Grants $50,000 to $100,000

Market NY - RC5 - EDF

Plattsburgh Chamber of Commerce Working Capital

NSF

This project aims to serve the national interest by improving students’ understanding of how social, cultural, economic, and political systems can inform their engineering designs. Traditionally, engineering design courses have focused primarily on the technical specifications and requirements of a design without full consideration of the wider context in which they will be used. There is also a lack of engineering design courses in the middle two years of a typical engineering curriculum. This leads to a gap in learning design principles and skills for students between their first design course and the final capstone design course. The project plans to develop, implement, refine, and evaluate engineering design courses for second and third year students at the University of Illinois-Chicago and the University of Texas-San Antonio. The courses should help students learn how to conduct in-depth analyses of the implications of design choices for society using historical and current examples of engineering design. This project intends to develop an instructional framework for the middle years of engineering programs' courses along with instructional materials for those courses. Using interviews and student surveys, the project will assess the impact of the courses on students. Project results will be disseminated to engineering educators through professional development workshops. The goal of this project is to help students learn how to consider social, cultural, economic, and political factors during the engineering design process. This project is based on Critical Consciousness theory which suggests that students need to develop an awareness of inequitable societal conditions that can occur due to engineering design decisions. A new teaching framework for engineering design in the middle years will be developed and refined so that engineering design includes a contextual perspective. The goals for the new engineering design course are: (1) to empower students to be more socially and critically driven engineers, (2) to help students learn about the engineering design process from a Critical Consciousness perspective, and (3) to help students become validated in their aspirations to pursue an engineering career. An inter-group dialogue approach will be used in the course to establish communication relationships, facilitate dialogue, and encourage collaborations between students. This study will address two research questions: (1) What teaching strategies are most helpful in developing students’ critical consciousness through an engineering design course? (2) How and in what ways do the programmatic features of the project impact students’ learning of engineering design and engineering identity development? The project team will use quantitative and qualitative methods to analyze data from classroom observations, course artifacts, and student surveys. The NSF IUSE: EHR Program supports research and development projects to improve the effectiveness of STEM education for all students. Through the Engaged Student Learning track, the program supports the creation, exploration, and implementation of promising practices and tools. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

Thomas Darling House Woodbridge Historical Society

Historic Restoration Fund

NYC Department of Cultural Affairs

Electronic Arts Intermix

NIAID - National Institute of Allergy and Infectious Diseases

ABSTRACT Autophagy, a conserved catabolic process mediated by autophagy-related (ATG) proteins, is integral for maintaining cellular homeostasis. Recent studies suggest that ATG proteins are also necessary for immune responses through processes that are mechanistically distinct from autophagy. The Cadwell lab has identified a novel innate immune mechanism whereby cells produce extracellular vesicles of endosomal origin (exosomes) which can neutralize bacterial toxins and inhibit viral pathogens in the lung and bloodstream. The production of these extracellular vesicles, termed “defensosomes”, is induced by innate immune sensors such as Toll-like receptor 9 (TLR9) and dependent on the ATG protein ATG16L1. However, the molecular pathways governing defensosome biogenesis and release remain elusive. This study aims to elucidate these mechanisms, focusing on the roles of ATG16L1 and its binding partner TMEM59 in defensosome biogenesis, as well as how TLR9 signaling modulates RAB7 activity for defensosome release. We will use a tractable lung epithelial cell culture system and measure defensosomes released in the blood stream of mice as an in vivo readout. Aim 1 investigates how the interaction between ATG16L1 and TMEM59 promotes defensosome biogenesis. We hypothesize that ATG16L1 and TMEM59 facilitate the formation of intraluminal vesicles within multivesicular endosomes. Aim 2 examines how TLR9 signaling influences RAB7 activity to incite defensosome release. We propose that TLR9 activation leads to the upregulation of a GTPase-activating protein, promoting RAB7 dissociation from MVEs, thus diverting defensosome precursors to the plasma membrane for release. This research will provide critical insights into the molecular mechanisms of defensosome biogenesis and release, while also enhancing the understanding of the intersection between autophagy and innate immunity, additionally contributing to the broader field of extracellular vesicle biology.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY Mitochondria serve as a central signaling hub for innate immune responses. Disruption of mitochondrial function is a hallmark for various infections and chronic inflammatory diseases. The overall objective of this proposal is to define the molecular mechanisms that regulate mitochondrial membrane homeostasis and determine how membrane disruption promotes inflammatory cell death. Mutations in leucine-rich repeat kinase 2 (LRRK2) are associated with disrupted mitochondrial integrity and increased reactive oxygen species. They are also associated with increased susceptibility to hormonal breast cancer, Crohn's disease, and mycobacterial infection, strongly suggesting a role in immune function. Recently, the Watson lab set out to investigate LRRK2's role in peripheral innate immunity, focusing on a gain of function mutation, Lrrk2G2019S. Mitochondrial stress conferred by the Lrrk2G2019S mutation increases demand on the electron transport chain, which leads to excessive ROS production. This increased ROS triggers a new type of cell death where a protein canonically associated with pyroptosis, gasdermin D (GSDMD), can associate with mitochondrial membranes and cause necroptotic cell death. Aim 1 of this proposal will identify the minimal domain within GSDMD that targets mitochondrial membranes, enabling a better understanding of the molecular mechanisms underlying GSDMD's newly described role in necroptosis. Aim 2 will investigate the contribution of various aspects of mitochondrial dysfunction to GSDMD mitochondrial targeting and necroptosis, providing new insights into connections between the disruption of mitochondrial homeostasis and GSDM relocalization. With the goal of understanding how mitochondria are impacted by genetic mutations and/or stress, Aim 3 will measure relocalization of the mitochondrial inner membrane phospholipid cardiolipin in WT and Lrrk2G2019S macrophages and catalog mitochondrial lipids in WT vs. Lrrk2G2019S macrophages. Defining the molecular mechanisms that drive inflammation in the face of specific mitochondrial mutations will help enable therapeutic interventions designed to correct specific aspects of mitochondrial dysfunction associated with a variety of inflammatory, infectious, cardiac, and neurological disorders.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY The consequences of maternal immune activation (MIA) are currently being explored and already show that inflammation during gestation can influence offspring behavior and immunity. However, the influence of paternal immune activation (PIA) on offspring is an understudied topic. Limited studies have shown that paternal immune activation, either through pathogen exposure or stimulation of the immune system with synthetic molecules before conception results in altered offspring behavior. Moreover, these events influence the epigenetic information within the sperm. Yet, we lack understanding of how immune stimulation alters the expression of epigenetic molecules within germ cells and if PIA can also shape offspring immune development. With this in mind, I sought to understand how immune stimulation due to viral infection in male parents could influence offspring immunity. First, I evaluated the impact of viral infection (Zika virus) or immune stimulation with a viral mimetic (Poly (I:C)) on the small RNAs, a carrier of epigenetic information, within sperm. I determined that immune stimulation, with or without viral replication resulted in an altered sperm small RNA profile. I have explored the impact of these modifications have on offspring at two developmental stages: during embryonic development and in juvenile aged offspring. Using sperm from Poly (I:C) treated males to fertilize naïve eggs, I found that gene expression in morula stage embryos was altered, indicating the sperm small RNA profile established by PIA influences early embryonic development. Moreover, when juvenile offspring sired by Poly (I:C) treated males were infected with influenza, I found these offspring had a higher survival rate and fewer signs of infection. Importantly, this is the first evidence that PIA may confer resistance to viral infection to offspring. Taken together, my central hypothesis is that proinflammatory cytokines induced during PIA lead to changes in the sperm small RNA profile which program early embryonic gene expression and confer lasting viral resistance to offspring through altered immune function. For my first aim, I will evaluate if this resistance to infection lasts into adult hood and what differences in offspring immunobiology allow for this resistance. For my second aim, I will determine if paternal immune activation modifies the sperm small RNA profile though cytokine signaling and if these cytokines induced changes are responsible for the offspring phenotypes I have observed. Completion of this proposal will provide new understanding into how paternal immune activation can impact offspring immunity. This study will provide key insights to developmental immunobiology and directly challenge our current interpretation of the evolutionary arms race between host and pathogen.

NICHD - Eunice Kennedy Shriver National Institute of Child Health and Human Development

Abstract Complex multicellular organisms develop through hundreds of precisely orchestrated genetic interactions that can lead to deleterious phenotypes like disease when disrupted. Many genes require conservation in their expression to perform their specific functions and activities in concert with other genes. Yet, developmental programs and regulatory networks are frequently modified to produce beneficial, novel, or alternate phenotypes. The most obvious example of this is the evolution of adaptive sexual dimorphisms within and across species. This suggests a degree of flexibility in highly conserved developmental systems that can be accessed by genetic variation. However, the molecular mechanisms by which genetic variation relates to developmental pathway architecture remains poorly understood. A powerful and natural system in which to investigate this is phenotypic polymorphism, defined as the co-occurrence of multiple distinct phenotypes within a species. Alternate phenotypes, including adaptive polymorphisms and disease states, arise from distinct developmental programs that are shaped by underlying genetic variation, often at just one or a few genes. My proposed work aims to characterize the link between genetic variation and developmental variation by using a natural system of female- limited polymorphism in Papilio butterflies. In several species, the gene doublesex controls the development of female-limited wing pattern polymorphism. Ancestrally, doublesex (dsx) regulates sex differentiation by controlling the development of multiple secondary sexual traits across insects. In addition to performing its role in sexual differentiation, alternate dsx alleles cause females to develop either a derived mimetic or an ancestral non-mimetic color pattern. Both female forms co-occur, with males developing a single phenotype. My proposed work will uncover and experimentally test what mutation(s) caused dsx to be reused to control the color pattern development switch, and characterize the cis-regulatory architecture controlling allele-specific expression. Additionally, I will characterize how dsx integrates with or modifies the wing development program by identifying the direct targets of DSX and associated gene expression changes. Collectively, these experiments will tease apart genetic interactions underlying developmental program modifications at a novel level of precision. Moreover, by contrasting results from closely-related species with and without female-limited polymorphism, my proposed work will highlight the common and unique elements of wing development programs that have been modified by the insertion of the same reused gene. This will bear directly on our understanding of how shared genes and programs are (re)deployed across diverse genetic and genomic backgrounds to produce new phenotypes.

NIDCD - National Institute on Deafness and Other Communication Disorders

Neural circuits allow animals to gather various types of sensory information from the complex environment and integrate this information to produce the appropriate behavioral responses. To decide whether to ingest potential food substances, animals must discriminate between nutrients and toxins. To this end, they integrate sensory information, such as taste, smell, texture, temperature, and visual cues, with internal states, such as hunger and satiety. It is well established that the integration of taste and smell, perceived as flavor in humans, is especially important for food discrimination. However, the precise points of integration between the taste and smell circuits remain unknown in humans due to the complexity of the nervous system. Studies monitoring feeding behavior upon smell stimulation in the fruit fly, Drosophila melanogaster, suggest that the taste and smell circuits also integrate in the fly. Since the neural circuits in fruit flies are simpler than those in humans, flies are an ideal organism for evaluating the anatomical and functional connections between taste and smell. Our laboratory has developed trans-Tango, a method for neural circuit mapping and manipulation in fruit flies. Using trans-Tango, we mapped the first and second-order neurons in the taste and smell circuits, showing that gustatory receptor neurons, which detect tastants, and olfactory receptor neurons, which detect odors, relay information to gustatory and olfactory projection neurons, respectively. Some of these projection neurons target the same higher-order brain areas, suggesting the possibility that shared neurons exist that integrate sensory inputs from both systems to influence feeding behavior. This proposal takes a two-pronged approach to investigate the integration of the gustatory and olfactory circuits. First, I will test how olfactory inputs affect feeding by activating, or silencing, olfactory projection neurons tuned to food-derived odors. In these studies, I will use the OptoPAD paradigm to measure feeding. I hypothesize that activating neurons tuned to attractive odors would enhance feeding, while activating those tuned to aversive odors would suppress it. Second, I will identify neurons in the lateral horn that receive inputs from both gustatory and olfactory projection neurons and integrate these inputs to produce the appropriate feeding responses. To this end, I have been developing trans-Tango(hub), a tool for identifying circuit nodes of integration. My experiments will determine whether these nodes maintain the valence of the stimuli. Further, since the gustatory and olfactory systems in insects and mammals are functionally homologous, identifying the mechanisms of this multisensory integration in fruit flies will provide insight into how the perception of flavor is formed in humans. This is crucial for understanding the pathologies associated with olfactory deficiencies, such as anosmia and hyposmia, and gustatory deficiencies, such as ageusia. Finally, this research program is at the core of a training plan that includes activities to develop professional skills for preparing Angel Okoro for a career in academic research.

NICHD - Eunice Kennedy Shriver National Institute of Child Health and Human Development

Abstract This study seeks to elucidate the impact of early postnatal oxytocin signaling on cerebellar development and function. The cerebellum shapes motor behavior, cognition, and emotion and, in consequence, its functional disruption during early development is one of the highest risk factors for neurodevelopmental disorders. Due to its protracted development, anatomical location, and dense vascularization, the cerebellum is highly susceptible to perinatal insults. The process of parturition for instance exposes the infant cerebellum to high levels neurohormones like oxytocin. Previous studies suggest that neonatal oxytocin exposure contributes to functional maturation of cortical and hippocampal circuits, but the impact of those hormone surges on cerebellar development remain unknown. Recent transcriptomic profiling of the developing mouse cerebellar cortex revealed that GABAergic progenitors transiently express oxytocin receptor mRNA, peaking around postnatal day 4, and then steeply declining. My goal is to determine the role of oxytocin signaling in the development of GABAergic neurons and cerebellar circuit formation. I will use a combination of molecular, functional, and genetic approaches in mice. In aim 1, I will use gene expression analysis and receptor labeling to determine the timing of Oxtr and OTR expression in molecularly identified cell types. In aim 2, I will use in vitro electrophysiology to identify the impact of oxytocin on the activity of immature GABAergic neurons and the GABA functional switch from excitatory to inhibitory. In aim 3, I will probe how oxytocin regulates emerging excitatory and inhibitory connectivity across development and probe the long-term impact of oxytocin on synaptic plasticity. Completion of this study will provide mechanistic insight into how transient oxytocin receptor signaling shapes circuit development.

NIAID - National Institute of Allergy and Infectious Diseases

Project Abstract: Significant advances in antiretroviral therapy (ART) has enabled functional suppression of HIV, allowing individual individuals on ART to live relatively normal lives. However, viral rebound from a persistent reservoir of provirus-harboring cells following ART cessation demands a lifelong reliance on ART. ART-free control of this persistent reservoir therefore represents the current focus of HIV cure strategies. Recent work from our group and others has identified HIV-infected CD4 T cells that exhibit cell-intrinsic resistance to CD8 cytotoxicity, despite antigen expression and recognition. While identification of resistance factors previously annotated in cancer (such as overexpression of the granzyme inhibitor SERPINB9 and the anti-apoptotic protein BCL-2) has provided some insight concerning underlying mechanisms, the factors leading to this resistance phenotype remain largely unknown. To address this, we have developed a novel genome-wide screening platform that allows identification of genes involved in primary human CD4 T cell resistance and susceptibility to CD8 T cell cytotoxicity. In this proposal, we aim to maximize the potential of this newly developed CD4 resistance screening model (CRSM). Additional screens in different contexts will be conducted to further enrich the dataset. We will also employ a methodical pipeline for hit validation and therapeutic characterization. This pipeline will begin with promising hits identified in the initial screen, including the lipid metabolism-associated PPAR family proteins. This proposal will produce a robust functional genomic dataset thoroughly characterizing the mechanisms underlying CD4 resistance to CD8 killing. With these datasets, we envision the nomination of a range of promising targets for small-molecule sensitization of HIV-infected CD4s to CD8 killing.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY/ABSTRACT Chronic Hepatitis B Virus (HBV) infection affects nearly 300 million individuals and is a leading cause of hepatocellular carcinoma worldwide. HBV persistence relies on the establishment of 1) the viral covalently closed circular DNA (cccDNA) minichromosome which serves as that viral transcriptional template and 2) the expression of the regulatory protein HBx which prevents cccDNA silencing. Due to limited in vitro and animal models that recapitulate transient events like cccDNA formation early during infection, the mechanisms through which HBx recognizes and regulates cccDNA remain poorly understood. The David lab strives to better understand the molecular mechanisms of chromatin regulation in disease. Towards this end, we have pioneered novel platforms to study HBV biology with the first methods to reconstitutes scarless cccDNA with infectious capacity and full- length HBx protein for biochemical and biophysical studies. We discovered that HBx directly binds to chromatin to induce decompaction in vitro and found that cccDNA histone occupancy is required for HBx expression early during infection. My proposal extends these findings by seeking to elucidate host epigenetic regulation of HBV transcription (Aim 1) and HBx-mediated cccDNA recognition (Aim 2). My working hypothesis is that HBx directly recognizes and decompacts the viral minichromosome to allow host epigenetic machinery to distinctly control viral transcription. We have shown that host histone incorporation onto cccDNA is important for HBV transcription. Importantly, we found that pharmacological disruption of viral chromatin assembly leads to reductions in HBV transcription, antigen secretion, and viral replication in hepatocyte models of infection. The proposed Aim 1 will identify host histone variants and chromatin remodelers that influence HBV transcriptional control. First, I will probe for the presence of active transcription- associated histone variants deposited onto cccDNA during infection using chromatin immunoprecipitation. I will then determine the effect on viral chromatin assembly and transcription of the loss of host epigenetic regulators implicated in cccDNA chromatin regulation using a limited shRNA screen and auxin-inducible degron tagging. In Aim 2, I will use biophysical approaches, such as biolayer interferometry, to define the determinants of HBx- nucleosome binding. I will then investigate changes in viral chromatin compaction due to this interaction through Mg2+ precipitation and MNase protection assays. If validated in vitro, I will expand to testing this phenomenon in cellulo using dCas9 to fluorescently label cccDNA and ATAC-see to measure and visualize host- and cccDNA- specific changes in chromatin accessibility in the absence and presence of HBx. Overall, Aim 2 will reveal new insights into HBx’s role in viral chromatin regulation. Successful completion of this project will reveal mechanisms for how HBx modulates HBV chromatin and uncover novel therapeutic targets for silencing or eradicating cccDNA, improving epigenetic strategies to develop a functional cure for chronic HBV infection.