Donald E Ingber

Boston Children's Hospital, Boston, Massachusetts, United States

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Publications (363)2517.61 Total impact

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    ABSTRACT: Tumor vessels are characterized by abnormal morphology and hyperpermeability that together cause inefficient delivery of chemotherapeutic agents. Although vascular endothelial growth factor has been established as a critical regulator of tumor angiogenesis, the role of mechanical signaling in the regulation of tumor vasculature or tumor endothelial cell (TEC) function is not known. Here we show that the mechanosensitive ion channel transient receptor potential vanilloid 4 (TRPV4) regulates tumor angiogenesis and tumor vessel maturation via modulation of TEC mechanosensitivity. We found that TECs exhibit reduced TRPV4 expression and function, which is correlated with aberrant mechanosensitivity towards extracellular matrix stiffness, increased migration and abnormal angiogenesis by TEC. Further, syngeneic tumor experiments revealed that the absence of TRPV4 induced increased vascular density, vessel diameter and reduced pericyte coverage resulting in enhanced tumor growth in TRPV4 knockout mice. Importantly, overexpression or pharmacological activation of TRPV4 restored aberrant TEC mechanosensitivity, migration and normalized abnormal angiogenesis in vitro by modulating Rho activity. Finally, a small molecule activator of TRPV4, GSK1016790A, in combination with anticancer drug cisplatin, significantly reduced tumor growth in wild-type mice by inducing vessel maturation. Our findings demonstrate TRPV4 channels to be critical regulators of tumor angiogenesis and represent a novel target for anti-angiogenic and vascular normalization therapies.Oncogene advance online publication, 13 April 2015; doi:10.1038/onc.2015.83.
    Oncogene 04/2015; DOI:10.1038/onc.2015.83 · 8.56 Impact Factor
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    ABSTRACT: Blood flow promotes emergence of definitive hematopoietic stem cells (HSCs) in the developing embryo, yet the signals generated by hemodynamic forces that influence hematopoietic potential remain poorly defined. Here we show that fluid shear stress endows long-term multilineage engraftment potential upon early hematopoietic tissues at embryonic day 9.5, an embryonic stage not previously described to harbor HSCs. Effects on hematopoiesis are mediated in part by a cascade downstream of wall shear stress that involves calcium efflux and stimulation of the prostaglandin E2 (PGE2)-cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) signaling axis. Blockade of the PGE2-cAMP-PKA pathway in the aorta-gonad-mesonephros (AGM) abolished enhancement in hematopoietic activity. Furthermore, Ncx1 heartbeat mutants, as well as static cultures of AGM, exhibit lower levels of expression of prostaglandin synthases and reduced phosphorylation of the cAMP response element-binding protein (CREB). Similar to flow-exposed cultures, transient treatment of AGM with the synthetic analogue 16,16-dimethyl-PGE2 stimulates more robust engraftment of adult recipients and greater lymphoid reconstitution. These data provide one mechanism by which biomechanical forces induced by blood flow modulate hematopoietic potential. © 2015 Diaz et al.
    Journal of Experimental Medicine 04/2015; 145(5). DOI:10.1084/jem.20142235 · 13.91 Impact Factor
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    ABSTRACT: Background: Mechanical compression of cells during mesenchymal condensation triggers cells to undergo odontogenic differentiation during tooth organ formation in the embryo. However, the mechanism by which cell compaction is stabilized over time to ensure correct organ specific cell fate switching remains unknown. Results: Here, we show that mesenchymal cell compaction induces accumulation of collagen VI in the extracellular matrix (ECM), which physically stabilizes compressed mesenchymal cell shapes and ensures efficient organ-specific cell fate switching during tooth organ development. Mechanical induction of collagen VI deposition is mediated by signaling through the actin-p38MAPK-SP1 pathway, and the ECM scaffold is stabilized by lysyl oxidase (LOX) in the condensing mesenchyme. Moreover, perturbation of synthesis or cross-linking of collagen VI alters the size of the condensation in vivo. Conclusions: These findings suggest that the odontogenic differentiation process that is induced by cell compaction during mesenchymal condensation is stabilized and sustained through mechanically-regulated production of collagen VI within the mesenchymal ECM. This article is protected by copyright. All rights reserved. ©2015. Wiley Periodicals, Inc.
    Developmental Dynamics 02/2015; DOI:10.1002/dvdy.24264 · 2.67 Impact Factor
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    ABSTRACT: The stability and longevity of surface-stabilized lubricant layers is a critical question in their application as low- and non-fouling slippery surface treatments in both industry and medicine. Here, we investigate lubricant loss from surfaces under flow in water using both quantitative analysis and visualization, testing the effects of underlying surface type (nanostructured versus flat), as well as flow rate in the physiologically-relevant range, lubricant type, and time. We find lubricant losses on the order of only ng/cm2 in a closed system, indicating that these interfaces are relatively stable under the flow conditions tested. No notable differences emerged between surface type, flow rate, lubricant type, or time. However, exposure of the lubricant layers to an air/water interface did significantly increase the amount of lubricant removed from the surface, leading to disruption of the layer. These results may help in the development and design of materials using surface-immobilized lubricant interfaces for repellency under flow conditions.
    Chemistry of Materials 02/2015; 27(5):150204154038006. DOI:10.1021/cm504652g · 8.54 Impact Factor
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    ABSTRACT: The ultimate goal of most biomedical research is to gain greater insight into mechanisms of human disease or to develop new and improved therapies or diagnostics. Although great advances have been made in terms of developing disease models in animals, such as transgenic mice, many of these models fail to faithfully recapitulate the human condition. In addition, it is difficult to identify critical cellular and molecular contributors to disease or to vary them independently in whole-animal models. This challenge has attracted the interest of engineers, who have begun to collaborate with biologists to leverage recent advances in tissue engineering and microfabrication to develop novel in vitro models of disease. As these models are synthetic systems, specific molecular factors and individual cell types, including parenchymal cells, vascular cells, and immune cells, can be varied independently while simultaneously measuring system-level responses in real time. In this article, we provide some examples of these efforts, including engineered models of diseases of the heart, lung, intestine, liver, kidney, cartilage, skin and vascular, endocrine, musculoskeletal, and nervous systems, as well as models of infectious diseases and cancer. We also describe how engineered in vitro models can be combined with human inducible pluripotent stem cells to enable new insights into a broad variety of disease mechanisms, as well as provide a test bed for screening new therapies.
    Annual Review of Pathology Mechanisms of Disease 01/2015; 10(1):195-262. DOI:10.1146/annurev-pathol-012414-040418 · 22.13 Impact Factor
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    ABSTRACT: Trans-epithelial electrical resistance (TEER) measurements are widely used as real-time, non-destructive, and label-free measurements of epithelial and endothelial barrier function. TEER measurements are ideal for characterizing tissue barrier function in organs-on-chip studies for drug testing and investigation of human disease models; however, published reports using this technique have reported highly conflicting results even with identical cell lines and experimental setups. The differences are even more dramatic when comparing measurements in conventional Transwell systems with those obtained in microfluidic systems. Our goal in this work was therefore to enhance the fidelity of TEER measurements in microfluidic organs-on-chips, specifically using direct current (DC) measurements of TEER, as this is the most widely used method reported in the literature. Here we present a mathematical model that accounts for differences measured in TEER between microfluidic chips and Transwell systems, which arise from differences in device geometry. The model is validated by comparing TEER measurements obtained in a microfluidic gut-on-a-chip device versus in a Transwell culture system. Moreover, we show that even small gaps in cell coverage (e.g., 0.4%) are sufficient to cause a significant (~80%) drop in TEER. Importantly, these findings demonstrate that TEER measurements obtained in microfluidic systems, such as organs-on-chips, require special consideration, specifically when results are to be compared with measurements obtained from Transwell systems.
    Lab on a Chip 11/2014; 15(3). DOI:10.1039/c4lc01219d · 5.75 Impact Factor
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    ABSTRACT: Obstruction of normal blood flow, which occurs in a variety of diseases, including thromboembolism in stroke and atherosclerosis, is a leading cause of death and long-term adult disability in the Western world. This review focuses on a novel nanotherapeutic drug-delivery platform that is mechanically activated within blood vessels by high-fluid shear stresses to selectively target drugs to sites of vascular obstruction. In vitro and in vivo studies have shown that this approach can be used to efficiently lyse clots using a significantly lower amount of thrombolytic drug than is required when administered in a soluble formulation. This nanotherapeutic strategy can potentially improve both the efficacy and safety of thrombolytic drugs, particularly in patients who are at high risk for brain hemorrhage, and thus provide a new approach for the treatment of many life-threatening and debilitating vascular disorders.
    JAMA Neurology 11/2014; 72(1). DOI:10.1001/jamaneurol.2014.2886 · 7.01 Impact Factor
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    ABSTRACT: Thrombosis and biofouling of extracorporeal circuits and indwelling medical devices cause significant morbidity and mortality worldwide. We apply a bioinspired, omniphobic coating to tubing and catheters and show that it completely repels blood and suppresses biofilm formation. The coating is a covalently tethered, flexible molecular layer of perfluorocarbon, which holds a thin liquid film of medical-grade perfluorocarbon on the surface. This coating prevents fibrin attachment, reduces platelet adhesion and activation, suppresses biofilm formation and is stable under blood flow in vitro. Surface-coated medical-grade tubing and catheters, assembled into arteriovenous shunts and implanted in pigs, remain patent for at least 8 h without anticoagulation. This surface-coating technology could reduce the use of anticoagulants in patients and help to prevent thrombotic occlusion and biofouling of medical devices.
    Nature Biotechnology 10/2014; DOI:10.1038/nbt.3020 · 39.08 Impact Factor
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    ABSTRACT: Here we describe a blood-cleansing device for sepsis therapy inspired by the spleen, which can continuously remove pathogens and toxins from blood without first identifying the infectious agent. Blood flowing from an infected individual is mixed with magnetic nanobeads coated with an engineered human opsonin-mannose-binding lectin (MBL)-that captures a broad range of pathogens and toxins without activating complement factors or coagulation. Magnets pull the opsonin-bound pathogens and toxins from the blood; the cleansed blood is then returned back to the individual. The biospleen efficiently removes multiple Gram-negative and Gram-positive bacteria, fungi and endotoxins from whole human blood flowing through a single biospleen unit at up to 1.25 liters per h in vitro. In rats infected with Staphylococcus aureus or Escherichia coli, the biospleen cleared >90% of bacteria from blood, reduced pathogen and immune cell infiltration in multiple organs and decreased inflammatory cytokine levels. In a model of endotoxemic shock, the biospleen increased survival rates after a 5-h treatment.
    Nature Medicine 09/2014; 20(10). DOI:10.1038/nm.3640 · 28.05 Impact Factor
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    ABSTRACT: Certain lower organisms achieve organ regeneration by reverting differentiated cells into tissue-specific progenitors that re-enter embryonic programs. During muscle regeneration in the urodele amphibian, post-mitotic multinucleated skeletal myofibers transform into mononucleated proliferating cells upon injury, and a transcription factor-msx1 plays a role in their reprograming. Whether this powerful regeneration strategy can be leveraged in mammals remains unknown, as it has not been demonstrated that the dedifferentiated progenitor cells arising from muscle cells overexpressing Msx1 are lineage-specific and possess the same potent regenerative capability as their amphibian counterparts. Here we show that ectopic expression of Msx1 reprograms post-mitotic, multinucleated, primary mouse myotubes to become proliferating mononuclear cells. These dedifferentiated cells reactivate genes expressed by embryonic muscle progenitor cells and generate only muscle tissue in vivo both in an ectopic location and inside existing muscle. More importantly, distinct from adult muscle satellite cells, these cells appear both to fuse with existing fibers and to regenerate myofibers in a robust and time-dependent manner. Upon transplantation into a degenerating muscle, these dedifferentiated cells generated a large number of myofibers that increased over time and replenished almost half of the cross-sectional area of the muscle in only 12 weeks. Our study demonstrates that mammals can harness a muscle regeneration strategy used by lower organisms when the same molecular pathway is activated. Stem Cells 2014.
    Stem Cells 09/2014; 32(9). DOI:10.1002/stem.1742 · 7.70 Impact Factor
  • Sangeeta N Bhatia, Donald E Ingber
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    ABSTRACT: An organ-on-a-chip is a microfluidic cell culture device created with microchip manufacturing methods that contains continuously perfused chambers inhabited by living cells arranged to simulate tissue- and organ-level physiology. By recapitulating the multicellular architectures, tissue-tissue interfaces, physicochemical microenvironments and vascular perfusion of the body, these devices produce levels of tissue and organ functionality not possible with conventional 2D or 3D culture systems. They also enable high-resolution, real-time imaging and in vitro analysis of biochemical, genetic and metabolic activities of living cells in a functional tissue and organ context. This technology has great potential to advance the study of tissue development, organ physiology and disease etiology. In the context of drug discovery and development, it should be especially valuable for the study of molecular mechanisms of action, prioritization of lead candidates, toxicity testing and biomarker identification.
    Nature Biotechnology 08/2014; DOI:10.1038/nbt.2989 · 39.08 Impact Factor
  • Javier G. Fernandez, Donald E. Ingber
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    ABSTRACT: Cover: A chess piece made of a chitosan‐based composite (queen) behind a commercial chess piece (king). The study on the correlation between molecular arrangement and mechanical characteristics of chitinous materials leads to use of the second most abundant biopolymer on earth for manufacture of several large‐scale 3D objects and composites. Further details can be found in the article by J. G. Fernandez and D. E. Ingber* on page 932.
    Macromolecular Materials and Engineering 08/2014; 299(8). DOI:10.1002/mame.201470022 · 2.78 Impact Factor
  • Javier G. Fernandez, Donald E. Ingber
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    ABSTRACT: Despite the urgent need for sustainable materials for mass‐produced commercial products, and the incredible diversity of naturally biodegradable materials with desired structural properties, the use of regenerated biomaterials in modern engineering remains extremely limited. Chitin is a prime example: although it is responsible for some of the most remarkable mechanical properties exhibited by natural materials, including nacre, insect cuticle, and crustacean shells, and it is the most abundant organic compound on earth after cellulose, it has not been utilized in manufacturing strategies for commercial applications. Here we describe how analysis of differences in the molecular arrangement and mechanical properties of chitosan polymer that result from different processing methods led to development of a scalable manufacturing strategy for production of large three‐dimensional (3D) objects of chitosan. This chitosan fabrication method offers a new pathway for large‐scale production of fully compostable engineered components with complex forms, and establishes chitosan as a viable bioplastic that could potentially be used in place of existing non‐degradable plastics for commercial manufacturing. Large‐scale functional components made of chitosan are fabricated. The mechanical properties of chitosan depend on the processing method, and their study result in new manufacturing methods to produce large objects of chitosan with mechanical properties similar to common synthetic polymers. These chitosan materials are completely recyclable, compostable, and their breakdown products support plant growth.
    Macromolecular Materials and Engineering 08/2014; 299(8). DOI:10.1002/mame.201300426 · 2.78 Impact Factor
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    ABSTRACT: Microfluidic water-in-oil droplets that serve as separate, chemically isolated compartments can be applied for single-cell analysis; however, to investigate encapsulated cells effectively over prolonged time periods, an array of droplets must remain stationary on a versatile substrate for optimal cell compatibility. We present here a platform of unique geometry and substrate versatility that generates a stationary nanodroplet array by using wells branching off a main microfluidic channel. These droplets are confined by multiple sides of a nanowell and are in direct contact with a biocompatible substrate of choice. The device is operated by a unique and reversed loading procedure that eliminates the need for fine pressure control or external tubing. Fluorocarbon oil isolates the droplets and provides soluble oxygen for the cells. By using this approach, the metabolic activity of single adherent cells was monitored continuously over time, and the concentration of viable pathogens in blood-derived samples was determined directly by measuring the number of colony-formed droplets. The method is simple to operate, requires a few microliters of reagent volume, is portable, is reusable, and allows for cell retrieval. This technology may be particularly useful for multiplexed assays for which prolonged and simultaneous visual inspection of many isolated single adherent or nonadherent cells is required.
    Proceedings of the National Academy of Sciences 07/2014; 111(31). DOI:10.1073/pnas.1404472111 · 9.81 Impact Factor
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Publication Stats

46k Citations
2,517.61 Total Impact Points

Institutions

  • 1987–2015
    • Boston Children's Hospital
      • Department of Pathology
      Boston, Massachusetts, United States
  • 1991–2014
    • Harvard University
      • • Wyss Institute for Biologically Inspired Engineering
      • • School of Engineering and Applied Sciences
      • • Department of Chemistry and Chemical Biology
      Cambridge, Massachusetts, United States
  • 1989–2014
    • Harvard Medical School
      • • Department of Surgery
      • • Department of Biological Chemistry and Molecular Pharmacology
      • • Department of Pathology
      Boston, Massachusetts, United States
  • 2013
    • University of Pennsylvania
      Philadelphia, Pennsylvania, United States
  • 2011
    • Karolinska Institutet
      • Department of Microbiology, Tumor and Cell Biology (MTC)
      Stockholm, Stockholm, Sweden
  • 2008–2011
    • The University of Calgary
      • Department of Biochemistry and Molecular Biology
      Calgary, Alberta, Canada
  • 2009
    • University of Michigan
      Ann Arbor, Michigan, United States
    • University of Illinois, Urbana-Champaign
      • Department of Mechanical Science and Engineering
      Urbana, IL, United States
  • 2007
    • Children's Hospital of Richmond
      Ричмонд, Virginia, United States
  • 1994–2004
    • Massachusetts Institute of Technology
      • • Department of Mechanical Engineering
      • • Department of Chemical Engineering
      • • Department of Materials Science and Engineering
      Cambridge, MA, United States
    • The Scripps Research Institute
      La Jolla, California, United States
  • 2000
    • Carnegie Mellon University
      • Department of Materials Science and Engineering
      Pittsburgh, PA, United States
  • 1999
    • Johns Hopkins University
      • Department of Biomedical Engineering
      Baltimore, MD, United States
  • 1991–1996
    • Brigham and Women's Hospital
      • Department of Pathology
      Boston, MA, United States
  • 1990
    • Wolfson Childrens Hospital
      Jacksonville, Florida, United States
  • 1981
    • Yale-New Haven Hospital
      New Haven, Connecticut, United States