Institute Affiliations


All Publications


  • A neurovascular-unit-on-a-chip for the evaluation of the restorative potential of stem cell therapies for ischaemic stroke. Nature biomedical engineering Lyu, Z., Park, J., Kim, K., Jin, H., Wu, H., Rajadas, J., Kim, D., Steinberg, G. K., Lee, W. 2021

    Abstract

    The therapeutic efficacy of stem cells transplanted into an ischaemic brain depends primarily on the responses of the neurovascular unit. Here, we report the development and applicability of a functional neurovascular unit on a microfluidic chip as a microphysiological model of ischaemic stroke that recapitulates the function of the blood-brain barrier as well as interactions between therapeutic stem cells and host cells (human brain microvascular endothelial cells, pericytes, astrocytes, microglia and neurons). We used the model to track the infiltration of a number of candidate stem cells and to characterize the expression levels of genes associated with post-stroke pathologies. We observed that each type of stem cell showed unique neurorestorative effects, primarily by supporting endogenous recovery rather than through direct cell replacement, and that the recovery of synaptic activities is correlated with the recovery of the structural and functional integrity of the neurovascular unit rather than with the regeneration of neurons.

    View details for DOI 10.1038/s41551-021-00744-7

    View details for PubMedID 34385693

  • Repurposing Disulfiram (Tetraethylthiuram Disulfide) as a Potential Drug Candidate against Borrelia burgdorferi In Vitro and In Vivo. Antibiotics (Basel, Switzerland) Potula, H. S., Shahryari, J., Inayathullah, M., Malkovskiy, A. V., Kim, K., Rajadas, J. 2020; 9 (9)

    Abstract

    Lyme disease caused by the Borrelia burgdorferi (Bb or B. burgdorferi) is the most common vector-borne, multi-systemic disease in the USA. Although most Lyme disease patients can be cured with a course of the first line of antibiotic treatment, some patients are intolerant to currently available antibiotics, necessitating the development of more effective therapeutics. We previously found several drugs, including disulfiram, that exhibited effective activity against B. burgdorferi. In the current study, we evaluated the potential of repurposing the FDA-approved drug, disulfiram for its borreliacidal activity. Our results indicate disulfiram has excellent borreliacidal activity against both the log and stationary phase B. burgdorferi sensu stricto B31 MI. Treatment of mice with disulfiram eliminated the B. burgdorferi sensu stricto B31 MI completely from the hearts and urinary bladder by day 28 post infection. Moreover, disulfiram-treated mice showed reduced expressions of inflammatory markers, and thus they were protected from histopathology and cardiac organ damage. Furthermore, disulfiram-treated mice showed significantly lower amounts of total antibody titers (IgM and IgG) at day 21 and total IgG2b at day 28 post infection. FACS analysis of lymph nodes revealed a decrease in the percentage of CD19+ B cells and an increase in total percentage of CD3+ T cells, CD3+ CD4+ T helpers, and naive and effector memory cells in disulfiram-treated mice. Together, our findings suggest that disulfiram has the potential to be repurposed as an effective antibiotic for treating Lyme disease.

    View details for DOI 10.3390/antibiotics9090633

    View details for PubMedID 32971817

  • Characterization of brain dysfunction induced by bacterial lipopeptides that alter neuronal activity and network in rodent brains. The Journal of neuroscience : the official journal of the Society for Neuroscience Kim, K., Zamaleeva, A. I., Woo Lee, Y., Ahmed, M. R., Kim, E., Lee, H., Raveendra Pothineni, V., Tao, J., Rhee, S., Jayakumar, M., Inayathullah, M., Sivanesan, S., Red-Horse, K., Palmer, T. D., Park, J., Madison, D. V., Lee, H., Rajadas, J. 2018

    Abstract

    The immunopathological states of the brain induced by bacterial lipoproteins have been well-characterized by employing biochemical and histological assays. However, these studies have limitations in determining functional states of damaged brains involving aberrant synaptic activity and network, which makes it difficult to diagnose brain disorders during bacterial infection. To address this, we investigated the effect of Pam3CSK4 (PAM), a synthetic bacterial lipopeptide, on synaptic dysfunction of female mice brains and cultured neurons in parallel. Our functional brain imaging using PET with [18F]-FDG and [18F]-FMZ revealed the brain dysfunction induced by PAM is closely aligned to disruption of neurotransmitter-related neuronal activity and functional correlation in the region of the limbic system rather than to decrease of metabolic activity of neurons in the injection area. This finding was verified by in vivo tissue experiments that analyzed synaptic and dendritic alterations in the regions where PET imaging showed abnormal neuronal activity and network. Recording of synaptic activity also revealed that PAM reorganized synaptic distribution and decreased synaptic plasticity in hippocampus. Further study using in vitro neuron cultures demonstrated that PAM decreased the number of presynapses and the frequency of mEPSC, which suggests PAM disrupts neuronal function by damaging presynapses exclusively. We also showed PAM caused aggregation of synapses around dendrites, which may have caused no significant change in expression level of synaptic proteins while synaptic number and function was impaired by PAM. New findings of this study could provide a useful guide for diagnosis and treatment of brain disorders specific to bacterial infection.SIGNIFICANCE STATEMENTIt is challenging to diagnose brain disorders caused by bacterial infection because neural damage induced by bacterial products involves non-specific neurological symptoms, which is rarely detected by laboratory tests with low spatiotemporal resolution. To better understand brain pathology, it is essential to detect functional abnormalities of brain over time. To this end, we investigated characteristic patterns of altered neuronal integrity and functional correlation between various regions in mice brains injected with bacterial lipopeptides by using PET with a goal to apply new findings to diagnosis of brain disorder specific to bacterial infection. In addition, we analyzed altered synaptic density and function using both in vivo and in vitro experimental models to understand how bacterial lipopeptides impair brain function and network.

    View details for PubMedID 30381406

  • In vitro and in vivo evaluation of cephalosporins for the treatment of Lyme disease DRUG DESIGN DEVELOPMENT AND THERAPY Pothineni, V., Parekh, M. B., Babar, M., Ambati, A., Maguire, P., Inayathullah, M., Kim, K., Tayebi, L., Potula, H. K., Rajadas, J. 2018; 12: 2915–21
  • PEG/Dextran Double Layer Influences Fe Ion Release and Colloidal Stability of Iron Oxide Nanoparticles. Scientific reports Mohammadi, M. R., Malkovskiy, A. V., Jothimuthu, P. n., Kim, K. M., Parekh, M. n., Inayathullah, M. n., Zhuge, Y. n., Rajadas, J. n. 2018; 8 (1): 4286

    Abstract

    Despite preliminary confidence on biosafety of polymer coated iron oxide nanoparticles (SPIONs), toxicity concerns have hampered their clinical translation. SPIONs toxicity is known to be due to catalytic activity of their surface and release of toxic Fe ions originating from the core biodegradation, leading to the generation of reactive oxygen species (ROS). Here, we hypothesized that a double-layer polymeric corona comprising of dextran as an interior, and polyethylene glycol (PEG) as an exterior layer better shields the core SPIONs. We found that ROS generation was cell specific and depended on SPIONs concentration, although it was reduced by sufficient PEG immobilization or 100 µM deferoxamine. 24 h following injection, PEGylated samples showed reduction of biodistribution in liver, heterogenous biodistribution profile in spleen, and no influence on NPs blood retention. Sufficient surface masking or administration of deferoxamine could be beneficial strategies in designing and clinical translation of future biomedical SPIONs.

    View details for PubMedID 29523826

  • From solvent-free microspheres to bioactive gradient scaffolds NANOMEDICINE-NANOTECHNOLOGY BIOLOGY AND MEDICINE Rasoulianboroujeni, M., Yazdimamaghani, M., Khoshkenar, P., Pothineni, V. R., Kim, K. M., Murray, T. A., Rajadas, J., Mills, D. K., Vashaee, D., Moharamzadeh, K., Tayebi, L. 2017; 13 (3): 1157-1169

    Abstract

    A solvent-free microsphere sintering technique was developed to fabricate scaffolds with pore size gradient for tissue engineering applications. Poly(D,L-Lactide) microspheres were fabricated through an emulsification method where TiO2 nanoparticles were employed both as particulate emulsifier in the preparation procedure and as surface modification agent to improve bioactivity of the scaffolds. A fine-tunable pore size gradient was achieved with a pore volume of 30±2.6%. SEM, EDX, XRD and FTIR analyses all confirmed the formation of bone-like apatite at the 14th day of immersion in Simulated Body Fluid (SBF) implying the ability of our scaffolds to bond to living bone tissue. In vitro examination of the scaffolds showed progressive activity of the osteoblasts on the scaffold with evidence of increase in its mineral content. The bioactive scaffold developed in this study has the potential to be used as a suitable biomaterial for bone tissue engineering and hard tissue regeneration.

    View details for DOI 10.1016/j.nano.2016.10.008

    View details for Web of Science ID 000401089100037

  • Screening of NCI-DTP library to identify new drug candidates for Borrelia burgdorferi. journal of antibiotics Pothineni, V. R., Wagh, D., Babar, M. M., Inayathullah, M., Watts, R. E., Kim, K., Parekh, M. B., Gurjarpadhye, A. A., Solow-Cordero, D., Tayebi, L., Rajadas, J. 2017; 70 (3): 308-312

    View details for DOI 10.1038/ja.2016.131

    View details for PubMedID 27826144

  • Attenuation of synaptic toxicity and MARK4/PAR1-mediated Tau phosphorylation by methylene blue for Alzheimer's disease treatment SCIENTIFIC REPORTS Sun, W., Lee, S., Huang, X., Liu, S., Inayathullah, M., Kim, K., Tang, H., Ashford, J. W., Rajadas, J. 2016; 6

    Abstract

    Alzheimer's disease (AD) is a neurodegenerative disease characterized by genotypic and phenotypic heterogeneity. Critical components of the two AD pathological pathways, Aβ-amyloidosis and Tauopathy, have been considered as therapeutic targets. Among them, much effort is focused on aberrant Tau phosphorylation and targeting Tau-phosphorylating kinases. Methylene blue (MB), a phenothiazine dye that crosses the blood-brain barrier, has been shown to hit multiple molecular targets involved in AD and have beneficial effects in clinical studies. Here we present evidence that microtubule affinity-regulating kinase (MARK4) is a novel target of MB. MB partially rescued the synaptic toxicity in Drosophila larva overexpressing PAR1 (MARK analog). In 293T culture, MB decreased MARK4-mediated Tau phosphorylation in a dose dependent manner. Further studies revealed a two-fold mechanism by MB including down-regulation of MARK4 protein level through ubiquitin-proteasome pathway and inhibition of MARK4 kinase activity in vitro. This study highlights the importance of MARK4 as a viable target for Tauopathy and provides fresh insight into the complex mechanism used by MB to treat AD.

    View details for DOI 10.1038/srep34784

    View details for PubMedID 27708431

  • Axon Outgrowth of Rat Embryonic Hippocampal Neurons in the Presence of an Electric Field ACS CHEMICAL NEUROSCIENCE Kim, K., Kim, S. Y., Palmore, G. T. 2016; 7 (10): 1325-1330

    Abstract

    Application of an electric field (EF) has long been used to induce axon outgrowth following nerve injuries. The response of mammalian neurons (e.g., axon length, axon guidance) from the central nervous system (CNS) to an EF, however, remains unclear, whereas those from amphibian or avian neuron models have been well characterized. Thus, to determine an optimal EF for axon outgrowth of mammalian CNS neurons, we applied a wide range of EF to rat hippocampal neurons. Our results showed that EF with either a high magnitude (100 mV/mm or higher) or long exposure time (10 h or longer) with low magnitude (10-30 mV/mm) caused a neurite collapse and cell death. We also investigated whether neuronal response to an EF is altered depending on the growth stage of neuron cultures by applying 30 mV/mm to cells from 1 to 11 days in vitro (DIV). Neurons showed the turnover of axon outgrowth pattern when electrically stimulated between 4-5 DIV at which point neurons have both axonal and dendritic formation. The findings of this study suggest that the developmental stage of neurons is an important factor to consider when using EF as a potential method for axon regeneration in mammalian CNS neurons.

    View details for DOI 10.1021/acschemneuro.6b00191

    View details for Web of Science ID 000385994000006

    View details for PubMedID 27529437

  • Identification of new drug candidates against Borrelia burgdorferi using high-throughput screening DRUG DESIGN DEVELOPMENT AND THERAPY Pothineni, V. R., Wagh, D., Babar, M. M., Inayathullah, M., Solow-Cordero, D., Kim, K., Samineni, A. V., Parekh, M. B., Tayebi, L., Rajadas, J. 2016; 10: 1307-1322

    Abstract

    Lyme disease is the most common zoonotic bacterial disease in North America. It is estimated that >300,000 cases per annum are reported in USA alone. A total of 10%-20% of patients who have been treated with antibiotic therapy report the recrudescence of symptoms, such as muscle and joint pain, psychosocial and cognitive difficulties, and generalized fatigue. This condition is referred to as posttreatment Lyme disease syndrome. While there is no evidence for the presence of viable infectious organisms in individuals with posttreatment Lyme disease syndrome, some researchers found surviving Borrelia burgdorferi population in rodents and primates even after antibiotic treatment. Although such observations need more ratification, there is unmet need for developing the therapeutic agents that focus on removing the persisting bacterial form of B. burgdorferi in rodent and nonhuman primates. For this purpose, high-throughput screening was done using BacTiter-Glo assay for four compound libraries to identify candidates that stop the growth of B. burgdorferi in vitro. The four chemical libraries containing 4,366 compounds (80% Food and Drug Administration [FDA] approved) that were screened are Library of Pharmacologically Active Compounds (LOPAC1280), the National Institutes of Health Clinical Collection, the Microsource Spectrum, and the Biomol FDA. We subsequently identified 150 unique compounds, which inhibited >90% of B. burgdorferi growth at a concentration of <25 µM. These 150 unique compounds comprise many safe antibiotics, chemical compounds, and also small molecules from plant sources. Of the 150 unique compounds, 101 compounds are FDA approved. We selected the top 20 FDA-approved molecules based on safety and potency and studied their minimum inhibitory concentration and minimum bactericidal concentration. The promising safe FDA-approved candidates that show low minimum inhibitory concentration and minimum bactericidal concentration values can be chosen as lead molecules for further advanced studies.

    View details for DOI 10.2147/DDDT.S101486

    View details for Web of Science ID 000373575000001

    View details for PubMedID 27103785

    View details for PubMedCentralID PMC4827596

  • Neuron Image Analyzer: Automated and Accurate Extraction of Neuronal Data from Low Quality Images SCIENTIFIC REPORTS Kim, K., Son, K., Palmore, G. R. 2015; 5: 17062

    Abstract

    Image analysis software is an essential tool used in neuroscience and neural engineering to evaluate changes in neuronal structure following extracellular stimuli. Both manual and automated methods in current use are severely inadequate at detecting and quantifying changes in neuronal morphology when the images analyzed have a low signal-to-noise ratio (SNR). This inadequacy derives from the fact that these methods often include data from non-neuronal structures or artifacts by simply tracing pixels with high intensity. In this paper, we describe Neuron Image Analyzer (NIA), a novel algorithm that overcomes these inadequacies by employing Laplacian of Gaussian filter and graphical models (i.e., Hidden Markov Model, Fully Connected Chain Model) to specifically extract relational pixel information corresponding to neuronal structures (i.e., soma, neurite). As such, NIA that is based on vector representation is less likely to detect false signals (i.e., non-neuronal structures) or generate artifact signals (i.e., deformation of original structures) than current image analysis algorithms that are based on raster representation. We demonstrate that NIA enables precise quantification of neuronal processes (e.g., length and orientation of neurites) in low quality images with a significant increase in the accuracy of detecting neuronal changes post-stimulation.

    View details for PubMedID 26593337

  • Borreliacidal activity of Borrelia metal transporter A (BmtA) binding small molecules by manganese transport inhibition DRUG DESIGN DEVELOPMENT AND THERAPY Wagh, D., Pothineni, V. R., Inayathullah, M., Liu, S., Kim, K., Rajadas, J. 2015; 9: 805-815

    Abstract

    Borrelia burgdorferi, the causative agent of Lyme disease, utilizes manganese (Mn) for its various metabolic needs. We hypothesized that blocking Mn transporter could be a possible approach to inhibit metabolic activity of this pathogen and eliminate the infection. We used a combination of in silico protein structure prediction together with molecular docking to target the Borrelia metal transporter A (BmtA), a single known Mn transporter in Borrelia and screened libraries of FDA approved compounds that could potentially bind to the predicted BmtA structure with high affinity. Tricyclic antihistamines such as loratadine, desloratadine, and 3-hydroxydesloratadine as well as yohimbine and tadalafil demonstrated a tight binding to the in silico folded BmtA transporter. We, then, tested borreliacidal activity and dose response of the shortlisted compounds from this screen using a series of in vitro assays. Amongst the probed compounds, desloratadine exhibited potent borreliacidal activity in vitro at and above 78 μg/mL (250 μM). Borrelia treated with lethal doses of desloratadine exhibited a significant loss of intracellular Mn specifically and a severe structural damage to the bacterial cell wall. Our results support the possibility of developing a novel, targeted therapy to treat Lyme disease by targeting specific metabolic needs of Borrelia.

    View details for DOI 10.2147/DDDT.S77063

    View details for Web of Science ID 000349239100003

    View details for PubMedID 25709405

  • The potential of apolipoprotein E4 to act as a substrate for primary cultures of hippocampal neurons BIOMATERIALS Kim, K., Vicenty, J., Palmore, G. R. 2013; 34 (11): 2694–2700

    Abstract

    The E4 isoform of apolipoprotein (apoE4) is known to be a major risk factor for Alzheimer's Disease (AD). Previous in vitro studies have shown apoE4 to have a negative effect on neuronal outgrowth when incubated with lipids. The effect of apoE4 itself on the development of neurons from the central nervous system (CNS), however, has not been well characterized. Consequently, apoE4 alone has not been pursued as a substrate for neuronal cultures. In this study, the effect of surface-bound apoE4 on developmental features of rat hippocampal neurons was examined. We show that apoE4 substrates elicit significantly enhanced values in all developmental features at day 2 of culture when compared to laminin (LN) substrates, which is the current substrate-of-choice for neuronal cultures. Interestingly, the adhesion of hippocampal neurons was found to be significantly lower on LN substrates than on glass substrates, but the axon lengths on both substrates were similar. In addition, this study demonstrates that the adhesion- and growth-enhancing effects of apoE4 substrates are not mediated by heparan sulfate proteoglycans (HSPGs), proteins that have been indicated to function as receptors or co-receptors for apoE4. In the absence of lipids, apoE4 appears to use an unknown pathway for up-regulating neuronal adhesion and neurite outgrowth. Our results indicate that apoE4 is better than LN as a substrate for primary cultures of CNS neurons and should be considered in the design of tissue engineered CNS.

    View details for PubMedID 23352042

  • A novel method for analyzing images of live nerve cells JOURNAL OF NEUROSCIENCE METHODS Kim, K., Kim, S., Minxha, J., Palmore, G. R. 2011; 201 (1): 98–105

    Abstract

    Analysis of images from live-cell experiments is a central activity to studying the effects of stimulation on neuronal behavior. Image analysis techniques currently used to study these effects rely for the most part on the salience of the neuronal structures within the image. In both fluorescent and electron microscopy, neuronal structures are enhanced and therefore easy to distinguish in an image. Unlike images obtained via fluorescent or electron microscopy, however, images produced via transmission microscopy (e.g., bright field, phase contrast, DIC) are significantly more difficult to analyze because there is little contrast between the object-of-interest and the image background. This difficulty is amplified when a time-dependent sequence of images are to be analyzed, because of the corresponding large data sets. To address this problem, we introduce a novel approach to the analysis of images of live cells captured via transmission microscopy that takes advantage of commercially available software and the Fourier transform. Specifically, our approach utilizes several morphological functions in MATLAB to enhance the contrast of the cells with respect to the background, which is followed by 2-D Fourier analysis to generate a spectrum from which the orientation and alignment of cells and their processes can be measured. We show that this method can be used to simplify the interpretation of complex structure in images of live neurons obtained via transmission microscopy and consequently, discover trends in neurite development following different types of stimulation. This approach provides a consistent and reliable tool for analyzing changes in cell structure that occurs during live-cell experiments.

    View details for PubMedID 21816174

  • Quantitative Control of Neuron Adhesion at a Neural Interface Using a Conducting Polymer Composite with Low Electrical Impedance ACS APPLIED MATERIALS & INTERFACES Kim, S., Kim, K., Hoffman-Kim, D., Song, H., Pamore, G. R. 2011; 3 (1): 16–21

    Abstract

    Tailoring cell response on an electrode surface is essential in the application of neural interfaces. In this paper, a method of controlling neuron adhesion on the surface of an electrode was demonstrated using a conducting polymer composite as an electrode coating. The electrodeposited coating was functionalized further with biomolecules-of-interest (BOI), with their surface concentration controlled via repetition of carbodiimide chemistry. The result was an electrode surface that promoted localized adhesion of primary neurons, the density of which could be controlled quantitatively via changes in the number of layers of BOI added. Important to neural interfaces, it was found that additional layers of BOI caused an insignificant increase in the electrical impedance, especially when compared to the large drop in impedance upon coating of the electrode with the conducting polymer composite.

    View details for PubMedID 21142128