Temple Univ Of The Commonwealth

NIH Research Projects · FY 2025 · 2021-06

Platelets play a crucial role in hemostasis and thrombosis, and more and more studies indicate their role in other disease states including inflammation, cancer, and atherosclerosis Platelets express a number of surface receptors, which through their activation, allow platelets to interpret their local environment and to detect vascular lesions and promote hemostasis. My group focuses on these signaling steps and how their interplay mediates platelet activation. Understanding signaling networks and their regulation has been my research focus for the past two decades and our group has made important contributions to the platelet-signaling field. My research goals are to identify novel signaling molecules that regulate main signaling pathways, characterize novel signaling pathways emanating from the same signaling molecule, and understand the differences in various tyrosine kinase pathways in platelets. My additional goals are to understand what changes occur in platelet composition, including miRNAs, with age and disease, such as diabetes, that make them more susceptible to thrombotic events. This work builds on our past contributions in the field and a host of reagents and genetic tools that we have amassed. In this proposal, we place particular emphasis on the intracellular interactions that regulate a signaling molecule. One of the novelties of the studies proposed is that the same protein kinase, through differential tyrosine phosphorylation, activates diverse signaling pathways, which have distinct roles in hemostasis. The studies proposed in this application will provide further insights into the regulation and identification of novel signaling pathways in platelets, which may be applicable to other cell systems expressing similar receptors and could form the basis for novel therapeutic targets to treat thrombosis and thrombocytopenia. In addition, understanding these signaling cascades in platelets will help us evaluate and predict possible implications of the therapeutic agents that could interfere with these pathways. For example, our studies anticipate that Ibrutinib, a Tec kinase inhibitor used for the treatment of chronic lymphocytic leukemia, will block the CLEC2 pathway in platelets and cause blood flow into lymphatic vessels. I would like to pursue these goals in the next decade with the same vigor and intensity that have employed in the past two decades. have been funded by NIH for about 22 years on the platelet signaling paradigms and have published over 180 papers (on an average of 8 papers a year). The OIA will alleviate the need to submit separate thematic grant applications to various agencies with coherent specific aims and will allow us to make significant contributions to the understanding of platelet signaling networks. Our overarching goal is to understand how the network of receptor- mediated signaling can be manipulated to control platelet function.

NIH Research Projects · FY 2025 · 2021-04

Project Summary Older adults are often victims of financial exploitation, which results in annual losses of at least $3 billion. Threat of exploitation may be greater in those at risk for Alzheimer's Disease and Related Dementias (ADRD). While most cases of financial exploitation are perpetrated by strangers, perpetration by individuals within the victim's social network (e.g., friends and family) is common. Although these observations highlight the social nature of financial exploitation, we know very little about how neural systems in older adults and those at risk for ADRD integrate information from the social domain to inform financial decision making. Tasks involving social information processing evoke activation in the temporal-parietal junction (TPJ) and dorsomedial prefrontal cortex; whereas tasks involving financial decision making and reward processing evoke activation in the striatum and ventromedial prefrontal cortex. It remains unclear whether responses within these neural systems—and connectivity between them and other regions—differ as a function of age and risk for financial exploitation. We recently demonstrated age differences in TPJ response during social decision-making in the context of financial exchange (e.g., trust and fairness). We have also shown that the influence of social context (e.g., presence of a peer) on striatal responses to reward is attenuated in older adults relative to younger adults. Building on these exciting preliminary data, the goal of the current proposal is to characterize neural responses during social decision making and reward processing across adulthood and quantify how these responses relate to risk for financial exploitation. We will recruit a large sample of adults (ages 21 to 80+ years) to participate in a neuroimaging experiment investigating social reward processing and the interplay between social context and financial decision making. We will also administer neuropsychological and health assessments, as well as questionnaires assessing socioemotional functioning and risk for financial exploitation. Our project will address four aims. We will examine how neural responses to social and nonsocial reward and decisions based on trust and fairness differ across the lifespan (Aim 1). Although neural responses evoked by our tasks may be associated with risk for financial exploitation, it is imperative to determine how such responses interact with sociodemographic factors, cognitive decline, socioemotional functioning, and health (Aim 2). Notably, our quantification of health status will leverage two understudied risk factors for ADRD—vascular health and white matter hyperintensities—and relate these factors to financial exploitation. Finally, we will re-test a subset of participants with mild cognitive impairment (MCI) two years after their baseline visit and assess changes in social reward processing and risk for financial exploitation (Aim 3). Overall, our project will generate new insights into the interplay between social and financial decision making across the lifespan and individuals at risk for ADRD and start the road to translation by characterizing risk factors for financial exploitation among vulnerable groups.

NIH Research Projects · FY 2025 · 2021-04

7. Project Summary/Abstract Adolescence is an “age of risk” for the emergence of 1st onset of major depressive disorder (MD). Despite its prevalence and public health significance, major unanswered questions exist regarding the mechanisms involved in vulnerability to MD. Depression (Dep) is associated with a reduced sensitivity to rewards and low reward-related brain function in cortico-striatal circuitry. However, research has not yet tested whether chronically low reward responsivity (RR) or attenuated RR development during adolescence predicts 1st onset of MD. A separate literature documents elevated peripheral inflammation in Dep. Yet, research also has not examined whether chronically elevated inflammation or increases in inflammation during adolescence predicts 1st onset of MD. Further, research on inflammation and RR mostly has proceeded in parallel. Recently, however, we and others have proposed neuroimmune network models of Dep. These models draw on work indicating that peripheral inflammatory mediators (e.g., cytokines) access the brain, where they lower RR. When dysregulated, this immune-to-brain signaling can lead to chronic and worsening low RR, which is reflected in dysphoria and anhedonia. This low RR is proposed to initiate unhealthy behaviors (substance use, poor diet), as well as sleep disruption and stress generation, which further heighten inflammation. Over time, dysregulation in RR and immune signaling may synergize in a positive feedback loop, whereby dysregulation in each system exacerbates dysregulation in the other. We propose that reward-immune dysregulation is a two-hit vulnerability for the 1st onset of MD and increases in Dep symptoms (Sxs) during adolescence. Moreover, childhood and adolescent adversity and recent stressors influence both RR and inflammation, and may set the foundation for reward-immune dysregulation. This proposal is the first systematic test of these hypotheses. We will use an innovative biobehavioral high-risk design to examine bidirectional relationships between peripheral inflammation and multiple indices and domains (monetary, social) of RR and their joint prediction of 1st onset of MD and increases in Dep Sxs, particularly anhedonia. Three hundred 14-15 year old participants (Ps) will complete a prospective 3-year longitudinal study. Ps with no prior MD will be selected along the entire dimension of self-reported RR, with oversampling at the low tail of the dimension in order to increase the likelihood of MD onsets. At Time 1 (T1), T3, and T5, each a year apart, Ps will complete blood draws to quantify inflammation, self-report and behavioral measures of RR, and fMRI scans of reward neural activity and functional connectivity. At T1-T5 (with T2 and T4 6 mo. between the yearly sessions), Ps also will complete diagnostic interviews, and measures of Dep Sxs, reward-relevant life events, and behaviors that increase inflammation. Adversity history will be assessed at T1 only. This proposal is an innovative integration of research on reward and inflammatory signaling in understanding 1st onset of MD in adolescence. It has the potential to facilitate novel neuroimmune and behavioral interventions to treat, and ideally prevent, MD.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMARY/ABSTRACT Despite antiretroviral therapy (ART), neurocognitive complications continue to be highly prevalent in people living with HIV (PLWH). One explanation could be the constant compromise of the blood brain barrier (BBB) driven by chronic inflammatory responses. The introduction of medicinal marijuana into HIV treatment practice appear to be beneficial for several virus associated complications (ranging from chronic pain to appetite stimulation). Yet, the effects and mechanisms of cannabis on HIV associated chronic inflammation, the endocannabinoid system, immune modulation and neurologic disorders are minimally understood. As indicated in the RFA, preclinical models can provide a rigorous in-depth analysis of the molecular and cellular mechanisms at the intersection between phytocannabinoids, HIV and ART. To this end, we propose a comprehensive evaluation of the two most used cannabinoid compounds (THC, CBD) on BBB function, immune-endothelial interactions and neuroinflammation. We will utilize state of the art chip microfluidics models of the neurovascular unit (NVU) and animal models for HIV (w/ w/o ART). Previously, we discovered that the brain endothelium upregulate CB2 in HIV infected human brain tissue. We have also found that modulation of CB2 affects indices of HIV pathology (in-vivo) and regulates the BBB. Our preliminary studies identify the diverse effects that phytocannabinoids can have on the different properties of the BBB. Specifically, cannabinoids (THC, CBD) alone can enhance the physical barrier, partially reduce endothelial activation and augment efflux transporter activity. Although some of these effects may appear beneficial, the presence of HIV and ART changes how the function of the BBB is regulated by cannabinoid substances. For example, the augmented transporter activity by THC has important considerations for altering ART-CNS penetrability. Thus, we hypothesize that phytocannabinoids differentially modulates BBB function that are both beneficial and deleterious in NeuroHIV. In Aim 1, using our latest tissue-engineered microfluidic NVU model, we will perform analyses of the kinetic changes in BBB permeability, transporter status and immune-endothelial interaction. Then, in Aim 2, we will compare outcomes between widely used routes of cannabinoid administration (oral vs. inhaled) in vivo using two relevant models of HIV infection (‘humanized’ mice and a model of aseptic meningitis/encephalitis). Experiments will evaluate changes in the BBB in the context of ART and cannabinoid exposure. Finally, we propose to identify novel crosstalk mechanisms that bridge cannabinoid receptor signaling to signals that control BBB maintenance (Aim 3). It’s clear that cannabinoids exert unknown cell specific effects that contribute to the tumultuous interpretation of how these compounds impact NeuroHIV. Using innovative preclinical tools, our studies will contribute significantly towards understanding the consequences of cannabinoid use on the BBB in the modern era of NeuroHIV.

NIH Research Projects · FY 2025 · 2021-04

SUMMARY Currently, ~5.8 million Americans suffer from Alzheimer's disease (AD)1 and there remain no approved therapeutics to lessen the associated neuronal dysfunction and cell death. Various AD clinical trials targeting the “amyloid cascade” have proven unsuccessful, suggesting a dire need to rethink AD disease progression. Alterations in cellular calcium (iCa2+) and mitochondrial function are reported as critical molecular contributors to AD pathogenesis. Our lab has previously shown that genetic modulation of either mCa2+ uptake or efflux is a powerful way to limit mitochondrial dysfunction and cell death in the context of cell stress featuring elevated iCa2+. As causal evidence of impaired mCa2+ exchange in AD, we generated multiple mutant mouse models to modulate neuronal NCLX expression, the primary mechanism for 2+ efflux in excitable cells, and mCa discovered that impaired mitochondrial calcium efflux proceeds neuropathology and memory decline in 3xTg- AD mouse model. In addition to alterations in mCa 2+ efflux, we also discovered significant proteomic remodeling of the mitochondrial calcium uniporter channel (mtCU). The mtCU is required for acute Ca2+ uptake into the mitochondrial matrix where it regulates mitochondrial function. The mtCU is a multiprotein channel, consisting of pore-forming, scaffold, and regulatory components. To date, there are no published in vivo studies using genetic models to define the contribution of mCa2+ uptake with AD disease progression and given our published findings regarding the role of mCa2+ efflux in AD pathogenesis17, it's imperative that we mechanistically define the mCa2+ uptake pathway prior to proceeding with therapeutic development. In this project, we hypothesize that mCa2+overload is a primary contributor to AD pathology by promoting metabolic dysfunction and neuronal cell death, and that reducing mtCU-dependent mCa2+ uptake will impede neurodegeneration and AD pathogenesis. Further, using complex mutant mouse models we will systematically define the mitochondrial metabolomic and proteomic changes associated with AD progression and alterations in mCa2+ flux. Optimally, these studies will identify new therapeutic targets for the treatment of AD.

NIH Research Projects · FY 2026 · 2021-02

Summary/Abstract Advances in molecular sequencing have led to the accumulation of vast datasets rich in information spanning multiple levels of biological organization, from individuals in populations to diverse species across the tree of life. These datasets offer unprecedented opportunities to develop biologically realistic models and uncover novel evolutionary patterns and processes. However, traditional methods of evolutionary analytics still rely on many simplifying assumptions that can limit biological insights and compromise the robustness of inferences. Integrating deep learning technologies with evolutionary methods provides a powerful complementary approach. Emerging foundation models (FMs), trained on extensive sequence datasets using deep learning with transformers, have shown an ability to implicitly learn structural, biochemical, and contextual properties within sequences. Unlike traditional models, FMs avoid assumptions about substitutional independence among sites and uniformity of substitution models across sites and species. Consequently, FMs employ data-driven learning to capture complex relationships of residues and bases in molecular sequences. Over the next five years, I propose to develop and explore FM models for molecular evolutionary and phylogenetic analysis. Our preliminary results have established the evolutionary premise and promise of FMs, which led to the proposal asking questions, such as: What evolutionary knowledge do foundation models capture? How can they be leveraged to uncover novel evolutionary patterns and processes? Answers will be pursued through applications in annotating protein sequence variants, polymorphisms, substitutions as well as molecular phylogenetics. Additionally, we will develop a novel FM for molecular phylogenetics, explore coding nucleotide FMs, and investigate transformer architectures that can handle longer sequences that would enable more powerful FMs. To maximize accessibility and impact, we will develop myFM, an open-source library, to implement our new approaches. We will link myFM functionalities with the widely used MEGA software to bring deep learning advances to more biological researchers, many of whom may have never applied them in their evolutionary datasets. Such an integration will democratize access, use, and tests of advanced technologies needed for their widespread evaluation and testing. Ultimately, the success of this project will significantly advance the study of molecular evolution, offering transformative tools to uncover evolutionary relationships and understand functional evolution.

NIH Research Projects · FY 2025 · 2021-01

PROJECT SUMMARY/ABSTRACT This application is a five-year plan to create a Mid-Atlantic Neuroscience Diversity Scholars (MiNDS) program to bolster the number of underrepresented minority (URM) students within the neuroscience academic pipeline and build a foundation for URM students to succeed in graduate school and beyond. The program will comprise a partnership between Temple University, Lincoln University, and University of Maryland – all institutions with a strong commitment to educating URM students and a commitment to building neuroscience research. Our program will recruit 9 scholars per year and provide them with the tools necessary for persistence within academia focusing on 6 elements: (1) integrated research experiences during the academic year, (2) immersive summer research experiences at R1 universities, (3) opportunities to build presentation skills at local and national meetings, (4) coursework to build technical excellence in Neuroscience, (5) professional skills training and mentoring to facilitate the transition to Neuroscience graduate programs, and (6) outreach activities to foster community and build teaching skills. Scholars will participate in a 2-year bridge program during their last two years of undergraduate study. Students in our MiNDS program will be provided with a comprehensive research training experience, including financial support for academic year research at their home institution, travel funds to present their research both at the MiNDS retreat, and at the Society for Neuroscience annual meeting, and stipend to engage in summer research at T32 funded institutions, Temple University or University of Maryland Baltimore, within the labs of faculty with exceptional behavioral or cognitive neuroscience research programs and extensive undergraduate mentoring experience. The program will provide MiNDS with a foundation of coursework and professional development to set the stage for the next step of their neuroscience research career. This will include one-on-one faculty mentoring in oral presentation skills, scientific writing, graduate school application review, and interview preparation. Additionally, students in MiNDS will be paired with senior graduate student mentors during this summer experience to gain further insight into the transition to PhD programs. The final core goal of the MiNDS initiative is to foster professional development of both students and mentors through community outreach. MiND scholars will team with faculty to develop outreach activities to engage 5th and 6th graders in neuroscience. These initiatives will position MiND scholars for success in a career in academic neuroscience research. To sure this success, the MiNDS program will be evaluated by our Advisory Board annually.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY No approved medications are available for cocaine abuse, and medications for opioid addiction are limited to agonist replacement therapies that are abuse liable themselves. Glutamate is one neurochemical system that has been recently for drug dependence and relapse. One of the most viable therapeutic targets within the glutamate system is glutamate transporter subtype 1 (GLT-1), a predominantly astrocytic protein that clears glutamate from the extracellular compartment in the CNS. GLT-1 transport and uptake mechanisms are dysregulated during COC or opioid exposure and facilitate the enhanced glutamate transmission in brain reward circuits that underlie dependence and relapse. Agents that enhance the expression of GLT-1 transporters do display efficacy in preclinical models of drug addiction and related CNS disorders, but an existing hurdle is the disappointing clinical translation. Part of the problem is the agents themselves, which are limited mostly to β-lactam antibiotics that suffer from a host of pharmacokinetic and pharmacodynamic issues, including parenteral administration, poor brain penetrability, chronic dosing, adverse effects, and a slow onset of CNS efficacy that is dependent on increased GLT-1 protein expression. In this proposal, we address both the lack of effective treatments for cocaine and opioid addiction and the limited diversity in the GLT-1 activator pipeline. We propose to characterize the efficacy of two non-β-lactam GLT-1 activators (e.g. troriluzole [TRLZ] and NA-014) in rat assays that model cocaine and opioid reinforcement, dependence, and relapse. TRLZ is a prodrug of riluzole (approved for ALS) that is already being tested in clinical trials for obsessive-compulsive disorder and spinal cerebella. TRLZ displays a unique pharmacodynamic profile in that it acts that acts through a dual mechanism to enhance cellular glutamate uptake and inhibit neuronal glutamate release. Despite a glutamate-based profile that is favorable for potentially treating drug abuse, the parent drug RLZ has only been assessed in few preclinical studies that have yielded mixed outcomes. Our interest in RLZ-like compounds for drug addiction was recently reignited by a 2018 study showing that RLZ reduces cocaine relapse in rats. RLZ itself, however, is an unlikely candidate for repurposing because of reduced efficacy and potency related to pharmacokinetic limitations, including high first-pass hepatic metabolism, elevated liver enzymes, a negative food effect, low aqueous solubility, and poor oral palatability. To mitigate limitations of RLZ, we designed, synthesized, and evaluated TRLZ as a third-generation prodrug with optimized in vitro and in vivo features. The second GLT-1 activator is NA-014, which directly activates GLT-1 through selective allosteric modulation of GLT-1 after a single exposure, making it different from β-lactams that rely on upregulation of the GLT-1 but only after repeated treatment with high doses. In summary, our results will offer the first comprehensive information about the efficacy of non-β-lactam GLT-1 activators in preclinical models of drug addiction, and, if positive, may pave the way for the development of safer and more effective GLT-1-based medications.

NIH Research Projects · FY 2023 · 2020-09

Project Summary / Abstract Homoharringtonine (HHT) is a clinically used protein translation inhibitor that is used to treat chronic myeloid leukemia. In addition to its FDA-approved role as a leukemia drug, HHT shows exciting promise for the treatment of other hematologic malignancies and tumors. Finally, it is a perfect molecule to use as a probe to investigate protein translation inhibition. HHT is currently prepared through semi-synthesis from naturally derived cephalotaxine. Cephalotaxine is obtained from Asian plum yew trees grown in China. The cost of HHT in particular, along with other leukemia treatments, has been described as “astronomical” and “harmful to patients” by a group of 100 leading cancer specialists. This cost results in part from a supply bottleneck reflecting its tree-based sourcing. This project will eliminate this supply problem, and provide cephalotaxine and HHT for our studies and those by other research groups. Natural products have been the source of the majority of drugs throughout history, and still are today. The field of chemical synthesis directly contributes to the application of natural products as medicines. Aromatic and heteroaromatic rings are indispensible motifs in biologically active compounds. Thus, chemical reactions that allow for the construction of molecular architectures containing substituted aromatic rings are particularly valuable to human health. Polycyclic nitrogenous molecules, exemplified by the Cephalotaxus alkaloids in this proposal, are also critically important as biologically active molecules and pharmaceuticals. However, polycyclic nitrogenous molecules, such as alkaloids, are notoriously difficult to prepare, often requiring arduous chemical syntheses for preparation. We will develop two new cascade reactions that efficiently prepare: 1. Substituted arenes, and 2. Complex polycyclic alkaloids. The innovativeness of this research is the strategic use of cascade reactions to assemble structures that previously required multiple steps to prepare. Specifically, we will prepare substituted: phenols, indoles, furans, and related structures. A distinguishing feature of this strategy is its inherent efficiency. Additionally, we will showcase these pericyclic cascades in syntheses of natural products and natural product analogs. More widely, this strategy will find immediate application in the preparation of biologically active molecules, such as HHT; chemical probes for biological systems, and related alkaloid natural products.

NIH Research Projects · FY 2024 · 2020-09

Abstract Alveolar type II (ATII) cells produce and secrete pulmonary surfactant and restore the epithelium after damage. Due to their energy-demanding functions and location in the lung, they are highly dependent on mitochondria. Aerosols produced from e-cigarette devices contain both the particulate and gas phases, nicotine and flavors. The effect of flavored constituents and electronic cigarette (e-cigarette) aerosols on ATII cells is unknown. Mitochondrial DNA (mtDNA) is susceptible to damage due to the lack of histones that serve as a barrier against damaging factors. MtDNA damage can lead to mitophagy and ATII cell death. Our preliminary results demonstrate the harmful effect of e-cigarette aerosols with fruit and dessert flavors on human and murine primary ATII cells. DJ-1 has a cytoprotective role and participates in transcriptional regulation and mitochondrial function. Our hypothesis is that flavored constituents and e-cigarette aerosols with fruit and dessert flavors cause mitochondrial dysfunction due to the impairment of DJ-1 function leading to ATII cell death. In SA#1, we will determine mitochondrial dysfunction induced by flavored constituents and e-cigarette aerosols in human primary ATII cells. The mechanism of DJ-1 cytoprotective function in ATII cells will be studied in SA#2. In SA#3, we will analyze the respiratory injury risk of flavored constituents and e-cigarette aerosols in wild-type and DJ-1 KO mice. The current project will fill the gap in our knowledge on the effect of this exposure on primary ATII cells and respiratory function and can provide novel therapeutic targets for lung regeneration.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY Spinal cord injury (SCI) causes life-long neurological impairment, and there is currently no effective treatment. The premise of this proposal is recent work demonstrating that afferent stimulation paired with treadmill training can enhance standing, stepping, and volitional control in humans and animal models. Therefore, it is critically important to understand the mechanisms by which afferent stimulation drives motor improvement. Tools that can identify which afferents are necessary and sufficient to enhance recovery, and that can facilitate characterization of the helpful neural plasticity, are urgently needed. Our long-term goal is to develop approaches for selective afferent modulation, and apply them to the dissection of the mechanisms underlying recovery from SCI. The objective of this grant is to identify which sets of afferents are important for recovery and how spinal circuits change to facilitate it. To achieve selective modulation of afferents and enable genetic tracing we will use Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) that can modulate excitability in specific populations of neurons. To accurately quantify improvement, we will use Deep Learning to analyze large kinematic data sets. Our preliminary data shows strong expression of DREADDs in large diameter DRG neurons, that their activation by CNO can excite or inhibit the H-reflex, and that activation of excitatory DREADDs during treadmill training post-SCI improves stepping. Our main hypothesis is that activation of large afferents by the excitatory DREADD (hM3Dq) during treadmill training will enhance recovery, whereas inhibitory DREADDs (hM4Di) will suppress recovery. Four sub- hypotheses will test whether recovery is mediated by increased afferent projection onto 1) motor neurons, or 2) inhibitory interneurons; or by sprouting of 3) reticulospinal and 4) propriospinal circuits. Our Specific Aims are to determine whether selective expression of DREADDs in (Aim 1) all large diameter (proprioceptive and tactile) neurons and (Aim 2) large proprioceptive afferents only can enhance recovery. The rationale for these aims is that afferent stimulation is hypothesized to work through selective excitation of large diameter sensory afferents (LDSA) that both drive motor pools locally and facilitate proprio- and surpraspinal input. To date, it has not been possible to definitively determine which afferents were recruited after electrical stimulation, or to select between afferents of similar diameter. The significance of this work lies in determining whether recovery is mediated exclusively by proprioceptive axons or a combination of proprioceptive and tactile afferents, and uncovering the mechanisms of functional plasticity in the spinal cord.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT A common feature of heart failure (HF) is excessive extracellular matrix deposition by a specialized and differentiated fibroblast population, known as myofibroblasts, in response to cardiac injury. While myofibroblasts help to maintain the structural integrity of the injured heart and prevent ventricular wall rupture, persistence of myofibroblasts results in excessive fibrosis and subsequent cardiac decompensation. Therefore, identifying molecular mechanisms of myofibroblast differentiation in cardiac fibrosis could yield novel clinical targets to delay or reverse the development of HF. Recent evidence suggests metabolism may drive cellular differentiation through the modulation of epigenetic-modifying enzymes that enhance or silence genes associated with cellular differentiation. Altered metabolism changes the concentration of metabolites that act as substrates for epigenetically modifying enzymes, such as the changing levels of acetyl-CoA that alter the activity of histone acetyltransferases (HAT). Our preliminary data indicate that increased glycolytic rate is a key feature driving myofibroblast differentiation. We identified metabolic regulation of histone demethylation as a feature of myofibroblast differentiation and we now turn our sights to histone acetylation as an epigenetic modification permissive of myofibroblast gene expression. This proposal hypothesizes that increased acetyl-CoA biosynthesis is necessary for histone lysine acetylation by HATs during differentiation for the transcriptional activation of the myofibroblast gene program. This study seeks to identify novel therapeutic targets to mitigate the consequences of fibrosis in HF.

NIH Research Projects · FY 2025 · 2020-09

Approximately 48 million people in the US are served by private, and frequently untreated, wells. Our best estimate is that 1.3 million cases of gastrointestinal illnesses (GI) per year are attributed to consuming water from untreated private wells in the US, but in reality, there are no robust epidemiological data that can be used to estimate cases of GI attributable to these sources. It is likely that well water-associated GI causes significant healthcare costs and lost work/school days, as well as increased risk for long term health complications. This impact is magnified when accounting for vulnerable populations such as children under the age of 5, the elderly, and the immunocompromised. We propose the first randomized controlled trial (RCT) to estimate the burden of GI associated with private well water. We will test if household treatment of private well water by ultraviolet light (UV) vs. sham (placebo inactive UV device) decreases the incidence of GI in children under 5. At present, there are no prior RCTs or studies that have sought causal links between GI and the consumption of untreated water from private wells despite the fact that pathogens have been recovered in groundwater, including deep aquifers. Under the guidance of an interdisciplinary advisory committee we will execute the following aims: Aim 1- Quantify the incidence rate of endemic childhood GI associated with consuming untreated private well water and compare that to the incidence rate of consuming well water treated by UV. Aim 1a- Construct a Quantitative Microbial Risk Assessment (QMRA) using water quality data we collect to estimate the risk of childhood GI associated with consuming untreated private well water and compare the incidence from the risk model to the incidence we calculate in Aim 1. Aim 2- Identify, quantify and compare viral, bacterial and protozoan pathogens in stool of children consuming UV treated or untreated (sham) private well water (including both asymptomatic and symptomatic cases). Aim 3- Explore the presence of pathogens in untreated well water and stool samples of children consuming untreated private well water (sham group only). These data will fill a knowledge gap on sporadic GI associated with federally-unregulated private water supplies in the US. Our results will test an affordable water treatment intervention and inform GI burden estimates and policy decisions for managing well water in the US and globally. Policy changes will help better protect rural families, especially children who are at highest risk for sporadic enteric infections.

NIH Research Projects · FY 2024 · 2020-08

Project Summary/Abstract Plasmodium falciparum is the deadliest of the five species of malaria parasites that infect humans. Annually, there are over 200 million symptomatic cases of falciparum malaria and over 400,000 deaths – the majority of which occur in African children under the age of five. The two major interventions that have had an effect on reducing malaria prevalence over the past twenty years are mosquito control and use of antimalarial drugs. The most important class of drugs in this effort has been the artemisinin derivatives, which since 2005 have been deployed as artemisinin combination therapies (ACTs) only, in order to reduce the probability of emergence and spread of artemisinin-resistant genotypes. Despite these efforts, artemisinin resistance did emerge, and we are now facing the dangerous prospect that these drug-resistant genotypes may spread to Africa, where most of the world’s malaria cases occur. In this proposal, we will introduce and evaluate a number of drug-resistance management strategies that are intended to prevent, delay, and slow down the spread of drug-resistant genotypes of P. falciparum. First, we will make a number of technical advances in our existing individual-based (agent-based) simulation that we already use to model the evolution and epidemiology of P. falciparum in human populations. We will add explicit grid- based spatial structure to make the model more realistic. Additionally, we will add a genotype-phenotype map and clinically-parameterized pharmacodynamic/pharmacokinetic sub-models to make the model’s drug- resistance component as realistic as current data allow. Second, we will evaluate strategies for how best to manage the population-level introduction of novel antimalarial therapies that will become available in the 2020s. The strategies will be aimed at minimizing the long-term risk of drug resistance to both the novel therapies and to currently used ACTs, in order to minimize the number of treatment failures in the long run. Finally, we will parameterize country scenarios for Cambodia, Zambia, and Burkina Faso to provide specific country-level advice in low, medium, and high transmission malaria settings on how best to preëmpt drug resistance or minimize its current spread. As Cambodia is the epicenter of the current wave of artemisinin resistance, the Cambodia- specific model will be used to provide advice on how to contain and eliminate currently circulating artemisinin- resistant genotypes of P. falciparum.

NIH Research Projects · FY 2024 · 2020-08

Abstract: Spinal cord injuries (SCI) affect approximately 100,000 patients each year and causes motor, sensory and autonomic dysfunction. Approximately 55% of all SCIs occur at the cervical level in human patients that effect forelimb. Patients effected by a cervical SCI desire recovery of hand and digit function to improve their current lifestyle. However, the majority of spinal cord injury models study the functional recovery associates with locomotion after thoracic lesions and not skilled forelimb patterning after cervical injuries. In general, skilled forelimb patterning is mediated through the corticospinal tract with support from indirect brainstem regions. Lesions that cut the corticospinal tract show significant loss of forelimb patterning; however, when the indirect pathways are preserved, rehabilitative training supports the recovery of the lost function. We show that this recovery is mediated by C3-C4 propriospinal neurons, a unique propriospinal neuron normally involved in the formation of an internal motor copy to correct movement errors during the movement. In this pathway, error correction is mediated by C3-C4 PN axons terminating onto neurons in the lateral reticular nucleus, a pre-cerebellar nucleus. Information is then routed through the cerebellum where the motor plan is compared to proprioceptive sensory information from the forelimb and back to brainstem motor control regions for execution. We hypothesize that after cervical injury this pathway undergoes rehabilitative adaptation to compensate for loss of the CST. Here we will take a systems approach to interrogate three components required for error correction and adaptation of this pathway. The first aim will examine the integration of information from several forelimb control pathways into the LRN and if modulation of LRN neuronal activity during rehabilitation augments recovery. The second aim will investigate the role of unconscious proprioceptive sensory information in rehabilitative recovery of injury. The third aim, will investigate the cerebellar efferent pathways connecting to know reticular regions involved in forelimb movements to determine their importance in driving recovery and error correction after cervical injury and during rehabilitation. Ultimately, this study will provide essential data to identify the pathways involved in rehabilitative recovery of skilled forelimb patterning after spinal cord injury and if recovery can be enhanced by activity dependent modulation during rehabilitative training.

NIH Research Projects · FY 2024 · 2020-07

ABSTRACT Obesity populations on environmental obesity impacts 2.7 million American children. Children from racial/ethnic minority and low-income have higher obesity prevalence, contributing to health disparities. Obesity reduction efforts focused individual health behavior change have seen limited success, resulting in calls to examine social and factors that contribute to obesity. A better understanding of complex risk factors for childhood can inform targeted interventions, clinical practice, and policy. 1 Adverse childhood experiences (ACEs) are one understudied complex risk factor for childhood obesity. ACEs are traumatic experiences that occur during childhood such as sexual abuse, family member incarceration, or witnessing violent crime. Almost half of children have experienced ACEs and populations with ACEs have higher obesity prevalence. ACEs can increase obesity risk via sequelae of chronic or severe stress or by hindering healthy behaviors, though the impact of ACEs on childhood obesity is not well understood. Multi-level research examining the influence of neighborhood environment on the ACEs-obesity relationships is particularly lacking. Neighborhood factors such as lack of access to healthcare and high crime may contribute to both ACEs and obesity via indirect and overlapping pathways. It is difficult to disentangle the impact of neighborhood factors because many co-occur within the broader context of neighborhood poverty and racial segregation; sophisticated approaches such as geospatial analysis, longitudinal research, and novel methods for quantifying neighborhood risk are necessary. This study will be the first to examine the relationship between neighborhood environment, ACEs, and childhood obesity using advanced statistical methods. Aims 1a and 1b entail development and testing of a neighborhood ACEs index using existing multi-level geospatial data. Aim 2 entails examining the relationship between the neighborhood ACEs index and childhood obesity incidence using existing longitudinal data. Aim 3 entails determining neighborhood factors most associated with ACEs exposure and childhood obesity in order to identify potential targets for intervention, clinical practice, and policy. This study has the potential to inform targeted efforts for improving the health of millions of children who experience ACEs and obesity. The associated Career Development Plan focuses on ACEs, advanced statistics, and geospatial analysis. The Plan will advance my ability to use diverse, multi-level, geospatial data to examine complex childhood obesity risk factors - a skillset rare among nurse scientists but directly relevant to my research on how social determinants of health impact obesity. The Plan includes expert mentorship, focused coursework, and training at the inaugural NICHD P50 Translational Center for Child Maltreatment Studies. Collectively, this study will advance knowledge about the impact of ACEs on childhood obesity and simultaneously further my career goal to establish a program of research on understanding and reducing childhood obesity disparities.

NIH Research Projects · FY 2026 · 2020-07

Project Summary Cocaine addiction exerts a high cost on society and individuals and to date no pharmacotherapies exist. Behavioral therapies are not effective at preventing relapse; indeed, 70-80% of cocaine users will experience relapse following therapy. Preventing relapse to cocaine use represents the primary challenge that exists for the treatment of cocaine dependent individuals. One of the many factors that contribute to relapse is the exceptionally strong associations that drugs of abuse, such as cocaine make between environmental contexts and the rewarding properties of the drug. Thus, understanding the neural mechanisms that are responsible for these drug-context associations and ways in which we can override them, is critical for the development of improved treatment options. The dorsal hippocampus (dHPC), well known for its role in learning and memory, is an important anatomical region involved in cocaine-context associations. Despite this knowledge, the role of this brain region in cocaine addiction remains understudied. Work from our laboratory has identified a novel role for the Cav1.2 L-type Ca2+ channel (LTCC) in the dHPC in extinction of cocaine-associated contextual memories, consistent with their well-known role in hippocampal-dependent synaptic plasticity underlying certain forms of learning/memory. Extinction learning involves a new form of learning that is capable of overriding original memories, particularly maladaptive memories. Thus Cav1.2 channels serve as a promising candidate for overriding drug-context associations. Using the cocaine conditioned place preference (CPP), a preclinical model used to study cocaine-associated contextual memories, we find that extinction of cocaine CPP increases synaptic levels of Cav1.2 and its phosphorylated form in the dorsal dentate gyrus (dDG), a hippocampal subregion, in a dopamine D1 receptor cell type-dependent manner. This is consistent with growing evidence for a role of dHPC dopamine for learning/memory mechanisms including cocaine contextual memories. Our molecular studies have identified that extinction increases key signaling molecules in the dDG. These include AKAP150 anchoring protein, PKA, NFATc3 and the GluA1 subunit of AMPA receptors. Thus, in this application we aim to capitalize on this knowledge to further explore dDG Cav1.2 channel mechanisms in extinction of cocaine-associated memories. We will test the central hypothesis that contextual extinction learning recruits Cav1.2 channel mechanisms at dDG synapses via recruiting dopamine D1R signaling. We will use a combination of genetic, pharmacological, electrophysiological, and in vivo calcium imaging techniques with behavioral testing for the proposed studies. Aim 1 will test the involvement of AKAP-PKA-Cav1.2 signaling. Aim 2 will address the involvement of AKAP-CaN-NFAT signaling and Aim 3 will examine the contribution of D1R signaling, in cocaine CPP extinction and dDG cell activity.

NIH Research Projects · FY 2026 · 2020-02

Project Summary/Abstract Mild functional difficulties are a strong predictor of future cognitive decline in older adults across the spectrum from healthy cognition to mild cognitive impairment (MCI) and precede impairment on traditional cognitive tests. Functional ability level is a criterion that distinguishes clinical stages of Alzheimer’s disease (AD) and AD related dementias (AD/ADRD) and is a highly meaningful outcome to patients and families in clinical trials. However, the current conventional approach to functional assessment includes questionnaires, which are useful and highly efficient, but often are atheoretical and limited by methodological drawbacks, including a range of biases, recall failures, and/or the availability of a knowledgeable informant. To address these needs and gaps in our original R01 proposal, we developed and validated an efficient performance-based measure called the Virtual Kitchen Challenge-Version 2 (VKC-2). The goal of this renewal is to evaluate the predictive validity of the VKC- 2 after 3-5 years, further develop the efficiency of the VKC in a new version with automated administration (VKC-3) and test a comprehensive framework for functional assessment using novel digital tools, such as smartphone digital phenotyping and ecological momentary assessment (EMA). This renewal includes three aims. Aim 1: Test the hypothesis that baseline VKC-2 automated scores will significantly predict cognitive abilities after 3-5 years (predictive validity). Our VKC-2 validation cohort will be re-evaluated after 3-5 years, and regression models with baseline VKC-2 scores as predictors and conventional cognitive measures as outcomes will be performed. A new version of the VKC with a participant-driven interface and no examiner input also will be piloted to further increase test efficiency. Aim 2: Test the hypothesis that VKC-2 automated scores will significantly predict everyday behaviors and routines (i.e., functional performance). Participants (N=300) from Aim 1 will download the open-source, mindLAMP app on their own smartphone to collect GPS sensor data and daily EMA responses about participants’ daily activities during a 4-week period. Two sets of regression models will include baseline VKC-2 (from the original R01) or concurrent VKC-3 scores as predictors and GPS/EMA measures of activity and activity variability as outcomes. Aim 3: Test the hypothesis that economic resources, participant motivation, and participant physical abilities will moderate the relations between functional capacity (VKC-2) and functional performance (GPS, EMA). Participants will complete validated questionnaires of economic resources, trait motivation, and physical abilities. Regression models will test moderation of the relation between VKC-2 scores and measures of functional performance (GPS, EMA). The proposed aims seek to validate a fully automated VKC-3 and a comprehensive framework for understanding everyday function, which together have the potential to address important functional assessment challenges in multiple contexts, including screening older adults for risk of functional decline and as a function endpoint in clinical trials of AD/ADRD treatments.

NIH Research Projects · FY 2025 · 2019-07

Principal Investigator/Program Director (Last, first, middle): Wang, Ross, E. Steric-free labeling strategies to study disease-related non-histone substrates of post- translational modifications The proposed research will utilize fluorine-based bioorthogonal reactions for steric-free labeling and chemical biology interrogation of disease-related post-translational modifications (PTMs) such as acetylation and O-linked N-acetylglucosamine. Despite recent advances in biomedical research, diseases are still haunting human beings. Many cancer types remain lethal, particularly at late stages, and are resistant to traditional therapy, while most inflammatory diseases result in chronic or long-term burdens, and there is a lack of selective immunosuppressives on the market. Thus, there is a need for new therapeutic approaches by exploiting novel proteins and protein-protein interactions (PPIs) as targets. Disease-related acetylation and O-GlcNAcylation have recently emerged as important biological pathways that could unravel such potential targets. These PTMs modify existing proteins with specific chemical functionalities to modulate protein function, and thereby mediate various cellular activities including activation, proliferation, and migration. Dysregulation of the related proteins has been reported to be key to certain human diseases such as cancer and inflammatory disorders. Yet, the identity of these non-histone proteins and PPIs has not been fully elucidated. Current research in this area is still at a preliminary stage, heavily relying on chemical proteomics, which directly tags target proteins within complex proteomes with alkyne or azide- modified pro-metabolites such as acetyl-CoA or glucosamine. These chemical tags, after their metabolic labeling onto protein substrates, can later be derivatized in situ through bio- orthogonal ‘click chemistry’ with a fluorophore for imaging or a biotin affinity probe for pull down and proteomics- based target identification. However, these alkyne/azide- based tags are bulky in length and size, and for many cases cannot be metabolized by PTM enzymes and were barely incorporated onto substrates, thereby limiting the related target identification. The proposed research will focus on developing an innovative chemical tagging approach which is steric free and can be broadly used for the global profiling of proteins and PPIs related to acetylation or O-GlcNAcylation. We hypothesize that unlike azides and alkynes, fluorine labeling can best mimic the intrinsic carbon-hydrogen bond and is thereby steric free and generally applicable to tag acetylation and O-GlcNAcylation. The first research thrust seeks to utilize the fluorine-displacement reaction toolkit to systematically interrogate acetylation- involved proteins and PPIs in breast cancer and T-cell activation. The second research thrust involves the new methodology development by combining the fluorine-displacement reaction toolkit with specific inhibitors of the acetyltransferase so that one can specifically label and profile potentially novel substrates and PPIs downstream of that isoform-specific acetylation enzyme. The fluorine displacement based chemical proteomics approach will also be tested on O-GlcNAc substrates to reveal novel PPIs related to prostate cancer. The final thrust will investigate new generations of fluorine-mediated chemical reactions for PTM labeling. Page 1

NIH Research Projects · FY 2025 · 2019-05

Project Summary/Abstract Our goal is to integrate structure and sequence-based approaches grounded in statistical mechanics to understand key features of molecular recognition by proteins, as well as protein fitness and function more generally. There are three aims. The first is focused on mapping complex conformational and fitness landscapes of proteins. We will integrate machine learning (ML) sequence co-variation and molecular dynamics structure- based approaches to analyze the sequence dependent conformational landscapes of proteins with a particular focus on the landscapes that govern the transitions of kinase family proteins from the active to functionally important inactive states. With our collaborators we will investigate the sequence dependent origin of the alternative binding modes for peptide substrates that TKs have compared with STKs. This work has important implications for the design of therapeutics targeting Src and other cytoplasmic TKs which appear to bind peptide substrates in an unusual way. Also, as part of our first aim, we will map the free energy landscape of the set of kinase P-loop active conformational states; this information is needed for efforts to develop anti-cancer therapeutics with higher specificity. The thrust of the second aim is to realize the power of the sequence- covariation ML methods we are developing to detect and decompose multi-residue allosteric interaction motifs within kinases and kinase protein complexes by evaluating connected mutational correlations that carry signatures of indivisible units of biological information flow. These methods are designed to identify allosteric pathways and will be generalizable to other protein targets we are working on including GPCRs and the HIV Intasome. Our third aim is to build on the structure-based molecular dynamics approaches we recently developed to determine the excess chemical potential of water molecules at the surface of proteins. The excess chemical potential provides quantitative information about position specific thermodynamic features of interfacial water molecules and their networks. Together with our experimental Cryo-EM collaborators we will use this information to refine solvent at the protein-solvent interface in Cryo-EM density distributions in an iterative and self-consistent way; this will substantially improve upon current methods for locating and refining solvent in Cryo- EM maps of proteins and their assemblies. This new refinement tool will be made available to the structural biology community. We will build on our recent development of classical density functional methods to evaluate how the displacement of specific solvent molecules located in protein binding sites affects the affinities and specificities of the small molecule ligands targeting these sites.

NIH Research Projects · FY 2025 · 2018-09

The Synergistic Partnership for Enhancing Excellence in Cancer Health (SPEECH) is a highly collaborative and mutually beneficial Partnership between Temple University/Fox Chase Cancer Center (TUFCCC) and Hunter College (HC) and the only NCI-funded U54 CPACHE in PA and NJ and one of two in NYC. Our established robust research infrastructure has made significant contributions to addressing substantial gaps in cancer research that affect patient health and outcomes across the United States. In the past five years, SPEECH directly supported 84 investigators, mentored 180 trainees, funded 56 cancer disparities projects, facilitated 93 publications and directly stimulated $45 million externally funded grants. Additionally, 14 ESIs received career advancement, we engaged 50 neighborhood-based organizations, trained 41 lay health workers and educated 1200 members of local neighborhoods to provide cancer prevention services at the local level. Leveraging this momentum of success our Vision is to “Promote excellence in cancer research, education, local engagement, and strengthen infrastructure and capacity building for long-term impact.” Through exceptional institutional commitments, two full projects, a pilot project and integrated cores of Administrative, Research Education, Biostatistics/Bioinformatics, Planning and Evaluation, and Outreach, we propose to achieve the following specific aims. Aim 1: Accelerate TUFCCC-HC Partnership transdisciplinary cancer research collaborations and advance cancer health across the spectrum of basic, clinical and population sciences. Aim 1a: build, strengthen and sustain cancer research capacities and infrastructure at HC. Aim 1b: advance cancer research to maximize impact at TUFCCC. Aim 1c: foster community-driven research to address unmet needs in cancer research and health across the PA-NJ-NYC region. Aim 2. Train the next generation of cancer scientists and workforce leaders, building a pipeline of students equipped to contribute to the nation’s cancer research efforts, providing them with multidisciplinary cancer research education experiences and mentorship, and career development opportunities through the implementation of enhanced Summer Cancer Research Institute and newly established Hunter/Temple HEAL Program. Aim 3. Recruit, mentor and retain ESIs, to enhance career development and facilitate their transition to independent investigators. Aim 4. Use bidirectional community-engaged approaches with local stakeholders to implement a robust outreach program that builds bridges between stakeholders and researchers to support: a) cancer education and interventions, b) cutting-edge impactful cancer research projects and c) key research competencies for investigators and trainees. Aim 5. Conduct a mixed-method evaluation aimed at monitoring progress, providing feedback and summative impact data on all Partnership programing for promoting sustainable solutions to improved cancer health excellence.

NIH Research Projects · FY 2025 · 2017-09

Summary For a complex organ like heart, well-organized interactions between different cell types are essential for its effective functioning. Cell-cell communication between these myriad cell types therefore appears to be key component of cardiac function and a better understanding of how different cells within injured myocardium communicate with each other during homeostasis and injury and elucidation of molecular events downstream of such communications is critical to identify new therapeutic targets for cardiac healing after injury. Increasing evidence suggest that extracellular vesicles (EVs) including exosomes are major paracrine conduits of cardiac cell-cell communication in homeostasis and disease conditions. However, a comprehensive understanding of how cardiac injury alters exosome-mediated cell-cell communications both locally within myocardium and at long distance organs such as bone marrow and adipose tissue and whether these exosome-mediated alterations in cell-cell communications influence overall cardiac structure and function and repair is not well understood. The overarching hypothesis of this renewal application is that cardiac injury alters exosome- mediated specific cell-cell communication both locally within the injured myocardium and distally with other organs and these intra-cellular and inter-organ communications influence overall myocardial responses to injury and repair processes. To test our hypothesis, we have assembled a team of highly accomplished scientists who have the potential to markedly advance the science and application of exosome biology for ischemic tissue repair. In particular, the design of this program, highlighted in inter-related yet unique individual projects, intends to capitalize on our existing synergy and assets. We will utilize the advantage of strategically designed scientific cores that will support each project. We will apply various techniques of molecular, cellular, and biochemical approaches in cell models and mice models of myocardial injury and repair. Project 1 (Kishore) examines the role of stressed cardiomyocyte-derived exosomes, particularly cardiomyocyte specific microRNAs, on post-injury endothelial and endothelial progenitor cell function and angiogenesis. Project 2 (Walter Koch) focuses upon the involvement of cardiac exosomes containing G-protein coupled receptor kinase 2 on adipocytes and cardiac cell function. Project 3 (Tilley) examines the role of beta-2 adrenergic receptor on myeloid cell exosomes and cardiac functions. All three projects involve in-depth molecular, cellular, and physiological studies comprising of mouse models of myocardial injury. Establishing the role of exosomes in post-injury cell to cell communications may identify new mechanisms and therapeutic targets for post-injury myocardial repair. The goal of this program will be to delineate exosome mediated signaling mechanisms and determine how they can be utilized to restore and enhance endogenous cellular repair processes that heal the damaged heart.

NIH Research Projects · FY 2026 · 2017-07

The Temple-led Emergency care research network (Temple-SIREN) is comprised of multiple community and academic medical centers and EMS providers throughout Philadelphia, New Jersey, Delaware, and Maryland to execute SIREN emergency care trials. To meet the needs for specific study enrollment, Temple-SIREN engaged the highest volume emergency and trauma centers and the busiest EMS systems in the region. Temple-SIREN Hub and spoke leaders are committed to emergency care research and are active in trials of acute cardiopulmonary and neurological illness and traumatic brain injury trials. Through collaboration and hub oversight, the Temple-SIREN group expects to significantly contribute SIREN studies' enrollment. In addition, we plan to stay ahead of the curve by translating preclinical work to the bedside and using our regional network for studies that address the needs of our local population. The Temple-SIREN Hub staff will work closely with participating site investigators and coordinators from multiple disciplines to efficiently execute SIREN studies. Mechanisms to enhance efficiency and quality of clinical research across sites include use of a common informatics system to identify potential study patients; frequent communication with investigators and coordinators via webinars and conference calls; diligent site management focused on good clinical research practices and compliance, and regular Hub-spoke meetings to provide additional training by study role. Temple-SIREN investigators and coordinators will participate in SIREN clinical coordinating center and data management center and ad hoc working groups to develop SIREN studies

NIH Research Projects · FY 2026 · 2017-07

Project Summary. In an ideal world, scientists devise interventions for human communication which are in turn translated and implemented by clinicians. Feedback from patients and caregivers is then used to optimize and fine-tune the treatment. Modern rehabilitation is a dynamic process, and its effectiveness requires coordination between numerous disciplines and stakeholders. Since 2006, the Eleanor M. Saffran Conference on the Cognitive Neuroscience and Rehabilitation of Communication Disorders has provided a hub for engaging basic science with implementation science. The conference attracts professionals and students with a common interest in language and cognitive disorders, including researchers in basic and clinical research, clinical practitioners, academic faculty, and students representing disciplines such as speech-language pathology, neuropsychology, cognitive neuroscience, and linguistics. In this renewal application, we will continue this mission and expand our coverage to include other key moderators (e.g., access to services) of successful rehabilitation of language and cognitive disorders. Effective evidence-based practice in the communication sciences and other health professions requires several stages of development from basic science to the clinic. Thus, we will retain our format that moves from basic science talks in the morning of Day 1 to applied clinical research in afternoon. The focus of Day 2 is on the challenges of implementing basic and translational research to the clinical or school setting, which stem in part from the practical differences between laboratory settings where interventions are developed and clinical settings where they are implemented. The second day provides a forum to address the practical considerations involved in translating laboratory developed diagnostic and treatment protocols to clinical practice. This component of the conference provides a unique opportunity for researchers and clinicians to participate together in the translation/implementation process. We are expanding this feature of the conference to include two other stakeholders in the successful translation of research to clinical implementation: individuals with communication disorders and care partners of people with communication disorders. These two groups will be involved as speakers, discussants and participants in ways that are fitting to the particular theme of a conference. We will also continue with our mission to train students and provide them with opportunities to meet and interact with world class researchers and clinicians . Students at all academic levels (undergraduate to postdoctoral) will attend the conference free of charge. Additionally, we will offer a competitive Student Scholar Award program open to students at the doctoral or post-doctoral level. The award includes travel, accommodations, hotel, and a chance for the students to present their research in a poster session at the conference. We will budget for ten students but accept as many qualified candidates as possible. This program has been enormously successful, with applicants coming from across the nation and worldwide. Finally, moving forward, the conference will be hybrid (in-person and streaming) to enable us to reach more people from a wider geographic area.

NIH Research Projects · FY 2026 · 2017-05

PROJECT SUMMARY Our laboratory has developed unique tools that have enabled further progress towards understanding molecular transport mechanisms involving three sub-cellular organelles in eukaryotic cells: the nucleus, cytoplasm, and primary cilium. Macromolecular trafficking among these compartments is suggested to be gated by two unique mechanisms. One is the nuclear pore complex (NPC) embedded in the nuclear envelope that mediates bidirectional trafficking of proteins and RNAs between the cytoplasm and the nucleus; the other is the transition zone (TZ) located at the base of primary cilium that regulates the entry of membrane and cytosolic proteins into the cilium. Progress is intimately linked to technical improvements in biophysical research putting us in the unique position to elucidate the fast kinetics and 3D transport routes of macromolecules as they transport through the sub-micrometer NPC or TZ in live cells. In the first four years of the current five-year MIRA grant, we have completed the development of single-point edge-excitation sub- diffraction (SPEED) microscopy and successfully applied the new method to solve several critical questions pertaining to nuclear transport and cytoplasmic-ciliary transport. We have included our major findings in thirty manuscripts and published them in prestigious scientific journals such as Nature Structural & Molecular Biology, Nature Communications, Protein Science, and Nature Protocols. Currently we are highly motivated to work on the following new research topics: 1) Development of a new optical imaging system termed enhanced photon microscopy aiming at doubling photon collection efficiency from fluorophores, which will significantly improve optical resolutions when integrated with epifluorescence microscopy, laser scanning confocal microscopy, SPEED microscopy and other super-resolution microscopy; 2) Mapping the spatial organization of dynamic disordered protein motifs in the NPC to distinguish between, and advance upon, FG- organizational models; and 3) Understanding the gating mechanism for the cytoplasmic-ciliary transport of membrane proteins. In summary, during the next funding period of the MIRA grant, we will continue to develop new microscopy imaging techniques and solve long-standing unanswered questions in both nucleocytoplasmic and cytoplasm-cilium transport mechanisms.