2009 — 2012 |
Wei, Jianning |
R15Activity Code Description: Supports small-scale research projects at educational institutions that provide baccalaureate or advanced degrees for a significant number of the Nation’s research scientists but that have not been major recipients of NIH support. The goals of the program are to (1) support meritorious research, (2) expose students to research, and (3) strengthen the research environment of the institution. Awards provide limited Direct Costs, plus applicable F&A costs, for periods not to exceed 36 months. This activity code uses multi-year funding authority; however, OER approval is NOT needed prior to an IC using this activity code. |
Regulation of Bimel Phosphorylation in the Pathogenesis of Huntington's Disease @ Florida Atlantic University
DESCRIPTION (provided by applicant): It is clear that the mutant huntingtin (mHtt) forms intracellular aggregates and induces apoptosis in Huntington's disease (HD). However, the underlying molecular mechanism remains elusive. The long-term goal of the project is to understand the molecular pathways that are altered by mHtt expression. Recent studies suggest a functional link between accumulation of mHtt and proteasomal dysfunction in context of mHtt-induced cell death in HD. However, it is unclear how mHtt-induced proteasomal dysfunction can be molecularly converted to neuronal death. In a series of pilot experiments, we began to discover that BimEL expression is significantly up-regulated in two different cell lines overexpressing mHtt. More interestingly, the up-regulation of BimEL is largely due to BimEL phosphorylation rather than the transcriptional regulation. Based on our preliminary findings, we specifically hypothesize that the pro-apoptotic BH3-only protein, BimEL, is the molecule that functionally links mHtt aggregates formation and apoptosis. In this application, we propose to test this hypothesis in the R6/2 mouse model of HD. In the first specific aim, we propose to confirm our preliminary findings in R6/2 mice. BimEL expression in the striatum of R6/2 and control mice will be examined at both transcriptional and post-translational levels using quantitative RT-PCR and western blot respectively. We next propose to determine the molecular mechanism that up- regulates BimEL expression in R6/2 mice with a special focus on the regulation of BimEL phosphorylation. BimEL phosphorylation and the related protein kinases activity will be examined by western blot analysis using specific phospho antibodies. Finally, we propose to determine the physiological relevance of BimEL phosphorylation and up-regulation in HD. mHtt-induced death in the presence or absence of silencing BimEL expression will be investigated by flow cytometry analysis. Moreover, Bax translocation to mitochondrial membrane will be analyzed by western blot analysis under different conditions that favor or prevent BimEL phosphorylation. Information derived from the proposed studies will improve the current understanding of the molecular pathways that are altered in response to mHtt in HD and may represent a universal mechanism in the pathogenesis of neurodegenerative diseases that are involved with protein misfolding and aggregation. PUBLIC HEALTH RELEVANCE: Huntington's disease (HD) is a devastating neurodegenerative disorder caused by an expanded polyglutamine repeat within the huntingtin protein. Today, approximately 30,000 Americans are living with HD. In addition, a staggering 200,000 more are at risk. Identifying the pathways that are altered in response to the mutant protein is crucial for understanding the cellular processes impacted by the disease as well as for the rational development of effective pharmacological interventions.
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2017 — 2020 |
Du, E Engeberg, Erik Hutchinson, Douglas T Tognoli, Emmanuelle (co-PI) [⬀] Wei, Jianning |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Sch: Int: Virtual Neuroprosthesis: Restoring Autonomy to People Suffering From Neurotrauma @ Florida Atlantic University
By reconnecting the previously severed sense of touch, the field of neuroprosthetics has tremendous potential to substantially improve the lives of millions of amputees and disabled people worldwide. However, the rate of progress to develop neuroprosthetic limbs has been comparatively slow relative to other areas of robotics for two primary reasons: research involving neural implants with human subjects is very expensive and a lengthy process is required to obtain FDA approval to implant electrodes in human subjects. Thus, the overall goal of this project is to develop a virtual neuroprosthesis in which a facsimile of a neural implant is externalized and housed in a well-controlled microfluidic chamber, thereby abating the intrinsic limitations of highly invasive studies with neural implants. Upper limb amputee subjects will be recruited to control a dexterous artificial hand and arm with electromyogram signals while electroencephalogram (EEG) signals are simultaneously measured. Robotic grip force measurements will be biomimetically converted into electrical pulses similar to those found in the peripheral nervous system to catalyze in. vitro nerve regeneration after neurotrauma. The synergistic contributions of this multidisciplinary project will lead to a transformative understanding of the symbiotic interaction of neural plasticity within human-robotic systems. Currently, there is no systematic understanding of how tactile feedback signals can contribute to the neural regeneration of afferent neural pathways to restore somatosensation and improve motor function in amputees fitted with neuroprosthetic limbs. Tackling this problem will be a significant breakthrough for the important field of neuroprosthetics. The proposed virtual neuroprosthesis will be much less expensive and vastly simpler to obtain IRB approval to conduct research with human subjects. Through this, the research team can conduct meaningful neuroprosthetic experiments with human subjects at a fraction of the cost while accumulating significant data much quicker.
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2019 |
Du, E Engeberg, Erik Tognoli, Emmanuelle (co-PI) [⬀] Wei, Jianning |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Diversity Supplement For Virtual Neuroprosthesis @ Florida Atlantic University
Project summary The key benefit of our virtual neuroprosthetic framework proposed in the parent R01 grant (1R01EB025819) is that multiple hypotheses can be tested simultaneously using dorsal root ganglia (DRG) cultures in a cost- effective manner. The overall goal of this supplement grant is to augment the haptic signaling pathways that are restored in the virtual neuroprosthetic platform from one to four, consistent with human anatomy. Compared to the parent R01 in which we proposed to model only one pathway for a type of slowly adapting mechanoreceptor in the fingertip (continuously signaling the grasp force applied by the robotic hand), we propose herein to explore multiple parallel channels to mimic the complex functional anatomy of the hand which contains at least four different sensory receptor types carrying information on different aspects of the haptic experience (pressure, vibrational information used to detect slippage, texture, hand conformation, etc). These signals provide important complementary haptic information for the fine control of movements. Therefore, neurostimulation of DRG cultures mimicking the signals from different mechanoreceptors will produce distinctive spatial, temporal and functional forms of DRG regeneration post-axotomy, and the resulting functional diversity of regenerated pathways will ultimately play a key role in the overall quality of somatosensory restoration after amputation, supporting better usage of haptic feedback-enabled hand protheses. Specifically, we propose to (1) design highly realistic electrical stimulation patterns from the four principal types of slowly adapting (SA) and rapidly adapting (RA) mechanoreceptors to electrically stimulate DRGs cultured in microfluidic chambers, (2) to study the contribution of those multiple pathways, individually and synergistically, to anatomical and functional restoration, and (3) to establish if this restoration scheme surpasses one with the lower attentional load but lesser informational richness obtained when the haptic feedback is derived from only one functional pathway, as is currently under investigation within the parent R01.
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2019 |
Capobianco, Enrico Wei, Jianning |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Dynamic Network Analysis of Huntingtin Interactome in Response to Cellular Stresses @ Florida Atlantic University
Neurons are selectively venerable with a low stressor-threshold in neurodegenerative diseases. At molecular levels, responses to cellular stresses are mediated by dynamic protein-protein interactions (PPI). Developing in-depth, dynamic PPI networks is therefore crucial to understand the pathogenesis of these diseases. Huntington?s disease (HD) is an inherited fatal neurodegenerative disorder caused by a mutation in the huntingtin (htt) gene. It is strongly suggested that Htt serves as a scaffold protein interacting with multiple protein complexes that are involved in diverse cellular functions. Our long-range goal is to understand the molecular functions of Htt and muHtt at various biological states. The objective of this multi- PI proposal is to map Htt/muHtt interactome under the proteotoxic stress. Our central hypothesis is that normal Htt remodels its interactome in response to cellular stresses and this capability is compromised in the presence of muHtt, causing accumulation of cellular damages overtime and eventually neurodegeneration. Specifically, the following two aims are proposed. Aim 1: Map the dynamic Htt/muHtt interactome in response to proteotoxic stress by unbiased quantitative proteomic and bioinformatic analyses. We will first establish an ascorbate peroxidase (APEX2)-based proximity labeling platform to spatiotemporally label Htt-interacting proteins in live cells. A striatal STHdhQ7 neuronal cell line stably expressing Htt-APEX2 will be subjected to three different conditions (normal, proteotoxic stress and stress recovery) followed by APEX2 labeling. Biotinylated proteins will then be identified by quantitative proteomics. The resulting protein list will be subjected to in-depth bioinformatic analyses. Aim 2: Quantitatively analyze the molecular responses of known Htt-interacting proteins to cellular stresses in normal and HD cells. We will focus on analyzing a signaling hub protein, p62, which directly interacts with Htt. Our working hypothesis is that Htt regulates the molecular responses of p62 to cellular stresses and the regulation is impaired in the presence of muHtt. To test this hypothesis, molecular changes of p62 to various stresses in normal and HD cells will be evaluated at (1) mRNA levels, (2) protein expression and (3) subcellular localization. The interaction between p62 and Htt/muHtt under various stresses will be quantified using the Htt-APEX2 platform. We are well-positioned to undertake the proposed study because our research team consists of uniquely qualified individuals with combined expertise in molecular neurobiology and large data analysis. Successful completion of these studies will contribute fundamental knowledge about molecular functions of normal and mutant Htt and the pathogenesis of HD. In a broader aspect, building dynamic interaction networks under diverse stress conditions could be the key to understand the molecular differences between healthy and any pathological states. The proposed research is highly innovative for its novel idea and approaches to study the dynamic nature of Htt interactome in response to stresses.
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2020 |
Du, E Engeberg, Erik Tognoli, Emmanuelle (co-PI) [⬀] Wei, Jianning |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Alzheimer's Disease Supplement For Virtual Neuroprosthesis @ Florida Atlantic University
Project Summary/Abstract This supplementary research proposal applies the expertise, techniques and hardware developed under the funded parent grant (R01EB025819) to study the contribution of neuronal activity to the pathogenesis of Alzheimer?s disease (AD). AD is a progressive multifactorial neurodegenerative disorder and the major type of dementia. Neuronal activity shows a complex relationship with AD, with (1) neurons and areas susceptible to more intense neuronal activity, clinically hyper-excitable or stimulated intensely exhibiting evidence of AD pathogeny in humans, animal models and cell cultures; and (2) neurons or synapses whose activity is reduced or putatively under-stimulated by lack of cognitive engagement also demonstrating altered prospects as the disease progresses. Since neuroinflammation is one of the central mechanisms in AD pathogenesis and an area currently under intensive research, elucidation of its systemic drivers at the neuronal level and pathogenic impact will provide mechanistic insights on disease progression and uncover intervention principles. We hypothesize that one of key mediators of this nonlinear relationship between neuronal activity and AD is neuroinflammation, because studies independently linked neuroinflammation to both sides of the hypothetical equation AD=f(neuronal activity). Specifically in this application, using an Alzheimer?s-in-a-dish model with neurons- microglia cocultures derived from induced pluripotent stem cells (iPSCs) of AD patients, we aim to determine the electrical parameters that modulate neuroinflammatory response and how this relates to AD progression. Our operational hypothesis is that different patterns of electrical stimulation will nonlinearly affect neuroinflammatory responses in AD neuron-microglia co-cultures, which in turn contributes to the pathogenesis of AD at the neurons? structural and functional levels. To test this hypothesis, we propose to deliver different patterns of electrical stimulation to Alzheimer?s-in-a-dish models and measure cytokine release using cytokine array (Exp. 1), analyze microglia migration behavior with impedimetric monitoring (Exp. 2) and investigate neuronal function electrophysiologically with multielectrode array (MEA) (Exp. 3). This plan is executed by an interdisciplinary team of 4 principle investigators whose skills cover broadly the needs of the research plan, including an expert in the molecular biology of neurodegeneration/regeneration, a roboticist expert in control systems; a biosensor and microfluidic nanoengineer and a complexity neuroscientist expert in electrophysiological spatiotemporal dynamics. This supplement grant is directly focused on investigating AD pathogenesis in the presence of different neuronal stimulation patterns, which resemble physiological or pathological neuroelectric events responding to environmental sources. Our proposed studies, thus well aligned with Milestone 2H outlined in the research implementation plans by National Institute of Aging (NIA), provide a novel in vitro platform with integrated electrical stimulation and cellular components and a broadly integrated analysis toolkit to gain more detailed mechanistic insights on neuronal activity-mediated AD progression.
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