Electrokinetic Microchip for label-free purification and characterization of Extracellular Vesicles
Esfandiari lab has been working on addressing some of these main challenges by developing a new device to extract sEVs from biofluids utilizing a label-free electrokintic based device while significantly reducing the number of processing steps and time between sample collection and sEVs isolation and maintaining the high yield and purity. Also, the device has the capability to characterize exosomes based on their unique dielectric properties and investigate their correlation with biochemical structures and functions.
The resulting device will also be able to detect and characterize other organelles including lysosomes and mitochondria, making this technology generalizable to studies investigating the role and composition of other sub-cellular compartments and viruses of similar size and structure. Technological innovation from this research will have applicability in a variety of medical fields and healthcare besides cancer diagnosis, including new EV-based therapeutics, monitoring the infectious and degenerative diseases.
The objective of this project is to develop a label-free and rapid Lab-on-a-Chip device to purify and characterize small extracellular vesicles (sEVs) from biofluids and cell culture media based on their unique biophysical properties. sEVs or exosomes are enclosed by a membrane and have a size ranging from 30 to 150 nm. They are released from all cell types into the extracellular space and are distributed in biofluids. Their composition and function depend on the originating cell type and they play an important role as a molecular cargo in cell-cell communication. Tumor-derived exosomes have potential use as circulating biomarkers in liquid biopsy for early stage diagnostics and routine clinical monitoring of cancer progression in difficult to access tumor sites. Rapid and efficient detection of exosomes is challenging owing to their heterogeneity and the complexity of biological samples. Current isolation methods are tedious and time-consuming, and exosomes isolated by current techniques often suffer from low purity and/or yield which causes inconsistencies in downstream proteomic and genomic analyses.
Smart Bioactive Material for Peripheral Nerve Regeneration
The goal of this project is to develop a biomimetic smart scaffolds and conduits for peripheral nerve regeneration. Severe peripheral nervous system (PNS) injuries which result in nerve gaps, require surgical repairs to replace lost nerve segments with a substitute or scaffold to enable functional recovery. The current clinical challenge is to improve biomaterial scaffolds. Factors that limit nerve regeneration and functional recovery include a lack of guidance cues from damaged extracellular matrix (ECM), and/or dysfunction of the native supporting Schwann cells (SCs). SCs are integral to facilitate axon guidance and injury disruption in ECM cues interferes with SC transition to a regenerative phenotype following injury that is required for axonal regeneration. In addition, delivering electric signals improves nerve regeneration, which can be done noninvasively and on demand by piezoelectric biomaterials. To overcome the limitations in current scaffold materials,
Esfandiari lab and collaborators have developed an electrospun piezoelectric nerve nanofiber scaffolds with controllable electrical stimulation and incorporating decellularized ECM. This multi-cue biomaterial will deliver signaling that can activate SCs and promote and guide axonal regeneration across injury gaps, thereby facilitating functional recovery. This innovative technology will address a critical need for deeper insights into multi-cue biomaterials and the response to such biomaterials in the field of regenerative medicine and tissue engineering.
3D Tumor Microenvironment for investigating the biomechanical and bioelectrical feedback loop in cancer progression and metastasis.
The aim of this project is to quantify the reciprocal interactions between biomechanical and bioelectrical cues in the cancer tumor microenvironment (TME). Dysregulation in the bioelectric signaling pathways within the tumor microenvironment (TME) is an important mechanism which disrupts tissue homeostasis in cancer. At the non-genetic level, this is primarily mediated by changes in ion channel expression, cell membrane potentials, and long-range propagation of oncogenic signals, e.g., through secretion of extracellular vesicles (EVs). The cancer TME is a complex entity comprised of a variety of cells, extracellular matrix (ECM) components and bioactive cues. Different aspects of the TME including biophysical, biochemical, and electrophysiological components are able to sense and influence one another, forming a feedback loop. However, understanding of the reciprocal interactions between mechanical and electrophysiological cues that influence carcinogenesis in the TME is limited. To begin to further this understanding,
Esfandiari Lab is developing and employing engineering tools to construct an in vitro spheroid culture platform capable of recapitulating the mechanical and electrical states of cells and the ECM in 3D. This work will lead to understanding and quantification of the mechanical-electrophysiological feedback loop in the TME. By better understanding the feedback loop, we will enable development of strategies capable of altering multiple aspects of the TME to better mimic those in vivo. This work will also result in a biomimetic model system capable of studying metastasis and recurrence upon treatments, hence advancing patient-specific prognosis. In the long-term, this research will enable development of non-invasive cancer therapeutics to regulate cues in the cancer microenvironment, hence lowering the likelihood of metastasis.
Engineering tools for real time monitoring and stimulation of spatially controlled Gastrointestinal Organogenesis
The goal of this proposal is to develop an improved system for differentiation of human pluripotent stem cells into functional intestinal organoids where tissue shape and function can be externally controlled and monitored in real time. Human organoids are multicellular 3D structures derived from adult or pluripotent stem cells and represent an incredibly exciting avenue of research aimed at human organ development, physiology, and disease. Organoids range in complexity from a few cell types to highly complex structures containing many diverse physiological functions approaching that of human tissue. Current methods to generate complex organoids result in tremendous variability in organoid size, shape, and functional maturity. In addition, current technologies lack the ability to experimentally manipulate organoid size, shape maturity and function, nor can one monitor cell responses to stimuli in real-time for precise control of outputs. Thus, a system to control spatial organoid formation with direct, real-time sensing and stimulation capabilities is needed for robust and reproducible studies of human tissue physiology and response to drugs.
Esfandiari Lab and collaborators at CuSTOM program at Cincinnati Children's Hospital seek to address this challenge by synthesizing tunable material with an implantable nanoelectronics to remotely monitor and define spatial organoid formation with a high degree of spatiotemporal resolution. In addition, biosensors will be utilized to continuously monitor secretion of organoid factors to ensure consistent development and function. By gaining far greater control of organogenesis and real-time sensing than previously accomplished, the team aims to develop differentiation protocols to gain a mechanistic insight into smooth muscle, neuronal and epithelial development, and function. Innovation from this research addresses the significant challenges currently faced in organoid development and will lead to more functional, consistently physiologically relevant models across healthcare in a wide range of tissue and disease.