Functional Biomaterials and Scaffolds

In the 1990’s, our lab worked with the inventors of the 3D Printing process at MIT to implement 3D printing for biomaterials, and commercialized this process, leading to clinically-approved scaffolds for bone regeneration that are still used in our regenerative medicine applications. The lab maintains an active program in solid-free-form fabrication, and has just completed building a custom projection microstereolithogrpahy machine in collaboration with the Nick Fang lab in Mechanical Engineering, which we are deploying to make high-resolution scaffolds for in vitro tissue engineered models.

High performance scaffolds require functionalization. A foundational research focus in the lab is design and synthesis of biomaterials that control receptor-mediated processes in highly targeted and biophysically-appropriate ways, e.g. the demonstration that nanoscale clustering of adhesion ligands influences cellular phenotypes compared to random presentation [1-4] and that bivalent growth factors bias signaling and cell phenotypes [5-7]. One major application area is in regenerative connective tissue engineering, where we are working with clinicians to implement a “tethered EGF” strategy for improving survival and function of bone marrow stem cells harvested and transplanted intraoperatively, based on extensive in vitro work demonstrating that soluble and tethered EGF signal very differently [8-10]. Another application is influencing epithelial morphogenesis and function, including liver cells, endometrial cells, and gut organoids [11]. An active area of research is designing new degradable materials for in vitro organogenesis of polarized epithelial structures, and epithelial-stromal constructs.

Integration of Systems Biology and Tissue Engineering for Drug Development

There is tremendous interest in using in vitro models, especially tissue engineered models, for understanding human disease processes and for efficacy and safety of drugs to treat diseases [1, 12-14]. A crucial aspect of designing and interpreting experiments with in vitro models is defining which facets of the human in vivo state are most important to model. We are especially interested in inflammation – how can we model inflammatory diseases, and how does inflammation influence safety and efficacy of drugs? At one end of the spectrum, we engage in clinical studies to understand how the complexity of inflammatory diseases in patients varies in different patient subpopulations, focusing on the cell-cell communication networks in inflammation (CGR research: http://web.mit.edu.ezproxy.canberra.edu.au/cgr/research.html) using a compendium of computational and experimental approaches that often involve highly multiplexed measurements. For example, by measuring the concentration of 50 different cytokines in the peritoneal fluid of endometriosis patients and analyzing the data in a multivariate way, we found a macrophage-driven immune network in a subset of patients and identified an intracellular kinase pathway controlling secretion of inflammatory cytokines [15]. We are now applying these approaches to parse immune networks in infertility. Endometriosis and adenomyosis are also invasive diseases driven in part by growth factors shed proteolytically from the cell surface. We developed a new combined experimental and computational approach to analyze a compendium of protease activities in the context of endometrial cell migration [16-17] and found that that ADAM-10 and -17 dynamically integrate numerous signaling pathways to direct endometrial cell motility and that growth-factor-driven ADAM-10 activity and MET shedding are jointly dysregulated in the peritoneal fluid of endometriosis patients [17]. To complement these systems biology studies on patient samples, we are building in vitro models of endometrium using synthetic biomaterials microenvironments that drive epithelial polarization in the context of a supporting stroma, and can be remodeled to accommodate invasion.

We have also shown how this systems biology approach can be used to analyze synergies between inflammation and drug metabolism in simple 2-D cultures of hepatocytes, as a model of idiosyncratic toxicity that shows up in late stage clinical trials [12] and shown that a very important aspect of bone marrow toxicity can be captured in vitro using erythroid progenitors induced to make erythrocytes [13].

Finally, we have applied this systems biology approach to understand how signaling cues early in osteogenic differentiation of bone marrow stem cells can predict later phenotypes [18], and are applying these approaches to understanding how survival and function in vivo might be predicted.

3D In Vitro Tissue Engineered Models and Microreactors for Drug Development

Most tissues and organs require blood flow both for distribution of nutrients and growth factors as well as mechanical stimulation and provision of trafficking of cells. We have developed and commercialized a microscale perfusion reactor to support 3D liver culture, including support of delicate liver sinusoidal endothelial cells [14, 19-22]. We are currently focused on creating more complex tissue microstructures through use of funcational biomaterials and projection microstereolithography, and applying the liver system to a variety of important physiology and drug development problems including liver inflammation and drug toxicity, and as a model of single cell cancer metastasis growth and response to chemotherapy.

The Griffith lab also leads a substantial program to build the “Human Physiome on a Chip”, funded by DARPA and NIH (http://physiomimetics.mit.edu.ezproxy.canberra.edu.au/). In this program, ten microphysiological systems, including liver, gut, lung, and reproductive systems, are interconnected in a physiologically relevant manner. In this project, a major focus is on quantitative in vitro in vivo correlation of responses to drugs and therapeutics, hence, we host a substantial Translational Systems Pharmacology core as part of this project.

Device and Software Development

Although not a central focus of the lab, work related to our core mission includes translation of basic science into the clinic through collaboration with device and software colleagues and clinicians. One collaborative project is the “cancer wand” to detect tumor cells at the margins of tumors intraoperatively, leveraging our experiences with proteases in inflammation (in collaboration with David Lee at the MIT Koch Institute, Moungi Bawendi in Chemistry, David Kirsch at Duke University, and Lumicell). Another collaborative project is development of software to manage endometriosis patients, in collaboration with John Guttag and Frans Kaashoek in the MIT EECS department, and Keith Isaason at Newton Wellesley Hospital http://web.mit.edu.ezproxy.canberra.edu.au/cgr .

References

[1] Griffith, L.G. and Swartz, M.A., “Capturing Complex 3D Tissue Physiology in vitro,” Nature Rev. Mol. Cell Biol., 7, 211-224 (2006).

[2] Maheshwari, G., Brown, G.L., Lauffenburger, D.A., Wells, A. and Griffith, L.G., “Cell Adhesion and Motility Depend on Nanoscale RGD Clustering,” J. Cell Sci., 113, 1677-1686 (2000).

[3] Griffith, L.G. and Lopina, S.T., “Micro-Distribution of Substratum-Bound Ligands Affects Cell Function: Hepatocyte Spreading on PEO-Tethered Galactose,” Biomaterials, 19, 979-986 (1998).

[4] Koo, L.Y., Irvine, D.J., Mayes, A.M., Lauffenburger, D.A. and Griffith, L.G., “Coregulation of Cell Adhesion by Nanoscale RGD Organization and Mechanical Stimulus,” J. Cell Sci., 115, 1423-1433 (2002).

[5] Jay, S.M., Murthy, A.C., Hawkins, J.F., Wortzel, J.R., Alvarez, L.M., Gannon, J., Macrae, C.A., Griffith, L.G and Lee, R.T., “An Engineered Bivalent Neuregulin Protects Against Doxorubicin-Induced Cardiotoxicity with Reduced Pro-Neoplastic Potential, Circulation,” Circulation, 128, 152-161 (2013).

[6] Jay, S.M., Kurtagic, E., Alvarez, L.M., de Picciotto, S., Sanchez, E., Hawkins, J.F., Prince, R.N, Guerrerro, Y., Treasure, C.L., Lee, R.T. and Griffith, L.G., “Engineered Bivalent EGF Receptor Family Ligands to Bias Signaling and Phenotypes,” J. Biol. Chem., 286, 27729-27740 (2011).

[7] Krueger, A.T., Kroll, C., Sanchez, E., Griffith, L.G. and Imperiali, B., “Tailoring Chimeric Ligands for Studying and Biasing ErbB Receptor Family Interactions,” Angew. Chem. Int. Ed., 53, 2662-2666 (2014).

[8] Fan, V.H., Tamama, K., Au, A., Littrell, R., Richardson, L.B., Wright, J.W., Wells, A. and Griffith, L.G., “Tethered EGF Provides a Survival Advantage to Mesenchymal Stem Cells” Stem Cells, 25, 1241-1251 (2007).

[9] Marcantonio, N.A., Boehm, C.A., Rozic, R., Au, A., Wells, A., Muschler, G.F., and Griffith, L.G., “Tethered Epidermal Growth Factor Increases Connective Tissue Progenitor Colony Formation,” Biomaterials, 30, 4629-4638 (2009).

[10] Platt, M.O., Roman, A.J., Wells, A., Lauffenburger, D.A. and Griffith, L.G., “Sustained Epidermal Growth Factor Receptor Levels and Activation by Tethered Ligand Binding Enhances Osteogenic Differentiation of Multi-Potent Marrow Stromal Cells,” J. Cell. Physiol., 221, 306-317 (2009).

[11] Mehta, G., Williams, C.M., Alvarez, L., Lesniewski, M., Kamm, R.D. and Griffith, L.G., “Synergistic Effects of Tethered Growth Factors and Adhesion Ligands on DNA Synthesis and Function of Primary Hepatocytes Cultured on Soft Synthetic Hydrogels,” Biomaterials, 31, 4657-4671 (2010).

[12] Cosgrove, B.D., Alexopoulos, L.G., Hang, T.C., Hendriks, B.S., Sorger, P.K., Griffith, L.G. and Lauffenburger, D.A., “Cytokine-Associated Drug Toxicity in Human Hepatocytes is Associated with Signaling Network Dysregulation,” Mol. Biosyst., 6, 1195-1206 (2010).

[13] Shuga, J., Zhang, J., Samson, L.D., Lodish, H.F. and Griffith, L.G., “In Vitro Erythropoiesis from Bone Marrow-Derived Progenitors Provides a Physiological Assay for Toxic and Mutagenic Compounds,” Proc. Natl. Acad. Sci., 104, 8737-42 (2007).

[14] Dash, A., Inman, W., Hoffmaster, K., Sevidal, S., Kelly, J., Obach, R.S., Griffith, L.G. and Tannenbaum, S.R., “Liver Tissue Engineering in the Evaluation of Drug Safety,” Expert Opin. Drug Metab. Toxicol., 5, 1159-1174 (2009).

[15] Beste, M.T., Pfaeffle-Doyle, N., Prentice, E., Lauffenburger, D.A., Isaacson, K.B. and Griffith, L.G., “Molecular Network Analysis of Endometriosis Reveals a Role for c-Jun-Regulated Macrophage Activation,” Sci. Transl. Med. 6, 222ra16 (2014).

[16] Miller, M.A., Barkal, L, Jeng, K., Griffith, L.G. and Lauffenburger, D.A., “Proteolytic Activity Matrix Analysis (PrAMA) for Simultaneous Determination of Multiple Protease Activities,” Integr. Biol., 3, 422-438 (2011).

[17] Miller, M., Meyer, A.S., Beste, M., Lasisi, Z., Reddy, S., Jeng., K., Chen, C.-H., Han, J., Isaacson, K.B., Griffith, L.G. and Lauffenburger, D.A., “Adam-10 and -17 Regulate Endometriotic Cell Migration via Concerted Ligand and Receptor Shedding Feedback on Kinase Signaling,” Proc. Natl. Acad. Sci., 110, E2074-E2083 (2013).

[18] Platt, M.O., Wilder, C.L., Wells, A., Griffith, L.G. and Lauffenburger, D.A., “Multi-Pathway Kinase Signatures of Multipotent Stromal Cells are Predictive for Osteogenic Differentiation,” Stem Cells, 27, 2804-2814 (2009).

[19] Hwa, A.J., Fry, R.C., Sivaraman, A., So, P.T., Samson, L.D., Stolz, D.B. and Griffith, L.G., “Rat Liver Sinusoidal Endothelial Cells Survive without Exogenous VEGF in 3D Perfused Co-Cultures with Hepatocytes,” FASEB J., 21, 2564-2579 (2007).

[20] Domansky, K., Inman, W., Serdy, J., Dash, A., Lim, M.H., and Griffith, L.G., “Perfused Multiwell Plate for 3D Liver Tissue Engineering,” Lab Chip, 7, 51-58 (2009).

[21] Sudo, R., Chung, S., Zervantonakis, I.K., Vickerman, V., Toshimitsu, Y., Griffith, L.G. and Kamm, R.D., “Transport-Mediated Angiogenesis in 3D Epithelial Coculture.” FASEB J., 23, 2155-2164 (2009).

[22] Yates, C., Shepard, C.R., Papworth, G., Dash, A., Beer Stolz, D., Tannenbaum, S., Griffith, L.G. and Wells, A., “Novel Three-Dimensional Organotypic Liver Bioreactor to Directly Visualize Early Events in Metastatic Progression,” Adv. Cancer Res., 97, 225-246 (2007).