Mark A. DeCoster - US grants

phospholipase A2; neurons;

This award provides funding for a 3 year continuing award to support a Research Experiences for Teachers (RET) Site program at Louisiana Tech University entitled, "NERO: Nanoscience Education and Research Outreach," under the direction of Dr. David Mills. The NERO program will link Louisiana Tech University engineers and scientists in the School of Biological Sciences, the Biomedical Engineering Program, and the Institute for Micromanufacturing with rural school systems in Louisiana, southern Arkansas and eastern Texas. The objectives of the program are to:
1) engage middle and high school teachers and their students in conducting research through internships and summer research experiences/technology workshops with follow-up through academic year programs, 2) provide guidance for teachers as they develop materials to translate their increased understanding of the research process into classroom learning experiences, 3) build lasting relationships among teachers, researchers, K-12 students, and graduate students, 4) improve communication of science and engineering and the scientific research process to teachers, students and the community, and 4) increase interest of K-12 students in pursuing STEM careers.

Co-funding is being provided by the NSF EPSCoR Program.

The PI will spend a year in the Program of Biomedical Engineering at her home institution, Louisiana Tech University, where she will work in two research areas while advising a PhD student in each area. The first project will integrate neuroscience, nano- and micro-technology engineering, mathematical modeling, and feedback control. Data sets of calcium ion dynamics will be the basis for mathematical analysis and modeling of brain cell network activity, to include development of a computational brain cell controller. As one of the most important secondary messengers in the body, calcium ions are involved in both intracellular and intercellular coordination. Mathematical model development of neuron dynamics will lay the foundation for design of a feedback controller to estimate calcium concentration in brain cells, which has the potential to further understanding of how the human body reacts to disorders of the brain and the predictive behavior of the brain.

The second project will integrate nanotechnology engineering, mathematical modeling, and feedback control design. The mathematical modeling involved will result in the development of an accurate prediction of the amount of a nano-therapeutic agent that reaches a tumor site based on real-time estimates of drug bioavailability. This, in turn, will lay the groundwork for the design of a computational feedback controller that would be used to noninvasively measure and regulate drug concentration in the body. Although the current treatment of cancerous tumors can be very effective, there is no way of monitoring the therapy as it is being administered. In fact, the only way the amount of chemotherapy to be administered is determined by the patient's body mass. By joining together mathematical modeling and feedback control techniques, this project seeks to provide a mechanism for prediction of the effects of the nano-particle chemotherapy drugs in a patient's body, thus leading to better models for drug regimen and administration.

The work will focus on models of computing with brain cells, more specifically the neurons and their major support cells in the brain, the glia. The major novelty of the project is the consideration of the glia cells in these models of computation. The interplay between glia and neurons has not been considered in the domain literature until now. Thus the project will not only look at how to perform computation taking inspiration from the organization of the neurons and glia into networks, but also explore topics such as investigating the power related to sequentiality/parallelism of such devices, synchronization, data processing and manipulation such as considering different types of output encoding for the computation. Several other topics considered already for neuronal systems such as universality/non-universality of networks with various restrictions, normal forms and consideration of the refractory period of neurons will also be investigated for the new devices.

The project will incorporate an experimental part in the second year of the timeline when with the help of the senior researcher on the project, Dr. Mark DeCoster (professor of Biomedical Engineering) the results of the first phase of the work will be validated in the lab. The work holds promise to significantly impact several research areas as well as the emerging area of computing with cells. In Computer Science, the research may yield new paradigms and new computing techniques (as has happened in the discovery of genetic algorithms and neural networks). In Biology, the project is likely to provide insight into the organization of brain cells networks, including the support cells for neurons, the glia.

One of the major objectives of this project is to gain more knowledge and insight into the extraordinary biological systems ? the cells and also to understand in a systemic way how the sub-cellular processes work and how the cell-cell communications can be useful for performing distributed computations. Algorithms, source codes and results of the project will be freely disseminated to the public through the project's website.

Cells are the basis of our whole bodies, and also communicate with each other. Therefore, cells contain many lessons about populations, change, and adaptations. Developing an artificial cell product that gains entrance into the classrooms of students will benefit their learning and provide expanded choices of teaching tools. More advance artificial cell platforms may impact applied and basic research sectors by also providing expanded choices for looking at dynamic processes. This project aims to commercialize artificial (manufactured) cells and cell platforms for educational, research, and industry needs. The technology will include the use of 3D printers to make these artificial cells, since these printers are becoming increasingly available and cost-effective for construction purposes. The technology developed in this project will be paired with educational and visualization software so that the artificial cells will provide hands-on experiments and testing to students, assisted by the software learning tools. Due to the complex nature and cost of living cells, this artificial cell technology is needed to cost-effectively teach students the dynamics of cellular processes and at the same time let them carry out experiments and test ideas.

The technological components of this project will utilize 3D printers to make artificial cells that are hollow and structured to allow for visualization and testing of the dynamic processes of cells in aqueous environments. Plastics will be the initial materials base, since they can be 3D printed, but other materials will also be incorporated into the platforms such as dyes and chemicals that will define the dynamic processes. The artificial cell kits will also include instructional and experimental software based on "predator-prey" and other dynamic processes that will guide experimentation and testing. Constructed artificial cells will be generated initially that are large enough to be held and visualized with digital cameras without the need of microscopes. Future generations of artificial cells will include dynamic processes for more advanced testing to include fluorescence- and microscopy- based measurements. Ultimately a technology "library" of artificial cells and cell components is envisioned for learning and research tools.

PI: DeCoster, Mark A.
Proposal Number: 1547693

One type of Biomanufacturing involves placing cells together in both two dimensions (2D) and three dimensions (3D). To better approximate what happens in the body in both healthy and diseased tissues, the growth of cells, as well as cell death must be understood. Much like pruning the limbs of a tree without killing it, the investigators of this project will use a controlled, natural process called apoptosis to prune groups of cells in both 2D and 3D environments to improve the function of the overall construct. These studies could enhance our understanding of how to control normal cell formations into tissues and how to control disease processes such as cancer. The containers for the cells to be studied in this project will include bioreactors generated using 3D printers.

The technological components of this project seek to address the challenges of generating and shaping assemblies of cells in both 2D and 3D environments. The project aims to understand the growth processes of both normal and cancer cells, with the goal of achieving better biomanufacturing strategies and insight into tumor growth. Beyond just growth, the investigators of this EAGER award will also apply apoptotic stimuli to cells using two types of high-aspect ratio structures (HARS), to prune away cells in a controlled manner. To facilitate imaging into thicker (>0.5 mm) 3D cell assemblies in this project, gradient index (GRIN) lenses combined with multi-photon microscopy will be used. The HARS materials used in this project include a hollow, non-degradable halloysite, and a novel, biodegradable biocomposite containing copper. Both HARS materials scale from the nano-dimension in diameter to the micro-dimension in length. The experiments carried out in this project will utilize bioreactors generated using 3D printers and functional outputs from the bioreactors will include detection of glutamate and pH dynamics using a fast-growing glioma cell line and slower growing (normal) astrocyte primary culture model to compare cellular outputs with growth before and after the pruning process of apoptosis. To better approximate dynamic processes in the brain, microglia will also be added to model recovery after apoptosis. A foundry of 3D printed bioreactors generated for the project will be established in the form of bioreactor images, .stl files, and animations, and will be tested for integration with commercially available millifluidic devices to detect, for example, chemical changes occurring in the bioreactors over time. Results from this project are anticipated to impact future biomanufacturing strategies and educational materials considering the increasing availability of 3D-printing technology and design software.

Non-technical Description
Twelve researchers across three institutions in three states (Louisiana, Arkansas, and Alabama) will collaborate on this brain research, and develop a foundation for the region as a hub for interdisciplinary, collaborative research activity in the neurosciences. This work will focus on understanding the initiation of epileptic brain seizures and longer-term impacts on brain function such as memory. Epileptic seizures directly impact roughly 1% of humans, and have indirect impacts on loved ones and caregivers as well as economic impacts on society. Epilepsy has been called a ?window to brain function? because the condition impairs different brain functions depending on the location of the seizure in the brain and the impacted network of neurons, and because it provides a unique opportunity to study an impaired brain?s function over time and space. The project will develop minimally invasive implantable sensors that can be used for monitoring before, during, and for several months following seizure events. Researchers will relate changes occurring during seizure events with those observed in the intervals between events. The project includes hiring four new faculty, design and purchase of equipment, development of new undergraduate and graduate courses, recruitment and training of a diverse student population to better reflect the regional community, and new student research and workforce opportunities.

Technical Description
The activities of the team across three institutions (Louisiana Tech University, the University of Arkansas, and the University of Alabama) are organized under four thrust areas that focus on recording and analysis of electrical activity, magnetic activity, neurochemical and optical signals, and memory function of the brain. A series of coordinated and synergistic investigations will be conducted using innovative methods and tools designed to probe brain function at the molecular, cellular, and macro levels in epileptic rats and human subjects. Patients with focal epilepsy, during their presurgical evaluation, will undergo implantation of intracranial electroencephalographic (iEEG) electrodes for subsequent long-term (days) recording and monitoring of their spontaneous seizures. Non-invasive magnetoencephalographic (MEG) imaging is also used to provide important complementary information on the electromagnetic activity of the brain. This collaboration will advance research and also result in the creation of unique databases of human and animal data from the participating institutions.