular calcium handling and signaling pathways. These myofibrils can be harvested from animal tissue, human muscle biopsies, or stem cell-derived ... ...

Abstract	Isolated myofibrils provide biomechanical data at the contractile organelle level that are independent of cellular calcium handling and signaling pathways. These myofibrils can be harvested from animal tissue, human muscle biopsies, or stem cell-derived striated muscle. Here we present our myofibril isolation and rapid solution switching protocols, which allow for precise measurements of activation (kinetics and tension generation) and a biphasic relaxation relationship (initial slow isometric relaxation followed by a fast exponential decay in tension). This experiment is generated on a custom-built myofibril apparatus utilizing a two-photodiode array to detect micron level deflection of our forged glass tip force transducers. A complete activation/relaxation curve can be produced from a single myofibril in under 30 minutes.
MeSH term(s)	Animals ; Humans ; Myofibrils/metabolism ; Myocytes, Cardiac/metabolism ; Induced Pluripotent Stem Cells/metabolism ; Myocardial Contraction/physiology ; Cardiomyopathies/metabolism ; Sarcomeres/metabolism ; Kinetics ; Calcium/metabolism
Chemical Substances	Calcium (SY7Q814VUP)
Language	English
Publishing date	2023-12-01
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3527-8_12
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Article ; Online: 3D curvature-instructed endothelial flow response and tissue vascularization.

Mandrycky, Christian / Hadland, Brandon / Zheng, Ying

Science advances

2020 Volume 6, Issue 38

Abstract: Vascularization remains a long-standing challenge in engineering complex tissues. Particularly needed is recapitulating 3D vascular features, including continuous geometries with defined diameter, curvature, and torsion. Here, we developed a spiral ... ...

Abstract	Vascularization remains a long-standing challenge in engineering complex tissues. Particularly needed is recapitulating 3D vascular features, including continuous geometries with defined diameter, curvature, and torsion. Here, we developed a spiral microvessel model that allows precise control of curvature and torsion and supports homogeneous tissue perfusion at the centimeter scale. Using this system, we showed proof-of-principle modeling of tumor progression and engineered cardiac tissue vascularization. We demonstrated that 3D curvature induced rotation and mixing under laminar flow, leading to unique phenotypic and transcriptional changes in endothelial cells (ECs). Bulk and single-cell RNA-seq identified specific EC gene clusters in spiral microvessels. These mark a proinflammatory phenotype associated with vascular development and remodeling, and a unique cell cluster expressing genes regulating vascular stability and development. Our results shed light on the role of heterogeneous vascular structures in differential development and pathogenesis and provide previously unavailable tools to potentially improve tissue vascularization and regeneration.
MeSH term(s)	Endothelial Cells ; Heart/physiology ; Humans ; Microvessels ; Neovascularization, Pathologic/genetics ; Tissue Engineering/methods ; Tissue Scaffolds
Language	English
Publishing date	2020-09-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2810933-8
ISSN	2375-2548 ; 2375-2548
ISSN (online)	2375-2548
ISSN	2375-2548
DOI	10.1126/sciadv.abb3629
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Article ; Online: Organ-on-a-chip systems for vascular biology.

Mandrycky, Christian J / Howard, Caitlin C / Rayner, Samuel G / Shin, Yu Jung / Zheng, Ying

Journal of molecular and cellular cardiology

2021 Volume 159, Page(s) 1–13

Abstract: Organ-on-a-chip (OOC) platforms involve the miniaturization of cell culture systems and enable a variety of novel experimental approaches. These range from modeling the independent effects of biophysical forces on cells to screening novel drugs in multi- ... ...

Abstract	Organ-on-a-chip (OOC) platforms involve the miniaturization of cell culture systems and enable a variety of novel experimental approaches. These range from modeling the independent effects of biophysical forces on cells to screening novel drugs in multi-organ microphysiological systems, all within microscale devices. As in living systems, the incorporation of vascular structure is a key feature common to almost all organ-on-a-chip systems. In this review we highlight recent advances in organ-on-a-chip technologies with a focus on the vasculature. We first present the developmental process of the blood vessels through which vascular cells assemble into networks and remodel to form complex vascular beds under flow. We then review self-assembled vascular models and flow systems for the study of vascular development and biology as well as pre-patterned vascular models for the generation of perfusable microvessels for modeling vascular and tissue function. We finally conclude with a perspective on developing future OOC approaches for studying different aspects of vascular biology. We highlight the fit for purpose selection of OOC models towards either simple but powerful testbeds for therapeutic development, or complex vasculature to accurately replicate human physiology for specific disease modeling and tissue regeneration.
MeSH term(s)	Animals ; Biology/methods ; Blood Vessels/physiology ; Guided Tissue Regeneration/methods ; Humans ; Lab-On-A-Chip Devices
Language	English
Publishing date	2021-06-09
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	80157-4
ISSN	1095-8584 ; 0022-2828
ISSN (online)	1095-8584
ISSN	0022-2828
DOI	10.1016/j.yjmcc.2021.06.002
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Article ; Online: The biochemically defined super relaxed state of myosin-A paradox.

Mohran, Saffie / Kooiker, Kristina / Mahoney-Schaefer, Max / Mandrycky, Christian / Kao, Kerry / Tu, An-Yue / Freeman, Jeremy / Moussavi-Harami, Farid / Geeves, Michael / Regnier, Michael

The Journal of biological chemistry

2023 Volume 300, Issue 1, Page(s) 105565

Abstract: The biochemical SRX (super-relaxed) state of myosin has been defined as a low ATPase activity state. This state can conserve energy when the myosin is not recruited for muscle contraction. The SRX state has been correlated with a structurally defined ... ...

Abstract	The biochemical SRX (super-relaxed) state of myosin has been defined as a low ATPase activity state. This state can conserve energy when the myosin is not recruited for muscle contraction. The SRX state has been correlated with a structurally defined ordered (versus disordered) state of muscle thick filaments. The two states may be linked via a common interacting head motif (IHM) where the two heads of heavy meromyosin (HMM), or myosin, fold back onto each other and form additional contacts with S2 and the thick filament. Experimental observations of the SRX, IHM, and the ordered form of thick filaments, however, do not always agree, and result in a series of unresolved paradoxes. To address these paradoxes, we have reexamined the biochemical measurements of the SRX state for porcine cardiac HMM. In our hands, the commonly employed mantATP displacement assay was unable to quantify the population of the SRX state with all data fitting very well by a single exponential. We further show that mavacamten inhibits the basal ATPases of both porcine ventricle HMM and S1 (K
MeSH term(s)	Animals ; Adenosine Triphosphatases/antagonists & inhibitors ; Adenosine Triphosphatases/metabolism ; Adenosine Triphosphate/analogs & derivatives ; Adenosine Triphosphate/metabolism ; Amino Acid Motifs ; Benzylamines/pharmacology ; Enzyme Assays/methods ; Enzyme Assays/standards ; Heart Ventricles/drug effects ; Heart Ventricles/enzymology ; Heart Ventricles/metabolism ; Myocardial Contraction ; Myosin Subfragments/chemistry ; Myosin Subfragments/metabolism ; Nonmuscle Myosin Type IIA/chemistry ; Nonmuscle Myosin Type IIA/metabolism ; ortho-Aminobenzoates/metabolism ; Swine ; Uracil/analogs & derivatives ; Uracil/pharmacology
Chemical Substances	3'-O-(N-methylanthraniloyl) ATP (85287-56-5) ; Adenosine Triphosphatases (EC 3.6.1.-) ; Adenosine Triphosphate (8L70Q75FXE) ; Benzylamines ; MYK-461 ; Myosin Subfragments ; Nonmuscle Myosin Type IIA (EC 3.6.1.-) ; ortho-Aminobenzoates ; Uracil (56HH86ZVCT)
Language	English
Publishing date	2023-12-14
Publishing country	United States
Document type	Journal Article
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1016/j.jbc.2023.105565
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Article: Tissue engineering toward organ-specific regeneration and disease modeling.

Mandrycky, Christian / Phong, Kiet / Zheng, Ying

MRS communications

2017 Volume 7, Issue 3, Page(s) 332–347

Abstract: Tissue engineering has been recognized as a translational approach to replace damaged tissue or whole organs. Engineering tissue, however, faces an outstanding knowledge gap in the challenge to fully recapitulate complex organ-specific features. Major ... ...

Abstract	Tissue engineering has been recognized as a translational approach to replace damaged tissue or whole organs. Engineering tissue, however, faces an outstanding knowledge gap in the challenge to fully recapitulate complex organ-specific features. Major components, such as cells, matrix, and architecture, must each be carefully controlled to engineer tissue-specific structure and function that mimics what is found in vivo. Here we review different methods to engineer tissue, and discuss critical challenges in recapitulating the unique features and functional units in four major organs-the kidney, liver, heart, and lung, which are also the top four candidates for organ transplantation in the USA. We highlight advances in tissue engineering approaches to enable the regeneration of complex tissue and organ substitutes, and provide tissue-specific models for drug testing and disease modeling. We discuss the current challenges and future perspectives toward engineering human tissue models.
Language	English
Publishing date	2017-07-31
Publishing country	United States
Document type	Journal Article
ZDB-ID	2645443-9
ISSN	2159-6867 ; 2159-6859
ISSN (online)	2159-6867
ISSN	2159-6859
DOI	10.1557/mrc.2017.58
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Article ; Online: Multiphoton-Guided Creation of Complex Organ-Specific Microvasculature.

Rayner, Samuel G / Howard, Caitlin C / Mandrycky, Christian J / Stamenkovic, Stefan / Himmelfarb, Jonathan / Shih, Andy Y / Zheng, Ying

Advanced healthcare materials

2021 Volume 10, Issue 10, Page(s) e2100031

Abstract: Engineering functional human tissues in vitro is currently limited by difficulty replicating the small caliber, complex connectivity, cellularity, and 3D curvature of the native microvasculature. Multiphoton ablation has emerged as a promising technique ... ...

Abstract	Engineering functional human tissues in vitro is currently limited by difficulty replicating the small caliber, complex connectivity, cellularity, and 3D curvature of the native microvasculature. Multiphoton ablation has emerged as a promising technique for fabrication of microvascular structures with high resolution and full 3D control, but cellularization and perfusion of complex capillary-scale structures has remained challenging. Here, multiphoton ablation combined with guided endothelial cell growth from pre-formed microvessels is used to successfully create perfusable and cellularized organ-specific microvascular structures at anatomic scale within collagen hydrogels. Fabrication and perfusion of model 3D pulmonary and renal microvascular beds is demonstrated, as is replication and perfusion of a brain microvascular unit derived from in vivo data. Successful endothelialization and blood perfusion of a kidney-specific microvascular structure is achieved, using laser-guided angiogenesis. Finally, proof-of-concept hierarchical blood vessels and complex multicellular models are created, using multistep patterning with multiphoton ablation techniques. These successes open new doors for the creation of engineered tissues and organ-on-a-chip devices.
MeSH term(s)	Ablation Techniques ; Endothelial Cells ; Humans ; Microvessels ; Perfusion ; Tissue Engineering ; Veins
Language	English
Publishing date	2021-02-15
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2649576-4
ISSN	2192-2659 ; 2192-2640
ISSN (online)	2192-2659
ISSN	2192-2640
DOI	10.1002/adhm.202100031
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Article: Full-length dystrophin deficiency leads to contractile and calcium transient defects in human engineered heart tissues.

Bremner, Samantha B / Mandrycky, Christian J / Leonard, Andrea / Padgett, Ruby M / Levinson, Alan R / Rehn, Ethan S / Pioner, J Manuel / Sniadecki, Nathan J / Mack, David L

Journal of tissue engineering

2022 Volume 13, Page(s) 20417314221119628

Abstract: Cardiomyopathy is currently the leading cause of death for patients with Duchenne muscular dystrophy (DMD), a severe neuromuscular disorder affecting young boys. Animal models have provided insight into the mechanisms by which dystrophin protein ... ...

Abstract	Cardiomyopathy is currently the leading cause of death for patients with Duchenne muscular dystrophy (DMD), a severe neuromuscular disorder affecting young boys. Animal models have provided insight into the mechanisms by which dystrophin protein deficiency causes cardiomyopathy, but there remains a need to develop human models of DMD to validate pathogenic mechanisms and identify therapeutic targets. Here, we have developed human engineered heart tissues (EHTs) from CRISPR-edited, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) expressing a truncated dystrophin protein lacking part of the actin-binding domain. The 3D EHT platform enables direct measurement of contractile force, simultaneous monitoring of Ca
Language	English
Publishing date	2022-08-17
Publishing country	England
Document type	Journal Article
ZDB-ID	2573915-3
ISSN	2041-7314
ISSN	2041-7314
DOI	10.1177/20417314221119628
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Article ; Online: Endothelial Responses to Curvature-Induced Flow Patterns in Engineered Cerebral Aneurysms.

Mandrycky, Christian J / Abel, Ashley N / Levy, Samuel / Marsh, Laurel M / Chassagne, Fanette / Chivukula, Venkat K / Barczay, Sari E / Kelly, Cory M / Kim, Louis J / Aliseda, Alberto / Levitt, Michael R / Zheng, Ying

Journal of biomechanical engineering

2022 Volume 145, Issue 1

Abstract: Hemodynamic factors have long been associated with clinical outcomes in the treatment of cerebral aneurysms. Computational studies of cerebral aneurysm hemodynamics have provided valuable estimates of the mechanical environment experienced by the ... ...

Abstract	Hemodynamic factors have long been associated with clinical outcomes in the treatment of cerebral aneurysms. Computational studies of cerebral aneurysm hemodynamics have provided valuable estimates of the mechanical environment experienced by the endothelium in both the parent vessel and aneurysmal dome walls and have correlated them with disease state. These computational-clinical studies have recently been correlated with the response of endothelial cells (EC) using either idealized or patient-specific models. Here, we present a robust workflow for generating anatomic-scale aneurysm models, establishing luminal cultures of ECs at physiological relevant flow profiles, and comparing EC responses to curvature mediated flow. We show that flow patterns induced by parent vessel curvature produce changes in wall shear stress (WSS) and wall shear stress gradients (WSSG) that are correlated with differences in cell morphology and cellular protein localization. Cells in higher WSS regions align better with the flow and display strong Notch1-extracellular domain (ECD) polarization, while, under low WSS, differences in WSSG due to curvature change were associated with less alignment and attenuation of Notch1-ECD polarization in ECs of the corresponding regions. These proof-of-concept results highlight the use of engineered cellularized aneurysm models for connecting computational fluid dynamics to the underlying endothelial biology that mediates disease.
MeSH term(s)	Endothelial Cells ; Endothelium/metabolism ; Hemodynamics/physiology ; Humans ; Hydrodynamics ; Intracranial Aneurysm ; Models, Cardiovascular ; Stress, Mechanical
Language	English
Publishing date	2022-07-11
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	243094-0
ISSN	1528-8951 ; 0148-0731
ISSN (online)	1528-8951
ISSN	0148-0731
DOI	10.1115/1.4054981
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Article ; Online: Endothelial-derived von Willebrand factor accelerates fibrin clotting within engineered microvessels.

Rayner, Samuel G / Scholl, Zackary / Mandrycky, Christian J / Chen, Junmei / LaValley, Karina N / Leary, Peter J / Altemeier, William A / Liles, W Conrad / Chung, Dominic W / López, José A / Fu, Hongxia / Zheng, Ying

Journal of thrombosis and haemostasis : JTH

2022 Volume 20, Issue 7, Page(s) 1627–1637

Abstract: Background: Von Willebrand factor (VWF) is classically associated with primary hemostasis and platelet-rich arterial thromboses, but recently has also been implicated in fibrin clotting and venous thrombosis. Direct interaction between fibrin and VWF ... ...

Abstract	Background: Von Willebrand factor (VWF) is classically associated with primary hemostasis and platelet-rich arterial thromboses, but recently has also been implicated in fibrin clotting and venous thrombosis. Direct interaction between fibrin and VWF may mediate these processes, although prior reports are conflicting. Objectives: We combined two complementary platforms to characterize VWF-fibrin(ogen) interactions and identify their potential physiologic significance. Methods: Engineered microvessels were lined with human endothelial cells, cultured under flow, and activated to release VWF and form transluminal VWF fibers. Fibrinogen, fibrin monomers, or polymerizing fibrin were then perfused, and interactions with VWF evaluated. Thrombin and fibrinogen were perfused into living versus paraformeldahyde-fixed microvessels and the pressure drop across microvessels monitored. Separately, protein binding to tethered VWF was assessed on a single-molecule level using total internal reflection fluorescence (TIRF) microscopy. Results: Within microvessels, VWF fibers colocalized with polymerizing fibrin, but not fibrinogen. TIRF microscopy showed no colocalization between VWF and fibrinogen or fibrin monomers in a microfluidic flow chamber across a range of shear rates and protein concentrations. Thrombin-mediated fibrin polymerization within living microvessels triggered endothelial VWF release, increasing the rate and amount of microvessel obstruction compared to fixed vessels with an inert endothelium. Conclusions: We did not identify specific binding between fibrin(ogen) and VWF at a single-molecule level. Despite this, our results suggest that rapid release of endothelial VWF during clotting may provide a physical support for fibrin polymerization and accelerate thrombosis. This interaction may be of fundamental importance for the understanding and treatment of human thrombotic disease.
MeSH term(s)	Endothelial Cells/metabolism ; Endothelium/metabolism ; Fibrin/chemistry ; Fibrinogen ; Humans ; Microvessels/metabolism ; Thrombin/chemistry ; Thrombosis ; von Willebrand Factor/metabolism
Chemical Substances	von Willebrand Factor ; Fibrin (9001-31-4) ; Fibrinogen (9001-32-5) ; Thrombin (EC 3.4.21.5)
Language	English
Publishing date	2022-04-11
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2112661-6
ISSN	1538-7836 ; 1538-7933
ISSN (online)	1538-7836
ISSN	1538-7933
DOI	10.1111/jth.15714
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Zs.A 5805: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article: Danicamtiv increases myosin recruitment and alters the chemomechanical cross bridge cycle in cardiac muscle.

bioRxiv : the preprint server for biology

2023

Abstract: Modulating myosin function is a novel therapeutic approach in patients with cardiomyopathy. Detailed mechanism of action of these agents can help predict potential unwanted affects and identify patient populations that can benefit most from them. ... ...

Abstract	Modulating myosin function is a novel therapeutic approach in patients with cardiomyopathy. Detailed mechanism of action of these agents can help predict potential unwanted affects and identify patient populations that can benefit most from them. Danicamtiv is a novel myosin activator with promising preclinical data that is currently in clinical trials. While it is known danicamtiv increases force and cardiomyocyte contractility without affecting calcium levels, detailed mechanistic studies regarding its mode of action are lacking. Using porcine cardiac tissue and myofibrils we demonstrate that Danicamtiv increases force and calcium sensitivity via increasing the number of myosin in the "on" state and slowing cross bridge turnover. Our detailed analysis shows that inhibition of ADP release results in decreased cross bridge turnover with cross bridges staying on longer and prolonging myofibril relaxation. Using a mouse model of genetic dilated cardiomyopathy, we demonstrated that Danicamtiv corrected calcium sensitivity in demembranated and abnormal twitch magnitude and kinetics in intact cardiac tissue. Significance statement: Directly augmenting sarcomere function has potential to overcome limitations of currently used inotropic agents to improve cardiac contractility. Myosin modulation is a novel mechanism for increased contraction in cardiomyopathies. Danicamtiv is a myosin activator that is currently under investigation for use in cardiomyopathy patients. Our study is the first detailed mechanism of how Danicamtiv increases force and alters kinetics of cardiac activation and relaxation. This new understanding of the mechanism of action of Danicamtiv can be used to help identify patients that could benefit most from this treatment.
Language	English
Publishing date	2023-02-03
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.01.31.526380
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