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Maine Arts Commission

The Maine Arts Commission awards project grants to nonprofit organizations for arts programming that increases public access to the arts, supports artists, and strengthens communities through creative engagement across Maine.

Maine Community Foundation — County Programs

Maine Community Foundation — County Programs ($700M in assets) supports nonprofits in ME through grants focused on education, environment, community development, arts. Awards range from $2,000 to $100,000.

Maine Community Foundation

Maine Community Foundation ($800M in assets) supports nonprofits in ME through grants focused on education, arts, environment, community development. Awards range from $1,000 to $100,000.

Rainier Avenue Radio

Rainier Avenue Radio will host a free event to celebrate music and to promote the activation of public spaces. Make Music Day will be held on June 21, 2019 from 11am - 8pm at Westlake Park located at 401 Pine St. The event will feature performance art and interactive musical activities for all ages and levels of musical ability.

Manitoba Arts Council

MAC provides grants to artists and arts organizations in Manitoba for creation, production, presentation, and community engagement across all art forms.

Maple Leaf Community Council

The Maple Leaf Summer Social 2023 is a free and public party in Maple Leaf Reservoir Park featuring food (including LOTS of free ice cream), games, music, art, and most importantly, the opportunity for participants to make vital post-pandemic connections with old and new neighbors.

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY Hormones, cytokines, and growth factors are essential signaling proteins in blood circulation that coordinate multi-organ functions to maintain physiological homeostasis. These “inter-organ” factors (e.g. Insulin) have inspired widely used therapeutics and biomarkers for diabetes, obesity (e.g. semaglutide), and cancer, owing to their systemic effects and accessibility in the blood. Despite decades of research, our understanding of the body’s inter-organ communication network remains limited. In contrast to systematic efforts to map the brain connectome, comprehensive mapping of inter-organ communication under both normal and disease conditions has lagged behind. This slow progress stems from the challenging qualities and complex roles of inter-organ factors, including their low abundance in the blood, regulated secretion, non-cell-autonomous functions, ligand- receptor binding, and distinct origin and target tissues. High-throughput approaches such as proteomics and transcriptomics struggle to capture this complexity, so most of our knowledge comes from laborious and unsystematic discovery efforts. This project will overcome these barriers by integrating state-of-the-art experimental and computational techniques to map inter-organ proteins in the blood. We will (1) identify and study inter-organ factors from our prior dataset of organ-secreted proteins, (2) identify disease-induced secreted proteins that influence pathology, (3) develop “receive-ome” tools to identify the tissue targets of circulating proteins, and (4) adapt AI-based tools to predict hormones and their receptors. These independent projects allow discoveries to be cross-validated across aims, increasing confidence in the results. In vivo experiments will be initially performed using Drosophila, an ideal model due to its conserved organ and endocrine systems, and then followed up in zebrafish and mice. Computational analysis will directly utilize human datasets. Our preliminary results demonstrate the feasibility of the proposed aims, leading our group to study novel circulating factors influencing body size and synaptic function. As technologies in the field evolve, we will expand to other signaling molecules including lipids and exosomes, as well as extracellular fluids such as cerebrospinal fluid (CSF). We will disseminate a variety of resources and datasets, including plasmids, transgenic animals, and detailed visualizations of inter-organ signaling networks. These resources will broadly benefit researchers investigating inter-organ communication, as well as those studying local cell-to-cell signaling and protein-protein interactions. Through these holistic efforts, we expect to find novel inter-organ ligands and receptors, unexpected tissue sources of circulating proteins, and potential targets for diagnostic markers and therapeutics.

Maryland State Arts Council

Maryland State Arts Council provides grants to support arts and cultural programming, artist fellowships, arts education, and community-based arts projects across MD.

RV Steering Committee

Over a six month period the Rainier Vista Steering Committee (RVSC) will facilitate 12 virtual workshops on subsequent Saturdays for youth ages 11-17. The project will begin January of 2021 and will run through June. Participants will share stories about their life experiences since Covid-19 using art, literature photography and spoken word. A booklet of reflections will be produced and shared with the community.

Massachusetts Cultural Council

Massachusetts Cultural Council provides grants to support arts and cultural programming, artist fellowships, arts education, and community-based arts projects across MA.

Massachusetts Cultural Council

The Cultural Investment Portfolio (CIP) provides general operating support grants to established Massachusetts cultural organizations, enabling them to deliver arts, humanities, and interpretive science programs of public benefit.

Massachusetts Cultural Council

The Local Cultural Council (LCC) Program distributes grants through 329 local cultural councils to support arts, humanities, and interpretive science programs that engage the public across Massachusetts communities.

NCI - National Cancer Institute

Project Summary/Abstract CANDIDATE: I am a postdoctoral fellow in Dr. Christopher Barbieri’s laboratory at Weill Cornell Medicine, where I investigate the molecular determinants of prostate cancer progression. My PhD training at Tianjin Medical University focused on androgen receptor (AR) signaling and therapy resistance. At Weill Cornell, I have advanced my expertise in high-throughput genomic and epigenomic analyses. My career development plan focuses on acquiring skills in single-cell transcriptomics and epigenomic profiling to prepare for an independent faculty position in translational prostate cancer research. RESEARCH: In the normal prostate, AR signaling maintains luminal identity and drives growth-suppressive programs; however, during tumorigenesis, AR is co- opted into oncogenic programs that drive malignancy. Preliminary data from my genome-wide approach with an unbiased CRISPR-screen have identified MBD4, a methyl-CpG binding protein, as a pivotal regulator of these opposing AR programs. Loss of MBD4 redirects AR activity away from growth suppression toward oncogenic programs by altering FOXA1 binding. In this proposal, I will (Aim 1) define the molecular mechanisms by which MBD4 loss modulates AR function through cell-based assays, organoids, and in vivo models; (Aim 2) elucidate MBD4’s role in maintaining prostate luminal specification using single-cell transcriptomic and functional studies; and (Aim 3) characterize the temporal dynamics of MBD4 downregulation during the transition to prostate cancer through integrated single-cell RNA and targeted methylation profiling. These studies will test whether MBD4 functions as a molecular switch controlling AR’s dual programs and will identify novel therapeutic strategies for precision AR-targeted therapy. ENVIRONMENT: Weill Cornell Medicine is internationally recognized for its excellence in prostate cancer research and provides cutting-edge genomic, epigenomic, and single-cell analysis platforms. Because of the bioinformatics relevance of the project, I have included Dr. Andrea Sboner as a co- mentor; his expertise in computational genomics and multi-omics analysis will be essential for guiding the study’s bioinformatics aspects. My advisory committee also includes Dr. Yu Chen from Memorial Sloan Kettering Cancer Center, whose expertise in prostate cancer organoid modeling and epigenetic regulation provides invaluable insight, and Dr. David Rickman, a leading authority in cellular transitions and lineage plasticity. The combined expertise of my mentors and advisors, along with the state-of-the-art resources at Weill Cornell Medicine, will strongly support my successful transition to an independent research career.

McKnight Foundation

The McKnight Foundation funds nonprofits, artists, and researchers in Minnesota and beyond in areas including arts and culture, Midwest climate and energy, Mississippi River, neuroscience, international programs, and regional grantmaking.

NIEHS - National Institute of Environmental Health Sciences

ABSTRACT The Measuring Micro- and Nanoplastics in Environmental Samples Workshop (“the Workshop”) will be held as part of a Great Lakes Microplastic and Health Symposium (June 17-18, 2026) in Rochester, New York. This Workshop aims to pool existing expertise, establish new collaborations, and prioritize future needs for characterizing micro- and nanoplastics (MNP) in complex environmental matrices relevant to health research in the Great Lakes ecosystem. This Workshop will bring together national experts on MNP analysis with researchers studying the interactions between Great Lakes ecosystems (particularly water and air) and health. The Workshop will be hosted by two multidisciplinary centers that are part of the NIH/NSF Centers for Oceans and Human Health program. The University of Rochester and Rochester Institute of Technology co-host the Lake Ontario MicroPlastics (LOMP) Center, a transdisciplinary hub for research, translation and engagement on how MNPs affect human health and the Great Lakes environment (particularly water and near shore air). The Great Lakes Center for Fresh Water and Human Health is based at the University of Michigan (the Great Lakes Center). Its central goal is to better understand the increasing risks that cyanobacterial harmful algal blooms (cHAB) pose to freshwater ecosystems and human health. One aspect of the Great Lakes Center’s research is to explore the association of aerosolized microplastics and cHABs. Fostering collaboration and scientific exchange between the Great Lakes Center, LOMP, and national experts studying MNPs is one example of how this workshop may have long-term impacts on regional MNP research collaborations that may yield results of global significance. The Workshop has three objectives: 1) Bring together scientists from multiple laboratories and institutions to present new findings, identify methodological challenges, and share analytical approaches to better understand environmental exposures to MNPs and their potential impact on human and environmental health; 2) Develop new collaborations between researchers and trainees across institutions, both regionally and nationally; and 3) Identify priorities and opportunities for analytical advances to support this rapidly growing field. The objectives will be met through a Workshop format with a limited number of participants (30-40) to encourage close interaction. The requested R13 conference funds will be used to pay for expenses for the June 17 Workshop, including travel expenses for trainees, invited experts, and researchers from other institutions. R-13 support will also enable these attendees to participate in the full Great Lakes Microplastic and Health Symposium, to be held on June 18 at the University of Rochester’s Memorial Art Gallery, that will include a larger number of researchers, trainees, and community partners (120-150) on a broad range of topics related to Microplastics and Health with a focus on the Great Lakes region.

NIH

Significance to the VA: Veterans experience strokes more frequently than non-Veterans. One of the most debilitating consequences of a stroke is aphasia, a chronic language disorder that affects one third of stroke survivors. While individuals experience a degree of spontaneous recovery within the first 6-12 months post- stroke, most individuals do not fully recover. Continued neuroplastic change to the language system is conferred via long-term behavioral speech-language treatment which is most effective at high doses. Unfortunately, Veterans receive considerably less treatment than best-practice guidelines. Motivation is likely a major contributor to this gap in clinical care. In fact, the VA Clinical Practice Guidelines for Management of Stroke Rehabilitation cite motivation as a barrier to post-stroke rehabilitation and recommend that clinicians assess motivation. However, there are no motivation-based measures with validity evidence in the context of aphasia rehabilitation, creating a critical gap in clinicians' ability to assess and ultimately intervene upon motivation to optimize the amount of treatment Veterans with aphasia receive. Innovation and Impact: This proposal aims to address this gap by modifying an existing measurement tool that, with appropriate adaptations, can identify causes of suboptimal motivation in the context of aphasia rehabilitation. The measure is grounded in self-determination theory, a well-studied theory of motivation that finds that optimal motivations (described as “autonomous”) lead to improvements in health behavior and well- being, and bolster cognitive functions associated with aphasia treatment response. Autonomous motivations are driven by contexts that support three basic psychological needs (BPNs). Measures of BPNs can identify underlying causes of suboptimal motivation and guide interventions that improve motivation – a critical variable that can increase the dosage of treatment and enhance cognitive skills key to treatment response. Specific Aims and Methodology: This proposal is guided by state-of-the-art guidelines for measure development. In Aim 1, speech pathologists and researchers (n = 30) will be presented with the unmodified scale via survey and identify irrelevant or missing items. An expert panel will modify or develop items using survey data. In Aim 2, Veterans with aphasia (n = 7-10) will be presented with the scale modified in Aim 1. Via cognitive interviewing, Veterans will identify items they find difficult to understand, irrelevant, or missing. An expert panel will modify or develop items using cognitive interview data. In Aim 3, Veterans with aphasia (n = 25) will complete the modified scale to generate validity evidence that can support its use in future research. Path to Translation/Implementation: This proposal constitutes the initial steps of a productive and programmatic line of research evaluating the effect of BPN-focused motivational interventions on aphasia treatment outcomes in a single-subject investigation (CDA-2) and large-scale efficacy research (MR Award). This work will enable clinicians to assess motivation as advised by VA Clinical Practice Guidelines. Existing frameworks of rehabilitation (e.g., the Rehabilitation Treatment Specification System) can aid researchers in using the scale to identify treatment ingredients that optimize motivation, a variable central to outcomes. Immediate and Long-term Career Goals: The applicant is a speech pathologist and an advanced fellow in geriatrics in the Geriatrics Research, Education, and Clinical Center at VAPHS, with experience in aphasia rehabilitation and research. During the CDA-1, mentorship and structured training in measurement theory, qualitative research, motivational science, and professional development will support the completion of the proposed CDA-1 and the applicant's career goals to become an independent VA clinician-scientist supported by VA Merit Review and NIH/NIDCD award mechanisms. The applicant's long-term research program will (1) identify neurocognitive and psychological mechanisms underlying aphasia recovery and (2) determine how treatment ingredients can be modified to maximize aphasia treatment outcomes at the individual level.

NIGMS - National Institute of General Medical Sciences

Predictions of drug disposition and toxicity from preclinical data, estimation of risk of drug-drug interactions and simulation of drug exposures in humans are essential for safe and effective drug development and clinical pharmacology. Despite decades of research, significant gaps still remain in translating preclinical findings to clinical drug development and practice. As a result, about 90% of drug candidates that enter Phase I clinical trials fail during development. In addition, uncertainty regarding the disposition characteristics of approved drugs often remains after drug approval leading to post marketing studies and label revisions. This wastes resources, leads to unnecessary risk to patients and limits access to life saving medications for specific patient populations. My laboratory works to bridge the knowledge gaps in translational science via mechanistic in vitro studies, mass spectrometry based proteomics experiments and state-of-the art in silico modeling. We develop novel proteomics methods to understand formation of drug adducts by small molecule drugs, evaluate how drug-protein and protein-protein interactions within a cell alter drug distribution and the activity of drug metabolizing enzymes, and assess how genetic variability and individual biology such as sex, disease and age alter drug exposures in tissues and in blood. Central questions of my research program include: What mechanisms cause significant under and overpredictions of drug exposures and drug-drug interactions in humans? Which individual factors define interindividual variability in drug-protein adduct formation making certain individuals highly susceptible? What mechanisms lead to changes in drug clearance in specific populations and between individuals? Answering these questions will advance developing individualized therapy and enable connecting observable patient specific factors with decisions of drug dosage adjustments. Our long term goal is to improve the preclinical, translational and computational methodologies used to predict and evaluate drug disposition and adverse drug reactions. Our current studies are focused on 1) development of innovative proteomic methods for discovery, characterization and quantification of protein adducts in simple and complex biological matrices; 2) evaluation of the role of fatty acid binding proteins in modulating tissue drug distribution and drug clearance and 3) developing novel physiologically based pharmacokinetic models to predict drug disposition in specific populations including vulnerable patients. Our studies advance innovative areas such as how intracellular binding proteins influence drug efficacy, and provide unprecedented insight into mechanisms of enzyme inactivation and quantitative relationships between adduct formation and metabolic activity in the liver. Our research will also provide open and accessible cutting-edge tools for high-dimensional proteomics data and novel PBPK models for prediction of drug disposition in specific patient populations. Our work promises to decrease failure rate during drug development and lead to expanded access to approved medications in specific patient groups through improved quantitative systems biology approaches.

NINDS - National Institute of Neurological Disorders and Stroke

Why do clinical trials fail? In our lab, we seek to understand some of the reasons for previous failures in translating preclinical assessments to clinical trial success. In 2 recent publications we elucidated a fundamental biological property of the neurovascular unit—we call this property differential vulnerability, and it means that although neurons are exquisitely sensitive to substrate deprivation and die rapidly, glia (astrocytes and microglia) are much more tolerant; endothelial cells and pericytes are intermediate. Neurons and glia tolerate ischemia on different time scales, suggesting to us that perhaps they need cytoprotection at different times. The molecular and cellular mechanisms underlying differential vulnerability are not clear. We believe that several mechanisms contribute to differential vulnerability, but the relative importance of each remains to be studied. Cerebral ischemia sets in motion a cascade of mechanisms resulting in cell death by one or more mechanisms including ferroptosis, necrosis, apoptosis, necroptosis, and parthanatos, among others. To explore mechanisms of astrocyte response during ischemia, we used regional transcriptomics after MCAo in rats. Agnostic Ingenuity Pathway Analysis of the astrocyte responses led to several candidate pathways and the greatest log-fold change was in the nuclear factor erythroid 2-related factor (Nrf2) mediated oxidative stress response. Other interesting pathways included the unfolded protein reasons and the NGF stimulated transcription response. We now propose a series of studies to test the role of Nrf2 mediated response underlying differential vulnerability. Oxidative injury results when free radicals oxidize and degrade proteins and nucleic acids and react with polyunsaturated fatty acids causing lipid peroxidation. Redox stress subsequently activates necrosis, apoptosis, and ferroptosis, which is also triggered by toxic lipid moieties. Neurons activate several defenses against oxidative stress and ferroptosis, but they depend heavily on astrocytes for both energy and resorption of toxic lipid moieties. These data lead to the central hypothesis to be tested in this project: neurons and astrocytes tolerate ischemia differently as a result of their different capacities to handle redox stress. We will test this hypothesis using state of the art genomic, pharmacological, and physiological manipulations. Specifically, the aims will test the relationship between glycogen metabolism, lipid peroxidation and stress response; test whether cell death mechanisms dependent on free radical stress differ between astrocytes and neurons; test the role of NRF2 in differential vulnerability; and finally look at whole animal stroke after optimized interventions timed and dosed to targeting free radical stress differently in neurons and astrocytes. CONFIDENTIAL

National Institutes of Health

-Purpose. This Funding Opportunity Announcement (FOA) issued by the National Institute on Alcohol Abuse and Alcoholism (NIAAA), the National Cancer Institute (NCI), and the Office of Dietary Supplements (ODS), National Institutes of Health (NIH), solicits grant applications from institutions/organizations that propose to employ an integrative approach using state-of-the-art technologies to gain insight into the molecular and biochemical mechanisms by which chronic alcohol consumption leads to the development of cancers of various organs such as oral cavity, pharynx, larynx, esophagus, stomach, large intestine, liver, and breast. This includes investigating roles of various factors such as acetaldehyde, cytochrome P450 2E1 (CYP2E1), vascular endothelial growth factor (VEGF), impaired immune function, and alcohol-induced impaired metabolism of s-adenosylmethionine (SAMe), folate, betaine, iron, and vitamin A. -Mechanism of Support. This FOA will utilize the NIH Research Project Grant (R01) award mechanism and runs in parallel with an FOA of identical scientific scope, PA-06-270, that solicits applications under the Exploratory/Developmental (R21) grant mechanism.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary Due to the progressive depletion of CD4+ T cells, HIV/AIDS patients are more susceptible to infection of Cryptococcus neoformans, an encapsulated fungus that is listed by WHO as a pathogen of critical priority. An estimated 112,000 people die from C. neoformans infections globally each year. C. neoformans is a leading cause of mortality among HIV/AIDS patients. Antiretroviral therapy (ART) can rapidly restore CD4+ T cells in HIV/AIDS patients, representing a major advance in the treatment of HIV-infections. However, the rapid recovery of CD4+ T cells in HIV/AIDS patients that co-infected with C. neoformans after initiation of ART often causes an exaggerated inflammatory response in the central nervous system (CNS), termed immune reconstitution inflammatory syndrome (IRIS). Cryptococcal IRIS is considered as a life-threating condition, as it can lead to a high mortality rate during ART among HIV/AIDS-associated IRIS patients who have a cryptococcal infection. Clinical studies have shown that cryptococcal IRIS is associated with a substantial CNS recruitment of immune cells and enhanced secretions of inflammatory cytokines. However, the cellular and molecular mechanisms involved in cryptococcal IRIS are poorly understood. In particular, it remains to be determined which leukocyte subset(s) and cytokine(s) mediate the death during cryptococcal IRIS and how CD4+ T cell responses are regulated during cryptococcal IRIS. These questions are hard to be answered without animal studies. Using a novel murine model of C. neoformans infection to closely mimic HIV/AIDS-associated cryptococcal IRIS, we will define the mechanisms of cryptococcal IRIS by addressing the following aims: 1) To identify the leukocyte subset(s) accounting for lethal cryptococcal IRIS; 2) To identify the cytokine(s) accounting for lethal cryptococcal IRIS; 3) To identify the factors regulating CD4+ T cells responses during cryptococcal IRIS. Successful completion of this study would gain novel insights into the cellular and molecular mechanisms that mediate lethal immune responses during cryptococcal IRIS and identify potential targets for treatment of HIV/AIDS-associated cryptococcal IRIS patients.

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY/ABSTRACT Dynamic interactions between intercellular membranes and the cortical cytoskeleton are critical for cell survival. During their normal functions, cells face physiological and environmental stresses that can lead to rupture of the plasma membrane and/or of the nuclear envelope. Rapid repair of such injuries, whether arising from daily activities or resulting from trauma, infection, or diseases such as cancer, is an active area of cell biology research. My lab has a long-standing track record in successfully identifying key molecules and elucidating their in vivo roles at the cell cortex necessary for the repair of plasma membrane and at the nuclear envelope necessary for repair of the nuclear cortex. The overall focus of the lab is to delineate how cells deal with such cell and/or nuclear cortex disruptions to efficiently and effectively repair the lesions. We have developed a robust inducible single cell repair model using the syncytial Drosophila embryo that has superb amenability for live imaging and genetic tractability that is unavailable in other cell wound repair models. We have also established a model for the newly appreciated nuclear export pathway (Nuclear Envelope budding) on the surface of Drosophila salivary gland and S2 cell nuclei that provides the same superb amenability for live imaging and genetic approaches. While both systems rely heavily on dynamic membrane and/or cytoskeleton/nucleoskeleton interactions, a major challenge in both of these systems is the absence of a molecular outline of the events occurring during the repair and/or export processes in any organism or system. Our goals during the proposed period are to establish the molecular framework underpinning these processes using a combination of state-of-the-art cell biological, genetic, developmental, biochemical, and high-resolution imaging approaches. Our studies are expected to be of significant medical relevance, as understanding the molecules, machineries, and pathways governing cell wound repair, NE-budding (nuclear export), and dynamic membrane-cytoskeleton/nucleoskeleton interactions will be extremely valuable for elucidating fundamental cellular mechanisms, as well as for developing new or enhancing existing strategies for treating conditions associated with cell/nuclear damage, and for disciplines such as regenerative medicine where cell based constructs are used to reconstruct tissues, clinical drug delivery systems where molecules cross cell membranes, and for virus nuclear egress.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY/ABSTRACT Mitral valve prolapse (MVP) is a common valvular disease, affecting 2-4% of the population. It is increasingly linked with life-threatening complex ventricular arrhythmias (CVA) and sudden cardiac death (SCD), even in the absence of significant mitral regurgitation. Despite this emerging association, the mechanisms underlying arrhythmogenesis remain poorly understood, limiting risk stratification and treatment strategies. New tools are needed to allow clinicians to identify patients at risk of devastating cardiac events from in vivo measurements of left ventricle (LV) function. This proposal aims to address the current gap in mechanistic understanding and clinical risk-assessment through the development of “personalized” computational cardiac models from in vivo imaging data. We hypothesize that stress-induced fibrosis in the myocardium is a substrate for altered mechanics, hemodynamics, and electrical disturbance in the LV; these patient-specific models can elucidate the relationship between these events. Previous work in our lab has leveraged subject-specific models of the heart to investigate the effects of biomechanical remodeling of cardiac function. We propose to use a similar framework to model the human heart and deepen our understanding of the biomechanical remodeling in MVP-associated arrhythmias. Building on these insights, we aim to elucidate image-derived biomarkers for the non-invasive prediction of arrhythmias in MVP patients. We will leverage multimodal imaging data - including fibrosis quantification, deformation measures, and flow dynamics - to predict arrhythmic risk and identify patients who may benefit from early intervention. Specifically, we intend to achieve the following aims: Determine the biomechanical mechanisms underlying fibrosis in arrhythmia MVP. We will use state-of-the-art in vivo imaging, image processing, and patient-specific 3-D computational modeling tools to characterize the biomechanical remodeling of the LV in MVP patients and link these events to arrhythmogenesis. Identify high-fidelity intracardiac hemodynamic characteristics of MVP patients not captured by traditional organ-level metrics. We will use established computational fluid dynamics (CFD) techniques to assess the patient-specific intraventricular hemodynamics in the LV of MVP patients and correlate altered flow with arrhythmia. We will integrate imaging-based biomechanical and hemodynamic biomarkers to reveal the predictive power of image-derived metrics to enable early identification of high-risk patients and guide personalized clinical management and intervention strategies.

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY Chronic pain affects one in five Americans, yet fewer than half find adequate relief from existing treatments. For severe pain, opioids often remain the primary option, but their overreliance has fueled the ongoing opioid crisis − a nationwide epidemic now in its third wave, according to the CDC. While effective pain therapies are lacking, promising therapeutic targets are abundant. Temperature receptors, such as TRPV1, act as gateway detectors of pain signals, making them ideal targets for developing analgesics that act directly at pain initiation sites. However, targeting these receptors in clinical settings has proven challenging, requiring mode-specific antagonists that selectively inhibit agonist sensitivity while preserving thermal functions. Understanding how these receptors respond to thermal and chemical stimuli is thus crucial for developing effective therapies. We have recently discovered that the gating of temperature receptors is uniquely complex, involving a “suicidal” mechanism where heat-induced opening is inherently coupled with protein unfolding. Their functional states are transient, lasting only milliseconds, posing significant challenges for structural studies. To overcome these obstacles, we propose developing a novel cryo-EM sample preparation device that enables time-resolved cryo-EM imaging to capture transiently active conformations. By integrating this technology with state-of-the-art methods – including ultrafast T- jump patch-clamp recording and ultrasensitive differential scanning calorimetry – we propose studies to elucidate the structural basis of the suicidal mechanism, uncover the molecular pathways underlying heat- induced activation within the receptors, and identify mode-specific conformational states triggered by heat and agonists. The successful completion of this project will advance both the methodology and understanding of thermal transduction and provide essential structural insights to support mode-specific antagonist development targeting temperature receptors. Furthermore, the innovative technology will have broad applications for studying protein dynamics and other temperature-driven biological processes.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary In most patients with HIV-1 infection, antiretroviral therapy (ART) successfully suppresses viral loads and restores CD4+ T cell numbers. However, lifelong therapy is required to maintain viral suppression, primarily due to a major latent reservoir in resting CD4+ T cells. The latent reservoir poses a great barrier to HIV cure and ensures viral persistence in patients. Knowledge about how such a latent reservoir is formed is limited. A more complete understanding of the mechanisms contributing to the establishment of the reservoir will impact the strategies in battling viral persistence. Microenvironment of the lymphoid tissue and cell-cell interactions in vivo played important roles in latency formation in resting CD4+ T cells. It was found that endothelial cells (ECs), which physiologically interact readily with T cells in the lymphoid tissues in vivo, promote both productive and latent HIV infection in resting CD4+ T cells and may play a significant role in latency formation in these cells in vivo. Having established the importance of ECs in HIV infection and latency formation, this study is to elucidate further the mechanisms involved in EC stimulation of T cells. The specific aims of the proposal are: 1. To investigate the roles of integrin on HIV infection of CD4+ T cells. 2. To examine the role of Bcl-3, Bcl-6, and SAMHD1 on HIV infection in EC-stimulated resting CD4+ T cells. 3. To identify T cell subtypes and signature molecules correlated with HIV infection in EC-stimulated CD4+ T cells. To achieve specific aim 1, blocking antibodies will be used to block integrins during EC stimulation of T cells. Chimeric antibodies will be used to stimulate integrins on T cells to see their effect on HIV infection, with or without IL-6. To achieve specific aim 2, intracellular staining will be used to determine whether there is correlation between expression of Bl-3, Bcl-6 and Phosphorylated SAMHD1 and HIV infection rates in EC stimulated resting CD4+ T cells. To achieve specific aim 3, intracellular staining for lineage-determining molecules will be used to identify potential subtypes of CD4+ T cells that have higher HIV infection rates upon EC stimulation. Single cell RNAseq method will also be used to identify novel subtypes and signature molecules that correlate with HIV infection in EC-stimulated resting T cells. The long-term objective of the study is to understand how endothelial cells impact HIV infection and latency formation in T helper cells. The knowledge gained from this study will significantly improve the understanding of HIV latent reservoir formation in a physiological context and will influence the strategies for battling viral persistence.