Grants

Public Services and Procurement Canada | Services publics et Approvisionnement Canada

At the end of each fiscal year, government-wide financial information is published in the Public Accounts. This dataset, based on the Volume III of the Public Accounts, provides a detailed listing of the transfer payments (i.e. cash payments and accrued charges) aggregating to $100,000 or over to a recipient (one individual or organization). A transfer payment is a grant, contribution or other payment made by the Government for which no goods or services are received. This detail shows the ministry, department, the name and location of the recipient, together with the total amount paid. Appendices referenced in the dataset can be found in the Official Record of Information. In cases where certain organizations have more than one place of business, their Head Office location was used for reporting purposes. In addition, this statement presents, for each class of recipients, the total amount of payments to a recipient aggregating to less than $100,000 and the total number of re

NINR - National Institute of Nursing Research

Abstract Eysz, Inc. is developing a mobile health (mHealth) platform, the Eysz HV Recorder, to remotely diagnose and monitor childhood absence epilepsy (CAE) to improve patient management and reduce the time to seizure freedom. Early treatment of CAE is critical, but absence seizures are currently challenging to identify, and diagnosis is frequently delayed. The gold standard for seizure detection is video EEG (VEEG), but VEEG has several drawbacks for treatment management of CAE, causing most clinicians to rely on self-reports, even though studies have shown that patients and caregivers report only about 6% and 14% of all absence seizures, respectively. Thus, there is an urgent need for an accurate remote monitoring to assist clinicians in assessing treatment response over time for patients with CAE. The Eysz HV Recorder is an mHealth app used to assist clinicians in the diagnosis of CAE and is currently being used in pediatric and child neurology clinics and undergoing a post-marketing study. The Eysz app guides patients through hyperventilation (HV) while giving continuous feedback to safely and effectively provoke absence seizures while video-recording the patient's face. The recording is then analyzed by a child neurologist and a report is sent to the treating physician. Our preliminary results from our post-marketing clinical study have shown that our video analyses have a sensitivity close to 100% and a specificity of 92% compared to VEEG for seizures greater than 7 seconds in duration. This Fast Track SBIR project seeks to develop and test a home-based version of the Eysz HV Recorder, enabling more frequent and convenient assessment of the patient’s treatment regimen. The FDA has requested additional testing prior to in-home use of the Eysz HV Recorder; therefore, this Fast Track proposal seeks to develop an in-home version of the app and validate safety protocols prior to launching an in-home study that will support FDA clearance of the app. Phase I Aim 1: Define and validate user and system requirements to ensure safety, usability, data quality, and regulatory compliance. Phase I Aim 2: Build and validate the software and training materials, using a standard risk-based software development approach. We will also develop training and educational materials for caregivers. Phase I Aim 3: Test safety procedures and app usability in a clinical setting. We will iteratively introduce the app and training materials to cohorts of users in a clinical setting to evaluate usability and data quality. After meeting our Go/No Go Milestones for Phase I, we will proceed to Phase II Aim 4: Perform an at-home study to assess usability, adherence, diagnostic accuracy and safety. This pivotal study will test the use of the app by caregivers in real-life home environments. This study will provide key data to support FDA clearance of the app and will set the stage for its future deployment as a tool for at-home monitoring and management of CAE.

ATSDR - Agency for Toxic Substances and Disease Registry

ABSTRACT Amyotrophic lateral sclerosis (ALS) is a rare, neurodegenerative condition that results in rapidly progressive muscle weakness and, ultimately, death from respiratory failure within 2-5 years of symptoms onset. The VCU Department of Neurology runs the largest ALS clinic in the region and, over the past 4 years, began witnessing an uptick in incident ALS cases. In partnership with the ALS Association, we analyzed the spatial distribution of ALS in Virginia. We identified apparent spatial ALS clustering with the highest relative risk of 3.75 for developing ALS in these “hotspots.” It should be mentioned that ALS has a global incidence of 2 per 100,000. There is mounting evidence that ALS stems from a combination of genetic susceptibility, age-related cellular damage, and environmental exposure. We, therefore, investigated whether local environmental factors contributed to increased ALS risk in these “hotspots.” Analysis of drinking water samples from households of ALS patients from this cluster identified elevated levels of a plasticizer dtBHQ (2,5-ditert-butylhydroquinone). At this point, it is unknown how dtBHQ appears in drinking water in these regions, but a reported application of dtBHQ is in the coatings of water pipes. Nonetheless, the presence of dtBHQ is intriguing, given some reports that at high doses, dtBHQ can cause muscle weakness and damage motor endplates in rats. Next, we investigated whether low-level exposure to dtBHQ could cause motor impairment in laboratory animals. Using a Zebrafish model system, we found that dtBHQ at nanomolar concentrations decreased touch-evoked response and caused shortening of primary motor axons and reduced branching indicative of motor circuit dysfunction. These preliminary results led us to propose a central hypothesis that the presence of a plasticizer dtBHQ in the environment of ALS clusters activates molecular mechanisms that cause motor neuron toxicity, contributing to the elevated risk of ALS in these hotspots. We are applying under Funding Option B and our proposal meets both of the RFA-TS-25-036 funding objectives by identifying a potential environmental risk factor for ALS and investigating how this risk factor contributes to the pathophysiology of ALS. Thus, for this investigation, in Aim 1, we will investigate the environmental and occupational risk factors in our ALS clusters through deep qualitative interviews. We will also compare this data with the National ALS Registry Epidemiological data of registrants from Central Virginia and the National cohort. In Aim 2, we will collect water specimens from households in ALS clusters and analyze them for dtBHQ levels and the presence of other toxicants. In Aim 3, we will investigate whether sustained, low-level exposure to dtBHQ activates neurotoxic pathways that cause motor neuron toxicity and lead to the expression of ALS-like phenotype using zebrafish and mice as model systems. This study will provide new data on the presence of a novel environmental toxicant and its role in elevating the risk of ALS in humans.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary / Abstract Bacillary dysentery or shigellosis is caused by bacteria of the Shigella species: S. dysenteriae, S. flexneri, S boydii, and S. sonnei. Infections range from mild or asymptomatic to severe bloody diarrhea, so reported prevalence grossly underestimates actual prevalence. Shigellosis is a global public health concern and antimicrobial resistance has compounded the problem. Moreover, shigellosis is also sexually transmitted in the United States, Europe, and other developed countries. Such outbreaks are driven by the emergence of antibiotic resistant strains of S. flexneri infecting men who have sex with men. The objective of this proof-of-concept study is to demonstrate that wastewater-based epidemiology (WBE) using digital PCR to detect pathogen molecular markers can provide a more accurate picture of community prevalence of bacterial pathogens than traditional case reporting. Wastewater surveillance for viral pathogens has been in use for decades and its use to monitor SARS-CoV-2 is now widely applied globally. Analogous methods for wastewater surveillance of bacterial pathogens by digital PCR has lagged. We will develop and validate methods to detect Shigella, an important bacterial agent of diarrheal disease, in wastewater. We will also field test the methods to assess shigellosis prevalence at community and building scale and link the data to the actual population residing in the wastewater collection area. We will provide actionable data to the local health department which can then develop targeted public health interventions. The independent but complementary aims in demonstrating the proof-of-concept are: 1. Develop and validate sensitive, specific, and reproducible WBE methods for detection of Shigella species in wastewater using digital PCR. Three subaims will: 1.1) optimize methods for extraction and detection of Shigella in wastewater using laboratory-grown strains of Shigella species to “spike” authentic wastewater and laboratory prepared “synthetic” wastewater; 1.2) validate molecular targets for differentiation of S. flexneri from S. sonnei in wastewater; and 1.3) culture of Shigella from wastewater to assess antibiotic resistance genotypes and phenotypes. 2. Assess proof-of-concept of WBE as a public health tool for Shigella at community and building scale with emphasis on high risk congregate populations, i.e., day care centers. Two subaims will: 2.1) measure the prevalence of Shigella species at community scale by sampling at the wastewater treatment plant intake and using wastewater flow rate as a population normalization marker; and 2.2) extend shigellosis surveillance to building scale targeting high risk congregate pediatric populations. Strengths of this proposal are the innovative application of WBE to a bacterial pathogen, our method for population normalization, differentiation of Shigella species, prevalence measurement of Shigella species at community and building scale, and the expertise of our multidisciplinary team.

Friends of Detective Cookie Chess Park

This project is to plan and design an outdoor chess park, and multi-functional community space. The space would honor the work and influence of Dectective Denis `Cookie` Bouldin's decade-old chess club an its positive impact on the youth of South Seattle.

Friends of Detective Cookie Chess Park

The Friends of Detective Cookie Chess Club and the Rainier Beach community will continue their work with a landscape architect to develop a site survey and 30% Street Improvement Permit (SIP) construction drawings for the 'Chess Park' located at 9100 Rainier Ave South. The work is scheduled to be completed in early 2018.

Friends of Detective Cookie Chess Park

The Friends of Detective Cookie Chess Club will continue working with a landscape architect, and civil engineer to develop the 60% Street Improvement Permit (SIP) construction drawings to submit to SDOT by late Spring 2019. This phase includes the permit review by SDOT. The resulting, permitted drawings will then be ready for the next phase of construction, separate from this proposal.

Friends of Detective Cookie Chess Park

The Friends of Detective Cookie Chess Park is planning Phase 4 of the project. The Detective Cookie Chess Park located at 9100 Rainer Ave S envisions a inter-generational, multicultural community space where neighbors can gather and play chess while honoring Detective Cookie, a local policewoman, who started a youth Chess Club over 12 years ago in Rainier Beach. Phase 4 will focus on initial construction of the park beginning with demo, grading and drainage, and utilities. This project is expected to be completed in the late Fall of 2021.

NSF

The National Science Foundation’s ground-breaking Advanced LIGO gravitational-wave observatories continue to explore the dark universe and have now measured the collision of over 250 black hole pairs. This award supports gravitational-wave detector technology research to improve the observational reach of the Advanced LIGO observatories. Their sensing is improved by controlling thermal distortion effects when operating at high laser power, and by lowering the thermal noise in their test mass mirror coatings. Specifically, this award explores technology to integrate solid-state laser cooling in optical mirror coatings to create radiation-balanced mirrors exhibiting no thermal deformations. It also supports investigating the suitability of much lower thermal noise crystalline optical mirror coatings. Both technologies can find much wider use in precision sensing and high-power laser applications, with examples including their use in laser fusion and defense. This award supports (i) investigating solid-state laser cooling to control thermal lensing in gravitational wave interferometers, to optimize ion-beam-sputtered materials for use in laser-cooled optical coatings. (ii) Characterizing the performance of low-noise crystalline coatings under high-power optical conditions found in gravitational-wave detectors. (iii) Preparing the LIGO Hanford detector for its O5 observation run, installing hardware to preheat optics so thermal transients during lock acquisition can be avoided. And (iv) designing test mass actuators to be used in the suspensions of the next-generation observatory Cosmic Explorer. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF

The project involvew research in two main directions in descriptive set theory, which is a branch of mathematics in which modern set-theoretic methods are used to develop the theory of subsets of the real line and related structures which are in turn the fundamental objects of mathematical analysis used throughout mathematics and its applications. One of the directions concerns developing the structural theory of sets in models of the axiom of determinacy. This is important as this axiom holds in various mathematical universes which interact with the ``true'' universe and also because it is giving us the theory of definable objects which is a central goal of descriptive set theory. A second direction concerns the theory of definable equivalence relations. This is a relatively recent area of descriptive set theory which has been investigated extensively the past several decades. This area interacts heavily with a number of different areas of mathematics and provides a unifying framework for them. There are a number of fundamental problems that remain open in this area concerning the structure of countable Borel equivalence relations in particular, but significant progress has been made in the last few years. The project will further develop these methods with a goal of attacking some of these problems. This project will involve graduate students. Specifically, the combinatorics of non-wellordered sets in models of determinacy is a main line of the first direction of the proposal. An example is Chan's recent proof of the ``ABCD'' conjecture describing the relation between cardinalities of the form A^B for ordinals A and B (the conjecture roughly speaking asserts that the only relations between these cardinalities are the obvious ones). The principal investigator along with Chan and Trang have isolated a general principle about infinity Borel sets which also proves the ABCD conjecture and seems likely to have other applications. There are, however, many fundamental questions concerning these non-wellordered cardinalities that remain open which the proposal plans to investigate. The theory of countable Borel equivalence relations has shown much progress in recent years. For example, one of the central questions is the hyperfiniteness question which asks which groups have the property that all of their Borel actions generate only hyperfinite (an increasing union of equivalence relations with finite classes) equivalence relations. Recent work has shown that this class extends to include the polycyclic groups, so, in particular contains finitely generated groups of exponential growth. How much further this extends is an open problem. Some new techniques have been introduced recently by the principal investigator and co-authors which will help answer several structuring questions for actions of Z^n and other fairly simple groups. This hopefully leads to marking the boundary of what types of Borel or continuous structurings can be done in an invariant manner. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIMH - National Institute of Mental Health

Project Abstract On a moment-by-moment basis, each animal must consider its environment, experience, and goals to choose future actions. The striatum, including its dorsal and ventral portions, is an evolutionarily old structure that helps animals choose their actions and update action plans based on experience. Within this structure, dopamine places a crucial function in these processes. Dopamine signals in two ways – with slowly changing “tonic” levels and rapidly changing “phasic” signals. It has been proposed that tonic dopamine has fundamentally different functions and represents different features of experience than phasic dopamine. We have developed methods to measure tonic and phasic levels of dopamine in real-time as mice behave in a variety of contexts. We will use this new technology to examine the mechanisms and function of tonic dopamine levels to understand how perturbations of dopamine contribute to the cause and treatment of human neuropsychiatric disease.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY Implementation of the RTS,S malaria vaccine in sub Saharan Africa has been recommended by the WHO and UNICEF has pledged $170 million for production of the first vaccine supply. Because the efficacy of RTS,S wanes within one year from the primary doses, the regimen includes a 4th booster dose. However, based on findings from the phase 3 trial, the magnitude of Ab responses induced by the booster dose is significantly lower than those induced by the primary regimen. The booster dose increased duration of efficacy marginally. The regimen of another leading vaccine candidate, R21, also requires a booster dose. Understanding the immune mechanisms underlying poor booster dose responses will be important to support the design of future implementation studies and optimize actual implementation of RTS,S and other malaria vaccines. The limited effect of the booster dose might be explained by repeated exposure to malaria compounded to exposure to other infections after the first months of life. Extensive epidemiological evidence supports this hypothesis including a recent trial that showed that efficacy of administration of RTS,S in combination with repeated chemoprophylaxis was higher than RTS,S or chemoprophylaxis alone. Repeated malaria exposure may negatively affect Ab magnitude and functional activity via effects on innate, memory cells, and T cells. Additionally, our data, and data from others suggest that deficiency of micronutrients, particularly iron and zinc, acquired after cessation of breast feeding may impact responses to the booster dose. A third determinant of reduced responses to the booster dose could be the suboptimal spacing between the primary and booster doses, whereby memory B cells induced by the primary doses have waned before the booster dose. Following a cohort of 600 Malawian children from the primary vaccination until eight months after the booster dose, we will undertake comprehensive immunologic analyses to investigate the mechanisms underpinning the poor responses to the booster. We will identify the specific Ab responses and functions that are suboptimal after the booster dose. Additionally, we will i) quantify innate cell responses with and without the stimulation with AS01, and assess epigenetic changes leading to tolerogenic responses in monocytes; ii) quantify subpopulations of vaccine specific CD4+ T follicular helper cells and regulatory cells and their effect on Ab and B cell responses; iii) quantify induction and duration of memory cells, that may be impairing Ab responses to the booster dose; iv) assess whether exposure to malaria parasites, and micronutrient deficiency may be affecting all these immune responses; v) evaluate whether children with dysfunctional immunity to RTS,S also have impaired immunity to other childhood vaccines associated with the same determinants. Data generated by this study will help elucidate the basis of the limited effect of the booster dose and, thus, will be crucial to inform strategies to optimize boosting of the RTS,S-induced response.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY By mentoring junior investigators from the United States and low-come countries (LICs) in patient- oriented research (POR) in global health, particularly in vaccine-preventable diseases, I hope to build local capacity, improve outcomes and reduce health disparities. Working within the resource-limited health systems in LICs is challenging, and the influences of co-infections (particularly parasitic infections), anemia, undernutrition, and various microorganisms on vaccine efficacy may be underappreciated. For these reasons, vaccines and vaccine campaigns developed for and assessed in high-income countries (HICs) cannot be assumed to be feasibly implemented or to perform similarly in LICs. For instance, SARS-CoV-2 vaccination with mRNA-based vaccines in sub-Saharan Africa has been limited by the cold-chain requirement of these vaccines. Cultural and socio-economic factors have led Malawians to generally choose receiving the single dose Ad26.COV2-S rather than the ChAdOx1 vaccine. Growing evidence suggests that performance of vaccines in LICs is lower than HICs and the protective immunity elicited by some vaccines is shorter than originally anticipated. In an interconnected world where diseases do not have borders, optimizing interventions to control transmittable diseases in LICs impacts the health of populations worldwide. The overall goal is to train junior investigators across disciplines to optimize vaccinations in Malawi, a sub-Saharan African country in which I have established a thriving research infrastructure. The first two aims of this application, supported by my R01AI164686-funded project, will assess i) the longevity of antibodies (magnitude and function) and memory cells induced by the Ad26.COV2-S SARS-CoV-2 vaccine; ii) the contribution of pre-existing changes in innate immunity to these responses; iii) the association of malaria, micronutrient deficiency, and microorganisms colonizing the nasopharynx with these responses. In the third aim, funded through this proposal, we will address the longevity of five childhood vaccines, focusing particularly on optimizing the timing of re-vaccination. The importance of this question became obvious in 2023 when three poliovirus cases were identified in Malawi. The Ministry of Health, with no relevant data available, decided to revaccinate all children with an arbitrary cutoff of 15 years of age. I have mentored several investigators in POR, particularly from Malawi, and have been conducting research in the immunoepidemiology of vaccines in LICs (including Malawi) since 2015. With this award, I would be able to expand my panel of trainees, devote more time to each of them, acquire training to further my mentorship skills as well as my own professional skillset, and expand the focus of the research to include other childhood vaccines. With the research infrastructure built in Malawi, I am now well-equipped to successfully train clinician scientists and other junior investigators in POR in infectious diseases, particularly in vaccines in LICS.

NIAID - National Institute of Allergy and Infectious Diseases

Summary Ebola virus (EBOV), a filovirus, causes periodic outbreaks with high fatality rates. To understand the basis for viral pathogenesis, emergence, and to devise control strategies, it is important to define the mechanisms that underlie EBOV assembly and release of infectious virus. The VP24 protein, produced by one of seven viral genes, is a 24 kDa multifunctional protein unique to the filovirus family. EBOV VP24 blocks cellular responses to interferons (IFN) by preventing nuclear translocation of tyrosine phosphorylated STAT1 (pSTAT1), a transcription factor central to antiviral IFN signaling. VP24 accomplishes this by interacting with importin α (IMPA) nuclear transport proteins, preventing binding and nuclear import of STAT1, a key transcription factor needed for IFN responses. That VP24 interacts with IMPA nuclear import factors suggests that VP24 can traffic into the nucleus. In addition, transfection studies identified a nuclear export signal (NES) at the C-terminus of EBOV VP24, further supporting nuclear its trafficking. However, whether EBOV VP24 undergoes nucleocytoplasmic trafficking and the functional significance of this trafficking remain to be determined. VP24 also plays a critical but incompletely understood role in viral genome packaging and production of infectious viral particles. Most notably, VP24, the EBOV nucleoprotein and EBOV VP35 together form filamentous nucleocapsid structures, called nucleocapsid- like structures (NCLS) that are morphologically indistinguishable from the nucleocapsids present in EBOV virions. The presence of VP24 also renders NCLS competent for actin-dependent transport. Fundamental questions remain including how VP24 facilitates actin-dependent transport and the specific molecular features required for NCLS formation and trafficking. Our Preliminary Data indicates that VP24 can traffic into and out of the nucleus and that disruption of IMPA binding significantly impairs virus growth at a late stage in the replication cycle. Disruption of nuclear export function completely abrogates infectious particle production. Consistent with these observations, an NES mutant VP24 recombinant virus could not be recovered. Providing mechanistic insight, initial studies on the NES mutant VP24 suggest that it is significantly impaired for actin-dependent trafficking. Finally, proteomic analysis suggests that VP24 recruits components of the Wave regulatory complex (WRC), a regulator of Arp2/3 dependent actin polymerization via its NES sequence. These data support roles for VP24 IMPA binding and NES sequences in virus production and suggest that the NES may recruit the WRC to promote NCLS trafficking. The proposed studies will determine how VP24 NES sequences recruits the WRC to promote actin-dependent trafficking of viral nucleocapsids and incorporation into viral particles, and it will define the IFN-dependent and IFN-independent consequences of VP24-IMPA interaction.

NCI - National Cancer Institute

ABSTRACT There is an urgent need to nominate biomarkers that are likely to predict the efficacy of radiotherapy and accelerate their clinical translation. Efforts thus far have been limited in large part because the omic features regulating tumor cell survival and their frequency across and within individual cancer types had not been studied. Also, radiation is one of the most utilized therapeutic modalities in cancer. Despite this, radiotherapy dose is still delivered using a ‘one size fits most’ paradigm and has not yet lent itself to personalization based on genetic features. Our group of investigators have expertise that spans several aspects of the study of translational lung radiotherapy including advanced functional genomics, use of pre-clinical models of human tumors, personalized therapeutic interventions, bioinformatics, computational biology, and clinical research. Collectively, we have: (1) completed the largest profiling effort of the genetic vulnerability of cancer cells to irradiation to date (533 cell lines); (2) completed an arrayed unary (one mutation per well) gene variant cellular profiling project that has interrogated ~400 common and rare genetic variants for response to ionizing radiation in cells; (3) developed new functional genomic tools to study genome-phenome associations at scale; (4) developed over 500 genetically and clinically annotated patient derived xenograft (PDXs), subset of ex vivo and 3D models for testing using a radiation platform and (5), developed new computational tools and frameworks to personalize radiation dose treatments. The major themes for this proposal are: (i) analyze matched pre- and post-radiotherapy genomes from patient tissue and using PDX to elucidate mechanisms of de novo and/or acquired resistance; (ii) develop predictive diagnostics related to multi-omic tumor variates in patients receiving radiotherapy for lung cancer; (iii) integrate multi-tiered omic data to accurately predict the probability of local failures after lung radiotherapy; and (iv) identify and validate functionally significant molecular lesions in tumors that impact radiotherapeutic efficacy. This project is explicitly designed to utilize patient and model systems data in iterative and complementary testing cycles. Due to our extensive and sustained efforts in this space, we have an established preclinical understanding of the most salient genetic determinants that regulate radiation sensitivity in lung cancer. Altogether, our highly integrated proposal will come together in an unprecedented effort to advance toward clinical integration new information and capabilities that will ultimately improve outcomes for the multitude of patients receiving lung radiotherapy each year in the US and abroad.

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY Autoimmune type 1 diabetes (T1D) is caused by the T cell-mediated destruction of insulin producing beta (β) cells in the pancreas. The incidence of T1D is rising globally. This is thought to be linked to early life exposure to microbes, environmental pollutants, and even diet. Viral infections in mouse models of diabetes have shown both acceleration and protection of diabetes, but these differences occur at different ages and degree of insulitis. The deciding factor in whether CD4+ T cells will prime the immune system to initiate T1D is the inflammatory context of the initial peripheral antigen encounter, particularly the timing of type I interferon (IFN-I) exposure triggered by microbes. Unfortunately, the vast majority of research has utilized specific pathogen free (SPF) mouse models examining activation in the absence of IFN-I. These conditions are very different from the environment in which humans live. At UMN, we have created a ‘dirty’ mouse model or normal microbial environment (NME) to study how the immune system responds or develops in the presence of microbes and viral pathogens, a more physiological environment driven by IFN-I production. Thus, we now have a diabetes model that we can study with NME conditions. Depending on the age and duration of time in NME, our NOD mice can be completely protected from diabetes. Our goal is to understand how infections impact immunity to either trigger or protect against diabetes. A better understanding of this process could provide therapies that prevent or treat human diabetes. We will test three specific aims; 1) Determine the role of IFN-I on naïve CD4 T cell fate following TCR activation in SPF conditions, 2) Determine the mechanism(s) by which NME prevents autoimmune diabetes, and 3) Develop autoimmune diabetes therapies. Our goal is to determine the role of IFN-I during initial T cell fate decision between Teff and Tregs. We hypothesize this fate decision leads to iTregs and will focus on induction, survival or enhanced suppressive function. Finally, we will explore IFN-I therapy and engineered TCR Tregs for autoimmune therapy.

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY (See instructions): Cell growth and division are the essential processes for tissue homeostasis, regeneration, and cancer development. Therefore, the study of cell growth regulation is important for understanding impaired tissue regeneration and tumorigenesis. Cell growth and cell division are coordinated via mutual regulation, and these two processes together determine a fundamental cellular property, the cell size. The molecular mechanisms underlying the linkages between these processes are largely unknown. Cell growth can trigger cell division by diluting the cell cycle inhibitor RB, which contributes to the maintenance of cell size homeostasis. The RB dilution mechanism relies on the cell cycle-dependent RB concentration dynamics, but the underlying mechanism is still unclear. On the other hand, cell cycle and cell size can feed back to regulate cell growth rate, of which the mechanisms are yet unknown. I found that the protein synthesis efficiency, a major determinant of cell growth efficiency, increases at the G1/S transition and decreases as cells grow larger within each cell cycle phase, suggesting that the global protein synthesis rate is actively regulated to facilitate cell cycle progression and cell size control. Here, in this proposal, I aim to address these gaps in our knowledge by determining the molecular mechanisms regulating RB concentration and global protein synthesis during cell cycle progression and cell growth. During the K99 phase, I’ve determined the molecular mechanisms underlying the RB concentration regulation during cell cycle progression (Aim1), which further revealed the molecular basis of how cell growth triggers cell division. During the R00 phase, I will continue to study how global protein synthesis is regulated during cell cycle progression (Aim2), which will help us understand how cell cycle regulates cell growth. These two aims link cell growth with cell division from the perspective of protein synthesis regulation. Then, the study on how protein synthesis is regulated during cell growth (Aim3), which will be pursued during R00 phase, explores the mechanism of how cell size feeds back to regulate growth rate. Thus, the three aims together set up a foundation towards a deeper understanding of the principles governing cell growth and proliferation. The completion of above aims will deepen our understanding of the fundamental molecular mechanisms regulating protein synthesis and cell growth rate, and therefore have broad impacts on cell and developmental biology. Moreover, this work will enhance our understanding of tissue regeneration deficiencies and tumorigenesis.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

PROJECT SUMMARY Monocarboxylate transporters (MCTs) mediate the transport of key metabolic intermediates, such as lactate, urate, and pyruvate, across cellular membranes. Despite their importance in metabolic homeostasis and their implication in a range of diseases—including cancer, cardiovascular disease, and neurodegeneration—the MCT family remains poorly understood. Novel approaches are needed to study the role of MCTs in physiology and disease. MCTs are highly conserved in the nematode Caenorhabditis elegans. C. elegans is a highly tractable genetic model, and its defined anatomy, simple diet, and optical transparency offers several advantages for studying conserved molecular and physiological processes. Through a genetic screen for mutants with defecation defects, I found that a point mutation in slcf-1 causes a shortened defecation cycle. slcf-1 is expressed in the intestine and regulates intestinal calcium waves (ICWs) that drive the defecation motor program (DMP). These findings implicate a novel role for MCTs, metabolism and Ca²⁺ homeostasis. slcf-1 shares homology with MCT9, which has been implicated in hyperuricemia and can contribute to cardiovascular disease, gout, and renal cell carcinoma. The overarching goal of this project is to define the role of slcf-1 in intestinal Ca2+ dynamics and metabolism. In Aim 1, I will analyze how mutations in slcf-1 affect Ca²⁺ waves. I will quantify ICW properties, such as intervals and rise and fall times, perform tissue-specific rescue, and examine SLCF-1::GFP localization. In Aim 2, I will determine SLCF-1 substrates and how SLCF-1 is dynamically regulated using HEK293T transport assays and mutant slcf-1 promotor constructs. Finally, in Aim 3, I will investigate the metabolic consequences of slcf-1 loss, assessing mitochondrial morphology, activity, and stress, alongside broader metabolic pathway alterations. These studies will provide greater insight into how conserved monocarboxylate transporters connect metabolism, calcium signaling, and intestinal motility, and promote a broader understanding of MCTs’ role in human health and disease.

NINDS - National Institute of Neurological Disorders and Stroke

Project Summary Denervation injuries lead to altered neural activity in sensory processing regions of the brain. Recovery from these injuries is achieved by learning new motor skills to perform tasks of daily living; this requires successful cortical remapping. This remapping is thought to be mediated in part by changes in thalamocortical (TC) connections from the thalamus to cortex. However, the reasons that some patients recover well while others do not is poorly understood. Microglia shape circuits during development by pruning synapses in an activity dependent manner. Neurons and other cells express “eat me” and “don’t eat me” signals that act through microglial receptors to guide removal of underused synaptic connections such as those arising from the denervated body part. Large scale activity changes in sensory circuits after denervation injury lead to altered signaling through these microglial receptors, altering the activity and function of synapse-associated microglia. However, it is poorly understood how microglia coordinate with neurons to direct adult cortical circuit reorganization after peripheral injury. The goals of this project are to determine the necessity of microglia for post-injury adult circuit reorganization, determine the impact of aberrant signaling from unpruned connections, and identify the molecular mechanisms microglia and neurons utilize to coordinate their activity with other cells in the brain. To understand how microglia mediate post-denervation circuit remodeling, we will use a mouse model of unilateral infraorbital nerve transection (IONX). IONX mimics many aspects of circuit remodeling that occurs after unilateral denervation injury. We hypothesize that IONX causes microglia-mediated TC remodeling in the deprived somatosensory cortex of adult mice. This remodeling is expected to cause behavioral adaptations, particularly related to orofacial sensory/motor function. We will address this hypothesis by 1) determining the necessity of microglia for TC synapse density changes after IONX. Using tissue collected from acute, subacute and chronic timepoints, we will quantify changes in thalamocortical input density within the deprived somatosensory cortex, and microglial association with and removal of TC synapses. We will then deplete microglia to establish if microglia are required for the changes in the deprived somatosensory cortex. We will assess orofacial function in sham and IONX mice to demonstrate the impact of microglial TC circuit reorganization on behavioral adaptations. We will also modulate the activity of TC connections in the deprived somatosensory cortex to establish that preserved, erroneous activity after IONX leads to orofacial dysfunction. 2) We will use gene expression to identify the mechanisms by which microglia and neurons influence adult circuit remodeling after injury, such as identification of inactive synapses and increased phagocytosis. The findings of this proposal will improve our mechanistic understanding of how cells in the adult brain adapt to peripheral denervation injury, and can be used to gain insight into healthy adult plasticity.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

Type 2 Diabetes (T2D) is estimated to affect over 30 million US adults and skeletal muscle (SkM) insulin resistance (IR) is one of the primary aberrations. SkM is a complex organ comprised of different cell types including: myofibers, endothelial cells (EC), smooth muscle cells, fibro-adipogenic progenitors, satellite cells and immune cells. Effective insulin-stimulated SkM glucose uptake is dependent on 1) insulin-stimulated activation of terminal arteriole EC to perfuse microvascular units, 2) transport of substrates across capillary ECs and 3) stimulation of myofiber insulin signaling, resulting in glucose uptake. Each of these steps can be dysregulated by pro-inflammatory cytokines released from localized immune cells in the SkM niche. Therefore, different cell types in SkM can impact insulin sensitivity. Previous research has identified a dysregulated transcriptional profile with SkM IR in the basal state and during insulin stimulation. A significant limitation of these prior studies is the restriction to whole SkM homogenates and therefore not identifying which cells or spatial area within the tissue the dysregulated signals originate from.  We have developed a pipeline for full-length amplification of single cells from SkM using the iCELL8 platform, resulting in 2-3 fold greater gene coverage than previous research, allowing us to investigate transcriptional aberrations that occur with SkM IR at a single cell resolution. Recent developments in spatial transcriptomics now permit transcriptional profiling on sectioned SkM tissue which identifies the location and proximity of dysregulated cells and how they relate to IR in SkM. This study will leverage a hyper-insulinemic euglycemic (HE) clamp with stable glucose isotope tracers and indirect calorimetry to assess insulin-stimulated glucose disposal (Rd) and non-oxidative glucose disposal (NOGD) in individuals with obesity with and without IR compared to an insulin sensitive control group. For the first time, the transcriptional responses to insulin in human SkM will be probed at a single cell and spatial resolution. We will identify which cells and areas of SkM respond to insulin and if they are spatially localized to each other and if their gene-network profiles correlate with a greater in vivo Rd and NOGD. This novel and innovative approach will reveal which cells (and importantly, where in SkM) the aberrations occur with SkM IR whilst dissociating the confounding effects of Obesity. Results from this proposal will identify specific molecular targets to alleviate SkM IR that will be targeted in future interventions.

NSF

Microbial energy and mass transfers have profound ecological and biogeochemical consequences, yet the controls directing microbial metabolic trade-offs remain poorly understood. The proposed project will study metabolic shifts in energy and mass transfers by microorganisms living through some of the earliest-evolving metabolisms known on Earth. By focusing efforts towards microbes driving primary production anaerobically with molecular hydrogen — through a metabolic process known as chemosynthesis — the project will test whether metabolic trade-offs can be predicted as a function of (i) growth temperature and/or (i) the amount of free energy associated with anaerobic respiration. State-of-the-art continuous cultivation techniques will be applied to compare changes in respiration rates, cell yields, and cell doubling times within and among chemosynthetic microbes under steady-state conditions. These experiments are key for integrating parameters describing microbial metabolic changes into mathematical and/or thermodynamic models of microbial growth and its biogeochemical consequence. Overall, this work will transform understanding of the underlying metabolic behavior of unicellular life at the base of the Universal Tree of Life and at the base of dark ecosystems. The objective of the proposed research is to determine whether microbial metabolic rate/yield trade-offs of hydrogenotrophic NO3--, Fe3+- and CO2-reducing microorganisms exhibit predictable adaptations to temperature and to the oxidation-reduction (redox) chemistry of catabolism during chemosynthesis. Specifically, we will test the hypotheses that (i) microbial metabolic rate-to-yield ratios will increase for all proposed microbial processes when at higher temperatures and that (ii) microbes relying on generally less exergonic catabolic redox reactions will systematically display higher anabolic efficiencies than those relying on more exergonic catabolic redox reactions. To test these hypotheses, the project will employ experimental microbiological and geochemical approaches to compare microbial respiration rates, cell (and total biomass) yields, and cell doubling times of different anaerobic chemosynthetic microorganisms during batch and continuous growth. The resulting metabolic and environmental data will demonstrate how microbial metabolic behaviors are shaped by their abiotic environments. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIDCD - National Institute on Deafness and Other Communication Disorders

Project Summary Hundreds of taste buds (TBs) are located in the epithelial trenches of the circumvallate taste papilla (CVP) at the posterior midline of the tongue. Each TB contains taste receptor cells (TRCs) that transduce taste information to the brain and are continually replaced from stem/progenitor cells outside of TBs. Cancer patients undergoing chemo- or radiotherapy often experience dysgeusia, or taste dysfunction, likely due to perturbation to TRC renewal. Patients experiencing dysgeusia have increased risk of depression, malnutrition, and poor treatment outcomes. Therefore, understanding the mechanisms regulating maintenance of taste epithelium and TRC renewal will inform development of treatments to prevent or restore taste loss in these patients. CVP homeostasis occurs through proliferation and differentiation of progenitor cells that generate both TRCs and the surrounding non-taste epithelium. Our lab has recently identified new, long-term SOX9+ progenitors that, as a population, generate TRCs and non-taste epithelium (NTE); however, the potential of individual SOX9+ progenitors is unexplored. SOX9+ progenitors are located distant from taste buds, in the epithelial junction linking CVP trenches to the ducts of the underlying von Ebner's minor salivary glands (VEG). Each CVP contains a dozen or more junctions containing SOX9+ cells, suggesting that multiple independent progenitor reservoirs contribute to taste epithelium. Additionally, in contrast to the highly proliferative CVP, the junction contains few proliferative cells, indicating SOX9+ cells are slow cycling. These findings lead to my hypothesis that individual SOX9+ progenitors from multiple junctions produce progeny that move into local CVP trench epithelium, become highly proliferative, and differentiate into TRCs and NTE. Thus, in Aim 1, I will use sparse SOX9 lineage trace, whole-tissue clearing, and EdU birth dating, to determine the lineage and proliferative potential of individual progenitors from multiple junctions. Our previous work has focused on SOX9+ progenitor contribution in adult homeostasis, but if and when these taste stem cells are established embryonically is unknown. Embryonic immunostaining reveals SOX9+ cells are present during initial formation of the CVP and have overlapping expression with Ptch, the Shh signaling receptor. Shh is necessary for anterior tongue taste bud development, and proper CVP trench invagination in the posterior tongue. These findings lead to my second hypothesis that embryonic Shh signaling is required to establish adult SOX9+ taste progenitors. In Aim 2, I will use embryonic lineage tracing and genetic deletion of Shh signaling in SOX9+ cells to determine when SOX9+ cells are established, begin contributing to TBs, and if Shh signaling is required in these processes. Overall, this work may lead to approaches to leverage concentrated pools of taste stem cells to restore taste function in patients.

NSF

This project explores how macrodomain proteins have evolved to balance their different multiple biochemical activities. Macrodomains are ancient enzymes that bind to and remove ADP-ribose, an important post-translation modification that is critical in several cellular stress responses, including DNA damage, ER stress, and virus infection. These enzymes are conserved through all domains of life, including archaea, bacteria, eukaryotes, and viruses, indicating that they are critical for multiple cellular processes. However, the function of macrodomain enzymes in cell biology and microbiology are just now being uncovered. Furthermore, recent research indicates that macrodomains have evolved to properly balance their biochemical functions depending on if they are expressed from a virus, a bacteria, or from a eukaryotic cell. This project will evaluate how viral macrodomains have evolved to develop the ideal biochemical properties that allow them function in the context of a virus infection. This project also has strong educational and community outreach components. Most notably, students at all training levels will participate in this project and will learn how to evaluate the evolution of proteins through workshops in phylogenetics. This project will provide new insights into the fundamental biology of macrodomain enzymes and could lead to new insights into antiviral drug-development. This project aims to define how the coronavirus macrodomain has evolved to best function in the context of a virus infection using the mouse coronavirus, murine hepatitis virus (MHV), as a model. The use of MHV, which is unable to infect humans, for creating macrodomain mutations eliminates the potential for gain-of-function research. Research over the last decade has demonstrated that the coronavirus macrodomain blocks innate immune responses, is critical for viral pathogenesis, and is a potential drug target. Furthermore, the macrodomain uses both its ADP-ribose binding and hydrolysis activities to promote virus replication, and they must be balanced for optimal replication. The PI will use recombinant proteins and a panel of recombinant viruses, developed using a bacterial artificial chromosome based reverse genetic system, to better understand how highly conserved amino acids and selective pressure induced mutations impact Mac1 biochemical activities, virus replication, and pathogenesis using well-established assays and model systems. Additionally, macrodomains from across the evolutionary spectrum will be expressed in the context of MHV infection to define the evolutionary limits of macrodomain divergence that is tolerated in MHV. This work will provide a deeper understanding of how macrodomains have evolved to counter various ADP-ribose dependent antiviral responses. This project is jointly funded by the Genetic Mechanisms Program and the Division of Molecular and Cellular Biosciences. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary Pneumonic plague is the deadliest form of disease caused by Yersinia pestis. Disease manifests as a rapidly progressing and highly lethal necrotic pneumonia that is typically fatal within seven days. If antibiotic treatment is not administered within 24 hours after the onset of symptoms, pneumonic plague is 100% fatal. The lethality of plague is due to the abrupt onset of a hyper-inflammatory response that compromises pulmonary function. Deletion of the Y. pestis inner membrane protein YbtX results in decreased neutrophil infiltration into the airways and decreased expression of select pro-inflammatory cytokines/chemokines during the later stages of pneumonic plague. YbtX is part of a zinc acquisition system by which the Y. pestis siderophore Yersiniabactin (Ybt) competes with host protein calprotectin to scavenges bioavailable zinc. YbtX acts as an inner membrane transporter that facilitates transport of Ybt-Zn into the bacterial cytosol. Calprotectin is also known to be a driver of host inflammatory responses. We hypothesize that YbtX-mediated zinc import contributes to the onset inflammation in the lung by activating calprotectin-dependent inflammatory responses. To test this, we will characterize how zinc depletion impacts inflammation during infection in the absence of YbtX. Further, we will alter the balance of host zinc/calprotectin in a murine intranasal infection model of pneumonic plague and characterize how this balance affects pulmonary inflammation. This work will determine how zinc and host calprotectin impact host inflammatory responses during pneumonic plague.