Tumor cell metastases to the brain are common in breast, skin, and lung cancer patients and are associated with high morbidity and poor prognosis. Blood-brain barrier (BBB) dysfunction is believed to occur in tumor metastasis, as well as infectious disease, stroke, multiple sclerosis, HIV, and Alzheimer’s disease. However, it is not understood how metastatic tumor cells are able to cross the BBB, given the BBB’s inherent function to tightly regulate flux of only select cellular and molecular materials into the brain’s interstitial fluid. A major hurdle preventing an understanding of the mechanisms of brain metastasis is the lack of physiologically-relevant BBB in vitro models. Dr. Stroka's team is working to develop a microsystem BBB-on-a-chip using microfabrication techniques and to use this model to understand brain endothelial cell (EC) biomechanics and the mechanobiology of brain metastasis. REU students participating in this project will have the opportunity to utilize the lab's BBB-microsystem to test hypotheses on the mechanisms of cancer metastasis. REU participants would conduct the majority of their work at the University of Maryland.
Over millions of years, the human kidney has evolved complex three-dimensional (3D) anatomy to filter waste products and excess fluid from the blood and regulate whole-body homeostasis. Unfortunately, these functions render the kidney highly susceptible to harmful side effects during the development of new therapeutic drugs. Ideally, the potential for a candidate medication to injure the kidney should be revealed in the early stages of drug screening (i.e., well before human trials). At present, however, state-of-the-art in vitro kidney testing systems primarily comprise flat (2D) surfaces and only one cell type, leading to poorly predictive results. Notably, this predominance of simplified kidney models is rooted not in the interests of science or medicine, but rather, in the limits of conventional micromanufacturing. REU students will work with Dr. Sochol and his lab to support their goal of better mimicking the 3D biophysical and organizational interactions of the kidney by leveraging recent breakthroughs for the additive nanomanufacturing or “nano3D printing” technology, direct laser writing (DLW). REU participants would conduct the majority of their work at the University of Maryland.
(Dr. Aldo Badano, Food and Drug Administration)
The presence of amyloid beta plaques in the human brain is considered one of the hallmark features of Alzheimer's disease (AD) and a promising biomarker for early diagnosis. In early AD stages, Aβ plaques are formed when Aβ peptides aggregate and accumulate in the brain to form higher-order fibrils with β-sheet structures. A structurally sensitive and high-resolution imaging method is required to detect plaques in the human brain and allow timely therapeutic interventions to prevent AD progression. To this end, we are designing a novel amyloid imaging system prototype based on small-angle x-ray scattering (SAXS) to quantify amyloid load without contrast agents based on the nanometer-scale structural features of plaques. REU student will have an opportunity to prepare human brain phantoms with amyloid plaque models and assist in testing the feasibility of detecting amyloid plaques using a newly designed amyloid imaging spectral SAXS prototype. REU participants would conduct the majority of their work at the Food and Drug Administration.
The human gastrointestinal (GI) microbiome, often viewed as a complex organ itself, influences homeostasis and is implicated in many human diseases. There are few methodologies that enable resolution of its signaling, cell growth, and chemical environments even at the macro length scale. In the last several years however, researchers have developed micro- and meso-systems for interrogating GI tract biology. At the microscale, organ- or animal-on-a-chip methodologies have been developed for studying the GI tract. They provide first-of-their-kind access to biological function in user-controlled conditions. However, current approaches lack the ability to interrogate and modulate molecular signaling at cellular length scales and in real time. We have developed methodologies to combine synthetic biology with biofabrication to create smart hydrogels for interrogating molecular space. We do this by embedding “rewired” bacteria to serve as information translators into electrodeposited hydrogels that are assembled onto the sidewalls of a microfluidic channel (see Fig. 4). As a demonstration, we have assembled engineered E. coli to express blue fluorescent protein in response to a signal molecule, in this case the gratuitous lac operon inducer IPTG (Fig. 4c). In Fig. 4d, we show that one can layer cells using electrodeposition in configurations that enable visual access as well as chemical access so that cells can respond to molecular cues. In this REU project, using the same electrodeposition methodologies we have already developed in large systems, REU students will test the hypothesis that biologically active bacterial signaling molecules can be detected by a hydrogel bilayer that contains (i) signal sensing bacteria and (2) a catechol-modified chitosan layer incorporated onto the side wall of the channels shown in Fig. 4a,b. The students will assess whether the assembled cells can convert signal molecules to redox mediators in a manner that enables electrochemical detection. A successful result would be the first case of a biofabricated glucose sensor in an animal on a chip device. Such tests will be the first examples of the electrochemical interrogation of interkingdom biological signaling in an in vitro system. REU participants would conduct the majority of their work at the University of Maryland.
Exosomes, a subset of cell-derived extracellular vesicles (EVs), have emerged as potential therapeutic vectors for a wide variety of applications. As alternatives to synthetic nanoparticles and cell therapies, exosomes have many potential benefits. Among these is the ability to transfer functional nucleic acids to recipient cells, as exosomes are physiological vectors for intercellular communication via nucleic acid transfer. However, exosomes also have shortcomings, especially their low potency due to containing relatively low amounts of nucleic acid cargo, especially non-coding RNAs such as microRNAs (miRNAs) and long non-coding RNAs (lncRNAs), which are critical to defining exosome bioactivity. Overcoming this limitation is crucial to enabling the therapeutic potential of exosomes. To this end, Dr. Jay and his group have studied how the biophysical culture microenvironment of exosome-producing cells can be manipulated to control non-coding RNA cargo and bioactivity of exosomes. This investigation is potentially transformative in identifying critical parameters that could be controlled towards development of rationally designed, scalable biomanufacturing approaches for therapeutic exosomes. REU students will have the opportunity to employ both 3D printing and microsystems design to test specific hypotheses related to the roles of biophysical parameters on exosome production and bioactivity. REU participants would conduct the majority of their work at the University of Maryland.
(Drs. Richard Gray & Pavel Takmakov, Food and Drug Administration)
Sudden cardiac death resulting from ventricular fibrillation is the leading cause of death in the industrialized world. A better understanding of heart physiology, including individualized function, has the potential to prevent such deaths. Multi-scale models comprised of equations governing sub-cellular and cellular biochemical processes and bioelectric phenomena on the level of tissue, organ, and whole body are well understood and used to study life threatening cardiac arrhythmias including fibrillation. However, we have reached a “crises of complexity” because over a hundred cell models have been developed each with multitude of variables parameters. PubMed, a publication database, returns more than 140,000 articles for “cardiac models” as a keyword. Unfortunately, these models are highly overdetermined and rarely properly validated which prevents appropriate evaluation for a specific context of use that is necessary for clinical applications. We are in the process of automating the cardiac cell model development process to make it transparent, reproducible, and which avoids over-determined models. Cardiac cell models have always been ‘data-driven’ with thousands of experimental studies in different formats providing the basis for the models. In this project, we are using data science approaches to develop an algorithm to extract quantitative data from these experimental studies and use machine learning for building a framework to translate these data into robust cardiac cell models with a training set consisting of manually generated database of over a hundred of such studies. REU participants would conduct the majority of their work at the Food and Drug Administration.
Lymph nodes are the immunological tissues that coordinate immune response. Thus, most vaccines and immunotherapies must reach these sites for efficacy. Engineered polymers offer new opportunities as carriers of vaccines and immunotherapies, as well as to actively control the specific types of immune responses that are generated. A broad challenge in this field is the inability to effectively target lymph nodes, and a knowledge gap of how the concentrations and combinations of signals that reach lymph nodes polarize systemic immune response. The objective of this project is to harness advanced manufacturing tools to recapitulate lymph node function in vitro. REU students will conduct research in one of two areas, defining the scope of the projects to ensure tangible progress can be made over the course of the summer REU period. Potential project areas include i) assessing the interactions of primary immune cells with polymer scaffolds designed to mimic key features of lymph nodes, and ii) building nanostructures entirely from immune signals to create rational platforms to control multiple immune pathways in parallel. These projects also offer opportunities to gain experience in translational research through the Jewell Lab’s multiple pre-clinical animal models of autoimmune disease and cancer. REU participants would conduct the majority of their work at the University of Maryland.
The constant generation and then movement of mucus from the lung airways to the throat, where it is then swallowed, is a key method by which the body protects against infection. However, in the airways of individuals with chronic lung diseases (e.g. asthma, chronic bronchitis, cystic fibrosis), build-up of mucus with abnormal biochemical and biophysical properties leads to airway obstruction and contributes to impaired lung function. Due to the natural heterogeneity of mucus derived from patients and animal models, we currently have a limited understanding of how mucus gel components impact the functional properties of mucus such as viscoelasticity for optimal mucus clearance and network architecture for pathogen/particulate capture. Toward this end, the goal of our NSF REU projects is to better understand the biophysical and biomolecular features of mucins that control their assembly into mucus. In our first REU student project, a bioengineered mucin-based hydrogel will be developed to mimic the physiological conditions under which the mucus gel network is assembled in health and disease. In a second project, an REU student will use CRISPR/Cas9 genetic engineering in order to design a tissue culture system with controlled airway mucus composition and function resembling that of a healthy and diseased airway. Together, this work will provide state-of-the art models that enable optimization and design of new therapies for chronic lung diseases. REU students engaged in this project will gain first-hand experience in biomaterial design, tissue culture, and/or genetic engineering. REU participants would conduct the majority of their work at the University of Maryland.
(Dr. Katherine Vorvokalos, Food and Drug Administration)
Tissue Engineering and Regenerative Medicine (TERM) products present enormous challenges and opportunities for basic science, manufacturing, standardization, regulation and clinical translation. Increasingly, engineering principles are emphasized to meet these challenges. For material scientists, one of the most interesting questions is that of multi-length-scale tissue quality (TQ) for engineered tissue meant to exhibit structural integrity. While the science of cell characterization is quite sophisticated (e.g., proteomics, transcriptomics, microscopy), there remains a chasm between these characterization methods and physiologically relevant markers. It becomes especially complex when resorbable materials are involved. This ongoing project attempts to bridge the chasm by examining the current TERM product landscape from a materials science perspective. The student would learn about aspects of the current TERM products marketed in the United States, FDA product databases, the international standards documents landscape surrounding TERM products and FDA guidance documents. Most importantly, they would examine the scientific literature to identify physiologically relevant markers – and gaps thereof – for success of TERM products. Coding experience would be beneficial. REU participants would conduct the majority of their work at the Food and Drug Administration.
(Dr. Shawn He, University of Maryland)
Stem cells are attracting ever-increasing attention for various biomedical applications including tissue engineering and regenerative medicine. However, the approaches used today for culturing stem cells in vitro are non-physiological and stem cells are banked in solutions containing highly toxic organic solvents (e.g., dimethylsulfoxide that is commonly known as DMSO). The former may induce genotypic and phenotypic changes in the stem cells, to compromise the quality of stem cells for tissue regeneration. The latter makes it indispensable to rigorously and tediously wash the stem cells after banking before implanting into patients, which may cause significant death or loss of the precious cells. The objective of this project is to develop 3D biomimetic culture of stem cells to minimize their spontaneous differentiation under in vitro culture, and further engineer the stem cells with micro and nanoscale biomaterials to achieve organic solvent-free banking so that they can be transplanted into patients without tedious washing. This will be achieved by utilizing the extensive nanotechnology approaches such as non-planar microfluidics and drug delivery with stimuli-responsive nanoparticles developed in the He Laboratory. REU students will learn how to fabricate microfluidic devices, use the devices to encapsulate stem cells for miniaturized 3D culture, use therapeutics-laden nanoparticles to engineer stem cells for banking, and make significant contributions to the innovative and impactful project. REU participants would conduct the majority of their work at the University of Maryland.